U.S. patent application number 14/915552 was filed with the patent office on 2016-08-04 for nutritional composition having lipophilic compounds with improved solubility and bioavailability.
This patent application is currently assigned to Abbott Laboratories. The applicant listed for this patent is Abbott Laboratories. Invention is credited to Daniel Albrecht, Stephen DeMichele, Paul Johns, Gary Katz, Todime Reddy, Mustafa Vurma.
Application Number | 20160219913 14/915552 |
Document ID | / |
Family ID | 51493121 |
Filed Date | 2016-08-04 |
United States Patent
Application |
20160219913 |
Kind Code |
A1 |
Johns; Paul ; et
al. |
August 4, 2016 |
NUTRITIONAL COMPOSITION HAVING LIPOPHILIC COMPOUNDS WITH IMPROVED
SOLUBILITY AND BIOAVAILABILITY
Abstract
Disclosed herein is a nutritional composition having at least
one protein, at least one fat, and at least one lipophilic
compound, the composition comprising at least one assembly
comprising at least one hydrophobic protein, monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C.
Inventors: |
Johns; Paul; (Columbus,
OH) ; Vurma; Mustafa; (Dublin, OH) ; Albrecht;
Daniel; (Columbus, OH) ; DeMichele; Stephen;
(Dublin, OH) ; Katz; Gary; (Columbus, OH) ;
Reddy; Todime; (New Albany, OH) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Abbott Laboratories |
Abbott Park |
IL |
US |
|
|
Assignee: |
Abbott Laboratories
Abbott Park
IL
|
Family ID: |
51493121 |
Appl. No.: |
14/915552 |
Filed: |
August 28, 2014 |
PCT Filed: |
August 28, 2014 |
PCT NO: |
PCT/US2014/053230 |
371 Date: |
February 29, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61871719 |
Aug 29, 2013 |
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61901207 |
Nov 7, 2013 |
|
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61920657 |
Dec 24, 2013 |
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61920666 |
Dec 24, 2013 |
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61920669 |
Dec 24, 2013 |
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61920675 |
Dec 24, 2013 |
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61949505 |
Mar 7, 2014 |
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62007037 |
Jun 3, 2014 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A23L 33/155 20160801;
A23C 9/1528 20130101; A23C 9/1526 20130101; A23L 33/40 20160801;
A23L 33/115 20160801; A23L 33/15 20160801; A23L 33/17 20160801;
A23L 33/19 20160801; A23V 2002/00 20130101; A61P 3/02 20180101;
A23V 2002/00 20130101; A23V 2250/182 20130101; A23V 2250/192
20130101; A23V 2250/211 20130101; A23V 2250/71 20130101 |
International
Class: |
A23L 1/302 20060101
A23L001/302; A23D 7/005 20060101 A23D007/005; A23L 1/303 20060101
A23L001/303 |
Claims
1. A method of preparing a nutritional composition having at least
one lipophilic compound having improved bioavailability, the method
comprising the steps of: providing a premix comprising an assembly
that comprises at least one hydrophobic protein, combination of
monoglycerides and diglycerides ("MDG") and at least one lipophilic
compound, wherein the assembly is (i) water soluble; and (ii) has a
size of 100 kD to 1000 kD; adding the premix to an aqueous solution
to form an activated premix; and adding the activated premix to the
nutritional composition.
2. The method of claim 1, wherein the assembly has a size of 60 kD
to 400 kD.
3. The method of claim 1 or 2, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof.
4. The method of claim 1, wherein the lipophilic compound is
lutein, vitamin A, vitamin D, vitamin E, vitamin K or any
combinations thereof.
5. The method of claim 1, wherein the premix is heated to a
temperature of 120.degree. F. for about 10 minutes.
6. A method of preparing a nutritional composition having at least
one lipophilic compound having improved bioavailability, the method
comprising the steps of: providing a premix comprising an assembly
that comprises at least one hydrophobic protein, a combination of
monoglycerides and diglycerides ("MDG") and at least one lipophilic
compound, wherein the assembly is (i) water soluble; and (ii) has a
size of 100 kD to 1000 kD; adding the premix to a fat-containing
solution to form a protected premix; and adding the protected
premix to the nutritional composition.
7. The method of claim 6, wherein the assembly has a size of 60 kD
to 400 kD.
8. The method of claim 6, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof.
9. The method of claim 8, wherein the lipophilic compound is
lutein, vitamin A, vitamin D, vitamin E, vitamin K or any
combinations thereof.
10. The method of claim 6, wherein the premix is heated to a
temperature of 120.degree. F. for about 10 minutes.
Description
RELATED APPLICATION INFORMATION
[0001] This application claims the benefit of U.S. Application No.
61/871,719, filed on Aug. 29, 2013, U.S. Application No.
61/901,207, filed on Nov. 7, 2013, U.S. Application No. 61/920,657,
filed on Dec. 24, 2013, U.S. Application No. 61/920,666, filed on
Dec. 24, 2013, U.S. Application No. 61/920,669 filed on Dec. 24,
2013, U.S. Application No. 61/920,675, filed on Dec. 24, 2013, U.S.
Application No. 61/949,505 filed on Mar. 7, 2014 and U.S.
Application No. 62/007,037 filed on Jun. 3, 2014, the contents of
each of which are herein incorporated by reference in its
entirety.
TECHNICAL FIELD
[0002] The present disclosure relates to compositions containing
lipophilic compounds with improved bioavailability. More
particularly, the compositions contain an activated premix or an
MDG-protected premix comprising monoglycerides and diglycerides.
These premixes may provide lipophilic compounds in a form that is
more available for absorption into the body upon consumption.
BACKGROUND
[0003] Adult, pediatric, and infant manufactured nutritional and
pharmaceutical products may comprise a variety of lipophilic
compounds such as carotenoids, lipid soluble vitamins, lipophilic
antioxidants or combinations thereof. Current nutritional
compositions comprising lipophilic compounds typically provide
limited bioavailability following consumption because lipophilic
compounds are unable to remain in stable, water soluble forms. As a
result, the products are often over-fortified with lipophilic
compounds to ensure that the desired nutritional or pharmaceutical
benefits from the compounds may be obtained. In some cases, the
over-fortification may be from about 2 times to about 10 times the
amount required to achieve the desired benefits.
[0004] These high fortification rates may lead to increased
production costs and potential complications in formulating the
final product without providing additional consumer benefits.
Furthermore, the over fortified formulations may overwhelm the
digestive tract with non-soluble lipophilic compounds leading to
indigestion.
[0005] Accordingly, there is a need for nutritional compositions
for infants, toddlers, children and adults that provide necessary
lipophilic compounds in a more stable soluble form.
SUMMARY OF THE INVENTION
[0006] In one aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C.
[0007] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD.
[0008] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG.
[0009] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG.
[0010] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG.
[0011] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C.
[0012] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation.
[0013] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof.
[0014] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C.
[0015] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature.
[0016] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0.
[0017] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof.
[0018] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD.
[0019] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof.
[0020] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof.
[0021] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof.
[0022] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition.
[0023] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition.
[0024] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition.
[0025] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition.
[0026] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w).
[0027] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w).
[0028] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w), wherein the
ratio of MDG to lipophilic compound is 4:1, 3:1 or 2:1.
[0029] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w), wherein the
ratio of MDG to lipophilic compound is 4:1, 3:1 or 2:1, comprising
about 1.0 g/L to about 100 g/L of hydrophobic protein based on
total weight of the composition.
[0030] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w), wherein the
ratio of MDG to lipophilic compound is 4:1, 3:1 or 2:1, comprising
about 1.0 g/L to about 100 g/L of hydrophobic protein based on
total weight of the composition, further comprising choline.
[0031] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w), wherein the
ratio of MDG to lipophilic compound is 4:1, 3:1 or 2:1, comprising
about 1.0 g/L to about 100 g/L of hydrophobic protein based on
total weight of the composition, further comprising choline,
comprising about 5 mg/kg to about 1 g/kg of choline based on total
weight of the composition.
[0032] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w), wherein the
ratio of MDG to lipophilic compound is 4:1, 3:1 or 2:1, comprising
about 1.0 g/L to about 100 g/L of hydrophobic protein based on
total weight of the composition, further comprising choline,
comprising about 5 mg/kg to about 1 g/kg of choline based on total
weight of the composition, further comprising lecithin.
[0033] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w), wherein the
ratio of MDG to lipophilic compound is 4:1, 3:1 or 2:1, comprising
about 1.0 g/L to about 100 g/L of hydrophobic protein based on
total weight of the composition, further comprising choline,
comprising about 5 mg/kg to about 1 g/kg of choline based on total
weight of the composition, further comprising lecithin, comprising
about 10 mg/kg to about 5 g/kg of lecithin based on total weight of
the composition.
[0034] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w), wherein the
ratio of MDG to lipophilic compound is 4:1, 3:1 or 2:1, comprising
about 1.0 g/L to about 100 g/L of hydrophobic protein based on
total weight of the composition, further comprising choline,
comprising about 5 mg/kg to about 1 g/kg of choline based on total
weight of the composition, further comprising lecithin, comprising
about 10 mg/kg to about 5 g/kg of lecithin based on total weight of
the composition, further comprising docosahexaenoic acid
("DHA").
[0035] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w), wherein the
ratio of MDG to lipophilic compound is 4:1, 3:1 or 2:1, comprising
about 1.0 g/L to about 100 g/L of hydrophobic protein based on
total weight of the composition, further comprising choline,
comprising about 5 mg/kg to about 1 g/kg of choline based on total
weight of the composition, further comprising lecithin, comprising
about 10 mg/kg to about 5 g/kg of lecithin based on total weight of
the composition, further comprising docosahexaenoic acid ("DHA"),
wherein the nutritional composition comprises about 10 mg/kg to
about 10 g/kg DHA based on total weight of the composition.
[0036] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w), wherein the
ratio of MDG to lipophilic compound is 4:1, 3:1 or 2:1, comprising
about 1.0 g/L to about 100 g/L of hydrophobic protein based on
total weight of the composition, further comprising choline,
comprising about 5 mg/kg to about 1 g/kg of choline based on total
weight of the composition, further comprising lecithin, comprising
about 10 mg/kg to about 5 g/kg of lecithin based on total weight of
the composition, further comprising docosahexaenoic acid ("DHA"),
wherein the nutritional composition comprises about 10 mg/kg to
about 10 g/kg DHA based on total weight of the composition, further
comprising high palmitic acid vegetable oil.
[0037] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w), wherein the
ratio of MDG to lipophilic compound is 4:1, 3:1 or 2:1, comprising
about 1.0 g/L to about 100 g/L of hydrophobic protein based on
total weight of the composition, further comprising choline,
comprising about 5 mg/kg to about 1 g/kg of choline based on total
weight of the composition, further comprising lecithin, comprising
about 10 mg/kg to about 5 g/kg of lecithin based on total weight of
the composition, further comprising docosahexaenoic acid ("DHA"),
wherein the nutritional composition comprises about 10 mg/kg to
about 10 g/kg DHA based on total weight of the composition, further
comprising high palmitic acid vegetable oil, wherein the
nutritional composition comprises about 0.5 g/kg to about 35 g/kg
of high palmitic acid vegetable oil based on total weight of the
composition.
[0038] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w), wherein the
ratio of MDG to lipophilic compound is 4:1, 3:1 or 2:1, comprising
about 1.0 g/L to about 100 g/L of hydrophobic protein based on
total weight of the composition, further comprising choline,
comprising about 5 mg/kg to about 1 g/kg of choline based on total
weight of the composition, further comprising lecithin, comprising
about 10 mg/kg to about 5 g/kg of lecithin based on total weight of
the composition, further comprising docosahexaenoic acid ("DHA"),
wherein the nutritional composition comprises about 10 mg/kg to
about 10 g/kg DHA based on total weight of the composition, further
comprising high palmitic acid vegetable oil, wherein the
nutritional composition comprises about 0.5 g/kg to about 35 g/kg
of high palmitic acid vegetable oil based on total weight of the
composition, wherein the nutritional composition comprises about
0.2 g/L to about 20 g/L of .beta.-casein based on total weight of
the composition.
[0039] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w), wherein the
ratio of MDG to lipophilic compound is 4:1, 3:1 or 2:1, comprising
about 1.0 g/L to about 100 g/L of hydrophobic protein based on
total weight of the composition, further comprising choline,
comprising about 5 mg/kg to about 1 g/kg of choline based on total
weight of the composition, further comprising lecithin, comprising
about 10 mg/kg to about 5 g/kg of lecithin based on total weight of
the composition, further comprising docosahexaenoic acid ("DHA"),
wherein the nutritional composition comprises about 10 mg/kg to
about 10 g/kg DHA based on total weight of the composition, further
comprising high palmitic acid vegetable oil, wherein the
nutritional composition comprises about 0.5 g/kg to about 35 g/kg
of high palmitic acid vegetable oil based on total weight of the
composition, wherein the nutritional composition comprises about
0.2 g/L to about 20 g/L of .beta.-casein based on total weight of
the composition, wherein the nutritional composition comprises a
carbohydrate.
[0040] In another aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 1% of the total MDG in the nutritional composition remains
in the aqueous phase after centrifugation at 100,000.times.g for 1
hour at 20.degree. C., wherein the assembly is (i) water soluble;
and (ii) has a size of 100 kD to 1000 kD, wherein MDG increases the
concentration of protein of the assembly by at least 2% compared to
a nutritional composition without MDG, wherein MDG increases the
bound lipophilic compound of the assembly by at least 2% compared
to a nutritional composition without MDG, wherein the lipophilic
compound of the composition has at least a 15% improvement in
bioavailability when administered to a subject, wherein the 15%
improvement in bioavailability is measured by lymphatic absorption
of the composition over a nutritional composition without MDG,
wherein the assembly is still water soluble after centrifugation at
about 31,000.times.g for at least 1 hour at 20.degree. C., wherein
at least 5% of the assembly remains in the aqueous phase after
centrifugation, wherein the protein is intact protein, hydrolyzed
protein or combinations thereof, wherein the assembly is stable for
at least 12 months at 23.degree. C., wherein the assembly is stable
for at least 24 months at room temperature, wherein the assembly
comprises a hydrophobic protein having a GRAVY value of about -0.5
to about 0, wherein the hydrophobic protein is 3-lactoglobulin,
bovine lactoferrin, .gamma..sub.2-casein, .beta.-casein,
.alpha.-lactalbumin, or combinations thereof, wherein the assembly
has a size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the fat is MDG, coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic safflower oil, sunflower oil, high
oleic sunflower oil, palm oil, palm kernel oil, palm olein, canola
oil, marine oils, cottonseed oil, flax seed oil, hemp seed oil,
peanut oil, borage oil, algal oils, fungal oils, MCT oil (medium
chain triglycerides) and combinations thereof, comprising about 145
mg/kg to about 890 mg/kg of MDG based on total weight of the
composition, comprising about 145 mg/kg to about 600 mg/kg of MDG
based on total weight of the composition, comprising about 1.0
.mu.g/kg to about 7.0 g/kg of a lipophilic compound based on total
weight of the composition, comprising about 147.4 mg/kg to about
589.6 mg/kg of MDG and about 1.12 mg/kg of a lipophilic compound
based on total weight of the composition, wherein the ratio of MDG
to lipophilic compound are selected from the group consisting of
12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1, 6000:1, 5000:1,
4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1,
400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1, 14:1,
13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and
1:1 (w/w), wherein the ratio of MDG to lipophilic compound
comprises a ratio of about 12000:1 to about 1:1 (w/w), wherein the
ratio of MDG to lipophilic compound is 4:1, 3:1 or 2:1, comprising
about 1.0 g/L to about 100 g/L of hydrophobic protein based on
total weight of the composition, further comprising choline,
comprising about 5 mg/kg to about 1 g/kg of choline based on total
weight of the composition, further comprising lecithin, comprising
about 10 mg/kg to about 5 g/kg of lecithin based on total weight of
the composition, further comprising docosahexaenoic acid ("DHA"),
wherein the nutritional composition comprises about 10 mg/kg to
about 10 g/kg DHA based on total weight of the composition, further
comprising high palmitic acid vegetable oil, wherein the
nutritional composition comprises about 0.5 g/kg to about 35 g/kg
of high palmitic acid vegetable oil based on total weight of the
composition, wherein the nutritional composition comprises about
0.2 g/L to about 20 g/L of .beta.-casein based on total weight of
the composition, wherein the nutritional composition comprises a
carbohydrate, wherein the carbohydrate is maltodextrin, hydrolyzed
or modified starch, hydrolyzed or modified cornstarch, glucose
polymers, corn syrup, corn syrup solids, rice-derived
carbohydrates, glucose, fructose, lactose, high fructose corn
syrup, honey, sugar alcohols, maltitol, erythritol, sorbitol, or
combinations thereof.
[0041] In one aspect, the disclosure may be directed to a
nutritional composition comprising at least one protein and vitamin
D, having improved bioavailability of vitamin D, wherein the
nutritional composition comprises: an assembly comprising a
combination of monoglycerides and diglycerides ("MDG") and vitamin
D, wherein the vitamin D has improved bioavailability when
administered to a subject.
[0042] In another aspect, the disclosure may be directed to a
nutritional composition comprising at least one protein and vitamin
D, having improved bioavailability of vitamin D, wherein the
nutritional composition comprises: an assembly comprising a
combination of monoglycerides and diglycerides ("MDG") and vitamin
D, wherein the vitamin D has improved bioavailability when
administered to a subject, comprising about 0.1 mg/kg to about 20
mg/kg of MDG based on total weight of the composition.
[0043] In another aspect, the disclosure may be directed to a
nutritional composition comprising at least one protein and vitamin
D, having improved bioavailability of vitamin D, wherein the
nutritional composition comprises: an assembly comprising a
combination of monoglycerides and diglycerides ("MDG") and vitamin
D, wherein the vitamin D has improved bioavailability when
administered to a subject, comprising about 0.1 mg/kg to about 20
mg/kg of MDG based on total weight of the composition, comprising
about 0.1 mg/kg to about 10 mg/kg of MDG based on total weight of
the composition.
[0044] In another aspect, the disclosure may be directed to a
nutritional composition comprising at least one protein and vitamin
D, having improved bioavailability of vitamin D, wherein the
nutritional composition comprises: an assembly comprising a
combination of monoglycerides and diglycerides ("MDG") and vitamin
D, wherein the vitamin D has improved bioavailability when
administered to a subject, comprising about 0.1 mg/kg to about 20
mg/kg of MDG based on total weight of the composition, comprising
about 0.1 mg/kg to about 10 mg/kg of MDG based on total weight of
the composition, comprising about 10 .mu.g/kg to about 30 .mu.g/kg
of vitamin D based on total weight of the composition.
[0045] In another aspect, the disclosure may be directed to a
nutritional composition comprising at least one protein and vitamin
D, having improved bioavailability of vitamin D, wherein the
nutritional composition comprises: an assembly comprising a
combination of monoglycerides and diglycerides ("MDG") and vitamin
D, wherein the vitamin D has improved bioavailability when
administered to a subject, comprising about 0.1 mg/kg to about 20
mg/kg of MDG based on total weight of the composition, comprising
about 0.1 mg/kg to about 10 mg/kg of MDG based on total weight of
the composition, comprising about 10 .mu.g/kg to about 30 .mu.g/kg
of vitamin D based on total weight of the composition, comprising
about 0.1 mg/kg to about 10 mg/kg of MDG and about 20 .mu.g/kg of
vitamin D based on total weight of the composition.
[0046] In one aspect, the disclosure may be directed to a
nutritional composition having at least one protein, at least one
fat, and at least one lipophilic compound, the composition
comprising: an activated assembly comprising at least one
hydrophobic protein, at least one combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
at least 15% of the total lipophilic compound in the nutritional
composition remains in the aqueous phase after centrifugation at
1,000.times.g for 1 hour at 20.degree. C.
[0047] In one aspect, the disclosure may be directed to a method of
preparing a nutritional composition having at least one lipophilic
compound having improved bioavailability, the method comprising the
steps of: providing a premix comprising an assembly that comprises
at least one hydrophobic protein, combination of monoglycerides and
diglycerides ("MDG") and at least one lipophilic compound, wherein
the assembly is (i) water soluble; and (ii) has a size of 100 kD to
1000 kD; adding the premix to an aqueous solution to form an
activated premix; and adding the activated premix to the
nutritional composition.
[0048] In another aspect, the disclosure may be directed to a
method of preparing a nutritional composition having at least one
lipophilic compound having improved bioavailability, the method
comprising the steps of: providing a premix comprising an assembly
that comprises at least one hydrophobic protein, combination of
monoglycerides and diglycerides ("MDG") and at least one lipophilic
compound, wherein the assembly is (i) water soluble; and (ii) has a
size of 100 kD to 1000 kD; adding the premix to an aqueous solution
to form an activated premix; and adding the activated premix to the
nutritional composition, wherein the assembly has a size of 60 kD
to 400 kD.
[0049] In another aspect, the disclosure may be directed to a
method of preparing a nutritional composition having at least one
lipophilic compound having improved bioavailability, the method
comprising the steps of: providing a premix comprising an assembly
that comprises at least one hydrophobic protein, combination of
monoglycerides and diglycerides ("MDG") and at least one lipophilic
compound, wherein the assembly is (i) water soluble; and (ii) has a
size of 100 kD to 1000 kD; adding the premix to an aqueous solution
to form an activated premix; and adding the activated premix to the
nutritional composition, wherein the assembly has a size of 60 kD
to 400 kD, wherein the lipophilic compound is a carotenoid, a lipid
soluble vitamin, a lipophilic antioxidant or combinations
thereof.
[0050] In another aspect, the disclosure may be directed to a
method of preparing a nutritional composition having at least one
lipophilic compound having improved bioavailability, the method
comprising the steps of: providing a premix comprising an assembly
that comprises at least one hydrophobic protein, combination of
monoglycerides and diglycerides ("MDG") and at least one lipophilic
compound, wherein the assembly is (i) water soluble; and (ii) has a
size of 100 kD to 1000 kD; adding the premix to an aqueous solution
to form an activated premix; and adding the activated premix to the
nutritional composition, wherein the assembly has a size of 60 kD
to 400 kD, wherein the lipophilic compound is a carotenoid, a lipid
soluble vitamin, a lipophilic antioxidant or combinations thereof,
wherein the lipophilic compound is lutein, vitamin A, vitamin D,
vitamin E, vitamin K or any combinations thereof.
[0051] In another aspect, the disclosure may be directed to a
method of preparing a nutritional composition having at least one
lipophilic compound having improved bioavailability, the method
comprising the steps of: providing a premix comprising an assembly
that comprises at least one hydrophobic protein, combination of
monoglycerides and diglycerides ("MDG") and at least one lipophilic
compound, wherein the assembly is (i) water soluble; and (ii) has a
size of 100 kD to 1000 kD; adding the premix to an aqueous solution
to form an activated premix; and adding the activated premix to the
nutritional composition, wherein the assembly has a size of 60 kD
to 400 kD, wherein the lipophilic compound is a carotenoid, a lipid
soluble vitamin, a lipophilic antioxidant or combinations thereof,
wherein the lipophilic compound is lutein, vitamin A, vitamin D,
vitamin E, vitamin K or any combinations thereof, wherein the
premix is heated to a temperature of 120.degree. F. for about 10
minutes.
[0052] In one aspect, the disclosure may be directed to a method of
preparing a nutritional composition having at least one lipophilic
compound having improved bioavailability, the method comprising the
steps of: providing a premix comprising an assembly that comprises
at least one hydrophobic protein, a combination of monoglycerides
and diglycerides ("MDG") and at least one lipophilic compound,
wherein the assembly is (i) water soluble; and (ii) has a size of
100 kD to 1000 kD; adding the premix to a fat-containing solution
to form a protected premix; and adding the protected premix to the
nutritional composition.
[0053] In another aspect, the disclosure may be directed to a
method of preparing a nutritional composition having at least one
lipophilic compound having improved bioavailability, the method
comprising the steps of: providing a premix comprising an assembly
that comprises at least one hydrophobic protein, a combination of
monoglycerides and diglycerides ("MDG") and at least one lipophilic
compound, wherein the assembly is (i) water soluble; and (ii) has a
size of 100 kD to 1000 kD; adding the premix to a fat-containing
solution to form a protected premix; and adding the protected
premix to the nutritional composition, wherein the assembly has a
size of 60 kD to 400 kD.
[0054] In another aspect, the disclosure may be directed to a
method of preparing a nutritional composition having at least one
lipophilic compound having improved bioavailability, the method
comprising the steps of: providing a premix comprising an assembly
that comprises at least one hydrophobic protein, a combination of
monoglycerides and diglycerides ("MDG") and at least one lipophilic
compound, wherein the assembly is (i) water soluble; and (ii) has a
size of 100 kD to 1000 kD; adding the premix to a fat-containing
solution to form a protected premix; and adding the protected
premix to the nutritional composition, wherein the assembly has a
size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof.
[0055] In another aspect, the disclosure may be directed to a
method of preparing a nutritional composition having at least one
lipophilic compound having improved bioavailability, the method
comprising the steps of: providing a premix comprising an assembly
that comprises at least one hydrophobic protein, a combination of
monoglycerides and diglycerides ("MDG") and at least one lipophilic
compound, wherein the assembly is (i) water soluble; and (ii) has a
size of 100 kD to 1000 kD; adding the premix to a fat-containing
solution to form a protected premix; and adding the protected
premix to the nutritional composition, wherein the assembly has a
size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof.
[0056] In another aspect, the disclosure may be directed to a
method of preparing a nutritional composition having at least one
lipophilic compound having improved bioavailability, the method
comprising the steps of: providing a premix comprising an assembly
that comprises at least one hydrophobic protein, a combination of
monoglycerides and diglycerides ("MDG") and at least one lipophilic
compound, wherein the assembly is (i) water soluble; and (ii) has a
size of 100 kD to 1000 kD; adding the premix to a fat-containing
solution to form a protected premix; and adding the protected
premix to the nutritional composition, wherein the assembly has a
size of 60 kD to 400 kD, wherein the lipophilic compound is a
carotenoid, a lipid soluble vitamin, a lipophilic antioxidant or
combinations thereof, wherein the lipophilic compound is lutein,
vitamin A, vitamin D, vitamin E, vitamin K or any combinations
thereof, wherein the premix is heated to a temperature of
120.degree. F. for about 10 minutes.
BRIEF DESCRIPTION OF THE FIGURES
[0057] FIG. 1 shows the hourly lymph flow rate over a period of 6
hours following lutein administration of different nutritional
composition batches.
[0058] FIG. 2 shows the hourly lutein output in lymph over a 6 hour
period after lutein administration of different nutritional
composition batches.
[0059] FIG. 3 shows cumulative lymphatic lutein absorption over a 6
hour period after lutein administration of different nutritional
composition batches.
[0060] FIG. 4 shows the percent change in lymphatic lutein
absorption of lutein with mono- and diglycerides versus control
over a period of 6 hours.
[0061] FIG. 5 shows the hourly lymph flow rate over a period of 8
hours following administration of variable nutritional compositions
containing lutein.
[0062] FIG. 6 shows the hourly lutein output in lymph over an 8
hour period after administration of variable nutritional
compositions containing lutein.
[0063] FIG. 7 shows cumulative lymphatic lutein absorption over a 6
hour and 8 hour period after administration of variable nutritional
compositions containing lutein.
[0064] FIG. 8 shows the percent change in lymphatic lutein
absorption over a period of 6 hours and 8 hours following
administration of variable nutritional compositions containing
lutein versus control.
[0065] FIG. 9 shows the hourly lymphatic triglyceride output over a
period of 6 hours following nutritional composition
administration.
[0066] FIG. 10 shows the hourly lymphatic phospholipid output over
6 hours following nutritional composition administration.
[0067] FIG. 11 shows the change in lymphatic absorption of ARA from
fasting, over a period of 6 hours following nutritional composition
administration
[0068] FIG. 12 shows the change in lymphatic absorption of DHA from
fasting, over a period of 6 hours following nutritional composition
administration
[0069] FIG. 13 shows presence of the high molecular weight lutein
assembly within the aqueous phase following centrifugations of
variable nutritional compositions, as a function of lutein
concentration.
[0070] FIGS. 14-16 show the effect of different centrifugal forces
on the presence of the FIG. 14: high molecular weight lutein
assembly; FIG. 15: protein greater than 137 kD; and FIG. 16: bound
lipophile within the aqueous fraction following centrifugation of
variable nutritional compositions.
[0071] FIG. 17 shows the effects that choline and lecithin addition
to the nutritional composition have on the formation of the high
molecular weight lutein assembly.
[0072] FIG. 18 shows the lymphatic output of triglycerides over a
period of 6 hours following infusion of variable nutritional
compositions.
[0073] FIG. 19 shows the lymphatic absorption of palmitic acid over
a 6 hour period following infusion of variable nutritional
compositions.
[0074] FIG. 20 shows the lymphatic absorption of linoleic acid over
a 6 hour period following infusion of variable nutritional
compositions.
[0075] FIG. 21 shows the lymphatic absorption of alpha-linolenic
acid over a period of 6 hours following infusion of variable
nutritional compositions.
[0076] FIG. 22 shows the change in lymphatic output of DHA over
time following the administration of variable nutritional
compositions.
[0077] FIG. 23 shows UV spectra of the aqueous fractions of
variable nutritional compositions. UF: ultrafiltration with
membrane of 10 kD or 100 kD; UC: centrifugation method C
(100,000.times.g); SMP: skimmed milk powder/nonfat dried milk; WPC:
whey protein concentrate 80.
[0078] FIG. 24 shows lymphatic triglyceride output over a 6 hour
period following administration of variable nutritional
compositions, wherein the compositions are stratified based on
length of time post-production.
DETAILED DESCRIPTION
[0079] The present invention relates to a nutritional composition
for the delivery of nutrients, including lipophilic compounds in a
more soluble form by using mono- and di-glycerides (hereafter
"MDG") and hydrophobic protein assembled with the lipophilic
protein. This assembly of the nutritional composition containing
MDG, hydrophobic protein and a lipophilic compound allows for a
chaperoning of the lipophilic compound into a more soluble form to
facilitate easy digestion. The formations of stable, high molecular
weight assemblies (MDG plus hydrophobic protein and lipophilic
protein at approximately 60 kD to approximately 400 kD) enable
maximum nutritional delivery and bioavailability of the lipophilic
compound due to the increase in these stable, soluble forms over
lipophilic compound combinations that do not contain MDG and
hydrophobic protein. Increased water soluble forms of the
lipophilic compound lead to increased bioavailability following
consumption.
[0080] Ultimately, this enables the nutritional composition to
deliver lipophilic compounds without over-fortification, which
decreases production costs, limits composition complications, and
results in a more desirable product. The nutritional composition
may be ingested by an infant, toddler, child or adult, thereby
providing the nutrients needed for proper development, growth,
and/or as a nutritional supplement. The nutritional composition may
further be consumed to deliver lipophilic compounds for the
treatment of numerous conditions, including bone health/growth, eye
health, cardiovascular health, etc.
2. DEFINITIONS
[0081] Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art. In case of conflict, the present
document, including definitions, will control. Preferred methods
and materials are described below, although methods and materials
similar or equivalent to those described herein may be used in
practice or testing of the nutritional composition. All
publications, patent applications, patents and other references
mentioned herein are incorporated by reference in their entirety.
The materials, methods, and examples disclosed herein are
illustrative only and not intended to be limiting.
[0082] The terms "comprise(s)," "include(s)," "having," "has,"
"can," "contain(s)," and variants thereof, as used herein, are
intended to be open-ended transitional phrases, terms, or words
that do not preclude the possibility of additional acts or
structures. The singular forms "a," "and" and "the" include plural
references unless the context clearly dictates otherwise. The
present disclosure also contemplates other embodiments
"comprising," "consisting of" and "consisting essentially of," the
embodiments or elements presented herein, whether explicitly set
forth or not.
[0083] The term "activated" refers to MDG that has been blended
with water or other aqueous liquid. The term "activation" refers to
the step of blending the MDG with the aqueous liquid. For example,
an "activated MDG component" means an aqueous liquid into which a
MDG has been blended. For the purpose of this disclosure, an
activated MDG is typically an aqueous liquid substantially free of
oils or fats other than the MDG.
[0084] The term "activated assembly" refers to an assembly that
comprises at least one MDG, at least one hydrophobic protein, and
at least one lipophilic compound, wherein the assembly may have
contacted an aqueous liquid.
[0085] The terms "adult formula" and "adult nutritional product" as
used herein, are used interchangeably to refer to formulas for
generally maintaining or improving the health of an adult, and
includes those formulas designed for adults who have, are
susceptible to, or are at risk of age-related macular
degeneration.
[0086] The term "bioavailable" as used herein, unless otherwise
specified, refers to the ability of a lipophilic compound to be
absorbed from the gastrointestinal tract into lymph which will then
enter into the bloodstream of an individual such that the substance
may be absorbed into organs and tissues in the body. As the degree
of bioavailability of a compound increases, the compound becomes
more likely to enter into and remain in the bloodstream where it
may be absorbed and used by the body. As the degree of
bioavailability of a compound decreases, the compound becomes less
likely to being absorbed into lymph from the gastrointestinal tract
and would be excreted from the body before entering the
bloodstream.
[0087] The term "casein(s)" as used herein, unless otherwise
specified, should be understood to refer to those proteins in
bovine milk that will precipitate from the milk at a solution pH of
4.6 (20.degree. C.); caseins typically make up about 80% of the
protein in whole, undiluted, bovine milk. Generally, any type of
milk-sourced protein source may be utilized for the nutritional
compositions as a source of bovine beta-casein, including whole
milk, nonfat dry milk, milk protein concentrate, total milk
protein, milk protein isolate, acid casein, calcium caseinate,
sodium caseinate, magnesium caseinate, purified beta-casein, and
combinations thereof, all of which will contain beta-casein.
Beta-casein comprises around 30% of the protein contained in cow's
milk, and may be present as one of several genetic variants, the
two most prevalent being genetic variants A1 and A2. Bovine
beta-casein has further been categorized into genetic variants
including A3, B, C, D, E, F, G, H1, H2, and I.
[0088] The term "composition" as used herein, unless otherwise
specified, refers to mixtures that are suitable for enteral
administration to a subject. Compositions may be in the form of
powders, solids, semi-solids, liquids, gels, and semi-liquids.
Compositions may further comprise vitamins, minerals, and other
ingredients.
[0089] The term "dry blended" as used herein, unless otherwise
specified, refers to the mixing of components or ingredients to
form a base nutritional powder or, to the addition of a dry,
powdered or granulated component or ingredient to a base powder to
form a powdered nutritional composition.
[0090] The terms "fat," "lipid," and "oil" as used herein, unless
otherwise specified, are used interchangeably to refer to lipid
materials derived or processed from plants or animals. These terms
also include synthetic lipid materials so long as such synthetic
materials are suitable for administration to subjects as defined
herein.
[0091] The term "glycerides" as used herein refer generally to
lipophilic compounds comprising a glycerol molecule bonded to fatty
acid groups. Monoglycerides are glycerol molecules bonded to a
single fatty acid group; diglycerides are glycerol molecules bonded
to two fatty acid groups; and triglycerides are glycerol molecules
bonded to three fatty acid groups. Fats and oils comprise
glycerides, and typical fats and oils from animal, fish, algae,
vegetable, or seed sources are comprised primarily of
triglycerides.
[0092] The terms "high palmitic acid vegetable oil" or "HPAV oil"
refer to vegetable oils comprising greater than about 22% palmitic
acid, as a percentage of the total fatty acids in the HPAV oil.
Examples of HPAV oil include, but are not limited to, palm oil and
palm olein. For the purpose of this disclosure, HPAV oil may come
from sources including, but not limited to, plants, genetically
modified plants, fungi, algae, bacteria, and other single-cell
organisms. For the purpose of this disclosure, the term "HPAV oil"
excludes oils and fats that come from multicellular animal sources,
such as beef tallow, chicken fat, fish oil, or milk butterfat.
[0093] The term "human milk fortifier" as used herein, unless
otherwise specified, refers to nutritional products suitable for
mixing with breast milk or preterm infant formula or infant formula
for consumption by a preterm or term infant.
[0094] The term "infant," as used herein, unless otherwise
specified, refers to a human about 12 months of age or younger. The
term "toddler," as used herein, unless otherwise specified, refers
to a human about 12 months of age to about 3 years of age. The term
"child," as used herein, unless otherwise specified, refers to a
human about 3 years of age to about 18 years of age. The term
"adult," as used herein, unless otherwise specified, refers to a
human about 18 years of age or older.
[0095] The terms "infant formula" or "infant nutritional product"
as used herein are used interchangeably to refer to nutritional
compositions that have the proper balance of macronutrients,
micro-nutrients, and calories to provide sole or supplemental
nourishment for and generally maintain or improve the health of
infants, toddlers, or both. Infant formulas preferably comprise
nutrients in accordance with the relevant infant formula guidelines
for the targeted consumer or user population, an example of which
would be the Infant Formula Act, 21 U.S.C. Section 350(a).
[0096] The term "lipophilic compound" as used herein refers to
components that have greater solubility in organic solvents such as
ethanol, methanol, ethyl ether, acetone, chloroform, benzene, or
lipids than they have in water. Vitamin D is one example of a
lipophilic nutrient. For the purpose of this disclosure, the term
"lipophilic nutrient" may be applied to other lipophilic compounds,
including but not limited to pharmaceutical compounds.
[0097] The terms "monoglyceride and diglyceride oil" or "MDG oil"
as used herein refer to a combination of both monoglycerides and
diglycerides that are present within a premix. The premix according
to the disclosure may comprise monoglycerides and diglycerides in
an amount of monoglycerides of at least 12%, including from about
12% to about 98%, and also including from about 20% to about 80%,
including from about 20% to about 60%, and including from about 25%
to about 50% by weight of the premix (excluding the weight of the
lipophilic compound). The premix may comprise at least 12%, about
12% to about 98%, about 20% to about 80%, about 20% to about 60% or
about 25% to about 50% by weight of the mixture of monoglycerides
and diglycerides; an amount of lipophilic compound as described
above; and impurities in an amount of less than about 20%, less
than about 15%, less than about 10%, or less than about 2% by
weight. It will be recognized by one skilled in the art based on
the disclosure herein that the premix may include a small
percentage of impurities such that the mixture of monoglycerides
and diglycerides is not 100% by weight solely monoglycerides and
diglycerides. In some embodiments, the impurity may be a
triglyceride and/or free glycerol. These impurities may typically
be less than about 20%, less than about 15%, less than about 10%,
or less than about 2% by weight. In embodiments in which the premix
comprises monoglycerides and impurities, the balance of the premix
(excluding the lipophilic compound) is diglycerides.
[0098] The term "protected premix" refers to a mixture comprising a
surfactant and a MDG. The surfactant may be present within, but is
not limited to, the premix, the activated premix, and other
components of the nutritional composition. Non-limiting examples of
suitable surfactants in a protected premix include lecithin,
polyglycerol esters, and combinations thereof. The protected premix
may be made by blending the surfactant and the MDG in the presence
of heat or at ambient temperature and, in some embodiments, with
agitation. The protected premix may be added to a fat-containing
solution to form an "MDG-protected component."
[0099] The term "nutritional composition" as used herein, unless
otherwise specified, refers to nutritional powders, solids,
semi-solids, liquids, gels, and semi-liquids that comprise at least
one of protein, carbohydrate, and lipid, and are suitable for
enteral administration to a subject. The nutritional composition
may further comprise vitamins, minerals, and other ingredients, and
represent sole, primary, or supplemental sources of nutrition.
[0100] The term "pharmaceutical composition" as used herein, may
encompass any suitable form for dosing to an individual.
Non-limiting examples of dosing means include enteral and
parenteral dosage.
[0101] The term "powdered nutritional composition" as used herein,
unless otherwise specified, refers to spray dried and/or dry
blended powdered and/or agglomerated nutritional compositions
comprising a lipid soluble nutrient, such as lutein, and a mixture
of monoglycerides and diglycerides, which are reconstitutable with
an aqueous liquid, and which are suitable for oral administration
to a human.
[0102] The term "shelf life" as used herein, unless otherwise
specified, refers to the stability of the assembly within a, but
not limited to, powder, premix, activated premix, protected
activated premix, suspension, liquid, concentrated liquid, mixture,
or combination thereof.
[0103] The term "subject" as used herein refers to a mammal,
including but not limited to a human (e.g., an infant, toddler,
child or adult), a domesticated farm animal (e.g., cow, horse, or
pig), or a pet (e.g., dog or cat), who ingests the composition.
[0104] The terms "susceptible to" or "at risk of" as used herein,
unless otherwise specified, are used interchangeably to mean having
little resistance to a certain condition or disease, including
being genetically predisposed, having a family history of, and/or
having symptoms of the condition or disease.
[0105] The term "vegetable oil" as used herein refers to a fat
derived from plant, seed, fungal, or algal sources (e.g., not from
a multicellular animal). A vegetable oil may be a solid or liquid
fat at room temperature. Examples of vegetable oils include, but
are not limited to, coconut oil, fractionated coconut oil, soy oil,
corn oil, olive oil, safflower oil, high oleic safflower oil,
sunflower oil, high oleic sunflower oil, palm oil, palm kernel oil,
palm olein, canola oil, cottonseed oil, flax seed oil, hemp seed
oil, peanut oil, borage oil, algal oils, fungal oils, and
combinations thereof.
[0106] To the extent that the terms "includes," "including,"
"contains," or "containing" are used herein, they are intended to
be inclusive in a manner similar to the term "comprising" as that
term is interpreted when employed as a transitional word in a
claim. Furthermore, to the extent that the term "or" is employed
(e.g., A or B) it is intended to mean "A or B or both." When the
applicants intend to indicate "only A or B but not both" then the
term "only A or B but not both" will be employed. Thus, use of the
term "or" herein is the inclusive, and not the exclusive use. Also,
to the extent that the terms "in" or "into" are used herein, they
are intended to additionally mean "on" or "onto."
[0107] All percentages, parts and ratios as used herein, are by
weight of the total composition, unless otherwise specified. All
such weights as they pertain to listed ingredients are based on the
active level and, therefore, do not include solvents or by-products
that may be included in commercially available materials, unless
otherwise specified.
[0108] All references to singular characteristics or limitations of
the present disclosure shall include the corresponding plural
characteristic or limitation, and vice versa, unless otherwise
specified or clearly implied to the contrary by the context in
which the reference is made.
[0109] All combinations of method or process steps as used herein
may be performed in any order, unless otherwise specified or
clearly implied to the contrary by the context in which the
referenced combination is made.
[0110] The various embodiments of the nutritional compositions of
the present disclosure may also be substantially free of any
optional or selected essential ingredient or feature described
herein, provided that the remaining composition still contains all
of the required ingredients or features as described herein. In
this context, and unless otherwise specified, the term
"substantially free" means that the selected compositions contain
less than a functional amount of the optional ingredient, typically
less than 0.1% by weight, and also including zero percent by weight
of such optional or selected essential ingredient.
[0111] The nutritional and pharmaceutical compositions and
corresponding manufacturing methods of the present disclosure may
comprise, consist of, or consist essentially of the essential
elements of the disclosure as described herein, as well as any
additional or optional element described herein or which is
otherwise useful in nutritional and pharmaceutical composition
applications.
[0112] For the recitation of numeric ranges herein, each
intervening number there between with the same degree of precision
is explicitly contemplated. For example, for the range of 6-9, the
numbers 7 and 8 are contemplated in addition to 6 and 9, and for
the range 6.0-7.0, the number 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6,
6.7, 6.8, 6.9, and 7.0 are explicitly contemplated.
3. NUTRIENT COMPOSITION
[0113] Provided herein is a nutritional composition. The
nutritional composition comprises at least one protein, at least
one fat, and at least one lipophilic compound. As explained below,
the nutritional composition includes assemblies of agglomerated
compounds that increase the bioavailability of the lipophilic
compounds following consumption. The nutritional composition may
include an assembly comprising at least one lipophilic compound, at
least one protein, and at least one fat. The combination of fat
(e.g., MDG) and protein (e.g., hydrophobic protein) interacts with
lipophilic compounds, such as lutein and Vitamin D, enhancing
solubility and stability of lipophilic compounds, compared to
assemblies without MDG and hydrophobic protein. The nutritional
composition may comprise a premix, an activated premix, and a
protected premix.
[0114] The nutritional composition may be used for consumption. The
nutritional composition may be ingested by an infant and thus,
provides the infant nutrients needed for proper development and
growth. The nutritional composition may also be ingested by a
toddler or child, for proper delivery of nutrients for continued
development and growth. The nutritional composition may also be
ingested by an adult, as a nutritional supplement. The nutritional
composition can be used to deliver lipophilic compounds for the
treatment of numerous conditions, including bone health/growth, eye
health, cardiovascular health, etc.
[0115] The composition may be in any useful form. Non-limiting
product forms include: solids, semi-solids, ready-to-drink liquids,
concentrated liquids, gels, and powders. In some embodiments, the
composition may be in the form of a flowable or substantially
flowable particulate composition. In some embodiments, the
composition may be easily scooped and measured with a spoon or
similar other device, such that the composition may easily be
reconstituted by the intended user with a suitable aqueous liquid,
such as water, to form a liquid nutritional or pharmaceutical
composition for immediate oral or enteral use. In this context,
"immediate" use generally means within about 48 hours, most
typically within about 24 hours, and in some embodiments,
immediately after reconstitution.
[0116] In some embodiments, the composition may include spray dried
powders, dry blended powders, agglomerated powders, combinations
thereof, or powders prepared by other suitable methods.
[0117] In some embodiments, the composition may be contained in a
dosage element suitable for oral consumption. Suitable dosage
elements include tablets, hard gelatin capsules, starch capsules,
cellulose-based capsules, softgel capsules, and elixirs.
[0118] In some embodiments, the powdered composition may be
compressed into a tablet. In some embodiments, the powdered
composition may be included in a capsule. Capsules comprise a shell
surrounding and containing the composition. The capsule shell
dissolves or disintegrates in the digestive tract after the capsule
is ingested, thereby releasing the composition to be absorbed by
the body of the individual ingesting the capsule. Capsule shells
are typically made of gelatin, starch, cellulose, or other
components that readily dissolve or disintegrate after ingestion,
and the composition, manufacture, and use of capsule shells are
well known in the art. In some embodiments, the dosage element for
the composition is a softgel capsule. Softgel capsules are
particularly suitable for containing liquid-based ingredients, such
as nutrients dissolved, dispersed or suspended in a carrier oil.
The shell of a softgel capsule is typically made of gelatin
plasticized with glycerin and water, although vegetarian softgel
capsules made from starch or carrageenan are also available.
Softgel capsule shells are typically made and filled with the
composition in continuous processes that are known in the art.
Softgel capsules are made by manufacturers such as Catalent Pharma
Solutions, LLC (Somerset, N.J.) and Captek Softgel International
(Cerritos, Calif.).
[0119] Capsules come in a wide range of sizes, and the capsule size
should be chosen to contain an appropriate volume or weight of the
composition and, hence, an appropriate dosage of the lipophilic
compound. The capsule size can be chosen to contain at least about
0.1 grams of the composition, including from about 0.1 grams to
about 30 grams, from about 0.2 grams to about 20 grams, from about
0.25 grams to about 15 grams, from about 0.25 grams to about 10
grams, from about 0.25 grams to about 5 grams, from about 0.25 to
about 3 grams, from about 0.25 grams to about 1.5 grams, or about
0.25 to about 1.0 grams of the composition. In some embodiments,
for use with humans, the capsules contain from about 0.1 grams to
about 1.5 grams or from about 0.2 grams to about 1.0 grams of the
composition, as these capsule sizes are most convenient for most
adults and children to swallow. In some embodiments, for use with
large animals, such as domesticated farm animals, the capsules
contain from about 0.1 grams to about 30 grams or from about 1.0
grams to about 30 grams of the composition.
[0120] The nutritional composition may be formulated with
sufficient kinds and amounts of nutrients so as to provide a sole,
primary, or supplemental source of nutrition, or to provide a
specialized nutritional composition for use in individuals
afflicted with specific diseases or conditions.
[0121] The nutritional composition may have a caloric density
tailored to the nutritional needs of the ultimate user, although in
most instances the nutritional composition may comprise from about
65 to about 800 kcal/240 mL. The nutritional composition, as
discussed herein, provides a method to easily and effectively
control caloric intake to an individual (e.g., infant, toddler,
child or adult). The ability to tightly control caloric intake is
important because different individuals have different caloric
needs. The nutritional composition may comprise a caloric density
of about 75 to about 700 kcal/240 mL. The nutritional composition
may comprise a caloric density of about 100 to about 650 kcal/240
mL. For example, the nutritional composition may comprise about 65
kcal/240 mL, 70 kcal/240 mL, 75 kcal/240 mL, 80 kcal/240 mL, 85
kcal/240 mL, 90 kcal/240 mL, 95 kcal/240 mL, 100 kcal/240 mL, 105
kcal/240 mL, 110 kcal/240 mL, 115 kcal/240 mL, 120 kcal/240 mL, 125
kcal/240 mL, 130 kcal/240 mL, 135 kcal/240 mL, 140 kcal/240 mL, 145
kcal/240 mL, 150 kcal/240 mL, 155 kcal/240 mL, 160 kcal/240 mL, 165
kcal/240 mL, 170 kcal/240 mL, 175 kcal/240 mL, 180 kcal/240 mL, 185
kcal/240 mL, 190 kcal/240 mL, 195 kcal/240 mL, 200 kcal/240 mL, 205
kcal/240 mL, 210 kcal/240 mL, 215 kcal/240 mL, 220 kcal/240 mL, 225
kcal/240 mL, 230 kcal/240 mL, 235 kcal/240 mL, 240 kcal/240 mL, 245
kcal/240 mL, 250 kcal/240 mL, 255 kcal/240 mL, 260 kcal/240 mL, 265
kcal/240 mL, 270 kcal/240 mL, 275 kcal/240 mL, 280 kcal/240 mL, 285
kcal/240 mL, 290 kcal/240 mL, 295 kcal/240 mL, 300 kcal/240 mL, 305
kcal/240 mL, 310 kcal/240 mL, 315 kcal/240 mL, 320 kcal/240 mL, 325
kcal/240 mL, 330 kcal/240 mL, 335 kcal/240 mL, 340 kcal/240 mL, 345
kcal/240 mL, 350 kcal/240 mL, 355 kcal/240 mL, 360 kcal/240 mL, 365
kcal/240 mL, 370 kcal/240 mL, 375 kcal/240 mL, 380 kcal/240 mL, 385
kcal/240 mL, 390 kcal/240 mL, 395 kcal/240 mL, 400 kcal/240 mL, 405
kcal/240 mL, 410 kcal/240 mL, 415 kcal/240 mL, 420 kcal/240 mL, 425
kcal/240 mL, 430 kcal/240 mL, 435 kcal/240 mL, 440 kcal/240 mL, 445
kcal/240 mL, 450 kcal/240 mL, 455 kcal/240 mL, 460 kcal/240 mL, 465
kcal/240 mL, 470 kcal/240 mL, 475 kcal/240 mL, 480 kcal/240 mL, 485
kcal/240 mL, 490 kcal/240 mL, 495 kcal/240 mL, 500 kcal/240 mL, 550
kcal/240 mL, 600 kcal/240 mL, 650 kcal/240 mL, 700 kcal/240 mL, 750
kcal/240 mL, or 800 kcal/240 mL.
[0122] Further, non-limiting examples of the composition include
human milk fortifiers, preterm infant formulas, infant formulas,
elemental and semi-elemental formulas, pediatric formulas, toddler
formulas, adult formulas, nutritional supplements, capsules,
suppositories, sprays, drops, lotions, ointments, microcapsules,
and liposomes.
[0123] The at least one lipophilic compound may be in the
nutritional composition at about 1 .mu.g/kg to about 10 g/kg of
lipophilic compound. The at least one lipophilic compound may be in
the nutritional composition at about 1 .mu.g/kg to about 7 g/kg of
lipophilic compound. The at least one lipophilic compound may be in
the nutritional composition at about 500 .mu.g/kg to about 5 g/kg
of lipophilic compound. The at least one lipophilic compound may be
in the nutritional composition at about 1 mg/kg to about 1 g/kg of
lipophilic compound. The at least one lipophilic compound may be in
the nutritional composition at about 100 mg/kg to about 1 g/kg of
lipophilic compound. For example, the lipophilic compound may be in
the nutritional composition at about 1 .mu.g/kg, 10 .mu.g/kg, 50
.mu.g/kg, 100 .mu.g/kg, 150 .mu.g/kg, 200 .mu.g/kg, 250 .mu.g/kg,
300 .mu.g/kg, 350 .mu.g/kg, 400 .mu.g/kg, 450 .mu.g/kg, 500
.mu.g/kg, 550 .mu.g/kg, 600 .mu.g/kg, 650 .mu.g/kg, 700 .mu.g/kg,
750 .mu.g/kg, 800 .mu.g/kg, 850 .mu.g/kg, 900 .mu.g/kg, 950
.mu.g/kg, 1 mg/kg, 10 mg/kg, 50 mg/kg, 100 mg/kg, 150 mg/kg, 200
mg/kg, 250 mg/kg, 300 mg/kg, 350 mg/kg, 400 mg/kg, 450 mg/kg, 500
mg/kg, 550 mg/kg, 600 mg/kg, 650 mg/kg, 700 mg/kg, 750 mg/kg, 800
mg/kg, 850 mg/kg, 900 mg/kg, 950 mg/kg, 1 g/kg, 0.5 g/kg, 1 g/kg,
1.5 g/kg, 2 g/kg, 2.5 g/kg, 3 g/kg, 3.5 g/kg, 4 g/kg, 4.5 g/kg, 5
g/kg, 5.5 g/kg, 6 g/kg, 6.5 g/kg, 7 g/kg, 7.5 g/kg, 8 g/kg, 8.5
g/kg, 9 g/kg, 9.5 g/kg, or 10 g/kg.
[0124] The at least one fat may be in the nutritional composition
at about 10 g/kg to about 100 g/kg. The at least one fat may be in
the nutritional composition at about 20 g/kg to about 80 g/kg. The
at least one fat may be in the nutritional composition at about 30
g/kg to about 70 g/kg. The at least one fat may be in the
nutritional composition at about 40 g/kg to about 60 g/kg. For
example, the at least one fat may be in the nutritional composition
at about 10 g/kg, 10.5 g/kg, 11 g/kg, 11.5 g/kg, 12 g/kg, 12.5
g/kg, 13 g/kg, 13.5 g/kg, 14 g/kg, 14.5 g/kg, 15 g/kg, 15.5 g/kg,
16 g/kg, 16.5 g/kg, 17 g/kg, 17.5 g/kg, 18 g/kg, 18.5 g/kg, 19
g/kg, 19.5 g/kg, 20 g/kg, 20.5 g/kg, 21 g/kg, 21.5 g/kg, 22 g/kg,
22.5 g/kg, 23 g/kg, 23.5 g/kg, 24 g/kg, 24.5 g/kg, 25 g/kg, 25.5
g/kg, 26 g/kg, 26.5 g/kg, 27 g/kg, 27.5 g/kg, 28 g/kg, 28.5 g/kg,
29 g/kg, 29.5 g/kg, 30 g/kg, 30.5 g/kg, 31 g/kg, 31.5 g/kg, 32
g/kg, 32.5 g/kg, 33 g/kg, 33.5 g/kg, 34 g/kg, 34.5 g/kg, 35 g/kg,
35.5 g/kg, 36 g/kg, 36.5 g/kg, 37 g/kg, 37.5 g/kg, 38 g/kg, 38.5
g/kg, 39 g/kg, 39.5 g/kg, 40 g/kg, 40.5 g/kg, 41 g/kg, 41.5 g/kg,
42 g/kg, 42.5 g/kg, 43 g/kg, 43.5 g/kg, 44 g/kg, 44.5 g/kg, 45
g/kg, 45.5 g/kg, 46 g/kg, 46.5 g/kg, 47 g/kg, 47.5 g/kg, 48 g/kg,
48.5 g/kg, 49 g/kg, 49.5 g/kg, 50 g/kg, 50.5 g/kg, 51 g/kg, 51.5
g/kg, 52 g/kg, 52.5 g/kg, 53 g/kg, 53.5 g/kg, 54 g/kg, 54.5 g/kg,
55 g/kg, 55.5 g/kg, 56 g/kg, 56.5 g/kg, 57 g/kg, 57.5 g/kg, 58
g/kg, 58.5 g/kg, 59 g/kg, 59.5 g/kg, 60 g/kg, 60.5 g/kg, 61 g/kg,
61.5 g/kg, 62 g/kg, 62.5 g/kg, 63 g/kg, 63.5 g/kg, 64 g/kg, 64.5
g/kg, 65 g/kg, 65.5 g/kg, 66 g/kg, 66.5 g/kg, 67 g/kg, 67.5 g/kg,
68 g/kg, 68.5 g/kg, 69 g/kg, 69.5 g/kg, 70 g/kg, 70.5 g/kg, 71
g/kg, 71.5 g/kg, 72 g/kg, 72.5 g/kg, 73 g/kg, 73.5 g/kg, 74 g/kg,
74.5 g/kg, 75 g/kg, 75.5 g/kg, 76 g/kg, 76.5 g/kg, 77 g/kg, 77.5
g/kg, 78 g/kg, 78.5 g/kg, 79 g/kg, 79.5 g/kg, 80 g/kg, 80.5 g/kg,
81 g/kg, 81.5 g/kg, 82 g/kg, 82.5 g/kg, 83 g/kg, 83.5 g/kg, 84
g/kg, 84.5 g/kg, 85 g/kg, 85.5 g/kg, 86 g/kg, 86.5 g/kg, 87 g/kg,
87.5 g/kg, 88 g/kg, 88.5 g/kg, 89 g/kg, 89.5 g/kg, 90 g/kg, 90.5
g/kg, 91 g/kg, 91.5 g/kg, 92 g/kg, 92.5 g/kg, 93 g/kg, 93.5 g/kg,
94 g/kg, 94.5 g/kg, 95 g/kg, 95.5 g/kg, 96 g/kg, 96.5 g/kg, 97
g/kg, 97.5 g/kg, 98 g/kg, 98.5 g/kg, 99 g/kg, 99.5 g/kg, or 100
g/kg.
[0125] The at least one protein may be in the nutritional
composition at about 10 g/kg to about 100 g/kg. The at least one
protein may be in the nutritional composition at about 20 g/kg to
about 80 g/kg. The at least one protein may be in the nutritional
composition at about 30 g/kg to about 70 g/kg. The at least one
protein may be in the nutritional composition at about 40 g/kg to
about 60 g/kg. For example, the at least one protein may be in the
nutritional composition at about 10 g/kg, 10.5 g/kg, 11 g/kg, 11.5
g/kg, 12 g/kg, 12.5 g/kg, 13 g/kg, 13.5 g/kg, 14 g/kg, 14.5 g/kg,
15 g/kg, 15.5 g/kg, 16 g/kg, 16.5 g/kg, 17 g/kg, 17.5 g/kg, 18
g/kg, 18.5 g/kg, 19 g/kg, 19.5 g/kg, 20 g/kg, 20.5 g/kg, 21 g/kg,
21.5 g/kg, 22 g/kg, 22.5 g/kg, 23 g/kg, 23.5 g/kg, 24 g/kg, 24.5
g/kg, 25 g/kg, 25.5 g/kg, 26 g/kg, 26.5 g/kg, 27 g/kg, 27.5 g/kg,
28 g/kg, 28.5 g/kg, 29 g/kg, 29.5 g/kg, 30 g/kg, 30.5 g/kg, 31
g/kg, 31.5 g/kg, 32 g/kg, 32.5 g/kg, 33 g/kg, 33.5 g/kg, 34 g/kg,
34.5 g/kg, 35 g/kg, 35.5 g/kg, 36 g/kg, 36.5 g/kg, 37 g/kg, 37.5
g/kg, 38 g/kg, 38.5 g/kg, 39 g/kg, 39.5 g/kg, 40 g/kg, 40.5 g/kg,
41 g/kg, 41.5 g/kg, 42 g/kg, 42.5 g/kg, 43 g/kg, 43.5 g/kg, 44
g/kg, 44.5 g/kg, 45 g/kg, 45.5 g/kg, 46 g/kg, 46.5 g/kg, 47 g/kg,
47.5 g/kg, 48 g/kg, 48.5 g/kg, 49 g/kg, 49.5 g/kg, 50 g/kg, 50.5
g/kg, 51 g/kg, 51.5 g/kg, 52 g/kg, 52.5 g/kg, 53 g/kg, 53.5 g/kg,
54 g/kg, 54.5 g/kg, 55 g/kg, 55.5 g/kg, 56 g/kg, 56.5 g/kg, 57
g/kg, 57.5 g/kg, 58 g/kg, 58.5 g/kg, 59 g/kg, 59.5 g/kg, 60 g/kg,
60.5 g/kg, 61 g/kg, 61.5 g/kg, 62 g/kg, 62.5 g/kg, 63 g/kg, 63.5
g/kg, 64 g/kg, 64.5 g/kg, 65 g/kg, 65.5 g/kg, 66 g/kg, 66.5 g/kg,
67 g/kg, 67.5 g/kg, 68 g/kg, 68.5 g/kg, 69 g/kg, 69.5 g/kg, 70
g/kg, 70.5 g/kg, 71 g/kg, 71.5 g/kg, 72 g/kg, 72.5 g/kg, 73 g/kg,
73.5 g/kg, 74 g/kg, 74.5 g/kg, 75 g/kg, 75.5 g/kg, 76 g/kg, 76.5
g/kg, 77 g/kg, 77.5 g/kg, 78 g/kg, 78.5 g/kg, 79 g/kg, 79.5 g/kg,
80 g/kg, 80.5 g/kg, 81 g/kg, 81.5 g/kg, 82 g/kg, 82.5 g/kg, 83
g/kg, 83.5 g/kg, 84 g/kg, 84.5 g/kg, 85 g/kg, 85.5 g/kg, 86 g/kg,
86.5 g/kg, 87 g/kg, 87.5 g/kg, 88 g/kg, 88.5 g/kg, 89 g/kg, 89.5
g/kg, 90 g/kg, 90.5 g/kg, 91 g/kg, 91.5 g/kg, 92 g/kg, 92.5 g/kg,
93 g/kg, 93.5 g/kg, 94 g/kg, 94.5 g/kg, 95 g/kg, 95.5 g/kg, 96
g/kg, 96.5 g/kg, 97 g/kg, 97.5 g/kg, 98 g/kg, 98.5 g/kg, 99 g/kg,
99.5 g/kg, or 100 g/kg.
[0126] The ratios and amounts between the different compounds are
critical in the resultant assemblies formed during composition. In
order to form stable, high molecular weight assemblies, the w/w
ratio of MDG to lipophilic compound may be in the nutritional
composition at about 12,000:1 to about 1:1. The w/w ratio of MDG to
lipophilic compound may be in the nutritional composition at about
10,000:1 to about 1:1. The w/w ratio of MDG to lipophilic compound
may be in the nutritional composition at about 1,000:1 to about
1:1. The w/w ratio of MDG to lipophilic compound may be in the
nutritional composition at about 100:1 to about 1:1. For example,
w/w ratio of MDG to lipophilic compound may be in the nutritional
composition at about 12000:1, 11500:1, 11000:1, 10500:1, 10000:1,
9500:1, 9000:1, 8500:1, 8000:1, 7500:1, 7000:1, 6500:1, 6000:1,
5500:1, 5000:1, 4900:1, 4800:1, 4700:1, 4600:1, 4500:1, 4400:1,
4300:1, 4200:1, 4100:1, 4000:1, 3900:1, 3800:1, 3700:1, 3600:1,
3500:1, 3400:1, 3300:1, 3200:1, 3100:1, 3000:1, 2900:1, 2800:1,
2700:1, 2600:1, 2500:1, 2400:1, 2300:1, 2200:1, 2100:1, 2000:1,
1900:1, 1800:1, 1700:1, 1600:1, 1500:1, 1400:1, 1300:1, 1200:1,
1100:1, 1000:1, 950:1, 900:1, 850:1, 800:1, 750:1, 700:1, 650:1,
600:1, 550:1, 500:1, 450:1, 400:1, 350:1, 300:1, 250:1, 200:1,
150:1, 100:1, 90:1, 80:1, 70:1, 60:1, 50:1, 40:1, 30:1, 20:1, 10:1,
9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, or 1:1.
[0127] One embodiment of the nutritional composition may comprise
about 145 mg/kg to about 600 mg/kg of MDG and about 1.12 g/kg of a
lipophilic compound.
[0128] In another embodiment, the nutritional composition may
comprise about 0.1 mg/kg to about 10 mg/kg of MDG and about 20
.mu.g/kg of vitamin D.
[0129] b. Assembly
[0130] The nutritional composition may comprise an assembly. The
assembly may comprise at least one fat, at least one protein, and
at least one lipophilic compound. The assembly may comprise MDG, at
least one hydrophobic protein, and at least one lipophilic
compound. It is well known in the art that lipophilic compounds
have limited aqueous solubility, resulting in their decreased
bioavailability when consumed as part of a nutritional composition.
The assembly increases the aqueous solubility of lipophilic
compounds. Accordingly, the assembly increases the bioavailability
of the lipophilic compounds, and their resultant absorption within
the gastrointestinal (GI) tract, relative to controls without MDG
and hydrophobic protein.
[0131] The assembly is water soluble, and the interactions between
MDG, the at least one hydrophobic protein and at least one
lipophilic compound allow for stability of the assembly to
withstand high-speed centrifugal forces. For example, the
nutritional composition may be centrifuged at 1.times.g,
50.times.g, 100.times.g, 500.times.g, 1000.times.g, 1500.times.g,
2000.times.g, 2500.times.g, 3000.times.g, 3500.times.g,
4000.times.g, 4500.times.g, 5000.times.g, 5500.times.g,
6000.times.g, 6500.times.g, 7000.times.g, 7500.times.g,
8000.times.g, 8500.times.g, 9000.times.g, 9500.times.g,
10000.times.g, 10500.times.g, 11000.times.g, 11500.times.g,
12000.times.g, 12500.times.g, 13000.times.g, 13500.times.g,
14000.times.g, 14500.times.g, 15000.times.g, 15500.times.g,
16000.times.g, 16500.times.g, 17000.times.g, 17500.times.g,
18000.times.g, 18500.times.g, 19000.times.g, 19500.times.g,
20000.times.g, 20500.times.g, 21000.times.g, 21500.times.g,
22000.times.g, 22500.times.g, 23000.times.g, 23500.times.g,
24000.times.g, 24500.times.g, 25000.times.g, 25500.times.g,
26000.times.g, 26500.times.g, 27000.times.g, 27500.times.g,
28000.times.g, 28500.times.g, 29000.times.g, 29500.times.g,
30000.times.g, 30500.times.g, 31000.times.g, 31500.times.g,
32000.times.g, 32500.times.g, 33000.times.g, 33500.times.g,
34000.times.g, 34500.times.g, 35000.times.g, 35500.times.g,
36000.times.g, 36500.times.g, 37000.times.g, 37500.times.g,
38000.times.g, 38500.times.g, 39000.times.g, 39500.times.g,
40000.times.g, 40500.times.g, 41000.times.g, 41500.times.g,
42000.times.g, 42500.times.g, 43000.times.g, 43500.times.g,
44000.times.g, 44500.times.g, 45000.times.g, 45500.times.g,
46000.times.g, 46500.times.g, 47000.times.g, 47500.times.g,
48000.times.g, 48500.times.g, 49000.times.g, 49500.times.g,
50000.times.g, 50500.times.g, 51000.times.g, 51500.times.g,
52000.times.g, 52500.times.g, 53000.times.g, 53500.times.g,
54000.times.g, 54500.times.g, 55000.times.g, 55500.times.g,
56000.times.g, 56500.times.g, 57000.times.g, 57500.times.g,
58000.times.g, 58500.times.g, 59000.times.g, 59500.times.g,
60000.times.g, 60500.times.g, 61000.times.g, 61500.times.g,
62000.times.g, 62500.times.g, 63000.times.g, 63500.times.g,
64000.times.g, 64500.times.g, 65000.times.g, 65500.times.g,
66000.times.g, 66500.times.g, 67000.times.g, 67500.times.g,
68000.times.g, 68500.times.g, 69000.times.g, 69500.times.g,
70000.times.g, 70500.times.g, 71000.times.g, 71500.times.g,
72000.times.g, 72500.times.g, 73000.times.g, 73500.times.g,
74000.times.g, 74500.times.g, 75000.times.g, 75500.times.g,
76000.times.g, 76500.times.g, 77000.times.g, 77500.times.g,
78000.times.g, 78500.times.g, 79000.times.g, 79500.times.g,
80000.times.g, 80500.times.g, 81000.times.g, 81500.times.g,
82000.times.g, 82500.times.g, 83000.times.g, 83500.times.g,
84000.times.g, 84500.times.g, 85000.times.g, 85500.times.g,
86000.times.g, 86500.times.g, 87000.times.g, 87500.times.g,
88000.times.g, 88500.times.g, 89000.times.g, 89500.times.g,
90000.times.g, 90500.times.g, 91000.times.g, 91500.times.g,
92000.times.g, 92500.times.g, 93000.times.g, 93500.times.g,
94000.times.g, 94500.times.g, 95000.times.g, 95500.times.g,
96000.times.g, 96500.times.g, 97000.times.g, 97500.times.g,
98000.times.g, 98500.times.g, 99000.times.g, 99500.times.g, or
100000.times.g.
[0132] In some embodiments, the assembly is still water soluble
after centrifugation at about 31,000.times.g for at least 1 hour at
20.degree. C. In some embodiments, at least 5% of the assembly
remains in the aqueous phase after centrifugation.
[0133] The nutritional composition may be spun down using a
centrifuge to assess the amount of fat, protein and lipophilic
compound remaining in the aqueous phase. It is hypothesized,
without being bound to any particular theory, that the fat and
lipophilic compound that remain in the aqueous phase following
centrifugation are likely associated with the assembly due to their
limited aqueous solubility. It should be noted that different
centrifugal speeds and times may be used to spin down the
nutritional composition. Accordingly, one skilled in the art would
appreciate that different centrifugal conditions may provide
different amounts of the assembly, and corresponding compounds,
within the aqueous phase, but will still allow quantification of
desired compound.
[0134] In some embodiments, the details of centrifugation are as
follows: speed of centrifugation: 1000.times.g, centrifugation
time: 15 min, centrifugation temperature: 20.degree. C., centrifuge
tubes: Cellstar tube 50 ml from Greiner bio-one, tube diameter: 23
mm, amount of product in tube: 45 ml, centrifuge: type 5810 R from
Eppendorf, and aqueous phase isolation by syringe through the cream
layer.
[0135] In some embodiments, the details of centrifugation are as
follows: speed of centrifugation: 4500.times.g, centrifugation
time: 15 min, centrifugation temperature: 20.degree. C., centrifuge
tubes: Cellstar tube 50 ml from Greiner bio-one, tube diameter: 23
mm, amount of product in tube: 45 ml, centrifuge: type 5810 R from
Eppendorf, and aqueous phase isolation by syringe through the cream
layer.
[0136] In some embodiments, the details of centrifugation are as
follows: speed of centrifugation: 100,000.times.g, centrifugation
time: 1 h, centrifugation temperature: 20.degree. C., centrifuge
tubes: Ultra Clear Centrifuge tubes from Beckmann Coulter, tube
size: 14.times.89 mm, amount of product in tube: 11.3 g,
centrifuge: type L-90K from Beckmann Coulter, aqueous phase
isolation by freezing the tube with content in liquid nitrogen and
cutting the part with the serum phase including the tube wall
followed by thawing the content of this part of the tube.
[0137] In some embodiments, the centrifuge procedure may be
performed as follows, approximately 35 g of nutritional composition
is transferred into a 50-mL polypropylene centrifuge tube (capable
of withstanding 100,000.times.g; e.g., VWR catalog no. 21007-290).
The tubes are capped and placed in a JA-20 rotor in a Beckman
Coulter Model Avanti J-E centrifuge. The temperature is set at
20.degree. C., and the tubes are centrifuged at 31,000.times.g
(16,000 rpm) for 1 to 4 hours. The tubes are removed from the
centrifuge, and approximately 0.6 to 1.0 g of supernatant is
transferred into a tared 10 mL volumetric flask, and the
supernatant sample weight is recorded. The supernatant sample in
the 10 mL volumetric flask is diluted to 7.0 g with Milli-Q Plus
water, and stirred for five minutes. The diluted supernatant sample
in the 10 mL volumetric flask is then diluted to 10 mL with
acetonitrile containing trifluoroacetic acid at 0.33% (v/v), and
stirred for an additional five minutes. The flask is stoppered, and
inverted repeatedly to thoroughly mix. A 1.0 to 1.5 mL aliquot is
transferred from the flask into an HPLC autosampler vial. The vial
is capped, and the aliquot is tested for the presence of different
variables within the aqueous phase, such as MDG, lutein,
hydrophobic protein, etc.
[0138] In one embodiment, the nutritional composition may be
centrifuged at 1,000.times.g for 1 hour at 20.degree. C. In one
embodiment, the nutritional composition may be centrifuged at
31,000.times.g for 1 hour at 20.degree. C. In one embodiment, the
nutritional composition may be centrifuged at 100,000.times.g for 1
hour at 20.degree. C.
[0139] MDG may be in the aqueous phase at about 0.001% to about 50%
of the nutritional composition, following centrifugation at
1,000.times.g for 1 hour at 20.degree. C. MDG may be in the aqueous
phase at about 0.5% to about 40% of the nutritional composition,
following centrifugation at 1,000.times.g for 1 hour at 20.degree.
C. MDG may be in the aqueous phase at about 5% to about 30% of the
nutritional composition, following centrifugation at 1,000.times.g
for 1 hour at 20.degree. C. MDG may be in the aqueous phase at
about 15% to about 25% of the nutritional composition, following
centrifugation at 1,000.times.g for 1 hour at 20.degree. C. For
example, MDG may be in the aqueous phase at about 0.001%, 0.05%,
0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%,
0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.25%,
1.5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%,
4.25%, 4.5%, 4.75%, 5%, 5.25%, 5.5%, 5.75%, 6%, 6.25%, 6.5%, 6.75%,
7%, 7.25%, 7.5%, 7.75%, 8%, 8.25%, 8.5%, 8.75%, 9%, 9.25%, 9.5%,
9.75%, 10%, 10.25%, 10.5%, 10.75%, 11%, 11.25%, 11.5%, 11.75%, 12%,
12.25%, 12.5%, 12.75%, 13%, 13.25%, 13.5%, 13.75%, 14%, 14.25%,
14.5%, 14.75%, 15%, 15.25%, 15.5%, 15.75%, 16%, 16.25%, 16.5%,
16.75%, 17%, 17.25%, 17.5%, 17.75%, 18%, 18.25%, 18.5%, 18.75%,
19%, 19.25%, 19.5%, 19.75%, 20%, 20.25%, 20.5%, 20.75%, 21%,
21.25%, 21.5%, 21.75%, 22%, 22.25%, 22.5%, 22.75%, 23%, 23.25%,
23.5%, 23.75%, 24%, 24.25%, 24.5%, 24.75%, 25%, 25.25%, 25.5%,
25.75%, 26%, 26.25%, 26.5%, 26.75%, 27%, 27.25%, 27.5%, 27.75%,
28%, 28.25%, 28.5%, 28.75%, 29%, 29.25%, 29.5%, 29.75%, 30%,
30.25%, 30.5%, 30.75%, 31%, 31.25%, 31.5%, 31.75%, 32%, 32.25%,
32.5%, 32.75%, 33%, 33.25%, 33.5%, 33.75%, 34%, 34.25%, 34.5%,
34.75%, 35%, 35.25%, 35.5%, 35.75%, 36%, 36.25%, 36.5%, 36.75%,
37%, 37.25%, 37.5%, 37.75%, 38%, 38.25%, 38.5%, 38.75%, 39%,
39.25%, 39.5%, 39.75%, 40%, 40.25%, 40.5%, 40.75%, 41%, 41.25%,
41.5%, 41.75%, 42%, 42.25%, 42.5%, 42.75%, 43%, 43.25%, 43.5%,
43.75%, 44%, 44.25%, 44.5%, 44.75%, 45%, 45.25%, 45.5%, 45.75%,
46%, 46.25%, 46.5%, 46.75%, 47%, 47.25%, 47.5%, 47.75%, 48%,
48.25%, 48.5%, 48.75%, 49%, 49.25%, 49.5%, 49.75%, or 50% of the
nutritional composition, following centrifugation at 1,000.times.g
for 1 hour at 20.degree. C.
[0140] MDG may be in the aqueous phase at about 0.001% to about 30%
of the nutritional composition, following centrifugation at
31,000.times.g for 1 hour at 20.degree. C. MDG may be in the
aqueous phase at about 0.1% to about 20% of the nutritional
composition, following centrifugation at 31,000.times.g for 1 hour
at 20.degree. C. MDG may be in the aqueous phase at about 1% to
about 5% of the nutritional composition, following centrifugation
at 31,000.times.g for 1 hour at 20.degree. C. For example, MDG may
be in the aqueous phase at about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%,
0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%,
0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.25%, 1.5%, 1.75%, 2%, 2.25%,
2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%, 4.25%, 4.5%, 4.75%, 5%,
5.25%, 5.5%, 5.75%, 6%, 6.25%, 6.5%, 6.75%, 7%, 7.25%, 7.5%, 7.75%,
8%, 8.25%, 8.5%, 8.75%, 9%, 9.25%, 9.5%, 9.75%, 10%, 10.25%, 10.5%,
10.75%, 11%, 11.25%, 11.5%, 11.75%, 12%, 12.25%, 12.5%, 12.75%,
13%, 13.25%, 13.5%, 13.75%, 14%, 14.25%, 14.5%, 14.75%, 15%,
15.25%, 15.5%, 15.75%, 16%, 16.25%, 16.5%, 16.75%, 17%, 17.25%,
17.5%, 17.75%, 18%, 18.25%, 18.5%, 18.75%, 19%, 19.25%, 19.5%,
19.75%, 20%, 20.25%, 20.5%, 20.75%, 21%, 21.25%, 21.5%, 21.75%,
22%, 22.25%, 22.5%, 22.75%, 23%, 23.25%, 23.5%, 23.75%, 24%,
24.25%, 24.5%, 24.75%, 25%, 25.25%, 25.5%, 25.75%, 26%, 26.25%,
26.5%, 26.75%, 27%, 27.25%, 27.5%, 27.75%, 28%, 28.25%, 28.5%,
28.75%, 29%, 29.25%, 29.5%, 29.75%, or 30% of the nutritional
composition, following centrifugation at 31,000.times.g for 1 hour
at 20.degree. C.
[0141] MDG may be in the aqueous phase at about 0.001% to about 20%
of the nutritional composition, following centrifugation at
100,000.times.g for 1 hour at 20.degree. C. MDG may be in the
aqueous phase at about 0.01% to about 15% of the nutritional
composition, following centrifugation at 100,000.times.g for 1 hour
at 20.degree. C. MDG may be in the aqueous phase at about 0.1% to
about 10% of the nutritional composition, following centrifugation
at 100,000.times.g for 1 hour at 20.degree. C. MDG may be in the
aqueous phase at about 1% to about 5% of the nutritional
composition, following centrifugation at 100,000.times.g for 1 hour
at 20.degree. C. For example, MDG may be in the aqueous phase at
about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%,
0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%,
0.95%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%,
2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%,
3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%,
4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5%, 5.1%, 5.2%, 5.3%,
5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6%, 6.1%, 6.2%, 6.3%, 6.4%,
6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%,
7.6%, 7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%,
8.7%, 8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%,
9.8%, 9.9%, 10%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%,
10.8%, 10.9%, 11%, 11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%, 11.7%,
11.8%, 11.9%, 12%, 12.1%, 12.2%, 12.3%, 12.4%, 12.5%, 12.6%, 12.7%,
12.8%, 12.9%, 13%, 13.1%, 13.2%, 13.3%, 13.4%, 13.5%, 13.6%, 13.7%,
13.8%, 13.9%, 14%, 14.1%, 14.2%, 14.3%, 14.4%, 14.5%, 14.6%, 14.7%,
14.8%, 14.9%, 15%, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%,
15.8%, 15.9%, 16%, 16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%, 16.7%,
16.8%, 16.9%, 17%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%,
17.8%, 17.9%, 18%, 18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%,
18.8%, 18.9%, 19%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%,
19.8%, 19.9%, or 20% of the nutritional composition, following
centrifugation at 100,000.times.g for 1 hour at 20.degree. C.
[0142] MDG may be in the assembly at about 0.001% to about 20% of
the nutritional composition, following centrifugation at
31,000.times.g for 1 hour at 20.degree. C. MDG may be in the
assembly at about 0.01% to about 15% of the nutritional
composition, following centrifugation at 31,000.times.g for 1 hour
at 20.degree. C. MDG may be in the assembly at about 0.1% to about
10% of the nutritional composition, following centrifugation at
31,000.times.g for 1 hour at 20.degree. C. MDG may be in the
assembly at about 1% to about 5% of the nutritional composition,
following centrifugation at 31,000.times.g for 1 hour at 20.degree.
C. For example, MDG may be in the assembly at about 0.001%, 0.05%,
0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%,
0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%, 1.2%,
1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%,
2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%,
3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%,
4.6%, 4.7%, 4.8%, 4.9%, 5%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%,
5.7%, 5.8%, 5.9%, 6%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%,
6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%,
7.9%, 8%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9%,
9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 10.1%,
10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11%, 11.1%,
11.2%, 11.3%, 11.4%, 11.5%, 11.6%, 11.7%, 11.8%, 11.9%, 12%, 12.1%,
12.2%, 12.3%, 12.4%, 12.5%, 12.6%, 12.7%, 12.8%, 12.9%, 13%, 13.1%,
13.2%, 13.3%, 13.4%, 13.5%, 13.6%, 13.7%, 13.8%, 13.9%, 14%, 14.1%,
14.2%, 14.3%, 14.4%, 14.5%, 14.6%, 14.7%, 14.8%, 14.9%, 15%, 15.1%,
15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16%, 16.1%,
16.2%, 16.3%, 16.4%, 16.5%, 16.6%, 16.7%, 16.8%, 16.9%, 17%, 17.1%,
17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%, 17.9%, 18%, 18.1%,
18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19%, 19.1%,
19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8%, 19.9%, or 20% of
the nutritional composition, following centrifugation at
31,000.times.g for 1 hour at 20.degree. C.
[0143] Lutein, a lipophilic compound discussed below, may be in the
aqueous phase at about 0.001% to about 50% of the nutritional
composition, following centrifugation at 1,000.times.g for 1 hour
at 20.degree. C. Lutein may be in the aqueous phase at about 0.5%
to about 40% of the nutritional composition, following
centrifugation at 1,000.times.g for 1 hour at 20.degree. C. Lutein
may be in the aqueous phase at about 5% to about 30% of the
nutritional composition, following centrifugation at 1,000.times.g
for 1 hour at 20.degree. C. Lutein may be in the aqueous phase at
about 15% to about 25% of the nutritional composition, following
centrifugation at 1,000.times.g for 1 hour at 20.degree. C. For
example, lutein may be in the aqueous phase at about 0.001%, 0.05%,
0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%,
0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.25%,
1.5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%,
4.25%, 4.5%, 4.75%, 5%, 5.25%, 5.5%, 5.75%, 6%, 6.25%, 6.5%, 6.75%,
7%, 7.25%, 7.5%, 7.75%, 8%, 8.25%, 8.5%, 8.75%, 9%, 9.25%, 9.5%,
9.75%, 10%, 10.25%, 10.5%, 10.75%, 11%, 11.25%, 11.5%, 11.75%, 12%,
12.25%, 12.5%, 12.75%, 13%, 13.25%, 13.5%, 13.75%, 14%, 14.25%,
14.5%, 14.75%, 15%, 15.25%, 15.5%, 15.75%, 16%, 16.25%, 16.5%,
16.75%, 17%, 17.25%, 17.5%, 17.75%, 18%, 18.25%, 18.5%, 18.75%,
19%, 19.25%, 19.5%, 19.75%, 20%, 20.25%, 20.5%, 20.75%, 21%,
21.25%, 21.5%, 21.75%, 22%, 22.25%, 22.5%, 22.75%, 23%, 23.25%,
23.5%, 23.75%, 24%, 24.25%, 24.5%, 24.75%, 25%, 25.25%, 25.5%,
25.75%, 26%, 26.25%, 26.5%, 26.75%, 27%, 27.25%, 27.5%, 27.75%,
28%, 28.25%, 28.5%, 28.75%, 29%, 29.25%, 29.5%, 29.75%, 30%,
30.25%, 30.5%, 30.75%, 31%, 31.25%, 31.5%, 31.75%, 32%, 32.25%,
32.5%, 32.75%, 33%, 33.25%, 33.5%, 33.75%, 34%, 34.25%, 34.5%,
34.75%, 35%, 35.25%, 35.5%, 35.75%, 36%, 36.25%, 36.5%, 36.75%,
37%, 37.25%, 37.5%, 37.75%, 38%, 38.25%, 38.5%, 38.75%, 39%,
39.25%, 39.5%, 39.75%, 40%, 40.25%, 40.5%, 40.75%, 41%, 41.25%,
41.5%, 41.75%, 42%, 42.25%, 42.5%, 42.75%, 43%, 43.25%, 43.5%,
43.75%, 44%, 44.25%, 44.5%, 44.75%, 45%, 45.25%, 45.5%, 45.75%,
46%, 46.25%, 46.5%, 46.75%, 47%, 47.25%, 47.5%, 47.75%, 48%,
48.25%, 48.5%, 48.75%, 49%, 49.25%, 49.5%, 49.75%, or 50% of the
nutritional composition, following centrifugation at 1,000.times.g
for 1 hour at 20.degree. C.
[0144] Lutein may be in the aqueous phase at about 0.001% to about
30% of the nutritional composition, following centrifugation at
31,000.times.g for 1 hour at 20.degree. C. Lutein may be in the
aqueous phase at about 0.1% to about 20% of the nutritional
composition, following centrifugation at 31,000.times.g for 1 hour
at 20.degree. C. Lutein may be in the aqueous phase at about 1% to
about 5% of the nutritional composition, following centrifugation
at 31,000.times.g for 1 hour at 20.degree. C. For example, lutein
may be in the assembly at about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%,
0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%,
0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.25%, 1.5%, 1.75%, 2%, 2.25%,
2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%, 4.25%, 4.5%, 4.75%, 5%,
5.25%, 5.5%, 5.75%, 6%, 6.25%, 6.5%, 6.75%, 7%, 7.25%, 7.5%, 7.75%,
8%, 8.25%, 8.5%, 8.75%, 9%, 9.25%, 9.5%, 9.75%, 10%, 10.25%, 10.5%,
10.75%, 11%, 11.25%, 11.5%, 11.75%, 12%, 12.25%, 12.5%, 12.75%,
13%, 13.25%, 13.5%, 13.75%, 14%, 14.25%, 14.5%, 14.75%, 15%,
15.25%, 15.5%, 15.75%, 16%, 16.25%, 16.5%, 16.75%, 17%, 17.25%,
17.5%, 17.75%, 18%, 18.25%, 18.5%, 18.75%, 19%, 19.25%, 19.5%,
19.75%, 20%, 20.25%, 20.5%, 20.75%, 21%, 21.25%, 21.5%, 21.75%,
22%, 22.25%, 22.5%, 22.75%, 23%, 23.25%, 23.5%, 23.75%, 24%,
24.25%, 24.5%, 24.75%, 25%, 25.25%, 25.5%, 25.75%, 26%, 26.25%,
26.5%, 26.75%, 27%, 27.25%, 27.5%, 27.75%, 28%, 28.25%, 28.5%,
28.75%, 29%, 29.25%, 29.5%, 29.75%, or 30% of the nutritional
composition, following centrifugation at 31,000.times.g for 1 hour
at 20.degree. C.
[0145] Lutein may be in the aqueous phase at about 0.001% to about
20% of the nutritional composition, following centrifugation at
100,000.times.g for 1 hour at 20.degree. C. Lutein may be in the
aqueous phase at about 0.01% to about 15% of the nutritional
composition, following centrifugation at 100,000.times.g for 1 hour
at 20.degree. C. Lutein may be in the aqueous phase at about 0.1%
to about 10% of the nutritional composition, following
centrifugation at 100,000.times.g for 1 hour at 20.degree. C.
Lutein may be in the aqueous phase at about 1% to about 5% of the
nutritional composition, following centrifugation at
100,000.times.g for 1 hour at 20.degree. C. For example, lutein may
be in the aqueous phase at about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%,
0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%,
0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%,
1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%,
2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%,
3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%,
4.9%, 5%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6%,
6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%,
7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%,
8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%,
9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 10.1%, 10.2%, 10.3%,
10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11%, 11.1%, 11.2%, 11.3%,
11.4%, 11.5%, 11.6%, 11.7%, 11.8%, 11.9%, 12%, 12.1%, 12.2%, 12.3%,
12.4%, 12.5%, 12.6%, 12.7%, 12.8%, 12.9%, 13%, 13.1%, 13.2%, 13.3%,
13.4%, 13.5%, 13.6%, 13.7%, 13.8%, 13.9%, 14%, 14.1%, 14.2%, 14.3%,
14.4%, 14.5%, 14.6%, 14.7%, 14.8%, 14.9%, 15%, 15.1%, 15.2%, 15.3%,
15.4%, 15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16%, 16.1%, 16.2%, 16.3%,
16.4%, 16.5%, 16.6%, 16.7%, 16.8%, 16.9%, 17%, 17.1%, 17.2%, 17.3%,
17.4%, 17.5%, 17.6%, 17.7%, 17.8%, 17.9%, 18%, 18.1%, 18.2%, 18.3%,
18.4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19%, 19.1%, 19.2%, 19.3%,
19.4%, 19.5%, 19.6%, 19.7%, 19.8%, 19.9%, or 20% of the nutritional
composition, following centrifugation at 100,000.times.g for 1 hour
at 20.degree. C.
[0146] Lutein may be in the assembly at about 0.001% to about 20%
of the nutritional composition, following centrifugation at
31,000.times.g for 1 hour at 20.degree. C. Lutein may be in the
assembly at about 0.01% to about 15% of the nutritional
composition, following centrifugation at 31,000.times.g for 1 hour
at 20.degree. C. Lutein may be in the assembly at about 0.1% to
about 10% of the nutritional composition, following centrifugation
at 31,000.times.g for 1 hour at 20.degree. C. Lutein may be in the
assembly at about 1% to about 5% of the nutritional composition,
following centrifugation at 31,000.times.g for 1 hour at 20.degree.
C. For example, lutein may be in the assembly at about 0.001%,
0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%,
0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%,
1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%,
2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%,
3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%,
4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5%, 5.1%, 5.2%, 5.3%, 5.4%,
5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%,
6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%,
7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%,
8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%,
9.9%, 10%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%,
10.9%, 11%, 11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%, 11.7%, 11.8%,
11.9%, 12%, 12.1%, 12.2%, 12.3%, 12.4%, 12.5%, 12.6%, 12.7%, 12.8%,
12.9%, 13%, 13.1%, 13.2%, 13.3%, 13.4%, 13.5%, 13.6%, 13.7%, 13.8%,
13.9%, 14%, 14.1%, 14.2%, 14.3%, 14.4%, 14.5%, 14.6%, 14.7%, 14.8%,
14.9%, 15%, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%,
15.9%, 16%, 16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%, 16.7%, 16.8%,
16.9%, 17%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%,
17.9%, 18%, 18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%,
18.9%, 19%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8%,
19.9%, or 20% of the nutritional composition, following
centrifugation at 31,000.times.g for 1 hour at 20.degree. C.
[0147] MDG plays a critical role in the formation and stability of
the compounds associated with the assembly, wherein MDG increases
both the concentration of protein and bound lipophile that remains
in the aqueous phase following centrifugation, relative to controls
without MDG. MDG in the nutritional composition may increase the
concentration of protein in the aqueous phase by about 0.001% to
about 20%, following centrifugation at 31,000.times.g for 1 hour at
20.degree. C., compared to controls without MDG. MDG in the
nutritional composition may increase the concentration of protein
in the aqueous phase by about 0.01% to about 15%, following
centrifugation at 31,000.times.g for 1 hour at 20.degree. C.,
compared to controls without MDG. MDG in the nutritional
composition may increase the concentration of protein in the
aqueous phase by about 0.1% to about 10%, following centrifugation
at 31,000.times.g for 1 hour at 20.degree. C., compared to controls
without MDG. MDG in the nutritional composition may increase the
concentration of protein in the aqueous phase by about 1% to about
5%, following centrifugation at 31,000.times.g for 1 hour at
20.degree. C., compared to controls without MDG. For example
protein may be increased in the aqueous phase by about 0.001%,
0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%,
0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%,
1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%,
2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%,
3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%,
4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5%, 5.1%, 5.2%, 5.3%, 5.4%,
5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%,
6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%,
7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%,
8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%,
9.9%, 10%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%,
10.9%, 11%, 11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%, 11.7%, 11.8%,
11.9%, 12%, 12.1%, 12.2%, 12.3%, 12.4%, 12.5%, 12.6%, 12.7%, 12.8%,
12.9%, 13%, 13.1%, 13.2%, 13.3%, 13.4%, 13.5%, 13.6%, 13.7%, 13.8%,
13.9%, 14%, 14.1%, 14.2%, 14.3%, 14.4%, 14.5%, 14.6%, 14.7%, 14.8%,
14.9%, 15%, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%,
15.9%, 16%, 16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%, 16.7%, 16.8%,
16.9%, 17%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%,
17.9%, 18%, 18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%,
18.9%, 19%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8%,
19.9%, or 20%, following centrifugation at 31,000.times.g for 1
hour at 20.degree. C., compared to controls without MDG.
[0148] MDG in the nutritional composition may increase the bound
lipophilic compound in the aqueous phase by about 0.001% to about
20%, following centrifugation at 31,000.times.g for 1 hour at
20.degree. C., compared to controls without MDG. MDG in the
nutritional composition may increase the bound lipophilic compound
in the aqueous phase by about 0.01% to about 15%, following
centrifugation at 31,000.times.g for 1 hour at 20.degree. C.,
compared to controls without MDG. MDG in the nutritional
composition may increase the bound lipophilic compound in the
aqueous phase by about 0.1% to about 10%, following centrifugation
at 31,000.times.g for 1 hour at 20.degree. C., compared to controls
without MDG. MDG in the nutritional composition may increase the
bound lipophilic compound in the aqueous phase by about 1% to about
5%, following centrifugation at 31,000.times.g for 1 hour at
20.degree. C., compared to controls without MDG. For example, the
bound lipophilic compound may be increased in the aqueous phase and
the assembly by about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%,
0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%,
0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%,
1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%,
2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%,
3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5%,
5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6%, 6.1%,
6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%,
7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%,
8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%,
9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 10.1%, 10.2%, 10.3%, 10.4%,
10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11%, 11.1%, 11.2%, 11.3%, 11.4%,
11.5%, 11.6%, 11.7%, 11.8%, 11.9%, 12%, 12.1%, 12.2%, 12.3%, 12.4%,
12.5%, 12.6%, 12.7%, 12.8%, 12.9%, 13%, 13.1%, 13.2%, 13.3%, 13.4%,
13.5%, 13.6%, 13.7%, 13.8%, 13.9%, 14%, 14.1%, 14.2%, 14.3%, 14.4%,
14.5%, 14.6%, 14.7%, 14.8%, 14.9%, 15%, 15.1%, 15.2%, 15.3%, 15.4%,
15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16%, 16.1%, 16.2%, 16.3%, 16.4%,
16.5%, 16.6%, 16.7%, 16.8%, 16.9%, 17%, 17.1%, 17.2%, 17.3%, 17.4%,
17.5%, 17.6%, 17.7%, 17.8%, 17.9%, 18%, 18.1%, 18.2%, 18.3%, 18.4%,
18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19%, 19.1%, 19.2%, 19.3%, 19.4%,
19.5%, 19.6%, 19.7%, 19.8%, 19.9%, or 20%, following centrifugation
at 31,000.times.g for 1 hour at 20.degree. C., compared to controls
without MDG.
[0149] The size of the assembly is dependent on components that
make up the nutritional powder, such as the MDG, lipophilic
compound(s), protein(s), etc. The size of the assembly may be
critical in the bioavailability of the lipophilic compound. For
example, lutein incorporation would increase with size of the
assembly. However, assembly sizes greater than outside of the range
of the invention, may result in complications with solubility and
digestibility following consumption. The assembly size may be
measured via size exclusion chromatography techniques know within
the art and then compared to a known calibration curve. The
assembly size may also be measured by size analysis techniques
known within the art that include, but are not limited to, laser
diffraction, dynamic light scattering, sieve separation analysis
and image analysis (e.g., using a microscopic method such as light
microscopy or scanning electron microscopy). The assembly size may
be measured by a Malvern Zetasizer Nano NS. In some embodiments,
the nutritional composition may be centrifuged as described above,
and then the aqueous fraction may be analyzed by dynamic light
scattering machine. Additionally, the size of the assembly may be
measured via size exclusion chromatography (SEC), following the
centrifugation step as described above. This is performed by using
reference proteins and their SEC elution times and molecular
weight. This then can be used to estimate the size of the assembly.
By extrapolation from a reference protein plot. The size of the
assembly may be about 15 kD to about 3000 kD. The size of the
assembly may be about 100 kD to 1000 kD. The size of the assembly
may be about 100 kD to about 500 kD. The size of the assembly may
be about 60 kD to about 400 kD. For example, the size of the
assembly may be about 15 kD, 20 kD, 25 kD, 30 kD, 35 kD, 40 kD, 45
kD, 50 kD, 55 kD, 60 kD, 65 kD, 70 kD, 75 kD, 80 kD, 85 kD, 90 kD,
95 kD, 100 kD, 105 kD, 110 kD, 115 kD, 120 kD, 125 kD, 130 kD, 135
kD, 140 kD, 145 kD, 150 kD, 155 kD, 160 kD, 165 kD, 170 kD, 175 kD,
180 kD, 185 kD, 190 kD, 195 kD, 200 kD, 205 kD, 210 kD, 215 kD, 220
kD, 225 kD, 230 kD, 235 kD, 240 kD, 245 kD, 250 kD, 255 kD, 260 kD,
265 kD, 270 kD, 275 kD, 280 kD, 285 kD, 290 kD, 295 kD, 300 kD, 305
kD, 310 kD, 315 kD, 320 kD, 325 kD, 330 kD, 335 kD, 340 kD, 345 kD,
350 kD, 355 kD, 360 kD, 365 kD, 370 kD, 375 kD, 380 kD, 385 kD, 390
kD, 395 kD, 400 kD, 405 kD, 410 kD, 415 kD, 420 kD, 425 kD, 430 kD,
435 kD, 440 kD, 445 kD, 450 kD, 455 kD, 460 kD, 465 kD, 470 kD, 475
kD, 480 kD, 485 kD, 490 kD, 495 kD, 500 kD, 505 kD, 510 kD, 515 kD,
520 kD, 525 kD, 530 kD, 535 kD, 540 kD, 545 kD, 550 kD, 555 kD, 560
kD, 565 kD, 570 kD, 575 kD, 580 kD, 585 kD, 590 kD, 595 kD, 600 kD,
605 kD, 610 kD, 615 kD, 620 kD, 625 kD, 630 kD, 635 kD, 640 kD, 645
kD, 650 kD, 655 kD, 660 kD, 665 kD, 670 kD, 675 kD, 680 kD, 685 kD,
690 kD, 695 kD, 700 kD, 705 kD, 710 kD, 715 kD, 720 kD, 725 kD, 730
kD, 735 kD, 740 kD, 745 kD, 750 kD, 755 kD, 760 kD, 765 kD, 770 kD,
775 kD, 780 kD, 785 kD, 790 kD, 795 kD, 800 kD, 805 kD, 810 kD, 815
kD, 820 kD, 825 kD, 830 kD, 835 kD, 840 kD, 845 kD, 850 kD, 855 kD,
860 kD, 865 kD, 870 kD, 875 kD, 880 kD, 885 kD, 890 kD, 895 kD, 900
kD, 905 kD, 910 kD, 915 kD, 920 kD, 925 kD, 930 kD, 935 kD, 940 kD,
945 kD, 950 kD, 955 kD, 960 kD, 965 kD, 970 kD, 975 kD, 980 kD, 985
kD, 990 kD, 995 kD, 1000 kD, 1010 kD, 1020 kD, 1030 kD, 1040 kD,
1050 kD, 1060 kD, 1070 kD, 1080 kD, 1090 kD, 1100 kD, 1110 kD, 1120
kD, 1130 kD, 1140 kD, 1150 kD, 1160 kD, 1170 kD, 1180 kD, 1190 kD,
1200 kD, 1210 kD, 1220 kD, 1230 kD, 1240 kD, 1250 kD, 1260 kD, 1270
kD, 1280 kD, 1290 kD, 1300 kD, 1310 kD, 1320 kD, 1330 kD, 1340 kD,
1350 kD, 1360 kD, 1370 kD, 1380 kD, 1390 kD, 1400 kD, 1410 kD, 1420
kD, 1430 kD, 1440 kD, 1450 kD, 1460 kD, 1470 kD, 1480 kD, 1490 kD,
1500 kD, 1510 kD, 1520 kD, 1530 kD, 1540 kD, 1550 kD, 1560 kD, 1570
kD, 1580 kD, 1590 kD, 1600 kD, 1610 kD, 1620 kD, 1630 kD, 1640 kD,
1650 kD, 1660 kD, 1670 kD, 1680 kD, 1690 kD, 1700 kD, 1710 kD, 1720
kD, 1730 kD, 1740 kD, 1750 kD, 1760 kD, 1770 kD, 1780 kD, 1790 kD,
1800 kD, 1810 kD, 1820 kD, 1830 kD, 1840 kD, 1850 kD, 1860 kD, 1870
kD, 1880 kD, 1890 kD, 1900 kD, 1910 kD, 1920 kD, 1930 kD, 1940 kD,
1950 kD, 1960 kD, 1970 kD, 1980 kD, 1990 kD, 2000 kD, 2010 kD, 2020
kD, 2030 kD, 2040 kD, 2050 kD, 2060 kD, 2070 kD, 2080 kD, 2090 kD,
2100 kD, 2110 kD, 2120 kD, 2130 kD, 2140 kD, 2150 kD, 2160 kD, 2170
kD, 2180 kD, 2190 kD, 2200 kD, 2210 kD, 2220 kD, 2230 kD, 2240 kD,
2250 kD, 2260 kD, 2270 kD, 2280 kD, 2290 kD, 2300 kD, 2310 kD, 2320
kD, 2330 kD, 2340 kD, 2350 kD, 2360 kD, 2370 kD, 2380 kD, 2390 kD,
2400 kD, 2410 kD, 2420 kD, 2430 kD, 2440 kD, 2450 kD, 2460 kD, 2470
kD, 2480 kD, 2490 kD, 2500 kD, 2510 kD, 2520 kD, 2530 kD, 2540 kD,
2550 kD, 2560 kD, 2570 kD, 2580 kD, 2590 kD, 2600 kD, 2610 kD, 2620
kD, 2630 kD, 2640 kD, 2650 kD, 2660 kD, 2670 kD, 2680 kD, 2690 kD,
2700 kD, 2710 kD, 2720 kD, 2730 kD, 2740 kD, 2750 kD, 2760 kD, 2770
kD, 2780 kD, 2790 kD, 2800 kD, 2810 kD, 2820 kD, 2830 kD, 2840 kD,
2850 kD, 2860 kD, 2870 kD, 2880 kD, 2890 kD, 2900 kD, 2910 kD, 2920
kD, 2930 kD, 2940 kD, 2950 kD, 2960 kD, 2970 kD, 2980 kD, 2990 kD,
or 3000 kD.
[0150] The nutritional composition may include an assembly that
increases the bioavailability of the lipophilic compound from about
5% to about 80%, relative to controls without MDG and hydrophobic
protein. The nutritional composition may include an assembly that
increases the range of bioavailability of the lipophilic compound
from about 40% to about 55%, relative to controls without MDG and
hydrophobic protein. The nutritional composition may include an
assembly that increases the range bioavailability of the lipophilic
compound from about 70% to about 75%, relative to controls without
MDG and hydrophobic protein. The bioavailability of the lipophilic
compound is dependent on the formation of the assemblies of the
nutritional composition, wherein MDG and hydrophobic protein are
comprised within the assembly. The bioavailability of the assembly
may be assessed via lymphatic analysis in rats. Specifically, rats
may be fasted overnight prior to surgery, and under anesthesia, a
laparotomy may be performed, and the intestinal lymph duct may be
cannulated according to the procedure of Tso et al., "The
Absorption of Lipid and Lipoprotein Synthesis," Lipid Research
Methodology, Chapter 5: 191-216 (1984) Alan R. Liss, Inc., NY,
N.Y., hereby incorporated by reference to the extent consistent
herewith. The superior mesenteric artery is isolated, but not
occluded. A silicon infusion tube (1.6 mm OD) is placed in the
stomach for future infusion of a test sample. The fundic incision
is closed by a purse string suture. Rats are allowed to recover for
24 hours before infusion begins. Animals are intragastrically
infused 24 hours after surgery with 3 ml of nutritional composition
that delivers lipophilic compound. Lymph are collected in
pre-cooled tubes 1 hour before the nutritional composition
infusions (fasting) and then hourly for 8 hours after initiation of
infusion. At the end of the 8 hour infusion, rats are sacrificed by
exsanguination.
[0151] The lymph lipids were extracted and analyzed for lutein
concentration using high-performance liquid chromatography with
programmed wavelength ultraviolet detection (Craft Technologies,
Wilson, N.C.). For example, the assembly may increase the
bioavailability of the lipophilic compound by about 5%, 5.5%, 6%,
6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%, 11.5%, 12%,
12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, 16.5%, 17%, 17.5%,
18%, 18.5%, 19%, 19.5%, 20%, 20.5%, 21%, 21.5%, 22%, 22.5%, 23%,
23.5%, 24%, 24.5%, 25%, 25.5%, 26%, 26.5%, 27%, 27.5%, 28%, 28.5%,
29%, 29.5%, 30%, 30.5%, 31%, 31.5%, 32%, 32.5%, 33%, 33.5%, 34%,
34.5%, 35%, 35.5%, 36%, 36.5%, 37%, 37.5%, 38%, 38.5%, 39%, 39.5%,
40%, 40.5%, 41%, 41.5%, 42%, 42.5%, 43%, 43.5%, 44%, 44.5%, 45%,
45.5%, 46%, 46.5%, 47%, 47.5%, 48%, 48.5%, 49%, 49.5%, 50%, 50.5%,
51%, 51.5%, 52%, 52.5%, 53%, 53.5%, 54%, 54.5%, 55%, 55.5%, 56%,
56.5%, 57%, 57.5%, 58%, 58.5%, 59%, 59.5%, 60%, 60.5%, 61%, 61.5%,
62%, 62.5%, 63%, 63.5%, 64%, 64.5%, 65%, 65.5%, 66%, 66.5%, 67%,
67.5%, 68%, 68.5%, 69%, 69.5%, 70%, 70.5%, 71%, 71.5%, 72%, 72.5%,
73%, 73.5%, 74%, 74.5%, 75%, 75.5%, 76%, 76.5%, 77%, 77.5%, 78%,
78.5%, 79%, 79.5%, or 80%.
[0152] (1) Activated Assembly
[0153] The assembly may be an activated assembly. The activated
assembly may be comprised in a powder, suspension, solution,
emulsion or combinations thereof. The activated assembly may
comprise MDG, at least one lipophilic compound, and at least one
hydrophobic protein. The activated assembly may be in, but is not
limited to, a premix, protected premix, and an activated premix.
The activated assembly may contact an aqueous liquid as part of the
activated premix.
[0154] (2) Premix/Protected Premix
[0155] The premix may or may not comprise the assembly. The
protected premix may or may not comprise the assembly.
[0156] (3) Activated Premix Assembly
[0157] The activated premix may comprise the assembly. The assembly
may be in a protected activated premix. The assembly in the
activated premix may comprise MDG, at least one lipophilic
compound, and at least one hydrophobic protein. The assembly in the
protected activated premix may comprise MDG, at least one
lipophilic compound, and at least one hydrophobic protein. The
activated premix comprising the assembly may be added to additional
components of the nutritional composition. The protected activated
premix comprising the assembly may be added to additional
components of the nutritional composition.
[0158] In some embodiments, the premix is contacted to a
protein-in-water slurry, to form an activated premix. This is then
added to the additional components of the nutritional composition.
In some embodiments, the premix comprising the assembly is
contacted to a protein-in-oil slurry, to form an activated premix.
This is then added to the additional components of the nutritional
composition. In some embodiments, the premix comprising the
assembly is contacted to a carbohydrate-mineral slurry and a
protein-in-oil slurry, to form an activated premix. This is then
added to the additional components of the nutritional composition.
In some embodiments, the premix comprising the assembly is
contacted to an oil slurry, to form an activated premix. This is
then added to the additional components of the nutritional
composition. In some embodiments, the premix comprising the
assembly is contacted to final blend slurry, to form an activated
premix. This is then added to the additional components of the
nutritional composition.
[0159] (4) Shelf-Life
[0160] The nutritional composition may include an assembly with an
enhanced shelf-life. The assembly may have an enhanced shelf-life
in a, but is not limited to, powder, premix, activated premix,
protected activated premix, suspension, liquid, emulsion mixture,
or combination thereof.
[0161] The nutritional composition may include an assembly that is
stable at 23.degree. C. for at least about 1 day to at least about
36 months. The nutritional composition may include an assembly that
is stable at 23.degree. C. for at least about 10 days to at least
about 36 months. The nutritional composition may include an
assembly that is stable at 23.degree. C. for at least about 6
months to at least about 12 months. The nutritional composition may
include an assembly that is stable at 23.degree. C. for at least
about 1 day to at least about 24 months. The stability of the
assembly is dependent on the combination of MDG and the hydrophobic
protein's ability to interact with the lipophilic compound, which
keeps the lipophilic compound in solution, thereby enhancing its
biocompatibility. The stability of the assembly may be investigated
by size exclusion chromatography techniques known within the art at
variable time points post formulating the nutritional composition.
The assembly may be stable at 23.degree. C. for at least about 1
day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9
days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16
days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23
days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30
days, 31 days, 2 months, 3 months, 4 months, 5 months, 6 months, 7
months, 8 months, 9 months, 10 months, 11 months, 12 months, 13
months, 14 months, 15 months, 16 months, 17 months, 18 months, 19
months, 20 months, 21 months, 22 months, 23 months, 24 months, 25
months, 26 months, 27 months, 28 months, 29 months, 30 months, 31
months, 32 months, 33 months, 34 months, 35 months, or 36
months.
[0162] c. Lipophilic Compound
[0163] The nutritional composition may comprise at least one
lipophilic compound. The lipophilic compound may be present within
the premix, activated premix, and/or the protected premix. The
lipophilic compound may be incorporated into the assembly.
Lipophilic compounds have poor aqueous solubility, resulting in
decreased bioavailability when incorporated within a nutritional
composition. However, by incorporating the lipophilic compound
within the aforementioned assembly, along with MDG and the at least
one hydrophobic protein, it is more soluble and stable. The
lipophilic compound may be a carotenoid, zeaxanthin, alpha- or
beta-cryptoxanthin, retinol, a ganglioside, a phosphoinositide, a
lipoprotein, a phospholipid, a glycolipid, a glycophospholipid, an
oil soluble vitamin such as synthetic and natural forms of vitamins
A, E, D, or K, lycopene or mixtures thereof. The lipophilic
compound may be a source of long chain polyunsaturated fatty acids
(LCPUFAs). Some examples of LCPUFAs include, but are not limited,
to eicosapentaenoic acid ("EPA"), arachidonic acid (ARA), linoleic
(18:2 n-6), .gamma.-linolenic (18:3 n-6), dihomo-.gamma.-linolenic
(20:3 n-6) acids in the n-6 pathway, .alpha.-linolenic (18:3 n-3),
stearidonic (18:4 n-3), eicosatetraenoic (20:4 n-3),
eicosapentaenoic (20:5 n-3), and DHA (22:6 n-3). The lipophilic
compound may be lutein. Additionally, the lipophilic compound may
be a mixture of different lipophilic compounds.
[0164] Suitable, non-limiting examples of lipophilic pharmaceutical
agents that may be included in the present composition may be
selected from: oil soluble drugs (e.g., immunosuppressive agents
such as Cyclosporin.TM., protease inhibitors such as Ritonavir.TM.
macrolide antibiotics and oil soluble anesthetics such as
Propofol.TM.); synthetic and natural forms of steroidal hormones
such as estrogens, estradiols, progesterone, testosterone,
cortisone, phytoestrogens, dehydroepiandrosterone (DHEA), and
growth hormones; oil soluble acids and alcohols (e.g., tartaric
acid, lactylic acid, butylated hydroxyanisole, butylated
hydroxytoluene, lignin, phytosterols, flavonoids such as quercetin
and resveratrol, and diallyl disulfides); and combinations thereof.
Additional lipophilic pharmaceutical agents may be found in U.S.
patent application Ser. No. 13/452,033, which was filed on Apr. 20,
2012 and is hereby incorporated by reference to the extent that it
is consistent with the present disclosure.
[0165] The lipophilic compound is a valuable source of nutrients
for infants, toddlers, children and adults. The lipophilic compound
may be used for treating or maintaining appropriate health, such
as, but is not limited to, bone, eye, cardiovascular, and cerebral
health.
[0166] (1) Lutein
[0167] The lipophilic compound of the nutritional composition may
be lutein. Lutein may be incorporated into the assembly.
Incorporation of lutein as part of the assembly enhances its
solubility, thus enhancing its bioavailability for treatment of
health conditions, relative to controls without MDG or hydrophobic
protein. The term "lutein" as used herein, unless otherwise
specified, refers to one or more of free lutein, lutein esters,
lutein salts, or other lutein derivatives or related structures as
described or otherwise suggested herein. Lutein or lutein sources
suitable for use in the nutritional composition of the present
disclosure include free lutein as well as esters, salts or other
derivatives or related structures thereof, including those that
conform to the formula:
##STR00001##
[0168] The above formula includes the general structure of lutein
and related derivatives or structures. Free lutein, for example,
corresponds to the formula wherein R.sub.1 and R.sub.2 are both
hydrogen, and includes cis and trans isomers thereof as well as
salts thereof, e.g., sodium, potassium.
[0169] Lutein esters suitable for use in the nutritional
composition of the present disclosure include any lutein ester of
the above formula wherein R.sub.1 and R.sub.2 are the same or
different, and are nutritionally acceptable monovalent salts,
hydrogen or an acyl residue of a carboxylic acid, provided that at
least one of R.sub.1 and R.sub.2 is an acyl residue of a carboxylic
acid. Suitable lutein esters include, as well, both cis and trans
isomers. The R.sub.1 and R.sub.2 moieties are residues of saturated
or unsaturated C.sub.1 to C.sub.22 fatty carboxylic acids,
non-limiting examples of which include formic, acetic, propionic,
butyric, valeric, caproic, caprylic, capric, lauric, myristic,
palmitic, stearic, and oleic acids.
[0170] Any natural or synthetic source of lutein is suitable for
use herein provided that such a source is also known for or
otherwise suitable for use in nutritional composition and is
compatible with the other selected ingredients within the
composition. Lutein sources may be provided as individual
ingredients or in any combination with other materials or sources,
including sources such as multivitamin premixes, mixed carotenoid
premixes, and pure lutein sources. In some embodiments, components
of the nutritional composition may inherently comprise lutein
(e.g., lutein rich oils, surfactants, etc.).
[0171] Non-limiting examples of suitable lutein sources for use
herein include FloraGLO.RTM. Crystalline Lutein, available from
Kemin Foods, Des Moines, Iowa, USA; and LUTIMAX Lutein Esters
provided by OmniActive, Mumbai, India.
[0172] In some embodiments, it may be desirable to blend lutein
with the MDG oil to form an MDG premix before blending the MDG oil
or lutein with other ingredients of the composition. To make the
MDG premix, lutein may be mixed with the MDG oil in the presence of
heat or at ambient temperature and, in some embodiments, with
agitation to allow the lutein to dissolve, disperse, or suspend
into the MDG oil.
[0173] In some embodiments, it may be desirable to form an
MDG-protected premix comprising lutein, a surfactant, and an MDG
oil before blending the MDG oil or lutein with other ingredients of
the composition. In these embodiments, lutein and a surfactant may
be mixed with the MDG oil in the presence of heat or at ambient
temperature and, in some embodiments, with agitation to allow the
lutein to dissolve, disperse, or suspend in the MDG oil.
[0174] Lutein is a good source of nutrient for infants, toddlers,
children and adults. Lutein may be used within the nutritional
composition to maintain and supplement healthy vision and eye care.
Lutein may be used within the nutritional composition to maintain
and supplement cerebral health and development.
[0175] Lutein may in the nutritional composition at about 0.01
mg/kg to about 50 mg/kg. Lutein may in the nutritional composition
at about 0.02 mg/kg to about 20 mg/kg. Lutein may in the
nutritional composition at about 0.1 mg/kg to about 20 mg/kg.
Lutein may in the nutritional composition at about 1 mg/kg to about
15 mg/kg. For example, lutein may be in the nutritional composition
at about 0.01 mg/kg, 0.02 mg/kg, 0.03 mg/kg, 0.04 mg/kg, 0.05
mg/kg, 0.1 mg/kg, 0.25 mg/kg, 0.5 mg/kg, 0.75 mg/kg, 1 mg/kg, 1.25
mg/kg, 1.5 mg/kg, 1.75 mg/kg, 2 mg/kg, 2.25 mg/kg, 2.5 mg/kg, 2.75
mg/kg, 3 mg/kg, 3.25 mg/kg, 3.5 mg/kg, 3.75 mg/kg, 4 mg/kg, 4.25
mg/kg, 4.5 mg/kg, 4.75 mg/kg, 5 mg/kg, 5.25 mg/kg, 5.5 mg/kg, 5.75
mg/kg, 6 mg/kg, 6.25 mg/kg, 6.5 mg/kg, 6.75 mg/kg, 7 mg/kg, 7.25
mg/kg, 7.5 mg/kg, 7.75 mg/kg, 8 mg/kg, 8.25 mg/kg, 8.5 mg/kg, 8.75
mg/kg, 9 mg/kg, 9.25 mg/kg, 9.5 mg/kg, 9.75 mg/kg, 10 mg/kg, 10.25
mg/kg, 10.5 mg/kg, 10.75 mg/kg, 11 mg/kg, 11.25 mg/kg, 11.5 mg/kg,
11.75 mg/kg, 12 mg/kg, 12.25 mg/kg, 12.5 mg/kg, 12.75 mg/kg, 13
mg/kg, 13.25 mg/kg, 13.5 mg/kg, 13.75 mg/kg, 14 mg/kg, 14.25 mg/kg,
14.5 mg/kg, 14.75 mg/kg, 15 mg/kg, 15.25 mg/kg, 15.5 mg/kg, 15.75
mg/kg, 16 mg/kg, 16.25 mg/kg, 16.5 mg/kg, 16.75 mg/kg, 17 mg/kg,
17.25 mg/kg, 17.5 mg/kg, 17.75 mg/kg, 18 mg/kg, 18.25 mg/kg, 18.5
mg/kg, 18.75 mg/kg, 19 mg/kg, 19.25 mg/kg, 19.5 mg/kg, 19.75 mg/kg,
20 mg/kg, 20.25 mg/kg, 20.5 mg/kg, 20.75 mg/kg, 21 mg/kg, 21.25
mg/kg, 21.5 mg/kg, 21.75 mg/kg, 22 mg/kg, 22.25 mg/kg, 22.5 mg/kg,
22.75 mg/kg, 23 mg/kg, 23.25 mg/kg, 23.5 mg/kg, 23.75 mg/kg, 24
mg/kg, 24.25 mg/kg, 24.5 mg/kg, 24.75 mg/kg, 25 mg/kg, 25.25 mg/kg,
25.5 mg/kg, 25.75 mg/kg, 26 mg/kg, 26.25 mg/kg, 26.5 mg/kg, 26.75
mg/kg, 27 mg/kg, 27.25 mg/kg, 27.5 mg/kg, 27.75 mg/kg, 28 mg/kg,
28.25 mg/kg, 28.5 mg/kg, 28.75 mg/kg, 29 mg/kg, 29.25 mg/kg, 29.5
mg/kg, 29.75 mg/kg, 30 mg/kg, 30.25 mg/kg, 30.5 mg/kg, 30.75 mg/kg,
31 mg/kg, 31.25 mg/kg, 31.5 mg/kg, 31.75 mg/kg, 32 mg/kg, 32.25
mg/kg, 32.5 mg/kg, 32.75 mg/kg, 33 mg/kg, 33.25 mg/kg, 33.5 mg/kg,
33.75 mg/kg, 34 mg/kg, 34.25 mg/kg, 34.5 mg/kg, 34.75 mg/kg, 35
mg/kg, 35.25 mg/kg, 35.5 mg/kg, 35.75 mg/kg, 36 mg/kg, 36.25 mg/kg,
36.5 mg/kg, 36.75 mg/kg, 37 mg/kg, 37.25 mg/kg, 37.5 mg/kg, 37.75
mg/kg, 38 mg/kg, 38.25 mg/kg, 38.5 mg/kg, 38.75 mg/kg, 39 mg/kg,
39.25 mg/kg, 39.5 mg/kg, 39.75 mg/kg, 40 mg/kg, 40.25 mg/kg, 40.5
mg/kg, 40.75 mg/kg, 41 mg/kg, 41.25 mg/kg, 41.5 mg/kg, 41.75 mg/kg,
42 mg/kg, 42.25 mg/kg, 42.5 mg/kg, 42.75 mg/kg, 43 mg/kg, 43.25
mg/kg, 43.5 mg/kg, 43.75 mg/kg, 44 mg/kg, 44.25 mg/kg, 44.5 mg/kg,
44.75 mg/kg, 45 mg/kg, 45.25 mg/kg, 45.5 mg/kg, 45.75 mg/kg, 46
mg/kg, 46.25 mg/kg, 46.5 mg/kg, 46.75 mg/kg, 47 mg/kg, 47.25 mg/kg,
47.5 mg/kg, 47.75 mg/kg, 48 mg/kg, 48.25 mg/kg, 48.5 mg/kg, 48.75
mg/kg, 49 mg/kg, 49.25 mg/kg, 49.5 mg/kg, 49.75 mg/kg, or 50
mg/kg.
[0176] (2) Vitamin D
[0177] The lipophilic compound of the nutritional composition may
be vitamin D. Vitamin D may be incorporated into the assembly.
Incorporation of vitamin D as part of the assembly enhances its
solubility, thus enhancing its bioavailability for treatment of
health conditions, relative to controls without MDG or hydrophobic
protein. "Vitamin D" refers to a group of lipophilic compounds, or
"vitamers," related to steroids. There are several vitamers
encompassed by the term "Vitamin D," but the most important of
these are ergocalciferol, also known as Vitamin D.sub.2:
##STR00002##
and cholecalciferol, also known as Vitamin D.sub.3:
##STR00003##
[0178] For the purpose of this disclosure, the term "Vitamin D" as
used herein, unless otherwise specified, encompasses all forms of
Vitamin D, whether as individual vitamers such as Vitamin D2 or
Vitamin D3, or as combinations of two or more vitamers.
[0179] Vitamin D may be ingested from the diet, and Vitamin D3 is
also synthesized in mammalian skin by the reaction of cholesterol
with UV radiation from sunlight. Once in the body, Vitamin D
vitamers are metabolized into other chemical forms that regulate
the concentration of calcium and phosphate in the bloodstream and
promote the healthy growth and maintenance of bone. Vitamin D may
be used within the nutritional composition to maintain and
supplement healthy bone care and maintenance. Vitamin D may be used
within the nutritional composition for functions to activate the
innate and dampen the adaptive immune systems and assist in
cognitive development.
[0180] Any natural or synthetic source of Vitamin D is suitable for
use in the composition disclosed herein, provided that such a
source is suitable for use in ingestible the composition and is
compatible with the other ingredients in the composition. Vitamin D
may be provided as an individual ingredient or in any combination
with other materials or sources, including sources such as
multivitamin premixes. For example, Vitamin D may be mixed with
other oil-soluble vitamins such as Vitamin A, Vitamin E, or Vitamin
K.
[0181] Non-limiting examples of suitable Vitamin D sources for use
herein include Vitamin D3 provided by BASF Corporation (Florham
Park, N.J., USA), Vitamin D3 provided by Fermenta Biotech Ltd.
(Kullu, Himachal Pradesh, India), or Quali.RTM.-D provided by DSM
Nutritional Products AG (Kaiseraugst, Switzerland).
[0182] Vitamin D may be in the nutritional composition at about 1
.mu.g/kg to about 30 .mu.g/kg. Vitamin D may be in the nutritional
composition at about 10 .mu.g/kg to about 30 .mu.g/kg. Vitamin D
may be in the nutritional composition at about 5 .mu.g/kg to about
25 .mu.g/kg. Vitamin D may be in the nutritional composition at
about 10 .mu.g/kg to about 20 .mu.g/kg. For example, vitamin D may
be in the nutritional composition at about 1 .mu.g/kg, 1.2
.mu.g/kg, 1.4 .mu.g/kg, 1.6 .mu.g/kg, 1.8 .mu.g/kg, 2 .mu.g/kg, 2.2
.mu.g/kg, 2.4 .mu.g/kg, 2.6 .mu.g/kg, 2.8 .mu.g/kg, 3 .mu.g/kg, 3.2
.mu.g/kg, 3.4 .mu.g/kg, 3.6 .mu.g/kg, 3.8 .mu.g/kg, 4 .mu.g/kg, 4.2
.mu.g/kg, 4.4 .mu.g/kg, 4.6 .mu.g/kg, 4.8 .mu.g/kg, 5 .mu.g/kg, 5.2
.mu.g/kg, 5.4 .mu.g/kg, 5.6 .mu.g/kg, 5.8 .mu.g/kg, 6 .mu.g/kg, 6.2
.mu.g/kg, 6.4 .mu.g/kg, 6.6 .mu.g/kg, 6.8 .mu.g/kg, 7 .mu.g/kg, 7.2
.mu.g/kg, 7.4 .mu.g/kg, 7.6 .mu.g/kg, 7.8 .mu.g/kg, 8 .mu.g/kg, 8.2
.mu.g/kg, 8.4 .mu.g/kg, 8.6 .mu.g/kg, 8.8 .mu.g/kg, 9 .mu.g/kg, 9.2
.mu.g/kg, 9.4 .mu.g/kg, 9.6 .mu.g/kg, 9.8 .mu.g/kg, 10 .mu.g/kg,
10.2 .mu.g/kg, 10.4 .mu.g/kg, 10.6 .mu.g/kg, 10.8 .mu.g/kg, 11
.mu.g/kg, 11.2 .mu.g/kg, 11.4 .mu.g/kg, 11.6 .mu.g/kg, 11.8
.mu.g/kg, 12 .mu.g/kg, 12.2 .mu.g/kg, 12.4 .mu.g/kg, 12.6 .mu.g/kg,
12.8 .mu.g/kg, 13 .mu.g/kg, 13.2 .mu.g/kg, 13.4 .mu.g/kg, 13.6
.mu.g/kg, 13.8 .mu.g/kg, 14 .mu.g/kg, 14.2 .mu.g/kg, 14.4 .mu.g/kg,
14.6 .mu.g/kg, 14.8 .mu.g/kg, 15 .mu.g/kg, 15.2 .mu.g/kg, 15.4
.mu.g/kg, 15.6 .mu.g/kg, 15.8 .mu.g/kg, 16 .mu.g/kg, 16.2 .mu.g/kg,
16.4 .mu.g/kg, 16.6 .mu.g/kg, 16.8 .mu.g/kg, 17 .mu.g/kg, 17.2
.mu.g/kg, 17.4 .mu.g/kg, 17.6 .mu.g/kg, 17.8 .mu.g/kg, 18 .mu.g/kg,
18.2 .mu.g/kg, 18.4 .mu.g/kg, 18.6 .mu.g/kg, 18.8 .mu.g/kg, 19
.mu.g/kg, 19.2 .mu.g/kg, 19.4 .mu.g/kg, 19.6 .mu.g/kg, 19.8
.mu.g/kg, 20 .mu.g/kg, 20.2 .mu.g/kg, 20.4 .mu.g/kg, 20.6 .mu.g/kg,
20.8 .mu.g/kg, 21 .mu.g/kg, 21.2 .mu.g/kg, 21.4 .mu.g/kg, 21.6
.mu.g/kg, 21.8 .mu.g/kg, 22 .mu.g/kg, 22.2 .mu.g/kg, 22.4 .mu.g/kg,
22.6 .mu.g/kg, 22.8 .mu.g/kg, 23 .mu.g/kg, 23.2 .mu.g/kg, 23.4
.mu.g/kg, 23.6 .mu.g/kg, 23.8 .mu.g/kg, 24 .mu.g/kg, 24.2 .mu.g/kg,
24.4 .mu.g/kg, 24.6 .mu.g/kg, 24.8 .mu.g/kg, 25 .mu.g/kg, 25.2
.mu.g/kg, 25.4 .mu.g/kg, 25.6 .mu.g/kg, 25.8 .mu.g/kg, 26 .mu.g/kg,
26.2 .mu.g/kg, 26.4 .mu.g/kg, 26.6 .mu.g/kg, 26.8 .mu.g/kg, 27
.mu.g/kg, 27.2 .mu.g/kg, 27.4 .mu.g/kg, 27.6 .mu.g/kg, 27.8
.mu.g/kg, 28 .mu.g/kg, 28.2 .mu.g/kg, 28.4 .mu.g/kg, 28.6 .mu.g/kg,
28.8 .mu.g/kg, 29 .mu.g/kg, 29.2 .mu.g/kg, 29.4 .mu.g/kg, 29.6
.mu.g/kg, 29.8 .mu.g/kg, or 30 .mu.g/kg.
[0183] (3) Vitamin E
[0184] The lipophilic compound of the nutritional composition may
be vitamin E. Vitamin E may be incorporated into the assembly.
Incorporation of vitamin E as part of the assembly enhances its
solubility, thus enhancing its bioavailability for treatment of
health conditions, relative to controls without MDG or hydrophobic
protein. Vitamin E may be used within the nutritional composition
to provide antioxidants and maintain and supplement cardiovascular
health. Vitamin E may also be referred to as tocopherol.
Tocopherols are available in four forms, alpha, beta, gamma, and
delta, which differ in the number and position of the methyl groups
on the chroman ring (see the table 1 below). Further, tocopherols
may exist in a number of stereoisomeric forms depending on the
chirality of the phytyl tail. Of the alpha tocopherols, RRR-alpha
tocopherol (also referred to as "natural vitamin E") has the
greatest biological activity and is reported to be the dominant
form of the alpha tocopherol in the brain. In one aspect, the
composition may comprise RRR-alpha tocopherol, which means that the
composition is either formulated with the addition of RRR-alpha
tocopherol or are otherwise prepared so as to contain RRR-alpha
tocopherol. As used herein, the term "RRR-alpha tocopherol" refers
to both exogenous sources and inherent sources of free RRR-alpha
tocopherol and RRR-alpha tocopherol esters such as RRR alpha
tocopherol acetate that are present in a composition. Inherent
sources include RRR-alpha tocopherol that is inherently present in
components that are present in a nutritional composition and may
include for example, various oils and fats. Exogenous sources of
RRR-alpha tocopherol include RRR-alpha tocopherol acetate that is
added to the nutritional composition not as part of another
component. Any source of RRR-alpha tocopherol is suitable for use
herein provided that the finished product contains RRR-alpha
tocopherol.
[0185] RRR-alpha tocopherol is a single stereoisomer whereas
synthetic vitamin E (all-rac-alpha tocopherol or tocopherol
acetate) is an equimolar mixture of eight isomers, only one of
which is RRR-alpha tocopherol. The fact that the dominant form of
alpha tocopherol is RRR alpha tocopherol (based on animal studies)
strongly suggests that the other seven chiral isomers must be
absorbed at a lower rate by the brain or oxidized at a faster rate.
Cholesterol is a major component of myelin, and it is likely that
stimulated cholesterol synthesis may stimulate newborn infant
neuron myelination.
##STR00004##
TABLE-US-00001 TABLE 1 Types of Tocopherols Compound R.sub.1
R.sub.2 R.sub.3 alpha-tocopherol Me Me Me beta-tocopherol Me H Me
gamma-tocopherol H Me Me delta-tocopherol H H Me
[0186] Vitamin E may be in the nutritional composition at about 5
mg/kg to about 75 mg/kg. Vitamin E may be in the nutritional
composition at about 10 mg/kg to about 60 mg/kg. Vitamin E may be
in the nutritional composition at about 20 mg/kg to about 40 mg/kg.
For example, vitamin E may be in the nutritional composition at
about 5 mg/kg, 5.5 mg/kg, 6 mg/kg, 6.5 mg/kg, 7 mg/kg, 7.5 mg/kg, 8
mg/kg, 8.5 mg/kg, 9 mg/kg, 9.5 mg/kg, 10 mg/kg, 10.5 mg/kg, 11
mg/kg, 11.5 mg/kg, 12 mg/kg, 12.5 mg/kg, 13 mg/kg, 13.5 mg/kg, 14
mg/kg, 14.5 mg/kg, 15 mg/kg, 15.5 mg/kg, 16 mg/kg, 16.5 mg/kg, 17
mg/kg, 17.5 mg/kg, 18 mg/kg, 18.5 mg/kg, 19 mg/kg, 19.5 mg/kg, 20
mg/kg, 20.5 mg/kg, 21 mg/kg, 21.5 mg/kg, 22 mg/kg, 22.5 mg/kg, 23
mg/kg, 23.5 mg/kg, 24 mg/kg, 24.5 mg/kg, 25 mg/kg, 25.5 mg/kg, 26
mg/kg, 26.5 mg/kg, 27 mg/kg, 27.5 mg/kg, 28 mg/kg, 28.5 mg/kg, 29
mg/kg, 29.5 mg/kg, 30 mg/kg, 30.5 mg/kg, 31 mg/kg, 31.5 mg/kg, 32
mg/kg, 32.5 mg/kg, 33 mg/kg, 33.5 mg/kg, 34 mg/kg, 34.5 mg/kg, 35
mg/kg, 35.5 mg/kg, 36 mg/kg, 36.5 mg/kg, 37 mg/kg, 37.5 mg/kg, 38
mg/kg, 38.5 mg/kg, 39 mg/kg, 39.5 mg/kg, 40 mg/kg, 40.5 mg/kg, 41
mg/kg, 41.5 mg/kg, 42 mg/kg, 42.5 mg/kg, 43 mg/kg, 43.5 mg/kg, 44
mg/kg, 44.5 mg/kg, 45 mg/kg, 45.5 mg/kg, 46 mg/kg, 46.5 mg/kg, 47
mg/kg, 47.5 mg/kg, 48 mg/kg, 48.5 mg/kg, 49 mg/kg, 49.5 mg/kg, 50
mg/kg, 50.5 mg/kg, 51 mg/kg, 51.5 mg/kg, 52 mg/kg, 52.5 mg/kg, 53
mg/kg, 53.5 mg/kg, 54 mg/kg, 54.5 mg/kg, 55 mg/kg, 55.5 mg/kg, 56
mg/kg, 56.5 mg/kg, 57 mg/kg, 57.5 mg/kg, 58 mg/kg, 58.5 mg/kg, 59
mg/kg, 59.5 mg/kg, 60 mg/kg, 60.5 mg/kg, 61 mg/kg, 61.5 mg/kg, 62
mg/kg, 62.5 mg/kg, 63 mg/kg, 63.5 mg/kg, 64 mg/kg, 64.5 mg/kg, 65
mg/kg, 65.5 mg/kg, 66 mg/kg, 66.5 mg/kg, 67 mg/kg, 67.5 mg/kg, 68
mg/kg, 68.5 mg/kg, 69 mg/kg, 69.5 mg/kg, 70 mg/kg, 70.5 mg/kg, 71
mg/kg, 71.5 mg/kg, 72 mg/kg, 72.5 mg/kg, 73 mg/kg, 73.5 mg/kg, 74
mg/kg, 74.5 mg/kg, or 75 mg/kg.
[0187] (4) Docosahexaenoic Acid (DHA)
[0188] The lipophilic compound of the nutritional composition may
be DHA. DHA may be incorporated into the assembly. Incorporation of
DHA as part of the assembly enhances its solubility, thus enhancing
its bioavailability for treatment of health conditions, relative to
controls without MDG or hydrophobic protein. DHA may be used within
the nutritional composition to maintain and supplement conditions
such as healthy brain function. DHA is an omega-3 fatty acid with
22 carbons in the lipophilic chain, typically found, for example,
in cold water fish and algae:
##STR00005##
[0189] DHA is a primary structural component of the human brain,
cerebral cortex, and retina, and is an essential fatty acid for
proper development of the eyes, brain, and nervous system of
infants and babies. Breast milk contains DHA, and many infant
formulas are enriched with DHA because of its crucial role in the
growth and development of these systems in infants. There is
increasing interest in providing supplemental DHA to pregnant
women. DHA is believed to lower the risk of developing lacrimal
keratoconjunctivitis, glaucoma, and macular degeneration. DHA is
also needed for healthy brain functioning in adults. DHA deficiency
may contribute to the decline in mental functioning of healthy
older adults, and preliminary studies indicate that early
intervention with DHA supplements may improve memory and learning
in adults over 50 years of age.
[0190] Non-limiting examples of suitable DHA sources for use herein
include Life's DHA.RTM., available from DSM Nutritional Products,
Kaiseraugst, Switzerland, and Maxomega.TM. DHA, available from BASF
Pharma Ltd., Callanish, Scotland.
[0191] DHA may be in the nutritional composition at about 10 mg/kg
to about 10 g/kg. DHA may be in the nutritional composition at
about 100 mg/kg to about 5 g/kg. DHA may be in the nutritional
composition at about 500 mg/kg to about 5 g/kg. DHA may be in the
nutritional composition at about 50 mg/kg to about 1 g/kg. For
example, DHA may be in the nutritional composition at about 10
mg/kg, 20 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg, 60 mg/kg, 70 mg/kg,
80 mg/kg, 90 mg/kg, 100 mg/kg, 110 mg/kg, 120 mg/kg, 130 mg/kg, 140
mg/kg, 150 mg/kg, 160 mg/kg, 170 mg/kg, 180 mg/kg, 190 mg/kg, 200
mg/kg, 210 mg/kg, 220 mg/kg, 230 mg/kg, 240 mg/kg, 250 mg/kg, 260
mg/kg, 270 mg/kg, 280 mg/kg, 290 mg/kg, 300 mg/kg, 310 mg/kg, 320
mg/kg, 330 mg/kg, 340 mg/kg, 350 mg/kg, 360 mg/kg, 370 mg/kg, 380
mg/kg, 390 mg/kg, 400 mg/kg, 410 mg/kg, 420 mg/kg, 430 mg/kg, 440
mg/kg, 450 mg/kg, 460 mg/kg, 470 mg/kg, 480 mg/kg, 490 mg/kg, 500
mg/kg, 510 mg/kg, 520 mg/kg, 530 mg/kg, 540 mg/kg, 550 mg/kg, 560
mg/kg, 570 mg/kg, 580 mg/kg, 590 mg/kg, 600 mg/kg, 610 mg/kg, 620
mg/kg, 630 mg/kg, 640 mg/kg, 650 mg/kg, 660 mg/kg, 670 mg/kg, 680
mg/kg, 690 mg/kg, 700 mg/kg, 710 mg/kg, 720 mg/kg, 730 mg/kg, 740
mg/kg, 750 mg/kg, 760 mg/kg, 770 mg/kg, 780 mg/kg, 790 mg/kg, 800
mg/kg, 810 mg/kg, 820 mg/kg, 830 mg/kg, 840 mg/kg, 850 mg/kg, 860
mg/kg, 870 mg/kg, 880 mg/kg, 890 mg/kg, 900 mg/kg, 910 mg/kg, 920
mg/kg, 930 mg/kg, 940 mg/kg, 950 mg/kg, 960 mg/kg, 970 mg/kg, 980
mg/kg, 990 mg/kg, 1 g/kg, 1.2 g/kg, 1.4 g/kg, 1.6 g/kg, 1.8 g/kg, 2
g/kg, 2.2 g/kg, 2.4 g/kg, 2.6 g/kg, 2.8 g/kg, 3 g/kg, 3.2 g/kg, 3.4
g/kg, 3.6 g/kg, 3.8 g/kg, 4 g/kg, 4.2 g/kg, 4.4 g/kg, 4.6 g/kg, 4.8
g/kg, 5 g/kg, 5.2 g/kg, 5.4 g/kg, 5.6 g/kg, 5.8 g/kg, 6 g/kg, 6.2
g/kg, 6.4 g/kg, 6.6 g/kg, 6.8 g/kg, 7 g/kg, 7.2 g/kg, 7.4 g/kg, 7.6
g/kg, 7.8 g/kg, 8 g/kg, 8.2 g/kg, 8.4 g/kg, 8.6 g/kg, 8.8 g/kg, 9
g/kg, 9.2 g/kg, 9.4 g/kg, 9.6 g/kg, 9.8 g/kg, or 10 g/kg.
[0192] (5) Combination of DHA and Lutein
[0193] In one embodiment, it has been unexpectedly found that when
DHA and lutein are present in a composition according to the
present disclosure, the bioavailability of DHA and lutein are
improved upon ingestion over nutrient composition without the
combination of DHA and lutein. The improved bioavailability allows
DHA and lutein to be more readily absorbed into the body of the
subject. Because the bioavailability of DHA and lutein may be
increased using the presently described processes, the level of DHA
and lutein fortification required in the composition to provide the
desired nutritional benefit may be reduced. DHA and lutein may be
incorporated into the nutritional composition as recited in the
values above.
[0194] c. Fat
[0195] The nutritional composition may comprise at least one fat.
The fat may be incorporated into the assembly. The fat may be
present within the premix, activated premix, and/or the protected
premix. The fat may be a monoglyceride, a diglyceride or
combination thereof (all hereafter "MDG"). Fat allows unique
interactions with lipophilic compounds and proteins to form a water
soluble, stable assembly. Accordingly, the presence of the fat aids
in assembly formation and subsequent stability of lipophilic
containing assemblies within the composition, relative to controls
without MDG and hydrophobic protein. The fat also enhances the
bioavailability of the lipophilic compound, relative to controls
without MDG or hydrophobic protein. Fat may also increase
protection of the nutritional composition from dissolved oxygen.
Non-limiting examples of suitable fats or sources thereof, in
addition to the mixture of monoglycerides and diglycerides
discussed above, for use in the nutritional composition described
herein may be selected from the group, but is not limited to,
coconut oil, fractionated coconut oil, soy oil, corn oil, olive
oil, safflower oil, high oleic coconut oil, fractionated coconut
oil, soy oil, corn oil, olive oil, safflower oil, high oleic
safflower oil, sunflower oil, high oleic sunflower oil, palm oil,
palm kernel oil, palm olein, canola oil, marine oils, cottonseed
oil, flax seed oil, hemp seed oil, peanut oil, borage oil, algal
oils, fungal oils, MCT oil (medium chain triglycerides) and
combinations thereof.
[0196] (1) Monoglycerides and Diglycerides (MDG)
[0197] The fat of the nutritional composition may by MDG. MDG is
the combination of monoglycerides and diglycerides. MDG may be
incorporated into the assembly. Incorporation of MDG as part of the
assembly enhances the solubility of lipophilic compounds, thus
enhancing its bioavailability for treatment of health conditions,
relative to controls without MDG or hydrophobic protein. It is
hypothesized, without being bound to a particular theory, that upon
consumption, the MDG in the activated premix or the MDG-protected
premix, associate with the lipophilic compound and the hydrophobic
protein making it easier for it to be incorporated into the
micelles which are formed in the gut. Thus, the lipophilic compound
may be more efficiently delivered to the brush boarder of the
intestine and absorbed into the body. Additionally, it is
hypothesized, without being bound to a particular theory, that MDG
may interact with the hydrophobic protein and increase access of
hydrophobic domains within said protein. Accordingly, the
lipophilic compound may form more stable interactions with the
newly accessible hydrophobic domains of said protein.
[0198] MDG is a component with limited water solubility. When MDG
is associated with the at least one hydrophobic protein and the at
least one lipophilic compound, as in the assembly, its water
solubility increase significantly. It is hypothesized, without
being bound to any particular theory, that the MDG that remains in
the aqueous phase following centrifugation are likely associated
with the assembly due to its limited aqueous solubility. The
increased aqueous solubility may investigated by centrifugation, as
listed above, wherein the aqueous phase following centrifugation
exhibits a greater presence of MDG compared to controls without
MDG, lipophilic compound and at least one hydrophobic protein.
[0199] Diglycerides, which are commonly referred to as
diacylglycerols (DAGs), are glycerides consisting of two fatty acid
chains covalently bonded to a glycerol molecule through ester
linkages. Monoglycerides are normal metabolites in the body formed
during the breakdown of triglycerides and diglycerides.
Non-limiting examples of commercial sources of monoglycerides and
diglycerides include natural sources (e.g., animal (cow- or
hog-derived) or vegetable, those which may be derived from
partially hydrogenated soy bean, sunflower, safflower and coconut
oil) or synthetic sources. In some embodiments, the mixture of
monoglycerides and diglycerides may be derived from the hydrolysis
of triglycerides in safflower oil. Monoglycerides may be in the MDG
of the nutritional composition at about 0.1 wt % to about 99.9 wt
%, wherein the balance of the MDG is diglycerides. Monoglycerides
may be in the MDG of the nutritional composition at about 10 wt %
to about 80 wt %, wherein the balance of the MDG is diglycerides.
Monoglycerides may be in the MDG of the nutritional composition at
about 25 wt % to about 50 wt %, wherein the balance of the MDG is
diglycerides. For example, monoglycerides may be in the MDG of the
nutritional composition at about 0.1 wt %, 0.5 wt %, 1 wt %, 1.5 wt
%, 2 wt %, 2.5 wt %, 3 wt %, 3.5 wt %, 4 wt %, 4.5 wt %, 5 wt %,
5.5 wt %, 6 wt %, 6.5 wt %, 7 wt %, 7.5 wt %, 8 wt %, 8.5 wt %, 9
wt %, 9.5 wt %, 10 wt %, 10.5 wt %, 11 wt %, 11.5 wt %, 12 wt %,
12.5 wt %, 13 wt %, 13.5 wt %, 14 wt %, 14.5 wt %, 15 wt %, 15.5 wt
%, 16 wt %, 16.5 wt %, 17 wt %, 17.5 wt %, 18 wt %, 18.5 wt %, 19
wt %, 19.5 wt %, 20 wt %, 20.5 wt %, 21 wt %, 21.5 wt %, 22 wt %,
22.5 wt %, 23 wt %, 23.5 wt %, 24 wt %, 24.5 wt %, 25 wt %, 25.5 wt
%, 26 wt %, 26.5 wt %, 27 wt %, 27.5 wt %, 28 wt %, 28.5 wt %, 29
wt %, 29.5 wt %, 30 wt %, 30.5 wt %, 31 wt %, 31.5 wt %, 32 wt %,
32.5 wt %, 33 wt %, 33.5 wt %, 34 wt %, 34.5 wt %, 35 wt %, 35.5 wt
%, 36 wt %, 36.5 wt %, 37 wt %, 37.5 wt %, 38 wt %, 38.5 wt %, 39
wt %, 39.5 wt %, 40 wt %, 40.5 wt %, 41 wt %, 41.5 wt %, 42 wt %,
42.5 wt %, 43 wt %, 43.5 wt %, 44 wt %, 44.5 wt %, 45 wt %, 45.5 wt
%, 46 wt %, 46.5 wt %, 47 wt %, 47.5 wt %, 48 wt %, 48.5 wt %, 49
wt %, 49.5 wt %, 50 wt %, 50.5 wt %, 51 wt %, 51.5 wt %, 52 wt %,
52.5 wt %, 53 wt %, 53.5 wt %, 54 wt %, 54.5 wt %, 55 wt %, 55.5 wt
%, 56 wt %, 56.5 wt %, 57 wt %, 57.5 wt %, 58 wt %, 58.5 wt %, 59
wt %, 59.5 wt %, 60 wt %, 60.5 wt %, 61 wt %, 61.5 wt %, 62 wt %,
62.5 wt %, 63 wt %, 63.5 wt %, 64 wt %, 64.5 wt %, 65 wt %, 65.5 wt
%, 66 wt %, 66.5 wt %, 67 wt %, 67.5 wt %, 68 wt %, 68.5 wt %, 69
wt %, 69.5 wt %, 70 wt %, 70.5 wt %, 71 wt %, 71.5 wt %, 72 wt %,
72.5 wt %, 73 wt %, 73.5 wt %, 74 wt %, 74.5 wt %, 75 wt %, 75.5 wt
%, 76 wt %, 76.5 wt %, 77 wt %, 77.5 wt %, 78 wt %, 78.5 wt %, 79
wt %, 79.5 wt %, 80 wt %, 80.5 wt %, 81 wt %, 81.5 wt %, 82 wt %,
82.5 wt %, 83 wt %, 83.5 wt %, 84 wt %, 84.5 wt %, 85 wt %, 85.5 wt
%, 86 wt %, 86.5 wt %, 87 wt %, 87.5 wt %, 88 wt %, 88.5 wt %, 89
wt %, 89.5 wt %, 90 wt %, 90.5 wt %, 91 wt %, 91.5 wt %, 92 wt %,
92.5 wt %, 93 wt %, 93.5 wt %, 94 wt %, 94.5 wt %, 95 wt %, 95.5 wt
%, 96 wt %, 96.5 wt %, 97 wt %, 97.5 wt %, 98 wt %, 98.5 wt %, 99
wt %, 99.5 wt %, or 99.9%, wherein the balance of the MDG is
diglycerides.
[0200] Non-limiting examples of suitable mixtures of monoglycerides
and diglycerides for use in the composition include Capmul GMO-40
(a MDG derived from high oleic safflower oil, available from Abitec
Corporation, Columbus, Ohio), and Kirnol CE1089 (a MDG derived from
corn oil, available from BASF, Ludwigshafen, Germany), RADIAMULS MG
F038 (a MDG derived from high oleic sunflower oil, available from
Oleon, Ertvelde, Belgium).
[0201] The MDG may be in the nutritional composition at about 140
mg/kg to about 900 mg/kg. The MDG may be in the nutritional
composition at about 145 mg/kg to about 890 mg/kg. The nutritional
composition may comprise MDG at about 140 mg/kg to about 700 mg/kg.
The MDG may be in the nutritional composition at about 145 mg/kg to
about 600 mg/kg. The MDG may be in the nutritional composition at
about 140 mg/kg to about 400 mg/kg. For example, the MDG may be in
the nutritional composition at about 140 mg/kg, 145 mg/kg, 150
mg/kg, 155 mg/kg, 160 mg/kg, 165 mg/kg, 170 mg/kg, 175 mg/kg, 180
mg/kg, 185 mg/kg, 190 mg/kg, 195 mg/kg, 200 mg/kg, 205 mg/kg, 210
mg/kg, 215 mg/kg, 220 mg/kg, 225 mg/kg, 230 mg/kg, 235 mg/kg, 240
mg/kg, 245 mg/kg, 250 mg/kg, 255 mg/kg, 260 mg/kg, 265 mg/kg, 270
mg/kg, 275 mg/kg, 280 mg/kg, 285 mg/kg, 290 mg/kg, 295 mg/kg, 300
mg/kg, 305 mg/kg, 310 mg/kg, 315 mg/kg, 320 mg/kg, 325 mg/kg, 330
mg/kg, 335 mg/kg, 340 mg/kg, 345 mg/kg, 350 mg/kg, 355 mg/kg, 360
mg/kg, 365 mg/kg, 370 mg/kg, 375 mg/kg, 380 mg/kg, 385 mg/kg, 390
mg/kg, 395 mg/kg, 400 mg/kg, 405 mg/kg, 410 mg/kg, 415 mg/kg, 420
mg/kg, 425 mg/kg, 430 mg/kg, 435 mg/kg, 440 mg/kg, 445 mg/kg, 450
mg/kg, 455 mg/kg, 460 mg/kg, 465 mg/kg, 470 mg/kg, 475 mg/kg, 480
mg/kg, 485 mg/kg, 490 mg/kg, 495 mg/kg, 500 mg/kg, 505 mg/kg, 510
mg/kg, 515 mg/kg, 520 mg/kg, 525 mg/kg, 530 mg/kg, 535 mg/kg, 540
mg/kg, 545 mg/kg, 550 mg/kg, 555 mg/kg, 560 mg/kg, 565 mg/kg, 570
mg/kg, 575 mg/kg, 580 mg/kg, 585 mg/kg, 590 mg/kg, 595 mg/kg, 600
mg/kg, 605 mg/kg, 610 mg/kg, 615 mg/kg, 620 mg/kg, 625 mg/kg, 630
mg/kg, 635 mg/kg, 640 mg/kg, 645 mg/kg, 650 mg/kg, 655 mg/kg, 660
mg/kg, 665 mg/kg, 670 mg/kg, 675 mg/kg, 680 mg/kg, 685 mg/kg, 690
mg/kg, 695 mg/kg, 700 mg/kg, 705 mg/kg, 710 mg/kg, 715 mg/kg, 720
mg/kg, 725 mg/kg, 730 mg/kg, 735 mg/kg, 740 mg/kg, 745 mg/kg, 750
mg/kg, 755 mg/kg, 760 mg/kg, 765 mg/kg, 770 mg/kg, 775 mg/kg, 780
mg/kg, 785 mg/kg, 790 mg/kg, 795 mg/kg, 800 mg/kg, 805 mg/kg, 810
mg/kg, 815 mg/kg, 820 mg/kg, 825 mg/kg, 830 mg/kg, 835 mg/kg, 840
mg/kg, 845 mg/kg, 850 mg/kg, 855 mg/kg, 860 mg/kg, 865 mg/kg, 870
mg/kg, 875 mg/kg, 880 mg/kg, 885 mg/kg, 890 mg/kg, 895 mg/kg, or
900 mg/kg.
[0202] In some embodiments, the nutritional composition may
comprise about 0.1 mg/kg to about 20 mg/kg of MDG and vitamin D. In
some embodiments, the nutritional composition may comprise about
0.1 mg/kg to about 10 mg/kg of MDG and vitamin D.
[0203] (2) Palmitic Acid
[0204] The fat of the nutritional composition of the present
invention may be palmitic acid. Palmitic acid may be incorporated
into the assembly. Incorporation of palmitic acid as part of the
assembly enhances its solubility, thus enhancing its
bioavailability for treatment of health conditions, relative to
controls without MDG or hydrophobic protein. Palmitic acid may be
used within the nutritional composition to maintain and supplement
healthy nutrition, especially within infants.
[0205] Palmitic acid is a saturated fatty acid with 16 carbons in
the lipophilic chain. Palmitic acid is an important energy source
for growing infants. It is also a precursor to longer fatty acids
that are synthesized in the body. These longer fatty acids, such as
docosahexaenoic acid, are vital structural components in the brain,
eyes, and central nervous system. As a consequence, palmitic acid
is an important nutrient in the healthy development of infants and
babies.
[0206] Palmitic acid may be provided by HPAV oil, which comprises
greater than about 22% palmitic acid, as a percentage of the total
fatty acids in the HPAV oil. Examples of HPAV oil include, but are
not limited to, palm oil and palm olein.
[0207] Palmitic acid is found in palm oil and palm olein,
comprising over 30% of the total fatty acids in each oil. Palmitic
acid is also found in cocoa butter, cottonseed oil, and the oil of
some other seed plants.
[0208] Palm oil is found in the pulp of the oil palm fruit.
Palmitic acid comprises from about 43% to about 45% of the fatty
acids found in palm oil. Palm oil also comprises from about 37% to
about 40% oleic acid, making it a good source of monounsaturated
fat. Palm oil also contains the essential fatty acid linoleic acid,
which comprises from about 5% to about 11% of the fatty acids in
palm oil. Palm oil should not be confused with palm kernel oil,
which comes from the kernel (seed or pit) of the oil palm fruit.
Palm kernel oil has a very different fatty acid profile, comprising
less than about 10% palmitic acid and primarily comprising
saturated fatty acids. Therefore, palm kernel oil is not considered
a HPAV oil for the purpose of this description.
[0209] During refinement, raw palm oil is fractionated by
crystallizing and separating the liquid fraction from the solid
fraction under controlled temperatures. The liquid fraction is
called palm olein. Palm olein comprises somewhat less palmitic acid
(from about 33% to about 40%) but more oleic acid (from about 42%
to about 48%) than does palm oil.
[0210] Palmitic acid may be provided by HPAV oil, which comprises
greater than about 22% palmitic acid, as a percentage of the total
fatty acids in the HPAV oil. Examples of HPAV oil include, but are
not limited to, palm oil and palm olein.
[0211] Suppliers of suitable HPAV oils, such as palm oil or palm
olein, include: Archer Daniels Midland, Decatur, Ill., USA; Fuji
Vegetable Oil, Inc., Savannah, Ga., USA; and California Oils
Corporation, Richmond, Calif., USA.
[0212] An HPAV oil may be provided in the nutritional composition
as an individual ingredient or in any combination with other
materials or sources. For example, a HPAV oil may be provided in a
mixture with other oils such as canola oil or corn oil.
[0213] In some embodiments, it may be desirable to blend a
protected premix comprising the HPAV oil, a surfactant, and the MDG
before blending the MDG oil or the HPAV oil with other ingredients
of the nutritional composition. In these embodiments, the HPAV oil
and a surfactant are mixed with the MDG oil in the presence of heat
or at ambient temperature and, in some embodiments, with agitation
to allow the HPAV oil to dissolve or disperse in the MDG oil.
[0214] In a protected premix that contains HPAV oil, the HPAV oil
is present in an amount relative to the amount of the MDG oil that
will provide effective amounts of the HPAV oil and the MDG oil in
the infant formula. For example, the HPAV oil may be present in an
amount of 15 g, about 20 g, about 30 g, about 50 g, about 75 g,
about 100 g, about 125 g, about 150 g, about 175 g, or about 200 g,
of the HPAV per 1 g of the MDG oil in the premix. Other ratios of
the HPAV oil to the MDG oil are within the scope of the present
disclosure, so long as the HPAV oil is completely or substantially
dissolved or dispersed in the resulting premix. Based on the
disclosure herein, one skilled in the art may calculate suitable
ratios that will allow effective amounts of the HPAV oil and the
MDG oil to be incorporated into the nutritional composition.
[0215] In human breast milk, about 70% of the palmitic acid is
found in the middle, or SN-2, location of the triglyceride. The
SN-2 position is relatively protected, and the palmitic acid
remains bonded to the glycerol backbone until late in the digestive
process. Palmitic acid in the SN-2 position is thus in a form that
is easily absorbed by the intestinal tract of the infant. However,
for vegetable oils such as palm oil or palm olein, only about 9% of
the palmitic acid is found in the SN-2 position. The rest of the
palmitic acid in these oils is found in the SN-1 or SN-3 locations
on the glycerol backbone. Fatty acids in the SN-1 or SN-3 position
are less protected, and are often cleaved off the glycerol backbone
early in the digestive process. These free fatty acids then react
with calcium in the gastric fluids, forming insoluble calcium-fatty
acid soap complexes. These insoluble soap complexes, which bind up
both fatty acids and calcium, cannot be absorbed in the intestinal
tract, and are instead excreted from the body. Research has shown
that infants who are fed formulas containing palm oil or palm olein
do not absorb as much fat or calcium as infants who are fed breast
milk.
[0216] Palmitic acid may be synthetically prepared from fats that
more closely mimic the fats in breast milk. These fats are
synthesized to have significantly more palmitic acid in the SN-2
position than is found in natural vegetable oils. The nutrient
composition may have palmitic acid in the SN-2 position at about
80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, 99.degree. A or 100% of the total
palmitic acid content of the nutrient composition.
[0217] Palmitic acid may be in the nutritional composition at about
0.5 g/kg to about 35 g/kg. Palmitic acid may be in the nutritional
composition at about 1 g/kg to about 30 g/kg. Palmitic acid may be
in the nutritional composition at about 1 g/kg to about 20 g/kg.
Palmitic acid may be in the nutritional composition at about 1 g/kg
to about 10 g/kg. For example, palmitic acid may be in the
nutritional composition at about 0.5 g/kg, 0.75 g/kg, 1 g/kg, 1.25
g/kg, 1.5 g/kg, 1.75 g/kg, 2 g/kg, 2.25 g/kg, 2.5 g/kg, 2.75 g/kg,
3 g/kg, 3.25 g/kg, 3.5 g/kg, 3.75 g/kg, 4 g/kg, 4.25 g/kg, 4.5
g/kg, 4.75 g/kg, 5 g/kg, 5.25 g/kg, 5.5 g/kg, 5.75 g/kg, 6 g/kg,
6.25 g/kg, 6.5 g/kg, 6.75 g/kg, 7 g/kg, 7.25 g/kg, 7.5 g/kg, 7.75
g/kg, 8 g/kg, 8.25 g/kg, 8.5 g/kg, 8.75 g/kg, 9 g/kg, 9.25 g/kg,
9.5 g/kg, 9.75 g/kg, 10 g/kg, 10.25 g/kg, 10.5 g/kg, 10.75 g/kg, 11
g/kg, 11.25 g/kg, 11.5 g/kg, 11.75 g/kg, 12 g/kg, 12.25 g/kg, 12.5
g/kg, 12.75 g/kg, 13 g/kg, 13.25 g/kg, 13.5 g/kg, 13.75 g/kg, 14
g/kg, 14.25 g/kg, 14.5 g/kg, 14.75 g/kg, 15 g/kg, 15.25 g/kg, 15.5
g/kg, 15.75 g/kg, 16 g/kg, 16.25 g/kg, 16.5 g/kg, 16.75 g/kg, 17
g/kg, 17.25 g/kg, 17.5 g/kg, 17.75 g/kg, 18 g/kg, 18.25 g/kg, 18.5
g/kg, 18.75 g/kg, 19 g/kg, 19.25 g/kg, 19.5 g/kg, 19.75 g/kg, 20
g/kg, 20.25 g/kg, 20.5 g/kg, 20.75 g/kg, 21 g/kg, 21.25 g/kg, 21.5
g/kg, 21.75 g/kg, 22 g/kg, 22.25 g/kg, 22.5 g/kg, 22.75 g/kg, 23
g/kg, 23.25 g/kg, 23.5 g/kg, 23.75 g/kg, 24 g/kg, 24.25 g/kg, 24.5
g/kg, 24.75 g/kg, 25 g/kg, 25.25 g/kg, 25.5 g/kg, 25.75 g/kg, 26
g/kg, 26.25 g/kg, 26.5 g/kg, 26.75 g/kg, 27 g/kg, 27.25 g/kg, 27.5
g/kg, 27.75 g/kg, 28 g/kg, 28.25 g/kg, 28.5 g/kg, 28.75 g/kg, 29
g/kg, 29.25 g/kg, 29.5 g/kg, 29.75 g/kg, 30 g/kg, 30.25 g/kg, 30.5
g/kg, 30.75 g/kg, 31 g/kg, 31.25 g/kg, 31.5 g/kg, 31.75 g/kg, 32
g/kg, 32.25 g/kg, 32.5 g/kg, 32.75 g/kg, 33 g/kg, 33.25 g/kg, 33.5
g/kg, 33.75 g/kg, 34 g/kg, 34.25 g/kg, 34.5 g/kg, 34.75 g/kg, or 35
g/kg.
[0218] d. Protein
[0219] The nutritional composition may comprise at least one
protein. The one protein may be incorporated into the assembly. The
protein may be present within the activated premix and/or the
protected premix. The protein uniquely interacts with the
lipophilic compound and fat in the nutritional composition to form
the water soluble, stable assembly discussed above. Accordingly,
the presence of the protein aids in assembly formation and
subsequent stability of lipophilic containing assemblies within the
composition. The protein also enhances the bioavailability of the
lipophilic compound, relative to controls without fat (e.g., MDG)
or hydrophobic protein.
[0220] The protein may be intact protein, hydrolyzed protein or
combinations thereof. The protein may be a hydrophobic protein.
Non-limiting examples of suitable protein or sources thereof for
use in the nutritional composition include partially hydrolyzed
(degree of hydrolysis less than 25%) or non-hydrolyzed proteins
(e.g., intact) or protein sources, which may be derived from any
known or otherwise suitable source such as milk (e.g., casein,
whey), animal (e.g., meat, fish), cereal (e.g., rice, corn) or
combinations thereof. Non-limiting examples of such proteins
include milk protein isolates, milk protein concentrates as
described herein, such as whey protein concentrates, casein protein
isolates, whey protein, caseinates, whole cow's milk, partially or
completely defatted milk, soy protein isolates, soy protein
concentrates, and so forth.
[0221] In some embodiments, the nutritional composition includes a
protein component that consists of only intact and/or partially
hydrolyzed protein; that is, the protein component is free of any
protein that has a degree of hydrolysis of about 25% or more. In
this context, the term "partially hydrolyzed protein" is used to
refer to proteins having a degree of hydrolysis of less than about
25%, including less than about 20%, including less than about 15%,
including less than about 10%, and including proteins having a
degree of hydrolysis of less than about 5%. The degree of
hydrolysis is the extent to which peptide bonds are broken by a
hydrolysis method. The degree of protein hydrolysis for purposes of
characterizing the partially hydrolyzed protein component of these
embodiments is easily determined by one of ordinary skill in the
composition arts by quantifying the amino nitrogen to total
nitrogen ratio (AN/TN) of the protein component of the selected
composition. The amino nitrogen component is quantified by USP
titration methods for determining amino nitrogen content, while the
total nitrogen component is determined by the Tecator Kjeldahl
method, all of which are well known methods to one of ordinary
skill in the analytical chemistry art.
[0222] (1) Hydrophobic Protein
[0223] The protein of the nutritional composition may comprise at
least one hydrophobic protein. The hydrophobic protein may be
incorporated into the assembly. Incorporation of the hydrophobic
protein as part of the assembly enhances the solubility of
lipophilic compounds, thus enhancing its bioavailability of the
lipophilic compounds for treatment of health conditions, relative
to controls without MDG or hydrophobic protein. It is hypothesized,
without being bound to any particular theory, that the lipophilic
compound's aqueous solubility is enhanced by associating within the
hydrophobic domains of said protein. The hydrophobic protein may
be, but is not limited to, 3-lactoglobulin, bovine lactoferrin,
.gamma..sub.2-casein, .beta.-casein, .alpha.-lactalbumin, or
combinations thereof. The hydrophobic protein may be in the
nutritional composition at about 1 g/L to about 100 g/L. The
hydrophobic protein may be in the nutritional composition at about
10 g/L to about 80 g/L. The hydrophobic protein may be in the
nutritional composition at about 20 g/L to about 60 g/L. The
hydrophobic protein may be in the nutritional composition at about
30 g/L to about 50 g/L. For example, the hydrophobic protein may be
in the nutritional composition at about 1 g/L, 1.5 g/L, 2 g/L, 2.5
g/L, 3 g/L, 3.5 g/L, 4 g/L, 4.5 g/L, 5 g/L, 5.5 g/L, 6 g/L, 6.5
g/L, 7 g/L, 7.5 g/L, 8 g/L, 8.5 g/L, 9 g/L, 9.5 g/L, 10 g/L, 10.5
g/L, 11 g/L, 11.5 g/L, 12 g/L, 12.5 g/L, 13 g/L, 13.5 g/L, 14 g/L,
14.5 g/L, 15 g/L, 15.5 g/L, 16 g/L, 16.5 g/L, 17 g/L, 17.5 g/L, 18
g/L, 18.5 g/L, 19 g/L, 19.5 g/L, 20 g/L, 20.5 g/L, 21 g/L, 21.5
g/L, 22 g/L, 22.5 g/L, 23 g/L, 23.5 g/L, 24 g/L, 24.5 g/L, 25 g/L,
25.5 g/L, 26 g/L, 26.5 g/L, 27 g/L, 27.5 g/L, 28 g/L, 28.5 g/L, 29
g/L, 29.5 g/L, 30 g/L, 30.5 g/L, 31 g/L, 31.5 g/L, 32 g/L, 32.5
g/L, 33 g/L, 33.5 g/L, 34 g/L, 34.5 g/L, 35 g/L, 35.5 g/L, 36 g/L,
36.5 g/L, 37 g/L, 37.5 g/L, 38 g/L, 38.5 g/L, 39 g/L, 39.5 g/L, 40
g/L, 40.5 g/L, 41 g/L, 41.5 g/L, 42 g/L, 42.5 g/L, 43 g/L, 43.5
g/L, 44 g/L, 44.5 g/L, 45 g/L, 45.5 g/L, 46 g/L, 46.5 g/L, 47 g/L,
47.5 g/L, 48 g/L, 48.5 g/L, 49 g/L, 49.5 g/L, 50 g/L, 50.5 g/L, 51
g/L, 51.5 g/L, 52 g/L, 52.5 g/L, 53 g/L, 53.5 g/L, 54 g/L, 54.5
g/L, 55 g/L, 55.5 g/L, 56 g/L, 56.5 g/L, 57 g/L, 57.5 g/L, 58 g/L,
58.5 g/L, 59 g/L, 59.5 g/L, 60 g/L, 60.5 g/L, 61 g/L, 61.5 g/L, 62
g/L, 62.5 g/L, 63 g/L, 63.5 g/L, 64 g/L, 64.5 g/L, 65 g/L, 65.5
g/L, 66 g/L, 66.5 g/L, 67 g/L, 67.5 g/L, 68 g/L, 68.5 g/L, 69 g/L,
69.5 g/L, 70 g/L, 70.5 g/L, 71 g/L, 71.5 g/L, 72 g/L, 72.5 g/L, 73
g/L, 73.5 g/L, 74 g/L, 74.5 g/L, 75 g/L, 75.5 g/L, 76 g/L, 76.5
g/L, 77 g/L, 77.5 g/L, 78 g/L, 78.5 g/L, 79 g/L, 79.5 g/L, 80 g/L,
80.5 g/L, 81 g/L, 81.5 g/L, 82 g/L, 82.5 g/L, 83 g/L, 83.5 g/L, 84
g/L, 84.5 g/L, 85 g/L, 85.5 g/L, 86 g/L, 86.5 g/L, 87 g/L, 87.5
g/L, 88 g/L, 88.5 g/L, 89 g/L, 89.5 g/L, 90 g/L, 90.5 g/L, 91 g/L,
91.5 g/L, 92 g/L, 92.5 g/L, 93 g/L, 93.5 g/L, 94 g/L, 94.5 g/L, 95
g/L, 95.5 g/L, 96 g/L, 96.5 g/L, 97 g/L, 97.5 g/L, 98 g/L, 98.5
g/L, 99 g/L, 99.5 g/L, or 100 g/L.
[0224] The hydrophobic protein may be characterized by its grand
average of hydropathicity index (GRAVY) value. Hydropathicity is
used as defined in Creighton TE, Proteins: structures and molecular
properties, 2nd edition, 1993, W.H. Freeman and Company, NY, NY,
page 160. The hydrophobic protein may have a GRAVY value of about
-0.5 to about 0. The hydrophobic protein may have a GRAVY value of
about -0.4 to about 0. The hydrophobic protein may have a GRAVY
value of about -0.2 to about 0. The GRAVY value of the hydrophobic
protein is a measure of hydrophobicity and water solubility, and
may be predictive of a protein's ability to bind a lipophilic
molecule such as lutein. The GRAVY value may be obtained from
protein data banks known within the art, as well as referenced in
Kyte et al., J. Mol. Biol. (1982) 157, 105-132, incorporated herein
by reference in its entirety. The GRAVY value of the hydrophobic
protein may be about -0.5, -0.498, -0.496, -0.494, -0.492, -0.49,
-0.488, -0.486, -0.484, -0.482, -0.48, -0.478, -0.476, -0.474,
-0.472, -0.47, -0.468, -0.466, -0.464, -0.462, -0.46, -0.458,
-0.456, -0.454, -0.452, -0.45, -0.448, -0.446, -0.444, -0.442,
-0.44, -0.438, -0.436, -0.434, -0.432, -0.43, -0.428, -0.426,
-0.424, -0.422, -0.42, -0.418, -0.416, -0.414, -0.412, -0.41,
-0.408, -0.406, -0.404, -0.402, -0.4, -0.398, -0.396, -0.394,
-0.392, -0.39, -0.388, -0.386, -0.384, -0.382, -0.38, -0.378,
-0.376, -0.374, -0.372, -0.37, -0.368, -0.366, -0.364, -0.362,
-0.36, -0.358, -0.356, -0.354, -0.352, -0.35, -0.348, -0.346,
-0.344, -0.342, -0.34, -0.338, -0.336, -0.334, -0.332, -0.33,
-0.328, -0.326, -0.324, -0.322, -0.32, -0.318, -0.316, -0.314,
-0.312, -0.31, -0.308, -0.306, -0.304, -0.302, -0.3, -0.298,
-0.296, -0.294, -0.292, -0.29, -0.288, -0.286, -0.284, -0.282,
-0.28, -0.278, -0.276, -0.274, -0.272, -0.27, -0.268, -0.266,
-0.264, -0.262, -0.26, -0.258, -0.256, -0.254, -0.252, -0.25,
-0.248, -0.246, -0.244, -0.242, -0.24, -0.238, -0.236, -0.234,
-0.232, -0.23, -0.228, -0.226, -0.224, -0.222, -0.22, -0.218,
-0.216, -0.214, -0.212, -0.21, -0.208, -0.206, -0.204, -0.202,
-0.2, -0.198, -0.196, -0.194, -0.192, -0.19, -0.188, -0.186,
-0.184, -0.182, -0.18, -0.178, -0.176, -0.174, -0.172, -0.17,
-0.168, -0.166, -0.164, -0.162, -0.16, -0.158, -0.156, -0.154,
-0.152, -0.15, -0.148, -0.146, -0.144, -0.142, -0.14, -0.138,
-0.136, -0.134, -0.132, -0.13, -0.128, -0.126, -0.124, -0.122,
-0.12, -0.118, -0.116, -0.114, -0.112, -0.11, -0.108, -0.106,
-0.104, -0.102, -0.1, -0.098, -0.096, -0.094, -0.092, -0.09,
-0.088, -0.086, -0.084, -0.082, -0.08, -0.078, -0.076, -0.074,
-0.072, -0.07, -0.068, -0.066, -0.064, -0.062, -0.06, -0.058,
-0.056, -0.054, -0.052, -0.05, -0.048, -0.046, -0.044, -0.042,
-0.04, -0.038, -0.036, -0.034, -0.032, -0.03, -0.028, -0.026,
-0.024, -0.022, -0.02, -0.018, -0.016, -0.014, -0.012, -0.01,
-0.008, -0.006, -0.004, -0.002, or 0.
[0225] Additionally, it has been surprisingly found that
hydrophobic protein associated with the assembly leads to a
corresponding increase in lipophilic compound within the assembly,
wherein hydrophobic protein comprises from about 10 wt % to about
95 wt % of the assembly. For example, the hydrophobic protein may
be in the assembly at about 10 wt %, 10.5 wt %, 11 wt %, 11.5 wt %,
12 wt %, 12.5 wt %, 13 wt %, 13.5 wt %, 14 wt %, 14.5 wt %, 15 wt
%, 15.5 wt %, 16 wt %, 16.5 wt %, 17 wt %, 17.5 wt %, 18 wt %, 18.5
wt %, 19 wt %, 19.5 wt %, 20 wt %, 20.5 wt %, 21 wt %, 21.5 wt %,
22 wt %, 22.5 wt %, 23 wt %, 23.5 wt %, 24 wt %, 24.5 wt %, 25 wt
%, 25.5 wt %, 26 wt %, 26.5 wt %, 27 wt %, 27.5 wt %, 28 wt %, 28.5
wt %, 29 wt %, 29.5 wt %, 30 wt %, 30.5 wt %, 31 wt %, 31.5 wt %,
32 wt %, 32.5 wt %, 33 wt %, 33.5 wt %, 34 wt %, 34.5 wt %, 35 wt
%, 35.5 wt %, 36 wt %, 36.5 wt %, 37 wt %, 37.5 wt %, 38 wt %, 38.5
wt %, 39 wt %, 39.5 wt %, 40 wt %, 40.5 wt %, 41 wt %, 41.5 wt %,
42 wt %, 42.5 wt %, 43 wt %, 43.5 wt %, 44 wt %, 44.5 wt %, 45 wt
%, 45.5 wt %, 46 wt %, 46.5 wt %, 47 wt %, 47.5 wt %, 48 wt %, 48.5
wt %, 49 wt %, 49.5 wt %, 50 wt %, 50.5 wt %, 51 wt %, 51.5 wt %,
52 wt %, 52.5 wt %, 53 wt %, 53.5 wt %, 54 wt %, 54.5 wt %, 55 wt
%, 55.5 wt %, 56 wt %, 56.5 wt %, 57 wt %, 57.5 wt %, 58 wt %, 58.5
wt %, 59 wt %, 59.5 wt %, 60 wt %, 60.5 wt %, 61 wt %, 61.5 wt %,
62 wt %, 62.5 wt %, 63 wt %, 63.5 wt %, 64 wt %, 64.5 wt %, 65 wt
%, 65.5 wt %, 66 wt %, 66.5 wt %, 67 wt %, 67.5 wt %, 68 wt %, 68.5
wt %, 69 wt %, 69.5 wt %, 70 wt %, 70.5 wt %, 71 wt %, 71.5 wt %,
72 wt %, 72.5 wt %, 73 wt %, 73.5 wt %, 74 wt %, 74.5 wt %, 75 wt
%, 75.5 wt %, 76 wt %, 76.5 wt %, 77 wt %, 77.5 wt %, 78 wt %, 78.5
wt %, 79 wt %, 79.5 wt %, 80 wt %, 80.5 wt %, 81 wt %, 81.5 wt %,
82 wt %, 82.5 wt %, 83 wt %, 83.5 wt %, 84 wt %, 84.5 wt %, 85 wt
%, 85.5 wt %, 86 wt %, 86.5 wt %, 87 wt %, 87.5 wt %, 88 wt %, 88.5
wt %, 89 wt %, 89.5 wt %, 90 wt %, 90.5 wt %, 91 wt %, 91.5 wt %,
92 wt %, 92.5 wt %, 93 wt %, 93.5 wt %, 94 wt %, 94.5 wt %, or 95
wt %. Additionally, the higher molecular weight of hydrophobic
protein associated with the assembly correlates with an increase in
lipophilic compound within the assembly, wherein the minimum
molecular weight of protein associated with the assembly may be
about 5 kD to about 15 kD. For example, the minimum molecular
weight of protein associated with the assembly may be about 5 kD,
5.2 kD, 5.4 kD, 5.6 kD, 5.8 kD, 6 kD, 6.2 kD, 6.4 kD, 6.6 kD, 6.8
kD, 7 kD, 7.2 kD, 7.4 kD, 7.6 kD, 7.8 kD, 8 kD, 8.2 kD, 8.4 kD, 8.6
kD, 8.8 kD, 9 kD, 9.2 kD, 9.4 kD, 9.6 kD, 9.8 kD, 10 kD, 10.2 kD,
10.4 kD, 10.6 kD, 10.8 kD, 11 kD, 11.2 kD, 11.4 kD, 11.6 kD, 11.8
kD, 12 kD, 12.2 kD, 12.4 kD, 12.6 kD, 12.8 kD, 13 kD, 13.2 kD, 13.4
kD, 13.6 kD, 13.8 kD, 14 kD, 14.2 kD, 14.4 kD, 14.6 kD, 14.8 kD, or
15 kD. In one embodiment, the minimum weight of protein associated
with the assembly is about 11.8 kD.
[0226] (a) .beta.-Casein
[0227] The hydrophobic protein of the nutritional composition may
comprise at least .beta.-casein. .beta.-casein may be incorporated
into the assembly. Incorporation of .beta.-casein as part of the
assembly enhances the solubility of lipophilic compounds, thus
enhancing its bioavailability of the lipophilic compounds for
treatment of health conditions, relative to controls without MDG or
hydrophobic protein. It has unexpectedly been shown that
.beta.-casein is implicated in the delivery of certain lipophilic
nutrients for absorption, digestion, or both. A2.beta.-casein is a
relatively more hydrophobic form of .beta.-casein (compared to A1
(.beta.-casein). The nutritional composition that replaces a
portion of the inherent .beta.-casein with genetic variant A2
demonstrate enhanced bioavailability of certain lipophilic
nutrients.
[0228] The majority of milk protein utilized in milk
protein-containing products in the United States is from the cattle
breed Holstein. The term "Holstein" as used herein should be
understood to encompass the Holstein breed, the Friesian breed and
cross-breeds of the two, so-called Holstein-Friesian cattle. Milk
from Holstein cattle includes genetic variant A1 as the predominant
genetic variant. In contrast, milk from the Bos indicus breed has
genetic variant A2 as the predominant genetic variant. Similarly,
milk from the Guernsey breed of Bos taurus has been shown to
express high levels of .beta.-casein variant A2 and low levels of
other .beta.-casein variants.
[0229] In certain embodiments, the nutritional composition may
comprise protein including 10 to 100% bovine .beta.-casein. The
bovine .beta.-casein comprises about 50 to about 100% genetic
variant A2. Thus, as a non-limiting example, in a nutritional
composition containing 10 grams of protein per serving, about 1-10
grams of that protein would be bovine-.beta.-casein, and about
0.5-10 grams of the protein would be genetic variant A2. In certain
exemplary embodiments the bovine .beta.-casein comprises about 60%
to about 100%, including about 70% to about 100%, including about
80% to about 100%, and including about 90% to about 100% by weight
bovine .beta.-casein.
[0230] The bovine .beta.-casein utilized in certain exemplary
embodiments may be from a single source, or may alternatively be
provided by a combination of sources. The bovine .beta.-casein
according to certain exemplary embodiments will generally be found
in milk protein isolates and milk protein concentrates, but may
also be found in other milk protein sources such as whole milk,
nonfat dry milk, milk protein concentrate, total milk protein, milk
protein isolate, acid casein, calcium caseinate, sodium caseinate,
magnesium caseinate, purified .beta.-casein, and combinations
thereof. It may be possible to purify a milk protein isolate (or
another milk protein source) containing unacceptably high levels of
less-desirable genetic variants (e.g., those other than A2).
Non-limiting examples of purification methods useful for reducing
unacceptably high levels of .beta.-casein genetic variants include:
a preparative chromatographic process (e.g., affinity
chromatography, ion exchange chromatography, reversed phase
chromatography) or by a selective salt precipitation (e.g.,
ammonium sulfate). Alternatively, a milk protein source (such as
milk protein isolate or milk protein concentrate) containing milk
sourced from primarily non-Holstein cattle, and hence, reduced
quantities of genetic variants other than A2 may also be utilized
in the nutritional composition and methods disclosed herein.
[0231] The term "milk protein concentrate" is generally used to
refer to a milk protein containing product that has had a
considerable amount of the inherent water from ordinary milk
removed and also has had inherent fat from the ordinary milk
removed. The term "milk protein isolate" is generally used to refer
to a type of milk-protein containing product that has not only had
a considerable amount of the inherent water from ordinary milk
removed and inherent fat but also a certain amount of inherent
lactose removed. In most instances, milk protein isolates may be
considered to be a type of further purified milk protein
concentrate. Certain manufacturers may use the term milk protein
concentrate to refer to milk-based protein products even if they
contain at least 85 weight % protein.
[0232] The protein in the nutritional composition according to
certain embodiments, may be provided by a single source of protein
or a combination of protein sources. As previously discussed, about
10 to about 100% by weight of the protein present in the
nutritional composition comprises bovine .beta.-casein. The
remaining portion of the protein (e,g, 0-90% by weight of the total
protein present in the nutritional composition) may be selected
from one or more other sources. As discussed in more detail below,
these additional sources of protein are not particularly limited
and may include one or more of soy protein, whey protein or any
other protein source, including but not limited to those discussed
herein. Furthermore, it should be understood that the source for
the remaining portion of the protein could also be selected from
milk protein isolate, milk protein concentrate, caseinates, or
non-fat dry milk that does not meet the previously discussed
requirements as long as these components are not present in amounts
so high as to violate other limitations on the amounts of genetic
variants discussed previously.
[0233] .beta.-casein may be in the nutritional composition at about
0.2 g/L to about 20 g/L. .beta.-casein may be in the nutritional
composition at about 0.5 g/L to about 15 g/L. .beta.-casein may be
in the nutritional composition at about 1 g/L to about 15 g/L.
.beta.-casein may be in the nutritional composition at about 5 g/L
to about 15 g/L. For example, .beta.-casein may be in the
nutritional composition at about 0.2 g/L, 0.4 g/L, 0.6 g/L, 0.8
g/L, 1 g/L, 1.2 g/L, 1.4 g/L, 1.6 g/L, 1.8 g/L, 2 g/L, 2.2 g/L, 2.4
g/L, 2.6 g/L, 2.8 g/L, 3 g/L, 3.2 g/L, 3.4 g/L, 3.6 g/L, 3.8 g/L, 4
g/L, 4.2 g/L, 4.4 g/L, 4.6 g/L, 4.8 g/L, 5 g/L, 5.2 g/L, 5.4 g/L,
5.6 g/L, 5.8 g/L, 6 g/L, 6.2 g/L, 6.4 g/L, 6.6 g/L, 6.8 g/L, 7 g/L,
7.2 g/L, 7.4 g/L, 7.6 g/L, 7.8 g/L, 8 g/L, 8.2 g/L, 8.4 g/L, 8.6
g/L, 8.8 g/L, 9 g/L, 9.2 g/L, 9.4 g/L, 9.6 g/L, 9.8 g/L, 10 g/L,
10.2 g/L, 10.4 g/L, 10.6 g/L, 10.8 g/L, 11 g/L, 11.2 g/L, 11.4 g/L,
11.6 g/L, 11.8 g/L, 12 g/L, 12.2 g/L, 12.4 g/L, 12.6 g/L, 12.8 g/L,
13 g/L, 13.2 g/L, 13.4 g/L, 13.6 g/L, 13.8 g/L, 14 g/L, 14.2 g/L,
14.4 g/L, 14.6 g/L, 14.8 g/L, 15 g/L, 15.2 g/L, 15.4 g/L, 15.6 g/L,
15.8 g/L, 16 g/L, 16.2 g/L, 16.4 g/L, 16.6 g/L, 16.8 g/L, 17 g/L,
17.2 g/L, 17.4 g/L, 17.6 g/L, 17.8 g/L, 18 g/L, 18.2 g/L, 18.4 g/L,
18.6 g/L, 18.8 g/L, 19 g/L, 19.2 g/L, 19.4 g/L, 19.6 g/L, 19.8 g/L,
or 20 g/L.
TABLE-US-00002 TABLE 2 Ratio of .beta.-casein to Lipophilic
Nutrients Table 1 Ratio of .beta.-casein to Concentration, .mu.M
lipophilic nutrient, molar, Nutrient (approximate) approx.
.beta.-casein (~2.40 g/L) 102 N/A vitamin A (2029 IU/L) 2.12 50
vitamin D (406 IU/L) 0.026 4000 vitamin E (10.1 IU/L) 0.016 6400
vitamin K (54 .mu.g/L) 0.120 900 Lutein (50 .mu.g/L) 0.088 1200 ARA
(150 mg/L) 500 0.20 DHA (65 mg/L) 200 0.50
[0234] e. Optional Ingredients
[0235] The nutritional composition may further comprise other
optional ingredients that may modify the physical, chemical,
hedonic or processing characteristics of the products or serve as
pharmaceutical or additional nutritional components when used in
the targeted population. Many such optional ingredients are known
or otherwise suitable for use in other nutritional or
pharmaceutical products and may also be used in the nutritional
composition described herein, provided that such optional
ingredients are safe and effective for oral administration and are
compatible with the essential and other ingredients in the selected
product form.
[0236] Non-limiting examples of such optional ingredients include
carbohydrates, preservatives, antioxidants, emulsifying agents,
buffers, pharmaceutical actives, additional nutrients as described
herein, colorants, flavors, thickening agents and stabilizers, and
so forth.
[0237] Non-limiting examples of suitable carbohydrates or sources
thereof for use in the nutritional composition described herein may
be selected from selected from the group of: maltodextrin,
hydrolyzed or modified starch or cornstarch, glucose polymers, corn
syrup, corn syrup solids, rice-derived carbohydrates, glucose,
fructose, lactose, high fructose corn syrup, honey, sugar alcohols
(e.g., maltitol, erythritol, sorbitol), and combinations
thereof.
[0238] The nutritional composition may further comprise vitamins or
related nutrients, non-limiting examples of which may be selected
from the group of: vitamin A, vitamin K, thiamine, riboflavin,
pyridoxine, vitamin B12, carotenoids (in addition to the lutein
discussed above), niacin, folic acid, pantothenic acid, biotin,
vitamin C, choline, inositol, salts, and derivatives thereof, and
combinations thereof.
[0239] The nutritional composition may further comprise minerals,
non-limiting examples of which may be selected from the group of:
phosphorus, magnesium, iron, zinc, manganese, copper, sodium,
potassium, molybdenum, chromium, selenium, chloride, and
combinations thereof.
[0240] In some embodiments, the nutritional composition may
comprise a compound selected from the group of: hydroxyl methyl
butyrate; leucine; beta alanine; epigallocatechin gallate; human
milk oligosaccharides; prebiotics; probiotics; and combinations
thereof.
[0241] The nutritional composition may also include one or more
masking agents to reduce or otherwise obscure bitter flavors and
after taste. Suitable masking agents may be selected from the group
of: natural and artificial sweeteners, sodium sources such as
sodium chloride, and hydrocolloids, such as guar gum, xanthan gum,
carrageenan, gellan gum, and combinations thereof. The amount of
masking agent in the composition may vary depending upon the
particular masking agent selected, other ingredients in the
composition, and other composition or product target variables.
Such amounts, however, most typically range from at least about
0.1%, including from about 0.15% to about 3.0%, and also including
from about 0.18% to about 2.5%, by weight of the composition.
[0242] The nutritional composition may also optionally include one
or more stabilizers. Appropriate stabilizers to use in formulation
for the nutritional products include, but are not limited to, gum
arabic, gum ghatti, gum karaya, gum tragacanth, agar, furcellaran,
locust bean gum, pectin, low methoxyl pectin, gelatin,
microcrystalline cellulose, CMC (sodium carboxymethylcellulose),
methylcellulose hydroxypropyl methyl cellulose, hydroxypropyl
cellulose, DATEM (diacetyl tartaric acid esters of mono- and
diglycerides), dextran, CITREM (citric acid esters of mono- and
diglycerides), and mixtures thereof.
[0243] (1) Choline
[0244] The optional ingredients of the nutritional composition may
comprise choline. Choline may act as both a nutrient within the
composition, as well as an agent that enhances assembly stability
and formation. Choline may also lead to an increase in lipophilic
concentration within the assembly. It is hypothesized, without
being bound to any particular theory, that the positive charge on
choline interacts with negative side chains of the hydrophobic
protein (e.g., aspartic acid residues), and alters the 3D
configuration of said protein. This may allow for more access to
the hydrophobic domains of the protein by the lipophilic compound.
Additionally, the use of choline may be lutein specific, in that
choline selectively increases the amount of lutein within the
assembly, relative to other lipophilic compounds.
[0245] Choline may be added at different stages of the composition.
The stage in which choline is added to the composition is critical
to the formation of stable, water soluble assemblies. Specifically,
it is more advantageous to add choline to the protein slurry,
relative directly to the premix/activated premix of MDG and
lipophilic compound.
[0246] Choline acts as a source of methyl groups for the
biosynthesis of other methylated products. It is the precursor of
the neurotransmitter acetylcholine. It has been proved that the
administration of choline is beneficial in patients suffering from
any disorder related to defective cholinergic
neurotransmission.
[0247] Choline is also a major component, along with lecithin, of
phospholipids and sphingomyelin. By virtue of its fundamental
functions in membrane structure, a choline deficiency causes a
whole range of phospholipid abnormalities which express themselves
clinically as fatty liver, kidney lesions (haemorrhagic renal
necrosis) and impairment of lipoprotein metabolism. With a diet
deficient in choline, cholesterol esters and fats accumulate in the
liver.
[0248] Choline chloride and bitartrate are mentioned in the US Code
of Federal Regulations as nutrition/dietary supplements which have
been accorded GRAS status (Generally Recognised As Safe).
[0249] Choline may be in the nutritional composition at about 5
mg/kg to about 1 g/kg. Choline may be in the nutritional
composition at about 50 mg/kg to about 1 g/kg. Choline may be in
the nutritional composition at about 100 mg/kg to about 1 g/kg.
Choline may be in the nutritional composition at about 500 mg/kg to
about 1 g/kg. For example, choline may be in the nutritional
composition at about 5 mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, 25
mg/kg, 30 mg/kg, 35 mg/kg, 40 mg/kg, 45 mg/kg, 50 mg/kg, 55 mg/kg,
60 mg/kg, 65 mg/kg, 70 mg/kg, 75 mg/kg, 80 mg/kg, 85 mg/kg, 90
mg/kg, 95 mg/kg, 100 mg/kg, 105 mg/kg, 110 mg/kg, 115 mg/kg, 120
mg/kg, 125 mg/kg, 130 mg/kg, 135 mg/kg, 140 mg/kg, 145 mg/kg, 150
mg/kg, 155 mg/kg, 160 mg/kg, 165 mg/kg, 170 mg/kg, 175 mg/kg, 180
mg/kg, 185 mg/kg, 190 mg/kg, 195 mg/kg, 200 mg/kg, 205 mg/kg, 210
mg/kg, 215 mg/kg, 220 mg/kg, 225 mg/kg, 230 mg/kg, 235 mg/kg, 240
mg/kg, 245 mg/kg, 250 mg/kg, 255 mg/kg, 260 mg/kg, 265 mg/kg, 270
mg/kg, 275 mg/kg, 280 mg/kg, 285 mg/kg, 290 mg/kg, 295 mg/kg, 300
mg/kg, 305 mg/kg, 310 mg/kg, 315 mg/kg, 320 mg/kg, 325 mg/kg, 330
mg/kg, 335 mg/kg, 340 mg/kg, 345 mg/kg, 350 mg/kg, 355 mg/kg, 360
mg/kg, 365 mg/kg, 370 mg/kg, 375 mg/kg, 380 mg/kg, 385 mg/kg, 390
mg/kg, 395 mg/kg, 400 mg/kg, 405 mg/kg, 410 mg/kg, 415 mg/kg, 420
mg/kg, 425 mg/kg, 430 mg/kg, 435 mg/kg, 440 mg/kg, 445 mg/kg, 450
mg/kg, 455 mg/kg, 460 mg/kg, 465 mg/kg, 470 mg/kg, 475 mg/kg, 480
mg/kg, 485 mg/kg, 490 mg/kg, 495 mg/kg, 500 mg/kg, 505 mg/kg, 510
mg/kg, 515 mg/kg, 520 mg/kg, 525 mg/kg, 530 mg/kg, 535 mg/kg, 540
mg/kg, 545 mg/kg, 550 mg/kg, 555 mg/kg, 560 mg/kg, 565 mg/kg, 570
mg/kg, 575 mg/kg, 580 mg/kg, 585 mg/kg, 590 mg/kg, 595 mg/kg, 600
mg/kg, 605 mg/kg, 610 mg/kg, 615 mg/kg, 620 mg/kg, 625 mg/kg, 630
mg/kg, 635 mg/kg, 640 mg/kg, 645 mg/kg, 650 mg/kg, 655 mg/kg, 660
mg/kg, 665 mg/kg, 670 mg/kg, 675 mg/kg, 680 mg/kg, 685 mg/kg, 690
mg/kg, 695 mg/kg, 700 mg/kg, 705 mg/kg, 710 mg/kg, 715 mg/kg, 720
mg/kg, 725 mg/kg, 730 mg/kg, 735 mg/kg, 740 mg/kg, 745 mg/kg, 750
mg/kg, 755 mg/kg, 760 mg/kg, 765 mg/kg, 770 mg/kg, 775 mg/kg, 780
mg/kg, 785 mg/kg, 790 mg/kg, 795 mg/kg, 800 mg/kg, 805 mg/kg, 810
mg/kg, 815 mg/kg, 820 mg/kg, 825 mg/kg, 830 mg/kg, 835 mg/kg, 840
mg/kg, 845 mg/kg, 850 mg/kg, 855 mg/kg, 860 mg/kg, 865 mg/kg, 870
mg/kg, 875 mg/kg, 880 mg/kg, 885 mg/kg, 890 mg/kg, 895 mg/kg, 900
mg/kg, 905 mg/kg, 910 mg/kg, 915 mg/kg, 920 mg/kg, 925 mg/kg, 930
mg/kg, 935 mg/kg, 940 mg/kg, 945 mg/kg, 950 mg/kg, 955 mg/kg, 960
mg/kg, 965 mg/kg, 970 mg/kg, 975 mg/kg, 980 mg/kg, 985 mg/kg, 990
mg/kg, 995 mg/kg, or 1 g/kg.
[0250] (2) Lecithin
[0251] The optional ingredients of the nutritional composition may
comprise lecithin. Lecithin may act as both a nutrient source
within the composition, as well as an agent that enhances assembly
stability and formation. Lecithin may also lead to an increase in
lipophilic concentration within the assembly. Lecithin is
predominantly a mixture of glycerol phospholipids (e.g.,
phosphatidylcholine, phosphatidylethanolamine and
phosphatidylinositol). Phosphatidylcholine is typically the major
glycerol phospholipid component. Lecithin may also contain other
compounds such as free fatty acids, monoglycerides, diglycerides,
triglycerides, glycolipids, and other lipid/fatty acid containing
compounds. Lecithins are sometimes classified as glycerol
phospholipids or phosphatides. This class of compounds has
amphiphilic properties and thus, emulsifying functionality.
[0252] Lecithin is typically added to liquid food products
(including nutritional compositions), so that the liquid products
remain homogeneous and does not separate. Lecithin is approved by
the United States Food and Drug Administration for human
consumption with the status "Generally Recognized As Safe."
Non-limiting examples of lecithin suitable for use herein include
egg lecithin, wheat lecithin, corn lecithin, soy lecithin, modified
lecithin, and combinations thereof. Lecithin may be provided in
deoiled or liquid form, or phosphatidylcholine enriched form.
Additionally, lecithin may be derived from sources that include,
but are not limited to organic soy, canola oil, nonfat dry milk, or
whey protein.
[0253] In one embodiment, the nutritional powder may comprise
deoiled lecithin, wherein the deoiled lecithin comprises 1% water;
3% triglycerides; 24% phosphatidylcholine; 20%
phosphatidylethanolamine; 14% phosphatidylinositol; 7% phosphatidic
acid; 8% minor phospholipids; 8% complexed sugars; and 15%
glycolipids.
[0254] In another embodiment, the nutritional powder may comprise
liquid lecithin, wherein the liquid lecithin comprises 1% water;
37% triglycerides; 16% phosphatidylcholine; 13%
phosphatidylethanolamine; 10% phosphatidylinositol; 5% phosphatidic
acid; 2% minor phospholipids; 5% complexed sugars; and 11%
glycolipids.
[0255] Lecithin suitable for use herein may be obtained from any
known or otherwise suitable nutrition source. Non-limiting examples
include soy lecithin from ADM Specialty Food Ingredients, Decatur,
Ill., USA; soy lecithin from Solae, LLC, St. Louis, Mo., USA; and
soy lecithin from American Lecithin Company, Oxford, Conn.,
USA.
[0256] Lecithin may be in the nutritional composition at about 10
mg/kg to about 5 g/kg. Lecithin may be in the nutritional
composition at about 50 mg/kg to about 5 g/kg. Lecithin may be in
the nutritional composition at about 100 mg/kg to about 1 g/kg.
Lecithin may be in the nutritional composition at about 500 mg/kg
to about 1 g/kg. For example, lecithin may be in the nutritional
composition at about 10 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30
mg/kg, 35 mg/kg, 40 mg/kg, 45 mg/kg, 50 mg/kg, 55 mg/kg, 60 mg/kg,
65 mg/kg, 70 mg/kg, 75 mg/kg, 80 mg/kg, 85 mg/kg, 90 mg/kg, 95
mg/kg, 100 mg/kg, 105 mg/kg, 110 mg/kg, 115 mg/kg, 120 mg/kg, 125
mg/kg, 130 mg/kg, 135 mg/kg, 140 mg/kg, 145 mg/kg, 150 mg/kg, 155
mg/kg, 160 mg/kg, 165 mg/kg, 170 mg/kg, 175 mg/kg, 180 mg/kg, 185
mg/kg, 190 mg/kg, 195 mg/kg, 200 mg/kg, 205 mg/kg, 210 mg/kg, 215
mg/kg, 220 mg/kg, 225 mg/kg, 230 mg/kg, 235 mg/kg, 240 mg/kg, 245
mg/kg, 250 mg/kg, 255 mg/kg, 260 mg/kg, 265 mg/kg, 270 mg/kg, 275
mg/kg, 280 mg/kg, 285 mg/kg, 290 mg/kg, 295 mg/kg, 300 mg/kg, 305
mg/kg, 310 mg/kg, 315 mg/kg, 320 mg/kg, 325 mg/kg, 330 mg/kg, 335
mg/kg, 340 mg/kg, 345 mg/kg, 350 mg/kg, 355 mg/kg, 360 mg/kg, 365
mg/kg, 370 mg/kg, 375 mg/kg, 380 mg/kg, 385 mg/kg, 390 mg/kg, 395
mg/kg, 400 mg/kg, 405 mg/kg, 410 mg/kg, 415 mg/kg, 420 mg/kg, 425
mg/kg, 430 mg/kg, 435 mg/kg, 440 mg/kg, 445 mg/kg, 450 mg/kg, 455
mg/kg, 460 mg/kg, 465 mg/kg, 470 mg/kg, 475 mg/kg, 480 mg/kg, 485
mg/kg, 490 mg/kg, 495 mg/kg, 500 mg/kg, 505 mg/kg, 510 mg/kg, 515
mg/kg, 520 mg/kg, 525 mg/kg, 530 mg/kg, 535 mg/kg, 540 mg/kg, 545
mg/kg, 550 mg/kg, 555 mg/kg, 560 mg/kg, 565 mg/kg, 570 mg/kg, 575
mg/kg, 580 mg/kg, 585 mg/kg, 590 mg/kg, 595 mg/kg, 600 mg/kg, 605
mg/kg, 610 mg/kg, 615 mg/kg, 620 mg/kg, 625 mg/kg, 630 mg/kg, 635
mg/kg, 640 mg/kg, 645 mg/kg, 650 mg/kg, 655 mg/kg, 660 mg/kg, 665
mg/kg, 670 mg/kg, 675 mg/kg, 680 mg/kg, 685 mg/kg, 690 mg/kg, 695
mg/kg, 700 mg/kg, 705 mg/kg, 710 mg/kg, 715 mg/kg, 720 mg/kg, 725
mg/kg, 730 mg/kg, 735 mg/kg, 740 mg/kg, 745 mg/kg, 750 mg/kg, 755
mg/kg, 760 mg/kg, 765 mg/kg, 770 mg/kg, 775 mg/kg, 780 mg/kg, 785
mg/kg, 790 mg/kg, 795 mg/kg, 800 mg/kg, 805 mg/kg, 810 mg/kg, 815
mg/kg, 820 mg/kg, 825 mg/kg, 830 mg/kg, 835 mg/kg, 840 mg/kg, 845
mg/kg, 850 mg/kg, 855 mg/kg, 860 mg/kg, 865 mg/kg, 870 mg/kg, 875
mg/kg, 880 mg/kg, 885 mg/kg, 890 mg/kg, 895 mg/kg, 900 mg/kg, 905
mg/kg, 910 mg/kg, 915 mg/kg, 920 mg/kg, 925 mg/kg, 930 mg/kg, 935
mg/kg, 940 mg/kg, 945 mg/kg, 950 mg/kg, 955 mg/kg, 960 mg/kg, 965
mg/kg, 970 mg/kg, 975 mg/kg, 980 mg/kg, 985 mg/kg, 990 mg/kg, 995
mg/kg, 1 g/kg, 1.2 g/kg, 1.4 g/kg, 1.6 g/kg, 1.8 g/kg, 2 g/kg, 2.2
g/kg, 2.4 g/kg, 2.6 g/kg, 2.8 g/kg, 3 g/kg, 3.2 g/kg, 3.4 g/kg, 3.6
g/kg, 3.8 g/kg, 4 g/kg, 4.2 g/kg, 4.4 g/kg, 4.6 g/kg, 4.8 g/kg, or
5 g/kg.
4. METHOD OF MANUFACTURING
[0257] To prepare the nutritional composition having improved
bioavailability of a lipophilic compound contained therein, a
method in accordance with the disclosure may be utilized. The
method may comprise the steps of: forming a premix; adding the
premix to a solution to form an activated premix or a protected
premix, and adding the activated premix to the composition, wherein
the assembly may be at least formed in the activated premix.
[0258] a. Premix
[0259] In some embodiments, a premix comprising MDG is provided.
The premix may comprise MGD alone, MGD and at least one lipophilic
compound, as well as other optional ingredients, as described
above. The premix may be activated. The premix may be heated prior
to being activated. In some emobiments, the premix is heated to
about 85.degree. F. for 30 min. In some embodiments, the premix is
heated to 120.degree. F. for about 10 minutes. One of skill would
appreciate that the premix composition may affect the temperature
and time that is needed to heat said premix. The premix may be
protected, for example by surfactants.
[0260] (1) Activated Premix
[0261] The premix may be added to an aqueous solution, for example
to a protein-in-water slurry, to form an "activated premix." The
activated premix is then added to the composition which may
comprise at least one lipophilic compound.
[0262] Notably, to form an "activated premix," the premix may not
be added to a quantity of oil that is sufficient to cause the MDG
to disassociate from the assembly in the oil prior to the addition
of the premix into the aqueous solution. In this instance, the
activate premix may be protected to limit dissociation of MDG and
lipophilic compound from each other.
[0263] When an "activated premix" is subsequently added to a
fat-containing solution, at least a portion of the MDG is not
disassociated from the assembly by the fat in the fat-containing
solution. To determine the amount of the monoglycerides and
diglycerides that are not disassociated in the fat, the
"Disassociated MDG (monoglyceride and diglyceride) Test" may be
utilized.
[0264] The Disassociated MDG Test is as follows. The fat-containing
solution is subjected to high speed centrifugation (31,000.times.g;
20.degree. C.; 4 h), as described above. The resulting aqueous
fraction is subjected to HPLC analysis for monoglycerides and
diglycerides content as described in "Determination of food
emulsifiers in commercial additives and food products by liquid
chromatography/atmospheric-pressure chemical ionization mass
spectrophotometry," by M. Suman et al., Journal of Chromatography
A, 1216 (2009) 3758-3766. The level of monoglycerides and
diglycerides detected in the aqueous fraction is compared to the
level that was added to the fat-containing solution to arrive at
the percentage of the mixture of monoglycerides and diglycerides
that is not disassociated by the fat.
[0265] Using the Disassociated MDG Test, one may determine the
impact of utilizing an activated premix versus a premix that has
not been activated in a composition. This may be accomplished by
comparing the amount of monoglycerides and diglycerides that is
present in the aqueous fraction of a composition comprising an
activated premix, with the amount of monoglycerides and
diglycerides that is present in the aqueous fraction of the same
composition in which the premix has not been activated. The aqueous
fraction of the composition comprising the activated premix will
typically contain a higher amount of monoglycerides and
diglycerides than the same composition in which the premix has not
been activated.
[0266] The difference between the content of monoglycerides and
diglycerides in the aqueous fraction of the composition comprising
the activated premix, and the content of monoglycerides and
diglycerides in the aqueous fraction of the same composition
comprising a premix that has not been activated, may be quantified.
In some embodiments, the content of monoglycerides and diglycerides
in the aqueous fraction of the composition comprising the activated
premix may be at least about 5%, 10% or 15% higher than in the same
composition that comprises a premix that has not been
activated.
[0267] (b) Variations of Manufacture
[0268] The premix, activated premix, and composition may comprise
different embodiments, as some are listed below. It should be noted
that the nutritional composition is not limited to the
representative embodiments listed below.
[0269] In some embodiments, a premix comprising the lipophilic
compound and MDG is provided. To make the premix, the lipophilic
compound may be mixed with the MDG in the presence of heat or at
ambient temperature, and in some embodiments, with agitation to
allow the lipophilic compound to dissolve into the monoglycerides
and diglycerides. The premix is added to an aqueous solution, for
example to a protein-in-water slurry, to form an activated premix,
which contains the assembly. The activated premix is then added to
the composition.
[0270] In some embodiments, a premix comprising the lipophilic
compound, surfactant, MDG is provided. To make the premix, the
lipophilic compound may be mixed with the MDG in the presence of
heat or at ambient temperature, and in some embodiments, with
agitation to allow the lipophilic compound to dissolve into the
mixture of monoglycerides and diglycerides. The premix is added to
a fat-containing solution, such as a protein-in-fat slurry, to form
an "protected premix", which contains the assembly. The "protected
premix" is then added to the composition.
[0271] Without wishing to be bound by theory, it is believed that
the surfactant combines with the lipophilic compound,
monoglycerides and diglycerides, to form a microemulsion when the
premix is added to the fat-containing solution. Thus, in a
"protected premix," the monoglycerides and diglycerides may be
protected from disassociating in the fat-containing solution.
[0272] The activated premix or protected premix may be added to any
other ingredients of the nutritional or pharmaceutical composition
at any useful point during the manufacturing process. In some
embodiments, the nutritional or pharmaceutical composition may then
be dried to form a powdered composition using any methods known in
the art. By way of example, nutritional powders may be prepared by
preparing at least two slurries that will later be blended together
(and further may be blended with the activated premix), heat
treated, standardized, heat treated a second time, evaporated to
remove water, and spray dried to form a powdered nutritional
composition.
[0273] Slurries to which the activated premix or protected premix
may be added include a carbohydrate-mineral (CHO-MIN) slurry and a
protein-in-oil (PIO) slurry. Initially, the CHO-MIN slurry may be
formed by dissolving selected carbohydrates (e.g., lactose,
galactooligosaccharides, etc.) in heated water with agitation,
followed by the addition of minerals (e.g., potassium citrate,
magnesium chloride, potassium chloride, sodium chloride, choline
chloride, etc.). The resulting CHO-MIN slurry may then be held with
continued heat and moderate agitation until it is later blended
with the other prepared slurries.
[0274] The protein-in-oil (PIO) slurry may be formed by heating and
mixing the oil (e.g., high oleic safflower oil, soybean oil,
coconut oil, monoglycerides, etc.) and emulsifier (e.g., soy
lecithin), and then adding oil soluble vitamins, protein (e.g.,
milk protein concentrate, milk protein hydrolysate, etc.),
carrageenan (if any), calcium carbonate or tricalcium phosphate (if
any), and arachidonic acid (ARA) oil and docosahexaenoic acid (DHA)
oil (if any) with continued heat and agitation. The resulting PIO
slurry may be held with continued heat and moderate agitation until
it is later blended with the other prepared slurries.
[0275] Water is heated and then combined with the CHO-MIN slurry,
nonfat milk (if any), and the PIO slurry under adequate agitation.
The pH of the resulting blend is adjusted to 6.6-7.0, and the blend
is held under moderate heated agitation. ARA oil and DHA oil is
added at this stage in some embodiments. In some embodiments, the
activated premix or protected premix is blended with one or more of
the heated water, CHO-MIN slurry, nonfat milk (if any), and the PIO
slurry. Alternatively, the activated premix or protected premix may
be blended into the composition after further processing and
immediately prior to drying of the composition.
[0276] The composition may then be subjected to high-temperature
short-time (HTST) processing, during which the composition is heat
treated, emulsified and homogenized, and then cooled. Water soluble
vitamins and ascorbic acid are added, the pH is adjusted to the
desired range if necessary, flavors (if any) are added, and water
is added to achieve the desired total solid level. In some
embodiments, the emulsions may then be further diluted, and
subsequently processed and packaged as ready-to-drink or
concentrated liquids. In some embodiments, the emulsions are
evaporated, heat treated and subsequently processed and packaged as
reconstitutable powder, e.g., spray dried, dry blended,
agglomerated.
[0277] The spray dried powdered nutritional composition or dry
blended powdered nutritional composition may be prepared by any
collection of known or otherwise effective techniques, suitable for
making and formulating a nutritional powder. For example, the spray
drying step may likewise include any spray drying technique that is
known for or otherwise suitable for use in the production of
nutritional powders. Many different spray drying methods and
techniques are known for use in the nutrition field, all of which
are suitable for use in the manufacture of the spray dried powdered
nutritional composition. Following drying, the finished powder may
be packaged into suitable containers.
5. Methods of Use
[0278] The nutritional composition may be packaged and sealed in
single or multi-use containers, and then stored under ambient
conditions or under refrigeration for up to 36 months or longer,
more typically from about 6 months to about 24 months. For
multi-use containers, these packages may be opened and then covered
for repeated use by the ultimate user. For liquid embodiments, the
opened and subsequently recovered package is typically stored under
refrigerated conditions, and the contents used within about 7 days.
For powdered embodiments, the opened and subsequently recovered
package may be typically stored under ambient conditions (e.g.,
avoid extreme temperatures) and the contents used within about one
month. Non-limiting examples of ways in which the present formulas
may be utilized include use as the following products, use as: a
beverage, e.g. a coffee beverage, a cocoa or chocolate beverage, a
malted beverage, a fruit or juice beverage, a carbonated beverage,
a soft drink, or a milk based beverage; a performance nutrition
product, e.g. a performance nutrition bar, powder or ready-to-drink
beverage; a medical nutrition product; a dairy product, e.g. a milk
drink, a yogurt or other fermented dairy product; an ice cream
product; a confectionary product, e.g. a chocolate product; a
functional food or beverage, e.g. a slimming product, a fat burning
product, a product for improving mental performance or preventing
mental decline, or a skin improving product. A beverage according
to the invention may be in the form of a powder or liquid
concentrate to be mixed with a suitable liquid, e.g. water or milk,
before consumption; or a ready-to-drink beverage.
[0279] In embodiments directed toward enhancing the absorption of
the lipophilic nutrient, the lipophilic nutrient is administered to
the subject in a nutritional composition comprising the lipophilic
nutrient and a lipophilic MDG carrier oil of the nutritional
composition. The absorption of the lipophilic nutrient may be
enhanced in one or more measurable ways. The absorption of the
lipophilic nutrient may be enhanced by increasing the maximum
plasma concentration of the nutrient in the blood of the ingesting
subject. The absorption of the lipophilic nutrient may be enhanced
by prolonging the plasma absorption of the nutrient in the blood of
the ingesting subject over a period of days or weeks. The
absorption of the lipophilic nutrient may be enhanced by increasing
the total plasma concentration of the nutrient in the blood of the
ingesting subject over a period of days or weeks.
[0280] In embodiments in which the lipophilic compound is lutein,
the powdered nutritional composition including the lutein may be
administered to improve age-related macular degeneration and other
retinal diseases and disorders. Although in some embodiments, the
methods of the present disclosure may be directed to individuals
who have age-related macular degeneration or other retinal diseases
and disorders, the methods of the present disclosure as described
herein are also intended in some embodiments to include the use of
such methods in "at risk" individuals, including individuals
unaffected by or not otherwise afflicted with age-related macular
degeneration or other retinal diseases or disorders, for the
purpose of preventing, minimizing, or delaying the development of
such diseases and conditions over time. For such prevention
purposes, the methods of the present disclosure preferably include
continuous, daily administration of the composition as described
herein. Such preventive methods may be directed at adults or
others, particularly older adults, who are susceptible to
developing age-related macular degeneration or other retinal
diseases and disorders due to hereditary considerations,
environmental considerations, and the like.
[0281] Composition including Vitamin D may be administered to
subjects to promote healthy bone development, reduce Vitamin D
deficiency, increase bone strength, preserve or increase bone
mineralization, and preserve or increase bone mineral density. The
composition including Vitamin D may also be administered to
subjects suffering from or at risk for suffering from diseases and
conditions associated with inadequate bone mineralization, such as
rickets, osteomalacia, osteoporosis, osteopenia, and bone
fractures. The subjects receiving the composition with Vitamin D
may include, but are not limited to, pregnant and post-natal women,
infants, children, adolescents, adults, post-menopausal women, and
older adults. In some embodiments, the methods of the present
disclosure are directed to pregnant or post-natal women, to promote
healthy bone development in the developing fetus or infant. In some
embodiments, the methods of the present disclosure are directed to
infants and children, to promote healthy bone development and
prevent rickets or bone fractures in the growing child. In some
embodiments, the methods of the present disclosure are directed to
adolescents and adults, to promote increased bone mineralization
and bone mineral density. In some embodiments, the methods of the
present disclosure are directed to post-menopausal women, to
prevent or slow the onset of osteoporosis. In some embodiments, the
methods of the present disclosure are directed to adults who have
osteomalacia, osteoporosis, osteopenia, or other bone diseases and
disorders. In some embodiments, the methods of the present
disclosure are directed to "at risk" individuals, including
individuals unaffected by or not otherwise afflicted by
osteomalacia, osteoporosis, osteopenia, loss of balance and
falling, bone fractures, or other bone diseases or disorders, for
the purpose of preventing, minimizing, or delaying the development
of such diseases and conditions over time. For such development,
maintenance, and prevention purposes, the methods of the present
disclosure preferably include continuous, daily administration of
the composition as described herein. Such development, maintenance,
and preventive methods are directed at subjects such as pregnant
and postnatal women, infants, children, adolescents, adults,
post-menopausal women, and older adults. Preventative methods are
particularly directed to older adults who are susceptible to
developing age-related osteoporosis, osteopenia, loss of balance or
falling, broken bones, or other diseases and disorders associated
with inadequate Vitamin D consumption or inadequate bone
mineralization.
[0282] In these embodiments, a subject desirably consumes servings
of the composition that provide an effective amount of Vitamin D
per day. Effective amounts of Vitamin D range from about 50 IU to
about 7,500 IU per day, including from about 100 IU per day to
about 5,000 IU per day, including from about 200 IU per day to
about 2,500 IU per day, including from about 250 IU per day to
about 1,500 IU per day, including from about 400 IU per day to
about 1,000 IU per day, and including from about 500 IU per day to
about 800 IU per day.
[0283] In embodiments in which the lipophilic nutrient is Vitamin
E, the nutritional composition including Vitamin E may be
administered to subjects to provide antioxidants, promote
cardiovascular health, and prevent or reduce the risks of some
types of cancer. The subjects receiving the nutritional composition
with Vitamin E may include, but are not limited to, pregnant and
post-natal women, infants, children, and adults. In some
embodiments, the methods of the present disclosure may be directed
to pregnant or post-natal women to promote healthy development in
the developing fetus or infant. In some embodiments, the methods of
the present disclosure may be directed to infants and children to
promote healthy development in the growing child. In some
embodiments, the methods of the present disclosure may be directed
to adults who have cardiovascular disease. In some embodiments, the
methods of the present disclosure are also directed to "at risk"
individuals, including individuals unaffected by or not otherwise
afflicted with cancer, cardiovascular disease, or other diseases or
disorders, for the purpose of preventing, minimizing, or delaying
the development of such diseases and conditions over time. For such
development, maintenance, and prevention purposes, the methods of
the present disclosure preferably include continuous, daily
administration of the nutritional composition as described herein.
Such development, maintenance, and preventive methods may be
directed at subjects such as pregnant and postnatal women, infants,
children, and adults, particularly older adults who are susceptible
to developing cancer, cardiovascular disease, or other diseases and
disorders due to hereditary considerations, environmental
considerations, and the like.
[0284] In embodiments in which the lipophilic nutrient is DHA, the
nutritional composition including DHA may be administered to
subjects to promote healthy eye, brain, and central nervous system
development, maintain overall brain health, prevent or reduce
age-related decline in mental functioning, and reduce
cardiovascular and other diseases and disorders. The subjects
receiving the nutritional composition with DHA may include, but are
not limited to, pregnant and post-natal women, infants, children,
and adults. In some embodiments, the methods of the present
disclosure may be directed to pregnant or post-natal women to
promote eye, brain, and central nervous system development in the
developing fetus or infant. In some embodiments, the methods of the
present disclosure may be directed to infants and children to
promote eye, brain, and central nervous system development in the
growing child. In some embodiments, the methods of the present
disclosure may be directed to adults to maintain overall brain
health, prevent or reduce age-related decline in mental
functioning, and reduce cardiovascular and other diseases and
disorders. In some embodiments, the methods of the present
disclosure are also directed to "at risk" individuals, including
individuals unaffected by or not otherwise afflicted with
age-related declines in mental functioning or cardiovascular
disease, for the purpose of preventing, minimizing, or delaying the
development of such diseases and conditions over time. For such
development, maintenance, and prevention purposes, the methods of
the present disclosure preferably include continuous, daily
administration of the nutritional composition as described herein.
Such development, maintenance, and preventive methods may be
directed at subjects such as pregnant and postnatal women, infants,
children, and adults, particularly older adults who are susceptible
to age-related declines in mental functioning, cardiovascular
disease, or other diseases and disorders due to hereditary
considerations, environmental considerations, and the like.
[0285] The nutritional composition including HPAV oil and an
activated MDG oil component or a protected premix may be
administered to infants to enhance palmitic acid absorbance,
promote healthy bone development, and maintain healthy bone mineral
density. The subjects receiving the composition with an HPAV oil
and an activated MDG oil component or a protected premix include,
but are not limited to, pre-term and term infants. In some
embodiments, the methods of the present disclosure may be directed
to infants, to promote healthy bone development and maintain
healthy bone mineral density in the growing infant. For such
development and maintenance purposes, the methods of the present
disclosure preferably include continuous, daily administration of
the nutritional composition as described herein. Such development
and maintenance methods may be directed at subjects such as preterm
and term infants.
[0286] In these embodiments, an infant desirably consumes servings
of the nutritional composition that provide effective amounts of
the HPAV oil per day. Effective amounts of the HPAV oil range from
about 0.6 g to about 18 g per day, including from about 1 g per day
to about 15 g per day, including from about 2.5 g per day to about
12.5 g per day, including from about 5 g per day to about 10 g per
day, and including from about 6 g per day to about 8 g per day.
[0287] The nutritional composition including DHA and lutein may be
administered to subjects to promote healthy eye and retina
development, maintain overall eye health, and prevent or reduce
age-related macular degeneration and other diseases and disorders
of the eye. The subjects receiving the nutritional composition with
DHA and lutein may include, but are not limited to, pregnant and
post-natal women, infants, children, and adults. In some
embodiments, the methods of the present disclosure may be directed
to pregnant or post-natal women, to promote eye and retina
development and healthy vision in the developing fetus or infant.
In some embodiments, the methods of the present disclosure may be
directed to infants, to promote eye and retina development and
visual acuity in the growing child. In some embodiments, the
methods of the present disclosure may be directed to children and
adults to improve visual acuity, improve photostress recovery, or
reduce sensitivity to glare. In some embodiments, the methods of
the present disclosure may be directed to adults who have
age-related macular degeneration, glaucoma, cataracts, lacrimal
keratoconjunctivitis, or other diseases and disorders of the eye.
In some embodiments, the methods of the present disclosure are also
directed to "at risk" individuals, including individuals unaffected
by or not otherwise afflicted with age-related macular
degeneration, glaucoma, cataracts, lacrimal keratoconjunctivitis,
diabetic retinopathy, or other diseases or disorders of the eye,
for the purpose of preventing, minimizing, or delaying the
development of such diseases and conditions over time. For such
development, maintenance, and prevention purposes, the methods of
the present disclosure preferably include continuous, daily
administration of the nutritional composition as described herein.
Such development, maintenance, and preventive methods may be
directed at subjects such as pregnant and postnatal women, infants,
children, and adults, particularly older adults who are susceptible
to developing age-related macular degeneration or other diseases
and disorders of the eye due to hereditary considerations,
environmental considerations, and the like.
[0288] In these embodiments, a subject desirably consumes servings
of the composition that provide effective amounts of DHA and lutein
per day. Effective amounts of DHA range from about 5 mg to about 10
g per day, including from about 10 mg per day to about 1 g per day,
including from about 20 mg per day to about 500 mg per day,
including from about 40 mg per day to about 200 mg per day, and
including from about 80 mg per day to about 150 mg per day.
Effective amounts of lutein range from about 5 .mu.g to about 10 mg
per day, including from about 10 .mu.g per day to about 5 mg per
day, including from about 25 .mu.g per day to about 1 mg per day,
including from about 50 .mu.g per day to about 500 .mu.g per day,
and including from about 100 .mu.g per day to about 250 .mu.g per
day.
[0289] In these embodiments, an individual desirably consumes at
least one serving of the nutritional composition daily, and in some
embodiments, may consume two, three, or even more servings per day.
Each serving is desirably administered as a single undivided dose,
although the serving may also be divided into two or more partial
or divided servings to be taken at two or more times during the
day. The methods of the present disclosure include continuous day
after day administration, as well as periodic or limited
administration, although continuous day after day administration is
generally desirable. The nutritional composition may be for use in
infants, children, and adults.
[0290] The nutritional composition has multiple aspects,
illustrated by the following non-limiting examples.
5. EXAMPLES
[0291] It is understood that the foregoing detailed description and
accompanying examples are merely illustrative and are not to be
taken as limitations upon the scope of the invention, which is
defined solely by the appended claims and their equivalents.
[0292] Various changes and modifications to the disclosed
embodiments will be apparent to those skilled in the art. Such
changes and modifications, including without limitation those
relating to the chemical structures, substituents, derivatives,
intermediates, syntheses, compositions, or methods of use of the
invention, may be made without departing from the spirit and scope
thereof.
[0293] The following examples illustrate specific embodiments
and/or features of the nutritional composition comprising an
activated premix and the composition comprising a protected premix.
The examples are given solely for the purpose of illustration and
are not to be construed as limitations of the present disclosure,
as many variations thereof are possible without departing from the
spirit and scope of the disclosure. All exemplified amounts are
weight percentages based upon the total weight of the composition,
unless otherwise specified.
[0294] The exemplified composition may be prepared in accordance
with the manufacturing methods described herein, such that each
exemplified nutritional or pharmaceutical composition has improved
lipophilic compound bioavailability.
[0295] The following details may be applied to the below-listed
Examples when appropriate. Changes outside of the details listed
below are recited within the specific Examples.
[0296] The size exclusion chromatography (SEC) system used in the
following Examples is as follows: Column: Superdex Peptide 10/300
GL, GE Healthcare P/N 17-5176-01; Mobile Phase: 700 mL Milli-Q Plus
water, 300 mL acetonitrile; 1.0 mL trifluoroacetic acid; Flow Rate:
0.4 mL/minute; Temperature: ambient (.about.21.degree. C.);
Detection: UV light at 214 nm, 280 nm; visible light at 476 nm;
Injection: 10 .mu.L; Run Time: 70 minutes; Sample Preparation:
dilute 0.6-1.1 g to 10 mL with Mobile Phase; Calibration: by 6
purified reference proteins, 14.2-160 kD.
[0297] The high molecular weight lutein assembly is determined as
follows: SEC peak eluting .ltoreq.20.0 minutes, using visible light
detection at 476 nm (a lutein absorbance maximum). The high MW
lutein assembly is a water dispersible or water soluble,
protein-dominant, macromolecular aggregate in which lutein is
bound. The size of the high MW lutein assembly lies approximately
in the 60-380 kD range (.about.60-95 .ANG.). The concentration of
the high MW lutein assembly is quantified as peak area (mAU-min. at
476 nm) per mg of sample injected. The values reported as high MW
lutein assembly are not absolute concentrations of bound lutein,
but are instead relative measures of the lutein bound in these high
MW but water dispersible/soluble, macromolecular aggregates.
[0298] The protein >137 kD is determined as follows: SEC peak
eluting .ltoreq.18.0 minutes, using UV detection at 214 nm (a
peptide bond and amino acid side chain signal). The peak consists
predominantly of aggregated proteins; e.g., its size is larger than
all major milk proteins (.about.14 to 25 kD), so that the protein
in this peak (>137 kD) is regarded as aggregated but still water
dispersible/soluble, and capable of encapsulating lutein and other
oil soluble vitamins/nutrients. It is quantified as g of protein
per kg of sample. Peak area of the protein is measured versus a
known concentration to provide the g/kg of said protein.
[0299] The bound lipophile index is determined as follows: a
non-specific indicator of lipophilic nutrients (including lutein
and any combination of other oil soluble vitamins) in the high MW
protein peak (described above). The value reported is the peak area
ratio, 280 nm/214 nm, for the SEC peak eluting .ltoreq.18.0
minutes, a relative marker of lipophilic nutrient concentration in
the high MW aggregate. The validity of the index is based on the
fact that the 280 nm/214 nm ratios for lipophilic nutrients is >
than that for proteins.
Examples 1-5
[0300] Examples 1-5 illustrate powdered infant formulas of the
present disclosure, the ingredients of which are listed in the
table below. All ingredient amounts are listed as kg per 1000 kg
batch, unless otherwise specified.
TABLE-US-00003 TABLE 3 Nutritional Composition Ingredients for
Examples 1-5 and other Exemplified Compositions. INGREDIENT EXAMPLE
1 EXAMPLE 2 EXAMPLE 3 EXAMPLE 4 EXAMPLE 5 Corn Syrup 504.1 504.1
504.1 504.1 504.1 Soy Protein Isolate - 5% DH 144.8 144.8 144.8
144.8 144.8 Sunflower Oil 112.5 112.5 101.3 112.5 112.5 Sucrose
98.3 98.3 98.3 98.3 98.3 Soy Oil 83.9 83.9 75.5 83.9 83.9 Coconut
Oil 75.6 75.6 68.0 75.6 75.6 Fructooligosaccharides 17.0 17.0 17.0
17.0 17.0 Potassium Citrate 16.5 16.5 16.5 16.5 16.5 Calcium
Phosphate 16.4 16.4 16.4 16.4 16.4 Sodium Chloride 3.8 3.8 3.8 3.8
3.8 ARA Oil 3.0 3.0 3.0 3.0 3.0 Magnesium Chloride 2.8 2.8 2.8 2.8
2.8 L-Methionine 1.7 1.7 1.7 1.7 1.7 Ascorbic Acid 1.1 1.1 1.1 1.1
1.1 DHA Oil 1.1 1.1 1.1 1.1 1.1 Mono-and Diglyceride/Lutein approx.
approx. approx. approx. approx. Blend 622.0 g 590.9 g 27.2 653.1 g
685.0 g Monodiglycerides and 621.0 g 590.3 g 27.0 652.7 g 683.5 g
Diglycerides Lutein 945.0 mg 566 mg 200 g 392 mg 1.5 g Choline
Chloride 507.7 g 507.7 g 507.7 g 507.7 g 507.7 g Taurine 457.5 g
457.5 g 457.5 g 457.5 g 457.5 g Inositol 353.0 g 353.0 g 353.0 g
353.0 g 353.0 g Ascorbyl Palmitate 347.5 g 347.5 g 347.5 g 347.5 g
347.5 g Ferrous Sulfate 319.2 g 319.2 g 319.2 g 319.2 g 319.2 g
Mixed Tocopherols 157.2 g 157.2 g 157.2 g 157.2 g 157.2 g
L-Carnitine 112.7 g 112.7 g 112.7 g 112.7 g 112.7 g Niacinamide
97.9 g 97.9 g 97.9 g 97.9 g 97.9 g d-alpha-tocopheryl acetate 78.8
g 78.8 g 78.8 g 78.8 g 78.8 g d-Calcium Pantothenate 58.7 g 58.7 g
58.7 g 58.7 g 58.7 g Zinc 56.0 g 56.0 g 56.0 g 56.0 g 56.0 g Iron
16.9 g 16.9 g 16.9 g 16.9 g 16.9 g Thiamine 15.2 g 15.2 g 15.2 g
15.2 g 15.2 g Vitamin A palmitate 14.8 g 14.8 g 14.8 g 14.8 g 14.8
g Copper 7.2 g 7.2 g 7.2 g 7.2 g 7.2 g Riboflavin 6.7 g 6.7 g 6.7 g
6.7 g 6.7 g Pyridoxine Hydrochloride 6.1 g 6.1 g 6.1 g 6.1 g 6.1 g
Folic Acid 2.1 g 2.1 g 2.1 g 2.1 g 2.1 g Potassium Iodide 1.1 g 1.1
g 1.1 g 1.1 g 1.1 g Phylloquinone 857.1 mg 857.1 mg 857.1 mg 857.1
mg 857.1 mg Vitamin D3 103.7 mg 103.7 mg 103.7 mg 103.7 mg 103.7 mg
Lycopene 980.0 mg 980.0 mg 980.0 mg 980.0 mg 980.0 mg Biotin 592.5
mg 592.5 mg 592.5 mg 592.5 mg 592.5 mg Beta-Carotene 215.6 mg 215.6
mg 215.6 mg 215.6 mg 215.6 mg Selenium 147.0 mg 147.0 mg 147.0 mg
147.0 mg 147.0 mg Cyanocobalamin 71.3 mg 71.3 mg 71.3 mg 71.3 mg
71.3 mg
Examples 6-10
[0301] Examples 6-10 illustrate powdered infant formulas of the
present disclosure, the ingredients of which are listed in the
table below. All ingredient amounts are listed as kg per 1000 kg
batch, unless otherwise specified.
TABLE-US-00004 TABLE 4 Nutritional Composition Ingredients for
Examples 6-10 and other Exemplified Compositions. INGREDIENT
EXAMPLE 6 EXAMPLE 7 EXAMPLE 8 EXAMPLE 9 EXAMPLE 10 Skim milk 2057.3
2057.3 2057.3 2057.3 2057.3 Lactose 414.1 414.1 414.1 414.1 414.1
Sunflower oil 111.1 88.9 111.1 111.1 111.1 Soy oil 83.3 66.6 83.3
83.3 83.3 Coconut oil 74.6 59.7 74.6 74.6 74.6 GOS 72.3 72.3 72.3
72.3 72.3 Whey Protein Concentrate 49.4 49.4 49.4 49.4 49.4 Whey
protein hydrolysate (<25DH) 5.6 5.6 5.6 5.6 5.6 Potassium
citrate 3.4 3.4 3.4 3.4 3.4 ARA 2.9 2.9 2.9 2.9 2.9 Calcium
carbonate 2.6 2.6 2.6 2.6 2.6 Potassium hydroxide 2.3 2.3 2.3 2.3
2.3 Soy lecithin 1.7 1.7 1.7 1.7 1.7 DHA 1.6 1.6 1.6 1.6 1.6
Ascorbic acid 1.5 1.5 1.5 1.5 1.5 Choline bitartrate 1.2 1.2 1.2
1.2 1.2 Magnesium chloride 1.2 1.2 1.2 1.2 1.2 Mono-and
diglyceride/Lutein blend approx. approx. approx. approx. approx.
1.10 53.8 0.99 1.155 1.21 Monoglycerides and Diglycerides 1.10 53.5
0.99 1.153 1.21 Lutein 2.65 g 320 g 568 mg 2.3 g 385 mg Sodium
chloride 1 1 1 1 1 Taurine 428 g 428 g 428 g 428 g 428 g
L-Tryptophan 277 g 277 g 277 g 277 g 277 g Cytidine
5'-monophosphate 257 g 257 g 257 g 257 g 257 g Myo-inositol 257 g
257 g 257 g 257 g 257 g Ascorbyl palmitate 200 g 200 g 200 g 200 g
200 g Disodium guanosine 5'- 187 g 187 g 187 g 187 g 187 g
monophosphate Disodium uridine 5'- 163 g 163 g 163 g 163 g 163 g
monophosphate Ferrous sulfate monohydrate 130.6 g 130.6 g 130.6 g
130.6 g 130.6 g RRR-.alpha.-Tocopheryl acetate 119.7 g 119.7 g
119.7 g 119.7 g 119.7 g Tocopherol-2 106.7 g 106.7 g 106.7 g 106.7
g 106.7 g Zinc sulfate monohydrate 104 g 104 g 104 g 104 g 104 g
Adenosine 5'-monophosphate 98 g 98 g 98 g 98 g 98 g Ferrous sulfate
88 g 88 g 88 g 88 g 88 g Mixed tocopherols 77 g 77 g 77 g 77 g 77 g
Niacinamide 59 g 59 g 59 g 59 g 59 g L-Carnitine 43 g 43 g 43 g 43
g 43 g Calcium d-pantothenate 42 g 42 g 42 g 42 g 42 g Retinyl
palmitate 13 g 13 g 13 g 13 g 13 g Copper sulfate 11 g 11 g 11 g 11
g 11 g Thiamin HCl 9 g 9 g 9 g 9 g 9 g Pyridoxine HCl 5 g 5 g 5 g 5
g 5 g Riboflavin 5 g 5 g 5 g 5 g 5 g Manganese sulfate monohydrate
3 g 3 g 3 g 3 g 3 g Folic acid 796 mg 796 mg 796 mg 796 mg 796 mg
.beta.-Carotene 770 mg 770 mg 770 mg 770 mg 770 mg Phylloquinone
618 mg 618 mg 618 mg 618 mg 618 mg Potassium iodide 550 mg 550 mg
550 mg 550 mg 550 mg d-Biotin 238 mg 238 mg 238 mg 238 mg 238 mg
Sodium selenate 117 mg 117 mg 117 mg 117 mg 117 mg Cholecalciferol
93 mg 93 mg 93 mg 93 mg 93 mg Cyanocobalamin 8 mg 8 mg 8 mg 8 mg 8
mg
Examples 11-15
[0302] Examples 11-15 illustrate powdered adult formulas of the
present disclosure, the ingredients of which are listed in the
table below. All ingredient amounts are listed as kg per 1000 kg
batch, unless otherwise specified.
TABLE-US-00005 TABLE 5 Nutritional Composition Ingredients for
Examples 11-15 and other Exemplified Compositions. INGREDIENT
EXAMPLE 11 EXAMPLE 12 EXAMPLE 13 EXAMPLE 14 EXAMPLE 15 Corn syrup
259.86 259.86 259.86 259.86 259.86 Corn maltodextrin 216.25 216.25
216.25 216.25 216.25 Sucrose 177.95 177.95 177.95 177.95 177.95
Corn oil 155.4 155.4 155.4 155.4 155.4 Sodium and calcium
caseinates 152.43 152.43 152.43 152.43 152.43 Acid Casein 132.97
132.97 132.97 132.97 132.97 Calcium Caseinate 19.46 19.46 19.46
19.46 19.46 Soy protein isolate 28.94 28.94 28.94 28.94 28.94
Mono-and diglyceride/Lutein approx. approx. approx. approx. approx.
Blend 15.53 15.6 2.3 7.8 5.91 Monoglycerides and diglycerides 15.50
15.57 2.3 7.8 5.91 Lutein 2.9 g 34.0 g 5.0 g 2.5 g 12.5 g
Artificial flavoring 14.55 14.55 14.55 14.55 14.55 Potassium
citrate 10.75 10.75 10.75 10.75 10.75 20% Potassium Citrate 3.22
3.22 3.22 3.22 3.22 Potassium Citrate 7.54 7.54 7.54 7.54 7.54
Magnesium chloride 7.53 7.53 7.53 7.53 7.53 Calcium phosphate 6.23
6.23 6.23 6.23 6.23 Sodium citrate 5.66 5.66 5.66 5.66 5.66
Potassium chloride 3.79 3.79 3.79 3.79 3.79 Soy lecithin 3.44 3.44
3.44 3.44 3.44 Ascorbic acid 1.91 1.91 1.91 1.91 1.91 Choline
chloride 1.64 1.64 1.64 1.64 1.64 Zinc sulfate 155.8 g 155.8 g
155.8 g 155.8 g 155.8 g dl-alpha-tocopheryl acetate 129.6 g 129.6 g
129.6 g 129.6 g 129.6 g Niacinamide 124.2 g 124.2 g 124.2 g 124.2 g
124.2 g Ferrous sulfate 121.7 g 121.7 g 121.7 g 121.7 g 121.7 g
Calcium pantothenate 80.3 g 80.3 g 80.3 g 80.3 g 80.3 g Manganese
sulfate 38.7 g 38.7 g 38.7 g 38.7 g 38.7 g Cupric sulfate 21.1 g
21.1 g 21.1 g 21.1 g 21.1 g Thiamine chloride hydrochloride 20.5 g
20.5 g 20.5 g 20.5 g 20.5 g Pyridoxine hydrochloride 19.8 g 19.8 g
19.8 g 19.8 g 19.8 g Riboflavin 16.0 g 16.0 g 16.0 g 16.0 g 16.0 g
Vitamin A palmitate 10.4 g 10.4 g 10.4 g 10.4 g 10.4 g Folic acid
2.8 g 2.8 g 2.8 g 2.8 g 2.8 g Biotin 2.4 g 2.4 g 2.4 g 2.4 g 2.4 g
Chromium chloride 1.3 g 1.3 g 1.3 g 1.3 g 1.3 g Sodium molybdate
1.0 g 1.0 g 1.0 g 1.0 g 1.0 g Potassium iodide 0.44 g 0.44 g 0.44 g
0.44 g 0.44 g Sodium selenate 0.43 g 0.43 g 0.43 g 0.43 g 0.43 g
Phylloquinone 0.22 g 0.22 g 0.22 g 0.22 g 0.22 g Cyanocobalamin
0.05 g 0.05 g 0.05 g 0.05 g 0.05 g Vitamin D3 0.03 g 0.03 g 0.03 g
0.03 g 0.03 g
Examples 16-18
[0303] Examples 16-18 illustrate liquid adult formulas of the
present disclosure, the ingredients of which are listed in the
table below. All ingredient amounts are listed as kg per 1000 kg
batch, unless otherwise specified.
TABLE-US-00006 TABLE 6 Nutritional Composition Ingredients for
Examples 16-18 and other Exemplified Compositions. Ingredient
Example 16 Example 17 Example 18 Water QS QS QS Corn Syrup 33 kg 33
kg 33 kg Maltodextrin 28 kg 28 kg 28 kg Sucrose 19.4 kg 19.4 kg
19.4 kg Mono- and 18.3 kg 6.1 kg 2.4 kg Diglyceride/DHA premix
Monoglycerides and 9.6 kg 5.1 kg 2 kg Diglycerides Docosahexaenoic
Acid Oil 8.7 kg 1 kg 0.4 kg (60% DHA in vegetable oil) Caseinate
8.7 kg 8.7 kg 8.7 kg High Oleic Safflower Oil -- 2 kg 3.3 kg Canola
Oil -- 2 kg 3.3 kg Soy Protein 3.7 kg 3.7 kg 3.7 kg Whey Protein
3.2 kg 3.2 kg 3.2 kg Caseinate 2.9 kg 2.9 kg 2.9 kg Corn Oil -- 1
kg 1.6 kg Tricalcium Phosphate 1.4 kg 1.4 kg 1.4 kg Potassium
Citrate 1.3 kg 1.3 kg 1.3 kg Magnesium Phosphate 952 gm 952 gm 952
gm Lecithin 658 gm 658 gm 658 gm Magnesium chloride 558 gm 558 gm
558 gm Vanilla Flavor 544 gm 544 gm 544 gm Sodium Chloride 272 gm
272 gm 272 gm Carrageenan 227 gm 227 gm 227 gm Choline chloride 218
gm 218 gm 218 gm UTM/TM Premix 165 gm 165 gm 165 gm Potassium
Chloride 146 gm 146 gm 146 gm Ascorbic Acid 145 gm 145 gm 145 gm
Sodium Citrate 119 gm 119 gm 119 gm Potassium Hydroxide 104 gm 104
gm 104 gm Docosahexaenoic Acid Oil 8.7 kg 1 kg 1 kg (60% DHA in
vegetable oil) WSV Premix 33 gm 33 gm 33 gm Vit DEK Premix 29 gm 29
gm 29 gm Vitamin A 3.7 gm 3.7 gm 3.7 gm Potassium Iodide 86 mcg 86
mcg 86 mcg
Example 19
[0304] In this Example, the effect of mixing lutein, a polar lipid
nutrient, in a combination of monoglycerides and diglycerides on
the lutein bioavailability is evaluated and compared to the
bioavailability of lutein that is mixed in typical
triglyceride-based oils.
[0305] Male Sprague Dawley rats weighing between 280 and 330 grams
were all fed a normal commercially available Purina Rat Chow for
one week. Rats were then fasted overnight prior to surgery, and
under anesthesia, a laparotomy was performed, and the intestinal
lymph duct was cannulated according to the procedure of Tso et al.,
"The Absorption of Lipid and Lipoprotein Synthesis," Lipid Research
Methodology, Chapter 5: 191-216 (1984) Alan R. Liss, Inc., NY, NY,
hereby incorporated by reference to the extent consistent herewith.
The superior mesenteric artery was isolated, but not occluded. A
silicon infusion tube (1.6 mm OD) was placed in the stomach for
future infusion of a test sample. The fundic incision was closed by
a purse string suture. The rats were allowed to recover for 24
hours before infusion began.
[0306] The test samples included the following solutions: (1)
lutein in safflower oil (control); (2) lutein in monoglycerides and
diglycerides derived from corn oil; and (3) lutein in
monoglycerides and diglycerides derived from high oleic acid
sunflower oil.
[0307] The rats were randomly assigned to three groups. The animals
were intragastrically infused 24 hours after surgery with 20 mg/kg
lutein from their respective lutein-containing solutions.
[0308] Lymph was collected in pre-cooled tubes 1 hour before the
lutein infusions (fasting) and then hourly for 6 hours after
initiation of infusion. At the end of the 6 hours infusion, the
rats were sacrificed by exsanguination.
[0309] The lymph lipids were extracted and analyzed for lutein
concentration using high-performance liquid chromatography with
programmed wavelength ultraviolet detection (Craft Technologies,
Wilson, N.C.).
[0310] FIG. 1 shows the hourly lymph flow rate over a period of 6
hours following lutein administration. FIG. 2 shows the hourly
lutein output in lymph over a 6 hour period after lutein
administration. FIG. 3 shows cumulative lymphatic lutein absorption
over a 6 hour period after lutein administration. FIG. 4 shows the
percent change in lymphatic lutein absorption of lutein with mono-
and diglycerides versus control over a period of 6 hours.
[0311] As shown in FIG. 1, the mean fasting lymph flow for all
three groups of rats varied between 2.3 and 2.6 mL/h. In all
groups, lymph flow increased significantly after lutein infusion
and reached a maximum output between 2.7 and 3.9 mL/h during hours
2-3 after lipid infusion. There were no differences in lymph flow
rates between the groups indicating that the use of lutein in mono-
and diglycerides did not significantly affect lymph flow rates
versus the lutein in triglyceride control. FIG. 2 shows the
lymphatic lutein output in .mu.g/h during the first 6 hours after
gastric feeding of the 3 different lutein samples. The lymphatic
output of lutein increased in all groups during the first 3 hours
and reached a steady state from hour 3 to hour 5 after lutein
administration. However, there was a significant increase in the
amounts of lymphatic lutein output 2-6 hours after lutein
administration with lutein given with mono and diglycerides (corn
and HOSO) as compared with lutein in the triglyceride control. FIG.
3 shows the above lutein absorption improvements with mono and
diglycerides by demonstrating a significantly higher cumulative
lutein absorption over the 6 hour absorption period (area under the
curve (AUC)) versus the lutein in the triglyceride control. When
this data is expressed as a percent change in lymphatic lutein
absorption, as shown in FIG. 4, there was a 78%-88% increase in
lymphatic lutein absorption of lutein with mono and diglycerides
versus the control over the 6 hour infusion period. This data
indicates that the polar lipid nutrient lutein has improved
bioavailability in a mixture of monoglycerides and diglycerides as
compared to triglyceride oils.
Example 20
[0312] In this Example, the effect of the addition of an activated
premix comprising lutein and the effect of the addition of a
protected premix on the bioavailability of the lutein in a
nutritional composition is evaluated and compared to the
bioavailability of lutein that is typically added to
triglyceride-based oils prior to addition to the nutritional
composition.
[0313] Male Sprague Dawley rats weighing between 280 and 330 grams
were all fed a normal commercially available Purina Rat Chow for
one week. Rats were then fasted overnight prior to surgery, and
under anesthesia, a laparotomy was performed, and the intestinal
lymph duct was cannulated according to the procedure of Tso et al.,
"The Absorption of Lipid and Lipoprotein Synthesis," Lipid Research
Methodology, Chapter 5: 191-216 (1984) Alan R. Liss, Inc., NY, NY,
hereby incorporated by reference to the extent consistent herewith.
The superior mesenteric artery was isolated, but not occluded. A
silicon infusion tube (1.6 mm OD) was placed in the stomach for
future infusion of a test sample. The fundic incision was closed by
a purse string suture. The rats were allowed to recover for 24
hours before infusion began.
[0314] The test samples included the following nutritional
compositions: (AET-C) lutein added to high oleic safflower oil at
120.degree. F. for 10 minutes, added to the protein-in-fat slurry
(control); (AET-1) lutein premixed with monoglycerides and
diglycerides at 120.degree. F. for 10 minutes, lecithin added to
the premix post heating and allowed to mix for 5 minutes, added to
the protein-in-fat slurry to form an MDG-protected premix prior to
addition to the remainder of the composition; (AET-2) lutein
premixed with monoglycerides and diglycerides at 120.degree. F. for
10 minutes, added to the protein-in-water slurry to form an
activated premix, prior to addition to the remainder of the
composition; (AET-3) lutein premixed with monoglycerides and
diglycerides at 120.degree. F. for 10 minutes added to the
protein-in-fat slurry prior to addition to the remainder of the
composition, and (AET-4) lutein premixed with monoglycerides and
diglycerides at 120.degree. F. for 10 minutes, choline-added as
choline chloride to the premix post heating and allowed to mix for
5 minutes, added to the protein-in-water slurry to form an
activated premix prior to addition to the remainder of the
composition. The compositions for each of the test samples are
provided in detail in the following table.
TABLE-US-00007 TABLE 7 Nutritional Composition Ingredients for
Example 20 and other Exemplified Compositions. AET-C AET-1 AET-2
AET-3 AET-4 Amount Amount Amount Amount Amount per per per per per
Ingredient Name: 1000 kg Unit 1000 kg Unit 1000 kg Unit 1000 kg
Unit 1000 kg Unit Ingredient Water Q.S. kg Q.S. kg Q.S. kg Q.S. kg
Q.S. kg Lactose 49.27 kg 49.27 kg 49.27 kg 49.27 kg 49.27 kg
Non-Fat Dry Milk 22.06 kg 22.06 kg 22.06 kg 22.06 kg 22.06 kg High
Oleic Safflower 13.44 kg 12.78 kg 12.78 kg 12.78 kg 12.78 kg Oil
Soy Oil 10.06 kg 9.57 kg 9.57 kg 9.57 kg 9.57 kg Coconut Oil 9.60
kg 9.13 kg 9.13 kg 9.13 kg 9.13 kg Galactooligosaccharides 9.40 kg
9.40 kg 9.40 kg 9.40 kg 9.40 kg Whey Protein 6.31 kg 6.31 kg 6.31
kg 6.31 kg 6.31 kg Concentrate Monoglyceride and 1.62 kg 1.62 kg
1.62 kg 1.62 kg Diglyceride Oil Ascorbic Acid 611.99 g 611.99 g
611.99 g 611.99 g 611.99 g Calcium Carbonate 559.70 g 559.70 g
559.70 g 559.70 g 559.70 g Potassium Citrate 545.40 g 545.40 g
545.40 g 545.40 g 545.40 g ARASCO 395.30 g 395.30 g 395.30 g 395.30
g 395.30 g Mortierella Alpina Oil KOH 45% 368.00 g 368.00 g 368.00
g 368.00 g 368.00 g Centrol (Lecithin) 346.99 g 346.99 g 346.99 g
346.99 g 346.99 g Myverol 346.99 g 346.99 g 346.99 g 346.99 g
346.99 g Nucleotide-Choline 314.15 g 314.15 g 314.15 g 314.15 g
314.15 g Premix DHASCO 208.80 g 208.80 g 208.80 g 208.80 g 208.80 g
Crypthecodinium Cohnii Oil Vit/Min/Taur Premix 176.00 g 176.00 g
176.00 g 176.00 g 176.00 g Seakem RLC 175.26 g 175.26 g 175.26 g
175.26 g 175.26 g Carrageenan Sodium Citrate 163.00 g 163.00 g
163.00 g 163.00 g 163.00 g Sodium Chloride 145.30 g 145.30 g 145.30
g 145.30 g 145.30 g Magnesium Chloride 126.50 g 126.50 g 126.50 g
126.50 g 126.50 g Potassium Phosphate 117.60 g 117.60 g 117.60 g
117.60 g 117.60 g Dibasic Ferrous sulfate 66.50 g 66.50 g 66.50 g
66.50 g 66.50 g Choline Chloride 44.97 g 44.97 g 44.97 g 44.97 g
44.97 g Vitamin A, D3, E, K1 35.80 g 35.80 g 35.80 g 35.80 g 35.80
g Magnesium Sulfate 27.40 g 27.40 g 27.40 g 27.40 g 27.40 g Zinc
Sulfate 17.90 g 17.90 g 17.90 g 17.90 g 17.90 g Lutein 12.35 g
12.35 g 12.35 g 12.35 g 12.35 g L-Carnitine 5.50 g 5.50 g 5.50 g
5.50 g 5.50 g Cupric Sulfate 1.90 g 1.90 g 1.90 g 1.90 g 1.90 g
Manganese Sulfate 110.00 mg 110.00 mg 110.00 mg 110.00 mg 110.00 mg
Citric Acid (Processing As As As As As aid) needed needed needed
needed needed Potassium Iodide As As As As As needed needed needed
needed needed Sodium Selenate As As As As As needed needed needed
needed needed Potassium Chloride As As As As As needed needed
needed needed needed
[0315] The rats were randomly assigned to five groups. The animals
were intragastrically infused 24 hours after surgery with 3 ml of
nutritional formula that delivered 5.8 .mu.g lutein per animal from
their respective lutein-containing solutions.
[0316] Lymph was collected in pre-cooled tubes 1 hour before the
nutritional composition infusions (fasting) and then hourly for 8
hours after initiation of infusion. At the end of the 8 hours
infusion, the rats were sacrificed by exsanguination.
[0317] The lymph lipids were extracted and analyzed for lutein
concentration using high-performance liquid chromatography with
programmed wavelength ultraviolet detection (Craft Technologies,
Wilson, N.C.).
[0318] FIG. 5 shows the hourly lymph flow rate over a period of 8
hours following administration of liquid infant compositions
containing lutein. FIG. 6 shows the hourly lutein output in lymph
over an 8 hour period after administration of liquid infant
compositions containing lutein. FIG. 7 shows cumulative lymphatic
lutein absorption over a 6 hour and 8 hour period after
administration of liquid infant compositions containing lutein.
FIG. 8 shows the percent change in lymphatic lutein absorption over
a period of 6 hours and 8 hours following administration of liquid
infant compositions containing lutein versus control.
[0319] As shown in FIG. 5, the mean fasting lymph flow for all
three groups of rats varied between 2.4 and 2.7 mL/h. In all
groups, lymph flow increased significantly after nutritional
composition infusion and reached a maximum output between 2.8 and
3.1 mL/h during hours 2-3 after nutritional composition infusion.
There were no differences in lymph flow rates between the groups
indicating that the use of a nutritional composition including
either an activated premix with lutein or an MDG-protected premix
with lutein, did not significantly affect lymph flow rates versus
the control nutritional composition including lutein in
triglyceride. FIG. 6 shows the lymphatic lutein output in .mu.g/h
during the first 8 hours after gastric feeding of the 5 different
nutritional composition samples. The lymphatic output of lutein
increased in all groups during the first 3 hours and reached a
steady state from hour 2 to hour 3 after nutritional formula
administration. However, there was a significant increase in the
amounts of lymphatic lutein output 2-8 hours after lutein
administration when lutein was given in the form of an activated
premix (AET-2, AET-4) or with lutein given in the form of an
MDG-protected premix (AET-1), as compared with lutein mixed
directly with the triglycerides control or non-activated premix
containing lutein, monoglycerides and diglycerides (AET-C and
AET-3, respectively). FIG. 7 shows the lutein absorption
improvements with an activated premix with lutein or an
MDG-protected premix with lutein by demonstrating significantly
higher cumulative lutein absorption over the 6 and 8 hour
absorption period (area under the curve (AUC)) versus the lutein in
the control and non-activated samples. When this data is expressed
as a percent change in lymphatic lutein absorption, as shown in
FIG. 8, there was a 106%-170% increase in lymphatic lutein
absorption of compositions comprising activated premix with lutein
or an MDG-protected premix with lutein versus the control and
non-activated sample over the 6 hour infusion period. There was a
121%-165% increase in lymphatic lutein absorption of the
compositions comprising an activated premix with lutein or an
MDG-protected premix with lutein versus the control and
non-activated sample over the 8 hour infusion period. These data
indicate that the lipophilic compound lutein has improved
bioavailability when in an activated premix or an MDG-protected
premix as compared to when it is in triglyceride oils.
Example 21
[0320] In this Example, the test samples evaluated in Example 20
are further evaluated for lutein that is not solubilized by the fat
in the sample nutritional compositions. The five samples, AET-C,
AET-1, AET-2, AET-3 and AET-4 were each subjected to high speed
centrifugation (31,000.times.g; 20.degree. C.; 4 h). The visible
absorbance of the resulting aqueous fractions (after 2:8 dilution
with water, followed by syringe filtration through a 0.45 .mu.m
PTFE membrane) was measured at 476 nm (a lutein absorbance
maximum). The absorbance readings (expressed as milliabsorbance
units per gram of aqueous fraction (mAU/g)) are reported in the
table below, along with a ranking of the in vivo lutein response
for the same variations tested in Example 20.
TABLE-US-00008 TABLE 8 Relationship between Lutein in the Aqueous
Fraction with in vivo Response Visible light absorbance, In vivo
ranking of lutein Sample ID mAU/g, 476 nm response AET-C 57.6 1
(lowest response) AET-1 62.7 4 AET-2 61.3 3 AET-3 59.4 2 AET-4 64.9
5 (highest response)
[0321] As may be seen from the table, the absorbance values are
correlated with the in vivo findings in that each of the sample
nutritional compositions comprising an activated premix (AET-2 and
AET-4) or an MDG-protected premix (AET-1) have higher lutein
absorbance values than those sample nutritional compositions that
do not comprise an activated premix (AET-C and AET-3). This means
that more lutein is present in the aqueous fraction of the sample
nutritional compositions comprising an activated premix or an
MDG-protected premix. Thus, at least in relation to the sample
nutritional compositions, the bioavailability of lutein was
accurately predicted in vitro utilizing this method.
[0322] In this Example, the effect of the addition of an activated
premix or an MDG-protected premix to a nutritional composition on
the bioavailability of other lipophilic compounds such as total
triglycerides, phospholipids, arachidonic acid (ARA) and
docosahexaenoic acid (DHA) is evaluated and compared to the
bioavailability of the lipophilic compounds that are consumed in
the absence of the activated premix or the MDG-protected
premix.
[0323] A similar protocol for animal infusions and lymph collection
as described in Example 20 was followed, including the use of the
same test samples. The lymph lipids were extracted and analyzed for
triglycerides, phospholipids, ARA and DHA concentration using
high-performance liquid chromatography.
[0324] FIG. 9 shows the hourly lymphatic triglyceride output over a
period of 6 hours following nutritional composition administration.
FIG. 10 shows the hourly lymphatic phospholipid output over 6 hours
following nutritional composition administration.
[0325] FIG. 11 and FIG. 12 show the change in lymphatic absorption
of ARA and DHA from fasting, respectively, over a period of 6 hours
following nutritional composition administration.
[0326] As shown in FIG. 9, the lymphatic output of triglycerides
increased in all groups during the 6 hours post administration of
nutritional composition samples. However, there was a significant
increase in the amounts of lymphatic triglyceride output 2-6 hours
after sample administration with an activated premix (AET-2, AET-4)
or an MDG-protected premix (AET-1) as compared with the samples
having the monoglycerides and diglycerides mixed directly with the
triglycerides in a non-activated premix or a non-MDG-protected
premix (AET-C and AET-3). Similarly, FIG. 10 shows that the
lymphatic output of phospholipids increased in all groups during
the 6 hours post sample administration. There was a significant
increase in the amount of lymphatic phospholipid output 2-6 hours
after sample administration with a given in an activated premix
(AET-2, AET-4) or an MDG-protected premix (AET-1) as compared with
the samples having the monoglycerides and diglycerides mixed
directly with the triglycerides in a non-activated premix or a
non-MDG-protected premix (AET-C and AET-3).
[0327] As shown in FIG. 11, there was a significant increase in
lymph levels of ARA from fasting during the 6 hours post
administration of the nutritional compositions containing an
activated premix (AET-2) or an MDG-protected premix (AET-1) as
compared with the non-activated or non-MDG-protected control. FIG.
12 shows similar results by demonstrating a significant increase in
lymph levels of DHA from fasting during the 6 hours post
administration of the nutritional formulas containing an activated
premix (AET-2) or an MDG-protected premix as compared with the
non-activated or non-MDG-protected control.
Example 22
[0328] A study was conducted to compare a commercially
representative nutritional powder (details of powder components are
listed below) with and without MDG. The nutritional powder batches
(control and MDG) were reconstituted (152 g/L), centrifuged
(31,000.times.g/20.degree. C./4 h), and analyzed as outlined below.
The metric used to analyze the bound lutein is the early eluting
peak of the SEC chromatogram, which corresponds to the high
molecular weight peak. The numerical value of this peak is the area
of the peak at 476 nm per microliter of sample injected.
Additionally, the cream layer, which is the upper layer obtained
upon the high speed centrifugation, is used to examine the
mechanism of action for enhanced lutein solubility, as well as to
determine which proteins are responsible for the bound lutein
assemblies.
[0329] The markers of bound lutein (high MW size exclusion
chromatography (SEC) peak area at 476 nm) were significantly higher
for MDG batches versus control. The cream layer protein comparison
and the attached chromatograms were consistent with MDG-facilitated
formation of high MW, but water dispersible (aqueous fraction), and
even water soluble (100 k ultrafiltrate), assemblies of native
proteins (casein, .alpha.-lactalbumin, .beta.-lactoglobulin) and
lutein.
TABLE-US-00009 TABLE 9 Nutritional Composition Ingredients for
Example 22 and other Exemplified Compositions. INGREDIENTS (MDG
INGREDIENTS (Control Incorporated Formula) AMOUNT PER 1000 KG
Formula) AMOUNT PER 1000 KG Non Fat Dry Milk 442 kg Non Fat Dry
Milk 442 kg Lactose 167.8 kg Lactose 167.8 kg Sucrose 111.9 kg
Sucrose 111.9 kg High oleic safflower oil 86.6 kg High oleic
safflower oil 89.9 kg Soy oil 67.4 kg Soy oil 67.4 kg Coconut Oil
64.4 kg Coconut Oil 64.4 kg Galactooligosaccharides (GOS) 36.2 kg
Galactooligosaccharides (GOS) 36.2 kg Whey protein concentrate
(75%) 35.7 kg Whey protein concentrate (75%) 35.7 kg Potassium
Citrate 6.8 kg Potassium Citrate 6.8 kg Capmul GM040 (MDG Oil) 3.29
kg Calcium Carbonate 2.7 kg Calcium Carbonate 2.7 kg Sodium
Chloride 1.7 kg Sodium Chloride 1.7 kg Nucleotide/Choline Premix
1.6 kg Nucleotide/Choline Premix 1.6 kg Choline bitartrate 805.5 g
Choline bitartrate 805.5 g Cytidine 5'-monophosphate 166.3 g
Cytidine 5'-monophosphate 166.3 g Disodium guanosine 121 g
5'-monophosphate Disodium guanosine 5'- 121 g Disodium uridine 5'-
105.4 g monophosphate monophosphate Disodium uridine 5'- 105.4 g
Adenosine 5'- 63.2 g monophosphate monophosphate Adenosine
5'-monophosphate 63.2 g Maltodextrin (premix diluent) q.s.
Maltodextrin (premix diluent) q.s. Lecithin, PCR negative, IP 1.4
kg Lecithin, PCR negative, IP 1.4 kg Ascorbic Acid 1.4 kg Ascorbic
Acid 1.4 kg Tricalcium phosphate 1.16 kg Tricalcium phosphate 1.16
kg Water Soluble Vitamin 869.7 g and Mineral Premix Water Soluble
Vitamin and Mineral 869.7 g Taurine 265.8 g Premix Taurine 265.8 g
m-Inositol 193.0 g m-Inositol 193.0 g Zinc Sulfate, Monohydrate
89.0 g Zinc Sulfate, Monohydrate 89.0 g Niacinamide 56.7 g
Niacinamide 56.7 g d-Calcium Pantothenate 34.0 g d-Calcium
Pantothenate 34.0 g Ferrous Sulfate, Dried 29.8 g (Monohydrate)
Ferrous Sulfate, Dried 29.8 g Copper Sulfate, Anhydrous 10.4 g
(Monohydrate) Copper Sulfate, Anhydrous 10.4 g Thiamine Chloride
8.80 g Hydrochloride Thiamine Chloride 8.80 g Riboflavin 3.88 g
Hydrochloride Riboflavin 3.88 g Pyridoxine Hydrochloride 3.56 g
Pyridoxine Hydrochloride 3.56 g Folic Acid 1.31 g Folic Acid 1.31 g
Manganese Sulfate, Monohydrate 1.01 g Manganese Sulfate,
Monohydrate 1.01 g Biotin 0.343 g Biotin 0.343 g Sodium Selenate,
Anhydrous 0.206 g Sodium Selenate, Anhydrous 0.206 g Cyanocobalamin
0.027 g Cyanocobalamin 0.027 g Corn Starch (modified) 27.4 g (0.1%
B12 spray dry) Corn Starch (modified) (0.1% 27.4 g Dextrose q.s.
B12 spray dry) Dextrose q.s. Docosahexaenoic acid (DHA) 759.2 g
Docosahexaenoic acid (DHA) 759.2 g Vitamin A, D3, E, K1 premix
386.9 g Vitamin A, D3, E, K1 premix 386.9 g d-.alpha.-tocopheryl
acetate 78.2 g d-.alpha.-tocopheryl acetate 78.2 g Vitamin A
palmitate 14.6 g Vitamin A palmitate 14.6 g Phylloquinone 851.2 mg
Phylloquinone 851.2 mg Vitamin D3 103.0 mg Vitamin D3 103.0 mg
Vegetable oil (premix diluent) q.s. Vegetable oil (premix diluent)
q.s. Choline Chloride 284 g Choline Chloride 284 g Magnesium
Chloride 261.1 g Magnesium Chloride 261.1 g Ferrous Sulfate 222.0 g
Ferrous Sulfate 222.0 g Arachidonic acid 205.2 g Arachidonic acid
205.2 g Ascorbyl Palmitate 164.4 g Ascorbyl Palmitate 164.4 g
Tocopherol-2 food grade antioxidant 87.8 g Tocopherol-2 food grade
antioxidant 87.8 g Mixed tocopherols 63.7 g Mixed tocopherols 63.7
g Vegetable oil (diluent) q.s. Vegetable oil (diluent) q.s. Lutein
20% 12.5 g Lutein 20% 12.5 g Lutein 2.5 g Lutein 2.5 g
Beta-Carotene (30%) 3.22 g Beta-Carotene (30%) 3.22 g
.beta.-carotene 966 mg .beta.-carotene 966 mg Potassium Hyrdoxide
as needed Potassium Hyrdoxide as needed
Example 23
[0330] A study was conducted to compare the bound lutein size
estimates within the aqueous fraction. Bound lutein size estimates
were investigated via SAXS, SEC, and MALDI-TOF.
[0331] In summary, the bound lutein may be placed into two or three
categories: (a) large (.about.100-1000 .ANG., or >250 kD) water
soluble/dispersible aggregates that are not discrete structures and
span a broad size range as determined by SAXS and SEC analysis; (b)
native protein/lutein complexes (.about.10-50 kD, .about.27 TO 57
.ANG.) that are generally discrete, well-defined structures, which
is supported by MALDI-TOF and SEC data; and (c) a relatively small
(.about.8 .ANG., or .about.3-4 kD) crystal. The small structure may
or may not comprise lutein.
Example 24
[0332] A study was conducted to examine the grand average of
hydropathicity index (GRAVY) of milk proteins. The GRAVY value is a
measure of hydrophobicity and water solubility, and may be
predictive of a protein's ability to bind a lipophilic molecule
such as lutein.
[0333] The lutein binding capacities of eight major milk proteins
were predicted from their GRAVY values using the plot listed below.
The GRAVY values of said milk proteins are listed in the table
below, as well as their predicated lutein binding capacity. It was
found that the milk proteins .gamma..sub.2-casein,
.beta.-lactoglobulin, .beta.-casein and .alpha.-lactalbumin had the
highest predicted lutein binding capacity of <2 .mu.M, which
correlates with MALDI-TOF data implicating these proteins as
binding participants in the lutein/protein complexes. Accordingly,
the GRAVY values were able to correlate accurate lutein binding
trends of proteins that have been shown to efficiently interact
with lutein within the nutritional product. This results in GRAVY
values potentially providing insight into other potential proteins,
and may be useful in optimizing the nutritional product system.
TABLE-US-00010 TABLE 10 GRAVY Value Analysis of Different Proteins
Protein MW, kD GRAVY Lutein K.sub.d, .mu.M K.sub.d Source StARD3
50.5 0.001 0.45 Published GSTP1 23.2 -0.131 1.35 Published HSA 66.5
-0.395 1.69 Published Milk proteins .gamma..sub.2-casein 11.8
-0.136 1.05 Predicted .beta.-lactoglobulin 18.3 -0.162 1.13
Predicted .beta.-casein 23.6 -0.355 1.68 Predicted
.alpha.-lactalbumin 14.2 -0.453 1.96 Predicted BSA 66.4 -0.475 2.02
Predicted .kappa.-casein 19.0 -0.557 2.26 Predicted
.alpha.-s.sub.1-casein 23.0 -0.704 2.68 Predicted .alpha.-s2-casein
24.3 -0.918 3.29 Predicted
Example 25
[0334] A study was conducted to investigate the size/mass of the
high molecular weight lutein assemblies. A SEC calibration curve
was determined to analyze the aforementioned high molecular weight
assemblies. These values were also compared to the data presented
in Example 3, regarding size of the high molecular weight lutein
assembly.
[0335] The reference proteins, their SEC elution times and
molecular weight are listed in the table below, and were used to
estimate the size of the high molecular weight lutein assembly. The
SEC analysis was performed as recited in the preceding Examples. By
extrapolation from a reference protein plot, the size of the high
molecular weight lutein assembly was found to be in the ranges of
60-380 kD, with a peak at 160 kD. These values correspond to
particle size diameters of 60-96 .ANG., with a peak at 78 .ANG..
The estimated values presented below, appear to be consistent with
SAXS data presented in Example 23, wherein it corresponds with the
intermediated size category of 10-100 .ANG.. It should be noted
that particles in the large category, presented in Example 23,
would be excluded from the ultrafiltrate, and there may be larger
lutein assemblies in the aqueous fraction prior to ultrafiltration.
Additionally, it is noted that the 214 nm/280 nm ratio of the high
molecular weight lutein assembly was in the range of the reference
proteins, suggesting a protein presence in the high molecular
weight assembly.
TABLE-US-00011 TABLE 11 Calibration of Assembly Size with Known
Protein Molecular Weights Reference Protein SEC Elution Time, 214
nm/280 nm (Sigma, bovine) MW, kD minutes peak area ratio
.alpha.-Lactalbumin 14.2 23.67 8.12 .beta.-Lactoglobulin 18.3 22.41
13.6 .beta.-Casein 23.6 21.17 19.1 BSA 66.4 19.01 17.3 Lactoferrin
76.1 19.08 10.6 IgG ~160 19.05 10.7 High MW lutein assembly Range
60 to 380 kD 15.7-20.0 10.6 (~60 to 95 .ANG.) Peak 160 kD 17.66 N/A
(~78 .ANG.)
Example 26
[0336] A study was conducted to examine the high molecular weight
lutein assemblies in commercially available nutritional powder
batches, both before and after centrifugation. The nutritional
powder batches (control and MDG incorporated) were reconstituted
(152 g/L), and were analyzed before and after centrifuged
(31,000.times.g/20.degree. C./4 h). The components of these powders
are listed in the table of Example 22. The table below displays the
high molecular weight lutein assembly analysis via SEC, and is
represented as mAU-min/mg injected. The MDG composition shows
higher initial lutein presence in both the whole product and the
aqueous fraction, wherein it shows even a higher difference in the
aqueous fraction relative to the control composition.
[0337] It was found that the high molecular weight lutein assembly
present within the aqueous fraction was 22% higher than in the
control batch. This indicates significantly enhanced solubility
and/or stability in the MDG composition. Additionally, the
chromatograms illustrate that some fraction (12.3% to 14.5%) of the
high molecular weight lutein assembly is recovered in the
supernatant after high speed centrifugation, which conveys that
these assemblies are sufficiently soluble and stable that they were
not removed or disassembled by the stresses associated with high
speed centrifugation.
TABLE-US-00012 TABLE 12 Analysis of Commercially Representative
Nutritional Composition comprising MDG Batch A545 (MDG Batch A544
(control) Incorporated) Whole product 1977 2070 (w/o
centrifugation) Aqueous Fraction 244 299 (after centrifugation) %
Recovery 12.3% 14.5%
Example 27
[0338] A study was conducted to evaluate high molecular lutein
assemblies facilitated by the presence of MDG vs. high oleic
safflower oil (HOSO). Bench scale model systems were prepared in
nonfat dry milk (NFDM), wherein the NFDM was used at 10% (w/w), a
lipid (HOSO or MDG), and lutein. Lutein was used at variable
amounts, and these compositions were then assessed for the
formation of high molecular weight assemblies. The different
compositions were centrifuged (31,000.times.g/20.degree. C./1 h) to
obtain aqueous fractions, which were compared by a pH 2.5 SEC as
detailed above, and lutein assemblies were analyzed by measuring a
visible light absorbance at 476 nm, and by measuring visible light
transmittance at 860 nm. Additionally, there was a group of
MDG-batches that were not centrifuged, so the whole product was
analyzed for the presence of lutein assemblies.
[0339] The results demonstrate that HOSO has little capacity for
facilitating the high molecular weight lutein assemblies at any of
the different lutein concentrations. In contrast, the MDG
compositions exhibited significant formation of high molecular
weight lutein assemblies, which may be seen with the higher
absorbance at 476 nm. Furthermore, this experiment revealed that
lutein concentration greater than 1.times., showed a decrease in
lutein-assembly formation. This data suggests that there is an
optimal ratio of lutein to MDG for the formation of high molecular
weight lutein assemblies. Additionally, the non-centrifuged samples
reveal that approximately 36% of the total assembly is recovered
following high-speed centrifugation.
Example 28
[0340] A study was conducted to analyze protein involvement with
the high molecular weight lutein assembly. These studies were
performed on batches that comprised MDG or HOSO. The different
batch variations were examined using the same techniques as listed
above. Specifically, bench scale model systems were prepared in
nonfat dry milk, wherein the NFDM was used at 10% (w/w), a lipid
(HOSO or MDG), and lutein. Lutein was used at variable amounts, and
these compositions were then assessed for the formation of high
molecular weight assemblies. The different compositions were
centrifuged (31,000.times.g/20.degree. C./1 h) to obtain aqueous
fractions, which were compared by a pH 2.5 SEC as detailed above.
Protein was analyzed to examine the different molecular weight
fractions and their association with lutein.
[0341] The results demonstrated that MDG enhances the solubility
and/or the stability of both molecular weight fractions, >57 kD
and >137 kD. This was especially enhanced in the 137 kD fraction
for the MGD batches. It should be noted that the density of the
protein assembly may be a factor in its centrifugation recovery, as
would be expected as the lipid content of the assembly
significantly increases, the assembly may be more vulnerable to
removal by high speed centrifugation. Additionally, as shown in the
prior Example, lutein incorporation for compositions with greater
than 1.times. lutein actually decreased assembly formation. This
further suggests that an optimal ratio/range of MDG and lutein are
needed.
TABLE-US-00013 TABLE 13 Model System Investigation of Assembly
Formation High MW lutein Tube Lipid Lutein Centrifugation assembly
1 HOSO 0X Yes 33.5 2 HOSO 1X Yes 33.9 3 HOSO 2X Yes 37.2 4 HOSO 4X
Yes 36.0 5 MDG 0X Yes 36.3 6 MDG 1X Yes 50.0 7 MDG 2X Yes 42.2 8
MDG 4X Yes 41.6 5 MDG 0X No 101 6 MDG 1X No 139 7 MDG 2X No 129 8
MDG 4X No 139
Example 29
[0342] A study was conducted to investigate the formation of high
molecular weight lutein assemblies at different concentrations of
MDG, HOSO and lutein. Specifically, lutein assembly was assessed by
measuring mAU-min/mg at 476 nm, and by determining the presence of
the >137 kD protein fraction present in the aqueous fraction
following centrifugation. These tests were performed as listed
above, with the different compositions being centrifuged
(31,000.times.g/20.degree. C./1 h) to obtain aqueous fractions,
which were analyzed by SEC as detailed above.
[0343] In summary, the MDG batches outperformed the HOSO batches at
both the presence of lutein high molecular weight assemblies, as
well as greater amounts of the >137 kD fraction being present
within the aqueous fraction. Additionally, it should be noted that
the size of the assembly varied, even though the MDG/lutein ratio
was held constant. Since the lutein assemblies were increased using
a combination of MDG and HOSO, their use within the composition may
be advantageous.
TABLE-US-00014 TABLE 14 Model System Investigation II of Assembly
Formation High MW lutein Aqueous assembly, Fraction HOSO, MDG,
Lutein, mAU-min/mg, protein >137 Variable g/L g/L .mu.g/L 476 nm
kD, g/kg 1 0 6.67 10.4 59.7 0.517 2 0 4.44 6.94 58.0 0.498 3 0 2.22
3.47 40.8 0.447 4 6.67 0 10.4 38.1 0.412 5 4.44 0 6.94 37.4 0.408 6
2.22 0 3.47 35.2 0.380 7 3.33 3.33 10.4 55.8 0.490 8 1.11 1.11 3.47
45.7 0.439
Example 30
[0344] A study was performed to assess the amino acid profiles of
the proteins that are involved in the high molecular weight lutein
assembly from the samples in Example 29. In order to compare the
SEC peak area, response to an independent measure of protein
concentration, each of the samples were fractionated by SEC, and
the individual fractions tested for amino acids. In each case,
10.times.4-min fractions were collected (manually), over the
elution range of the hydrolysate (from about 20 min to about 60
min), so that the volume of each fraction was 1.6 mL (i.e., 0.4
mL/min.times.4 min). Each fraction was acid-digested (6 M HCl,
110.degree. C., 22 h), and each digest was tested for eight amino
acids (arginine [R], serine [S], ASX, [B]=aspartic acid
[D]+asparagine [N], GLX [Z]=glutamic acid, [E]+glutamine [Q],
threonine [T], and glycine [G]) by reversed phase HPLC (RPLC) of
their 9-fluorenylmethoxycarbonyl (FMOC) derivatives, using a
modification of methods known in the art. The derivatised standard
and sample solutions were tested for the FMOC derivatives of the
six amino acids by RPLC, using an Agilent model 1100 HPLC system
(Agilent Technologies, Wilmington, Del., USA) with an Agilent model
G1321A fluorescence detector (FLD, Agilent Technologies,
Wilmington, Del., USA). The RPLC system was equipped with a
4.6.times.250 mm i.d., 5 .mu.m, 120 .ANG., YMC-Pack ODS-AM reversed
phase column (Waters, Milford, Mass., USA), maintained at
20.degree. C. by a model G1316A thermostatted column compartment
(Agilent Technologies, Wilmington, Del., USA). RPLC mobile phase A
was 65% (v/v) 0.05 M citric acid, pH 3.0 with NaOH, and 35% (v/v)
ACN; RPLC mobile phase B was 20% (v/v) 0.05 M citric acid, pH 3.0
with NaOH, and 80% (v/v) ACN. The flow rate was 0.5 mL/min, and the
elution program was as follows: 0.0-2.0 min, 0% B; 2.0-25.0 min, 0%
B to 50% B (linear); 25.1-48.0 min, 100% B; 48.1-63.0 min, 0% B.
The FLD excitation wavelength was 262 nm, the emission wavelength
was 310 nm, and the gain was set at 10. The injection volume was 4
.mu.L. The concentrations of R, S, B (D+N), Z (E+Q), T, and G in
the sample solutions (i.e., prepared from the SEC fractions) were
calculated by linear regression from the respective,
individually-constructed, standard curves.
[0345] The table listed below displays the different amino acid
ratios for different proteins and the experimental groups. For
example, NFDM, casein, whey, as well as the protein present in the
aqueous fraction and native protein. Of note, native protein refers
to monomeric unaggregated protein, which has a molecular weight of
approximately in the range of 14-25 kD. Tube 1 refers to the sample
with the highest amount of lutein assembly, while tube 6 refers to
the sample having the lowest. They were chosen to optimally
contrast the distribution of individual proteins within the
assembly.
[0346] The data presented, suggest that a greater presence of
casein is in the high molecular weight fraction than compared to
native protein.
TABLE-US-00015 TABLE 15 Analysis of Proteins Associated with
Assembly Amino acid ratio, w/w GLX/ASX ASX/Gly ASX/Ser ASX/Arg NFDM
2.76 3.59 1.39 2.10 Casein 3.20 3.90 1.23 1.90 Whey 1.58 5.96 2.09
4.42 .beta.-casein 3.92 1.02 0.44 1.72 Tube 1, high 2.47 1.22 0.53
2.59 MW protein Tube 1, native 1.77 1.41 1.05 5.51 protein Tube 6,
high 2.71 0.97 0.42 1.96 MW protein Tube 6, native 1.76 1.48 1.04
5.32 protein
Example 31
[0347] A study was conducted to evaluate the presence of specific
proteins that associate with the high molecular weight lutein
complex. Commercially representative nutritional powder batches
(control and MDG incorporated) were reconstituted (152 g/L), and
were analyzed before and after centrifugation
(31,000.times.g/20.degree. C./2 h). The components of these powders
are listed in the table of Example 22. Reversed phase HPLC was then
used to compare the distribution of individual proteins.
[0348] Determination of the proteins associated with lutein complex
was performed on an Agilent model 1100 HPLC system (Agilent
Technologies, Wilmington, Del., USA) with a model G1315A diode
array detection (DAD) system (Agilent Technologies). The system was
equipped with a 250 mm.times.4.6 mm I.D., 5 .mu.m, 120 .ANG.,
YMC-Pack ODS-AQ reversed-phase column (Waters, Milford, Mass.,
USA). The column was maintained at 40.degree. C. during analyses by
a model G1316A thermostatted column compartment
(AgilentTechnologies). Direct determinations of intact proteins
were performed on the same system equipped with a 250 mm.times.4.6
mm I.D., 5 .mu.m, 300 .ANG., Jupiter C18 reversed phase column
(Phenomenex, Torrance, Calif., USA), also with column temperature
maintained at 40.degree. C.
[0349] The reverse phase HPLC method used a binary gradient
elution, with vacuum-degassed mobile phases A (water, Milli-Q Plus)
and B (650 mL 0.02M KH2PO4, pH 2.9, +175 mL acetonitrile+175 mL
isopropanol). The injection volume was 2 .mu.L, and the elution
program was 0% B at 0.3 mL/min from 0.0 to 20.0 min, 16% B at 0.5
mL/min from 20.2 to 40.0 min, 100% B at 0.5 mL/min from 40.1 to
45.0 min, 0% B at 0.5 mL/min from 45.1 to 59.0 min, and 0% B at 0.3
mL/min from 59.5 to 60.0 min. The intact protein method also used a
binary gradient elution, with vacuum-degassed mobile phases A (800
mL Milli-Q Plus water+200 mL acetonitrile+0.500 mL trifluoroacetic
acid) and B (250 mL Milli-Q Plus water+750 mL acetonitrile+0.500 mL
trifluoroacetic acid). The elution program, with a constant flow
rate of 0.6 mL/min, was 0% B from 0.0 to 5.0 min 0 to 100% B from
5.0 to 40.0 min, 100% B from 40.0 to 45.0 min, and 0% B from 45.0
to 60.0 min. The detection wavelengths were 214 nm and 280 nm, with
reference at 590 nm. The injection volume was 10 .mu.L, and the run
time was 60 min per injection.
[0350] The results demonstrated that the whole product had a
significant decreased presence of native .alpha.-casein in the MDG
batches. This suggests that there may be enhanced calcium
solubility within the MDG composition. Additionally, the aqueous
fraction of MDG compositions had a significant increase in
.beta.-casein, which suggests that said protein may be involved in
the high molecular weight lutein assembly. The presence of
.beta.-casein was further confirmed by the decrease in presence
within the cream layer.
TABLE-US-00016 TABLE 16 Analysis of Proteins Associated with
Assembly for Commercially Representative Nutritional Composition
Sample Comparison Observation Interpretation Whole Product
Significantly Apparent increase in decreased presence of soluble
calcium in A545 native alpha-casein in (alpha-caseins are A545
especially sensitive to soluble calcium) Aqueous Fraction
Significantly Apparent increased (centrifugation increased presence
of incorporation of supernatant) native beta-casein in beta-casein
into water A545 soluble, high MW assemblies, by MDG Cream Layer
Slightly decreased Consistent with (extracted levels of native
interpretation of protein) beta-casein and Aqueous Fraction
beta-lactoglobulin in comparison A545
Example 32
[0351] A study was conducted to investigate the effects of variable
concentrations of MDG and HOSO, as well as the presence of
different proteins, on the formation of high molecular weight
lutein assemblies. Model systems of different proteins: sodium
caseinate, total milk protein, whey protein concentrate and whey
protein hydrolysate were incorporated into the composition at 3.7
wt % of protein in the system. The details of the amounts of MDG,
HOSO and lutein are detailed in the table below. Samples were then
centrifuged (31,000.times.g/20.degree. C./1 h), and the aqueous
fractions were tested by SEC. The mobile phase used for SEC was 70%
H.sub.2O, 30% acetonitrile, and 0.1% trifluoracetic acid.
[0352] The data demonstrated that MDG compositions increased lutein
assembly, and high molecular protein fractions across all groups
compared to HOSO compositions. The addition of whey protein
hydolysate showed the greatest amount of bound lutein. This
suggests that it may be advantageous to have whey protein
hydrolysate within an MDG-based composition. This finding is
interesting in that it may be whey protein hydrolysate's ability to
aggregate that leads to enhanced lutein assembly. Furthermore, the
SEC plots for sodium caseinate and total milk protein suggest the
presence of casein within the lutein complex.
TABLE-US-00017 TABLE 17 Model System Investigation III of Assembly
Formation High MW lutein Aqueous assembly, Fraction Protein HOSO,
MDG, Lutein, mAU-min/mg, protein >137 ingredient g/L g/L .mu.g/L
476 nm kD, g/kg Sodium 6.67 0 6.94 35.6 0.425 Caseinate 0 6.67 6.94
41.6 0.466 Total Milk 6.67 0 6.94 40.6 0.908 Protein 0 6.67 6.94
50.7 0.968 Whey 6.67 0 6.94 34.4 0.115 Protein Concentrate 0 6.67
6.94 53.0 0.253 Whey 6.67 0 6.94 65.9 0.299 Protein Hydrolysate 0
6.67 6.94 97.0 0.453
Example 33
[0353] A study was conducted to evaluate the effect of relative
centrifugal force (RCF) on analyzing the high molecular weight
lutein assembly in the aqueous phase. The components of these
powders are listed in the table of Example 22. Samples were
centrifuged, and the aqueous fractions were tested by SEC. The
mobile phase used for SEC was 70% H.sub.2O, 30% acetonitrile, 0.1%
trifluoracetic acid. The different RCFs used were 1.times.g,
484.times.g, 1935.times.g, 7741.times.g and 31,000.times.g. These
variables and others used in the compositions are listed in the
table below. The compositions were then assessed for high molecular
weight lutein assembly and the presence of protein >137 kD as
done above, as well as bound lipophile index. Bound lipophile
index, a relative marker for of lipophilic nutrients in the high
molecular weight assembly, was determined as the peak ratio, 280
nm/214 nm for protein >137 kD.
[0354] The results exhibit that the MDG compositions generated a
high molecular weight lutein assembly, across all RCFs, which has
significantly greater water solubility, and/or is more stable to
external stress compared to control compositions.
TABLE-US-00018 TABLE 18 Analysis of Variable Centrifugal Forces on
the Assembly RCF High MW Bound Nutritional (20.degree. C., lutein
Protein >137 lipophile Powder Batch 2 hrs) assembly kD, g/kg
index A544 1 .times. g 1839 5.23 0.299 A545 2056 5.34 0.318 A544
484 .times. g 1969 5.27 0.306 A545 2274 5.60 0.327 A544 1935
.times. g 1761 4.92 0.294 A545 2146 5.35 0.322 A544 7741 .times. g
729 2.74 0.232 A545 1163 3.50 0.277 A544 31,000 .times. g.sup. 244
1.07 0.184 A545 299 0.98 0.228
Example 34
[0355] A study was conducted to evaluate the effect of pH, during
the SEC process, on analyzing the presence of high molecular weight
lutein assemblies. The components of these powders are listed in
the table of Example 22. In this experiment, neutral pH SEC was
performed on MDG and control samples. After the samples were
reconstituted, they were analyzed via SEC under the conditions
listed below. No centrifugation was performed on samples prior to
running on SEC.
[0356] The results demonstrate that under these conditions, the
MDG-based compositions performed better than controls in the
formation of the high molecular weight lutein assembly, high
molecular weight protein, and bound lipophile index. Additionally,
this Example demonstrates that the difference between MDG samples
and control is not an artifact of the analytical techniques used to
assess the lutein assemblies.
TABLE-US-00019 TABLE 18 The Effect of pH on Assembly
Characterization A544 (control) A545 (MDG) High MW lutein assembly,
372 419 mAU-min/mg injected High MW protein, g/kg 3.61 3.73 Bound
lipophile index 0.216 0.225
Example 35
[0357] A study was performed to investigate the effects of choline
on the formation of high molecular weight lutein assemblies. Model
systems of a protein ingredient (whey protein concentrate or NFDM
at 3.7 wt. % of protein), MDG (4.4 g/L) and lutein (6.7 mg/L).
Additionally, different experimental groups also had either
lecithin (949 mg/L), choline (45 mg/L), or the combination thereof.
Samples were then centrifuged (31,000.times.g/20.degree. C./1 h),
and the aqueous fractions were tested by SEC for analysis on high
molecular weight lutein assembly, high molecular weight protein,
and bound lipophile index. The data is presented in the table
below.
[0358] In summary, choline had a significant positive effect on the
formation of high molecular weight lutein assemblies in the whey
protein concentrate assemblies. It is hypothesized, without being
bound to a particular theory, that choline being a basic compound
may interact with aspartic acid side chains, which are acidic.
These interactions may alter the 3-dimensional configuration of the
major whey protein .beta.-lactoglobulin such that the hydrophobic
domains are more accessible to lutein binding.
[0359] It should also be noted that choline may have a selective
enhancement of lutein incorporation, since the bound lipophile
index actually decreased, even though lutein increased, when
choline was present.
TABLE-US-00020 TABLE 19 Model System Investigation IV of Assembly
Formation Bound High MW lipophile lutein Aqueous index Protein
assembly, Fraction (280 ingredient mAU- protein > nm/214 (37 g
protein Lecithin, Choline, min/mg, 137 kD, nm Tube per kg) mg/L
mg/L 476 nm g/kg ratio) 1 WPC 0 0 31.2 0.105 0.157 2 NFDM 0 0 41.3
0.413 0.118 3 WPC 949 0 40.3 0.143 0.163 4 NFDM 949 0 47.0 0.441
0.123 5 WPC 0 45 58.6 0.295 0.151 6 NFDM 0 45 50.4 0.443 0.131 7
WPC 949 45 61.7 0.338 0.146 8 NFDM 949 45 48.2 0.451 0.126
Example 36
[0360] A study was conducted to examine variations of commercially
representative nutritional composition batches. Samples were
prepared as detailed above, with composition with and without MDG.
Samples were centrifuged (7,7741.times.g/20.degree. C./2 h), and
the aqueous fractions were tested by SEC for analysis on high
molecular weight lutein assembly, high molecular weight protein,
and bound lipophile index. The data is presented in the table
below.
[0361] In summary, the high molecular weight lutein assembly was
10-30% greater in MGD compositions relative to control
compositions. Additionally, the MDG compositions correlated with a
high molecular weight protein peak. The estimated mass of the high
molecular peak for these batches was in the 240 kD range. There is
also evidence that other lipophilic nutrients (e.g., Vitamin A, D,
E, K, etc.) may be present within the high molecular weight
assembly.
TABLE-US-00021 TABLE 20 Analysis of Commercially Representative
Nutritional Composition in Assembly Formation Batch 017-1 017-2
017-3 017-4 017-5 Lutein/MDG Control PIF, w/ PIW PIF PIW, w/
addition lecithin choline Total lutein, 2.18 1.99 2.06 1.80 1.88
mg/kg Lutein 2236 2464 2815 2844 2845 assembly, conc. Lutein 237 kD
240 kD 247 kD 237 kD 244 kD assembly, mass High MW 5.93 6.34 6.75
6.92 6.87 protein, g/kg Bound 0.301 0.311 0.329 0.327 0.329
lipophile index
Example 37
[0362] A study was conducted to investigate using a neutral pH as
the SEC running condition and its subsequent effect on the high
molecular weight lutein assemblies during separation Samples were
prepared as detailed above, with composition with and without MDG.
Samples were tested by SEC for analysis on high molecular weight
lutein assembly and bound lipophile index. SEC was performed using
0.05M HEPES, pH 7.0, and a Superdex Peptide column. The data is
presented in the table below.
[0363] The results indicate that the high molecular weight lutein
assembly and protein both increased in all MDG batches relative to
control batches. This Example supports the integrity of the
analytical techniques used for lutein assembly analysis.
TABLE-US-00022 TABLE 21 The Effect of pH on Assembly
Characterization in Commercially Representative Nutritional
Composition Batch 017-1 017-2 017-3 017-4 017-5 Lutein/MDG Control
PIF, w/ PIW PIF PIW, w/ addition lecithin choline Total lutein,
2.18 1.99 2.06 1.80 1.88 mg/kg Lutein 820 1170 1160 1190 1060
assembly, conc. Lipophile 7820 10,500 11,300 10,800 9510 assembly,
conc.
Example 38
[0364] A study was performed to evaluate the effects of choline on
the formation of high molecular weight lutein assemblies by
providing choline at variable points during the composition.
Mixtures of water, NFDM, MDG, lutein and choline chloride, wherein
the choline was either added to the MDG and lutein or to the NFDM
slurry. Samples were then centrifuged (31,000.times.g/20.degree.
C./1 h), ultrafiltrated through a 100 k filter, and the aqueous
fractions were tested by SEC for analysis on high molecular weight
lutein assembly, high molecular weight protein, and native protein
bound lutein. The supernatant was diluted 1:4 (v/v) prior to
running on the SEC, with the SEC mobile phase (0.05M KH2PO4, 0.15M
NaCl, pH 7.5). The different parameters examined were high
molecular weight lutein assembly, protein >137 kD, and middle
range protein, which corresponds to the native protein, bound
lutein. Additionally, ultrafiltrate analysis was performed on the
lutein assembly and high molecular weight protein metrics. The data
is presented in the table below.
[0365] The enhancing effect of choline on lutein incorporation
within high molecular weight assemblies appears to be process
dependent. Specifically, it was found that the bound lutein was
significantly higher when choline was added to the NFDM slurry, as
opposed to addition to the MDG and lutein.
TABLE-US-00023 TABLE 22 Model System Investigation V of Assembly
Formation Choline addition point Added to Added to MDG + lutein
NFDM slurry Aqueous Fraction, 214 nm 121 190 (protein metric)
Aqueous Fraction, 476 nm 0.265 0.330 (bound lutein metric) Aqueous
Fraction, 476 nm, 0.0346 0.0455 native protein range (native
protein bound lutein metric) 100K UF permeate, 214 nm 44.8 59.7
(protein metric) 100K UF permeate, 476 nm 0.0343 0.0387 (lutein
metric)
Example 39
[0366] A study was performed to evaluate the effects of variable
components on the formation of high molecular weight lutein
assemblies. The different compositions include various proteins
ingredients (NFDM, .beta.-lactoglobulin, or bovine lactoferrin);
varying amounts of MDG and HOSO, and lutein. The specific details
of the aforementioned variables are listed in the table below.
Samples were then centrifuged (31,000.times.g/20.degree. C./1 h),
and tested for bound lutein via SEC.
[0367] The results demonstrate that both lutein assembly and
protein >137 kD metrics are increased with the presence of MDG
within the composition. Interestingly, the combination of MDG and
HOSO exhibited the greatest amount of lutein assembly formation and
protein >137 kD, compared to MDG and HOSO alone. Additionally,
the use of .beta.-lactoglobulin, or bovine lactoferrin, alone, as
part of the composition showed the capability to form lutein
assemblies and protein >137 kD.
TABLE-US-00024 TABLE 23 Model System Investigation VI of Assembly
Formation Tube 1 2 3 4 5 6 7 8 Protein, 37 g NFDM NFDM NFDM NFDM
NFDM NFDM .beta.LG BLF protein/kg MDG, g/kg 6.64 6.62 3.31 1.66
0.826 0 6.62 6.62 HOSO, g/kg 0 0 3.31 4.96 5.79 6.62 0 0 Lutein,
mg/kg 5.05 10.1 10.1 10.1 10.1 10.1 10.1 10.1 MDG/lutein, 1315 655
328 164 81.8 0 655 655 w/w High MW 49.4 51.7 57.4 57.9 50.7 36.4
11.3 18.6 lutein assembly Protein > 137 0.413 0.407 0.412 0.416
0.393 0.343 0.012 0.018 kD, g/kg Bound 0.136 0.137 0.147 0.147
0.141 0.124 0.270 0.174 lipophile index
Example 40
[0368] A study was conducted to investigate the effects of variable
components on the formation of high molecular weight lutein
assemblies. The different compositions include different protein
ingredients (NFDM, .beta.-lactoglobulin, or bovine lactoferrin) at
37 g of protein per kg, MDG at 6.62 g/kg, and lutein 10.1 mg/kg.
The .beta.-lactoglobulin, and bovine lactoferrin were tested both
with and without centrifugation. Samples were then analyzed by SEC
for high molecular weight lutein assembly, high molecular weight
protein (>137 kD), either with centrifugation or without. If
samples were centrifuged, they were done so at
31,000.times.g/20.degree. C./1 h. Details of the different
experimental group's ability to produce high molecular weight
lutein assemblies are shown in the table below.
[0369] The data revealed that both .beta.-lactoglobulin and bovine
lactoferrin were capable of forming stable, high molecular weight
lutein assemblies. Furthermore, these assemblies were still present
after centrifugation, demonstrating the high solubility/stability
of the bound lutein assemblies.
TABLE-US-00025 TABLE 24 Effect of Different Proteins on Assembly
Formation in a Model System Protein Sample High MW Bound (37 g prep
lutein Protein >137 lipophile Tube protein/kg) for SEC assembly
kD, g/kg index 2 NFDM Aqueous 51.7 0.407 0.137 7 .beta.-LG Fraction
11.3 0.012 0.270 8 BLF 18.6 0.018 0.174 7 .beta.-LG Whole 66.4
0.144 0.324 8 BLF suspension 60.5 0.132 0.305
Example 41
[0370] A study was conducted to examine the effects of variable
components on the formation of high molecular weight lutein
assemblies. Samples were subjected to appropriate thermal
processing during which the composition is pre-heated to
165-185.degree. F. and then passed through homogenizers, first at
1000 psig and then at 4000 psig with NFDM at 37 g of protein/L,
with lutein at 10.1 mg/L, and with varying amounts of HOSO and/or
MDG. After homogenization the each variable is subjected to a high
temperature-short time (HTST) process of about 165-185.degree. F.
and a 16 second hold and then cooled to about 40.degree. F. All
samples were then tested by SEC, either following centrifugation
(31,000.times.g/20.degree. C./1 h) or without centrifugation. The
data is listed in the table below.
[0371] The results demonstrate that the combination of MDG and HOSO
led to the highest amount of lutein assembly formation, as well as
protein >137 kD prior to centrifugation.
TABLE-US-00026 TABLE 25 Model System Investigation VII of Assembly
Formation High MW Bound HOSO, MDG, lutein Protein >137 lipophile
Tube g/L g/L assembly kD, g/kg index 1 6.62 0 216 0.706 0.258 2
2.21 0 102 0.464 0.195 3 0 6.62 167 0.667 0.226 4 0 2.21 65 0.384
0.165 5 3.31 3.31 195 0.712 0.246 6 4.97 1.65 208 0.747 0.247
results of testing without centrifugation 1 6.62 0 612 2.06 0.258 2
2.21 0 242 1.18 0.198 3 0 6.62 648 2.27 0.246 4 0 2.21 260 1.66
0.165 5 3.31 3.31 970 2.96 0.286 6 4.97 1.65 888 2.57 0.297
Example 42
[0372] A study was conducted to examine the effects of variable
components on the formation of high molecular weight lutein
assemblies using a commercially available nutritional powder.
Samples were tested (without dilution or centrifugation) for bound
lutein and related attributes by SEC. The attributes are compared
below, and the bound lutein values (peak area at 476 nm, per mg
injected) are also compared in the table below.
[0373] The results demonstrate that the combination of MDG provided
the highest amount of the metrics listed below relative to
controls. Specifically, MDG/choline outperformed all other groups
across all metrics.
TABLE-US-00027 TABLE 26 Effect of Different Variables on Assembly
Formation in a Commercially Representative Nutritional Composition
Lutein, High MW Bound mg/kg lutein Protein >137 lipophile
Variation (as fed) assembly kD, g/kg index Control 2.47 6995 13.8
0.373 MDG/PIF 2.47 9189 16.3 0.419 MDG/PIW 2.35 10,566 17.8 0.441
MDG/lecithin 2.44 10,298 17.2 0.446 MDG/choline 2.11 10,937 18.1
0.448
Example 43
[0374] A study was conducted to examine the effects of variable
components on the formation of high molecular weight lutein
assemblies. Each variable was subjected to appropriate thermal
processing during which the composition is pre-heated to
165-185.degree. F. and then passed through homogenizers, first at
1000 psig and then at 4000 psig. After homogenization, each
variable was subjected to a high temperature-short time (HTST)
process of about 165-185.degree. F. and a 16 second hold and then
cooled to about 40.degree. F. Samples were homogenized and
pasteurized with the following parameters of protein ("P"=nonfat
dry milk, at 10% w/w), lutein ("L"), HOSO ("H"), MDG ("M"),
lecithin ("Le"), and/or choline ("C"). These samples were tested
for bound lutein and related attributes by size exclusion
chromatography (SEC). Each variable was tested both before and
after centrifugation (at 31,000.times.g/20 C/1 h).
[0375] The results demonstrate that the presence of MDG led to the
formation of lutein assembly formation and protein >137 kD.
TABLE-US-00028 TABLE 27 Model System Investigation VIII of Assembly
Formation High MW Bound lutein Protein >137 lipophile Variable
Sample assembly kD, g/kg index PLH Whole 779 2.21 0.293 PLH
Supernatant 328 (42%) 0.97 (44%) 0.281 PLM Whole 439 1.28 0.273 PLM
Supernatant 321 (73%) 0.90 (75%) 0.286 PLMLe Whole 845 3.03 0.253
PLMLe Supernatant 239 (28%) 1.13 (37%) 0.215 PLMC Whole 432 1.27
0.269 PLMC Supernatant 323 (75%) 0.98 (77%) 0.276
Example 44
[0376] A study was conducted to examine the effects of variable
components on the formation of high molecular weight lutein
assemblies. Samples were subjected to appropriate thermal
processing during which the composition was pre-heated to
165-185.degree. F. and then passed through homogenizers, first at
1000 psig and then at 4000 psig with NFDM at 37 g of protein/L,
with lutein at 10.1 mg/L, and with varying amounts of HOSO and/or
MDG. After homogenization each variable was subjected to a high
temperature-short time (HTST) process of about 165-185.degree. F.
and a 16 second hold, and then cooled to about 40.degree. F. All
samples were then tested by SEC, either following centrifugation
(31,000.times.g/20.degree. C./1 h) or without centrifugation. The
data is listed in the table below.
[0377] Samples prepared with lutein, a lipid (HOSO or MDG), and
varying levels of protein (nonfat dry milk), were tested for bound
lutein and related attributes by size exclusion chromatography
(SEC). Specifically, the parameters investigated were high
molecular weight lutein assembly, protein >137 kD, and bound
lipophile index. The results for samples prepared with and without
centrifugation (31,000.times.g; 20 C; 1 h) are compared below.
[0378] The results demonstrate that both of MDG and HOSO led to the
higher amounts of lutein assembly formation, protein >137 kD,
and bound lipophilic index with increasing amounts of protein.
Additionally, the presences of MDG resulted mostly in higher
amounts of the aforementioned metrics, relative to HOSO.
Interestingly, MDG outperformed HOSO more significantly as the
protein was increased via all metrics.
TABLE-US-00029 TABLE 28 Model System Investigation VIII of Assembly
Formation Protein High MW Bound (NFDM), Lipid, lutein Protein
>137 lipophile Tube g/kg 4.4 g/kg assembly kD, g/kg index 1 24.1
HOSO 107 2.17 0.103 2 24.1 MDG 122 2.11 0.107 3 48.2 HOSO 218 4.15
0.110 4 48.2 MDG 232 4.15 0.112 5 96.4 HOSO 506 10.1 0.121 6 96.4
MDG 616 11.6 0.125 Tested after centrifugation at 31,000 .times. g
and at 20.degree. C. for 1 hr 1 24.1 HOSO 49.8 0.884 0.096 2 24.1
MDG 53.8 1.02 0.098 3 48.2 HOSO 108 2.44 0.102 4 48.2 MDG 118 2.48
0.101 5 96.4 HOSO 287 5.31 0.119 6 96.4 MDG 309 5.47 0.125
Example 45
[0379] A study was conducted to examine the presence of tocopherol
within the high molecular weight assembly. The components of the
tested powders are listed in the table of Example 22. All samples
were formulated and then tested by SEC, following centrifugation
(31,000.times.g/20.degree. C./3 h). The data is listed in the table
below.
TABLE-US-00030 TABLE 29 Analysis of Tocopherol within the Assembly
Nutritional Tocopherols (.alpha.-, .gamma.-, and .delta.-) in
Powder Aqueous Fraction, approximate A544 (control) 1.51 mg/kg A545
(MDG) 1.69 mg/kg
[0380] The results demonstrated that the aqueous fractions of
commercially available nutritional powders with MDG contained
tocopherols are at a level that is 11% greater than control
compositions. The aqueous fraction concentrations (.about.1.69 and
.about.1.51 mg/kg, respectively) appear to represent an appreciable
proportion of the total tocopherol within the commercially
available nutritional composition (approximately 20-30%). The
tocopherol solubility provided by the MDG within the composition is
in stark contrast to the reported solubility limit of vitamin E in
water (20.9 mg/kg at 33.degree. C.). This suggests that the
assemblies comprising MDG enhance the aqueous solubility of
tocopherol.
Example 46
TABLE-US-00031 [0381] TABLE 30 Analysis of Casein Associated with
the Assembly Current commercial Protein system composition
Composition 64% nonfat dry milk NFDM from Holstein- NFDM from
Guernsey (NFDM), 36% WPC Friesian breeds breed Total protein 2.07 g
per 100 kcal 2.07 g per 100 kcal concentration Total .beta.-casein
0.36 g per 100 kcal 0.36 g per 100 kcal concentration Major genetic
variant A1 A2 of .beta.-casein
[0382] Table 31 below shows partial composition information
(protein content) for a commercial liquid nutritional composition
designed for individuals with diabetes, compared with an exemplary
liquid nutritional composition according to the embodiments
disclosed herein. Both illustrate liquid nutritional product with
varying sources of caseinates, and have a caseinate, SPC
combination. The beta-casein utilized in the embodiment on the
right is sourced from Bos indicus has A2 as the major genetic
variant.
TABLE-US-00032 TABLE 31 Partial Composition Information of
Commercially Available Nutritional Composition Current commercial
Protein system composition Composition 80% caseinates, Caseinates
from Caseinates from 20% SPC Bos taurus milk, Bos indicus milk
Holstein-Friesian breeds Total protein 76 g per liter 76 g per
liter concentration Total .beta.-casein 22 g per liter 22 g per
liter concentration Major genetic variant A1 A2 of
.beta.-casein
[0383] A nutritional composition intended for infants and suitable
for utilizing the protein disclosed herein is described in Table 32
below. The 14 grams of protein per liter of nutritional composition
comprises about 18% by weight bovine beta-casein.
TABLE-US-00033 TABLE 32 Nutritional Composition Ingredients for
Example 46 and other Exemplified Compositions Amount per Ingredient
Name 1000 Kg batch Kg/g/mg Ingredient Water Q.S. Kg Nonfat Milk
83.00 Kg Lactose 51.82 Kg High Oleic Safflower Oil 13.06 Kg Soy Oil
10.50 Kg Coconut Oil 9.256 Kg Galacto-oligosaccharides 8.630 Kg
Whey Protein Concentrate 6.075 Kg 5% Potassium Hydroxide 2.494 Kg
KOH 124.7 g Potassium Citrate 536.6 g Calcium Carbonate 449.9 g
Ascorbic Acid 446.9 g ARASCO Mortierella Alpina Oil 359.3 g Soy
Lecithin 339.7 g Myverol 18-06 339.7 g Nucleotide-Choline Premix
293.2 g Choline Bitartrate 51.75 g Cytidine 5'-Monoohosohate 30.49
g Disodium Guanosine 5'-Monophosphate 15.64 g Disodium Uridine
5'-Monophosphate 13.15 g Adenosine 5'-Monophosphate 11.60 g
Potassium Chloride 199.9 g Magnesium Chloride 154.0 g Vit/Min/Taur
Premix 149.9 g Taurine 45.83 g m-lnositol 33.28 g Zinc Sulfate
15.35 g Niacinamide 9.781 g Calcium Pantothenate 5.865 g Ferrous
Sulfate 5.131 g Cupric Sulfate 1.800 g Thiamine Chloride HCI 1.518
g Riboflavin 669.3 g Pvridoxine HCI 613.1 mg Folic Acid 206.1 mg
Manganese Sulfate 174.6 mg Biotin 59.21 mg Sodium Selenate 35.51 mg
Cyanocobalamin 4.722 mg DHASCO Crypthecodinium Cohnii Oil 131.0 g
Choline Chloride 123.7 g Seakem GP-359 120.0 g Ultra-Micronized
Tricalcium Phosphate 103.2 g Potassium Phosphate Monobasic 90.6 g
Vitamin A, D3, E, K1 Premix 64.7 g RRR Aloha-Tocophervl Acetate
9.063 g Vitamin A Palmitate 1.725 g Vitamin K1 (Phylloquinone)
100.3 mg Vitamin D3 14.06 mg Ferrous Sulfate 60.9 g Seakem RLC
Carrageenan 60.0 g Carotenoid Premix 57.1 g Lutein 119.9 mg
Lycopene 119.9 mg Beta- Carotene 25.98 mg Sodium Chloride 40.1 g
Citric Acid (Processing Aid) 29.8 g L-Carnitine 3.62 g Riboflavin
2.18 g
[0384] A nutritional composition suitable for utilizing the protein
disclosed herein is described in Table 33 below. The 34 grams of
protein per liter of nutritional composition comprises about 25% by
weight bovine beta-casein.
TABLE-US-00034 TABLE 33 Nutritional Composition Ingredients for
Example 46 and other Exemplified Compositions Ingredient Kg per
1000 Kg Water Q.S. Sucrose 75.0 Milk Protein Isolate (Alapro 4900-
Fonterra) 34.9 Soy Oil 8.0 Soy Protein Isolate 3.7 Potassium
Citrate 3.3 Phytosterol Ester 3.2 Avicel CL611 3.0 Magnesium
Phosphate Dibasic 1.92 N&A Vanilla Flavor 1.80 m-TCP 1.75
N&A Dairv Cream Flavor 32122 1.30 Soy Lecithin 1.0 Magnesium
Chloride 1.15 Sodium Chloride 0.75 Choline Chloride 0.53 Ascorbic
Acid 0.377 Potassium Chloride 0.309 Potassium Hydroxide 45%
(Processing Aid) 0.262 WSV Premix 0.185 Corn Starch (Processing
Aid) 0.0631143 Dextrose (Processing Aid) 0.062977 Niacinamide
0.0274725 d-Calcium Pantothenate 0.0177239 Thiamine Chloride
Hydrochloride 0.0045325 Pyridoxine Hydrochloride 0.0043625
Riboflavin 0.0035335 Folic Acid 0.0006739 Biotin 0.0005467
Cyanocobalamin 0.00006318 Carrageenan (Viscarin SA-359) 0.180
UTM/TM Premix 0.150 Zinc Sulfate, Monohydrate 0.0837855
Maltodextrin (Processing aid) 0.0346267 Manganese Sulfate,
Monohydrate 0.0207663 Citric acid, Anhydrous 0.009258 Chromium
Chloride, Hexahydrate 0.0007288 Sodium Molybdate, Dihydrate
0.0005882 Sodium Selenate, Anhydrous 0.0002465 Vitamin ADEK Premix
0.135 Coconut Oil (Processing Aid) 0.0799578 Vitamin E (dl-Alpha
Tocophervl Acetate) 0.0495827 Vitamin A Palmitate 0.0051266
Phylloquinone 0.000297 Vitamin D3 0.00003594 Sucralose Liquid (25%)
0.10 Seakem CM514 0.090 Acesulfame potassium 0.050 Potassium Iodide
0.00023
[0385] In certain exemplary embodiments disclosed herein, the
nutritional compositions are administered (or consumed) orally as
needed to provide the desired level of nutrition. In certain of
these embodiments, the nutritional compositions are administered
(or consumed) in the form of one to two servings daily or in one or
two or more divided doses daily. In certain embodiments, when the
nutritional composition is a liquid, the serving may be 150
milliliters to 500 milliliters. In certain other embodiments, when
the nutritional composition is a liquid, the serving is 237
milliliters (.about.8 fl. oz.). In other embodiments, when the
nutritional composition is a liquid, the serving is 177 milliliters
to 414 milliliters (.about.6 fl. oz. to .about.14 fl. oz.). In yet
other embodiments, when the nutritional composition is a liquid,
the serving is 207 milliliters to 266 milliliters (.about.7 fl. oz.
to .about.9 fl. oz.). Various calorie contents may be associated
with each serving of the nutritional compositions according to the
first, second and third embodiments disclosed herein, typically
from 25 to 500 Kcal, including 50 to 400 Kcal; 100 to 350 Kcal or
150 to 350 Kcal per serving. Alternatively, a serving may be
construed as any amount which is intended to be consumed in one
sitting or within one hour or less.
[0386] To the extent that the term "includes" or "including" is
used in the specification or the claims, it is intended to be
inclusive in a manner similar to the term "comprising" as that term
is interpreted when employed as a transitional word in a claim.
Furthermore, to the extent that the term "or" is employed (e.g., A
or B) it is intended to mean "A or B or both." When the applicants
intend to indicate "only A or B but not both" then the term "only A
or B but not both" will be employed. Thus, use of the term "or"
herein is the inclusive, and not the exclusive use. See Bryan A.
Garner, A Dictionary of Modern Legal Usage 624 (2d. Ed. 1995).
Also, to the extent that the terms "in" or "into" are used in the
specification or the claims, it is intended to additionally mean
"on" or "onto." Furthermore, to the extent the term "connect" is
used in the specification or claims, it is intended to mean not
only "directly connected to," but also "indirectly connected to"
such as connected through another component or components.
[0387] While the present application has been illustrated by the
description of embodiments thereof, and while the embodiments have
been described in considerable detail, it is not the intention of
the applicants to restrict or in any way limit the scope of the
appended claims to such detail. Additional advantages and
modifications will readily appear to those skilled in the art.
Therefore, the application, in its broader aspects, is not limited
to the specific details, the representative apparatus, and
illustrative examples shown and described. Accordingly, departures
may be made from such details without departing from the spirit or
scope of the applicant's general inventive concept.
Examples 47-53
[0388] Examples 47-49 illustrate soy-based powdered infant formulas
of the present disclosure, the ingredients of which are listed in
Table 34. All ingredient amounts are listed as kg per 1000 kg
batch, unless otherwise specified.
TABLE-US-00035 TABLE 34 Nutritional Composition Ingredients for
Examples 47-49 and other Exemplified Compositions Ingredient
Example 1 Example 2 Example 3 Corn Syrup 504.1 504.1 504.1 Soy
Protein Isolate (5% DH) 144.8 144.8 144.8 Sunflower Oil 112.5 112.5
112.5 Sucrose 98.3 98.3 98.3 Soy Oil 83.9 83.9 83.9 Coconut Oil
75.6 75.6 75.6 Fructooligosaccharides 17 17 17 Potassium Citrate
16.5 16.5 16.5 Calcium Phosphate 16.4 16.4 16.4 Sodium Chloride 3.8
3.8 3.8 Arachidonic Acid Oil 3 3 3 Magnesium Chloride 2.8 2.8 2.8
L-Methionine 1.7 1.7 1.7 Ascorbic Acid 1.1 1.1 1.1 Docosahexaenoic
Acid Oil 1.1 1.1 1.1 MDG Oil 621.0 g 590.3 g 558.9 g Lutein 945.0
mg 346 mg 1.1 g Choline Chloride 507.7 g 507.7 g 507.7 g Taurine
457.5 g 457.5 g 457.5 g Inositol 353.0 g 353.0 g 353.0 g Ascorbyl
Palmitate 347.5 g 347.5 g 347.5 g Ferrous Sulfate 319.2 g 319.2 g
319.2 g Mixed Tocopherols 157.2 g 157.2 g 157.2 g L-Carnitine 112.7
g 112.7 g 112.7 g Niacinamide 97.9 g 97.9 g 97.9 g
D-Alpha-Tocopheryl Acetate 78.8 g 78.8 g 78.8 g Calcium
D-Pantothenate 58.7 g 58.7 g 58.7 g Zinc 56.0 g 56.0 g 56.0 g Iron
16.9 g 16.9 g 16.9 g Thiamine 15.2 g 15.2 g 15.2 g Vitamin A
Palmitate 14.8 g 14.8 g 14.8 g Copper 7.2 g 7.2 g 7.2 g Riboflavin
6.7 g 6.7 g 6.7 g Pyridoxine Hydrochloride 6.1 g 6.1 g 6.1 g Folic
Acid 2.1 g 2.1 g 2.1 g Potassium Iodide 1.1 g 1.1 g 1.1 g
Phylloquinone 857.1 mg 857.1 mg 857.1 mg Vitamin D3 47 mg 103.7 mg
324 mg Lycopene 980.0 mg 980.0 mg 980.0 mg Biotin 592.5 mg 592.5 mg
592.5 mg Beta-Carotene 215.6 mg 215.6 mg 215.6 mg Selenium 147.0 mg
147.0 mg 147.0 mg Cyanocobalamin 71.3 mg 71.3 mg 71.3 mg
[0389] Examples 50-52 illustrate milk-based powdered infant
formulas of the present disclosure, the ingredients of which are
listed in Table 35. All ingredient amounts are listed as kg per
1000 kg batch, unless otherwise specified.
TABLE-US-00036 TABLE 35 Nutritional Composition Ingredients for
Examples 50-52 and other Exemplified Compositions Ingredient
Example 4 Example 5 Example 6 Skim Milk 2057.3 2057.3 2057.3
Lactose 414.1 414.1 414.1 Sunflower Oil 111.1 111.1 111.1 Soy Oil
83.3 83.3 83.3 Coconut Oil 74.6 74.6 74.6 Galactooligosaccharides
72.3 72.3 72.3 Whey Protein Concentrate 49.4 49.4 49.4 Whey Protein
Hydrolysate 5.6 5.6 5.6 (<25% DH) Potassium Citrate 3.4 3.4 3.4
Arachidonic Acid Oil 2.9 2.9 2.9 Calcium Carbonate 2.6 2.6 2.6
Potassium Hydroxide 2.3 2.3 2.3 Soy Lecithin 1.7 1.7 1.7
Docosahexaenoic Acid Oil 1.6 1.6 1.6 Ascorbic Acid 1.5 1.5 1.5
Choline Bitartrate 1.2 1.2 1.2 Magnesium Chloride 1.2 1.2 1.2 MDG
Oil 1.1 1.044 0.99 Lutein 2.65 g 1.5 g 364 mg Sodium Chloride 1 1 1
Taurine 428 g 428 g 428 g L-Tryptophan 277 g 277 g 277 g Cytidine
5'-Monophosphate 257 g 257 g 257 g Myo-Inositol 257 g 257 g 257 g
Ascorbyl Palmitate 200 g 200 g 200 g Disodium Guanosine 187 g 187 g
187 g 5'-Monophosphate Disodium Uridine 163 g 163 g 163 g
5'-Monophosphate Ferrous Sulfate Monohydrate 130.6 g 130.6 g 130.6
g RRR-Alpha-Tocopheryl Acetate 119.7 g 119.7 g 119.7 g Tocopherol-2
106.7 g 106.7 g 106.7 g Zinc Sulfate Monohydrate 104 g 104 g 104 g
Adenosine 5'-Monophosphate 98 g 98 g 98 g Ferrous Sulfate 88 g 88 g
88 g Mixed Tocopherols 77 g 77 g 77 g Niacinamide 59 g 59 g 59 g
L-Carnitine 43 g 43 g 43 g Calcium D-Pantothenate 42 g 42 g 42 g
Retinyl Palmitate 13 g 13 g 13 g Copper Sulfate 11 g 11 g 11 g
Thiamin Hydrochloride 9 g 9 g 9 g Pyridoxine Hydrochloride 5 g 5 g
5 g Riboflavin 5 g 5 g 5 g Manganese Sulfate Monohydrate 3 g 3 g 3
g Folic Acid 796 mg 796 mg 796 mg Beta-Carotene 770 mg 770 mg 770
mg Phylloquinone 618 mg 618 mg 618 mg Potassium Iodide 550 mg 550
mg 550 mg D-Biotin 238 mg 238 mg 238 mg Sodium Selenate 117 mg 117
mg 117 mg Vitamin D3 50 mg 93 mg 295 mg Cyanocobalamin 8 mg 8 mg 8
mg
[0390] Example 53 illustrates a ready-to-feed milk-based liquid
infant formula of the present disclosure, the ingredients of which
are listed in Table 36. All ingredient amounts are listed as kg per
1000 kg batch, unless otherwise specified.
TABLE-US-00037 TABLE 36 Nutritional Composition Ingredients for
Example 53 and other Exemplified Compositions Ingredient Example 7
Water Q.S. Lactose 48.2 Skim Milk 22.1 High Oleic Safflower Oil
13.3 Soy Oil 10.2 Coconut Oil 9.7 Galactooligosaccharides 9.4 Whey
Protein Concentrate 6.3 Potassium Hydroxide 166 g Calcium Carbonate
747 g Potassium Citrate 650 g Ascorbic Acid 610 g Arachidonic Acid
Oil 410 g Distilled Monoglycerides 347 g Lecithin 350 g Cytidine
5'-Monophosphate 32.8 g Disodium Guanosine 5'-Monophosphate 16.8 g
Disodium Uridine 5'-Monophosphate 14.1 g Adenosine 5'-Monophosphate
12.5 g Docosahexaenoic Acid Oil 225 g MDG Oil 210 g Potassium
Phosphate 185 g Taurine 57 g Myo-Inositol 48 g L-Tryptophan 19 g
Niacinamide 6 g Calcium D-Pantothenate 5 g Thiamine Hydrochloride
1.4 g Riboflavin 1.3 g Pyridoxine Hydrochloride 750 mg Folic Acid
145 mg D-Biotin 36 mg Cyanocobalamin 5.4 mg Carrageenan 175 g
Sodium Citrate 174 g Sodium Chloride 145 g Magnesium Chloride 126 g
Ferrous Sulfate 66.5 g Choline Chloride 45 g Vitamin E Acetate 21 g
Vitamin A Palmitate 2 g Phylloquinone 131.6 mg Vitamin D3 16 mg
Magnesium Sulfate 32 g Zinc Sulfate 17.9 g L-Carnitine 5.5 g Cupric
Sulfate 2 g Lutein 160 mg Beta-Carotene 80 mg Manganese Sulfate 110
mg
Example 54
[0391] In this Example, the effect of using an activated MDG oil
component or an MDG-protected premix on the absorbance of
triglycerides and fatty acids in a nutritional composition is
evaluated and compared to the absorbance of triglycerides and fatty
acids in a nutritional composition without MDG oil. The fatty acids
that were measured for this test included palmitic acid, linoleic
acid, and alpha-linolenic acid.
[0392] The following nutritional compositions were tested:
[0393] Triglyceride oil control (AET-C): high oleic safflower oil
was added to the protein-in-fat slurry prior to addition to the
remainder of the composition;
[0394] MDG-protected premix (AET-1): MDG oil was premixed with
lecithin for 5 minutes; this premix was added to the protein-in-fat
slurry to form an MDG-protected premix prior to addition to the
remainder of the composition;
[0395] Activated MDG oil component (AET-2): MDG oil was added to
the protein-in-water slurry to form an activated MDG oil component
prior to addition to the remainder of the composition;
[0396] MDG oil, not activated (AET-3): MDG oil was added to the
protein-in-fat slurry prior to addition to the remainder of the
composition; and
[0397] Activated MDG oil component with choline (AET-4): MDG oil
was premixed with choline for 5 minutes; this premix was added to
the protein-in-water slurry to form an activated MDG oil component
prior to addition to the remainder of the composition.
TABLE-US-00038 TABLE 37 Nutritional Composition Ingredients for
Example 54 and other Exemplified Compositions Sample ID: AET-C
AET-1 AET-2 AET-3 AET-4 Amount/ Amount/ Amount/ Amount/ Amount/
Ingredient Name: 1000 kg Unit 1000 kg Unit 1000 kg Unit 1000 kg
Unit 1000 kg Unit Water Q.S. kg Q.S. Kg Q.S. kg Q.S. kg Q.S. kg
Lactose 49.27 kg 49.27 Kg 49.27 kg 49.27 kg 49.27 kg Non-Fat Dry
Milk 22.06 kg 22.06 Kg 22.06 kg 22.06 kg 22.06 kg High Oleic
Safflower 13.44 kg 12.78 Kg 12.78 kg 12.78 kg 12.78 kg Oil Soy Oil
10.06 kg 9.57 Kg 9.57 kg 9.57 kg 9.57 kg Coconut Oil 9.60 kg 9.13
Kg 9.13 kg 9.13 kg 9.13 kg Galactooligosaccharides 9.40 kg 9.40 Kg
9.40 kg 9.40 kg 9.40 kg Whey Protein 6.31 kg 6.31 Kg 6.31 kg 6.31
kg 6.31 kg Concentrate MDG Oil 0 kg 1.62 Kg 1.62 kg 1.62 kg 1.62 kg
Ascorbic Acid 611.99 g 611.99 G 611.99 g 611.99 g 611.99 g Calcium
Carbonate 559.70 g 559.70 G 559.70 g 559.70 g 559.70 g Potassium
Citrate 545.40 g 545.40 G 545.40 g 545.40 g 545.40 g ARASCO 395.30
g 395.30 G 395.30 g 395.30 g 395.30 g Mortierella Alpina Oil
Potassium Hydroxide, 368.00 g 368.00 G 368.00 g 368.00 g 368.00 g
45% Lecithin 346.99 g 346.99 G 346.99 g 346.99 g 346.99 g Myverol
346.99 g 346.99 G 346.99 g 346.99 g 346.99 g Nucleotide-Choline
314.15 g 314.15 G 314.15 g 314.15 g 314.15 g Premix DHASCO 208.80 g
208.80 G 208.80 g 208.80 g 208.80 g Crypth. Cohnii Oil
Vitamin/Mineral/ 176.00 g 176.00 G 176.00 g 176.00 g 176.00 g
Taurine Premix Carrageenan 175.26 g 175.26 G 175.26 g 175.26 g
175.26 g Sodium Citrate 163.00 g 163.00 G 163.00 g 163.00 g 163.00
g Sodium Chloride 145.30 g 145.30 G 145.30 g 145.30 g 145.30 g
Magnesium Chloride 126.50 g 126.50 G 126.50 g 126.50 g 126.50 g
Potassium Phosphate 117.60 g 117.60 G 117.60 g 117.60 g 117.60 g
Dibasic Ferrous sulfate 66.50 g 66.50 G 66.50 g 66.50 g 66.50 g
Choline Chloride 44.97 g 44.97 G 44.97 g 44.97 g 44.97 g Vitamin A,
D3, E, K1 35.80 g 35.80 G 35.80 g 35.80 g 35.80 g Magnesium Sulfate
27.40 g 27.40 G 27.40 g 27.40 g 27.40 g Zinc Sulfate 17.90 g 17.90
G 17.90 g 17.90 g 17.90 g Lutein 12.35 g 12.35 G 12.35 g 12.35 g
12.35 g L-Carnitine 5.50 g 5.50 G 5.50 g 5.50 g 5.50 g Cupric
Sulfate 1.90 g 1.90 G 1.90 g 1.90 g 1.90 g Manganese Sulfate 110.00
mg 110.00 Mg 110.00 mg 110.00 mg 110.00 mg Citric Acid As As As As
As needed needed needed needed needed Potassium Iodide As As As As
As needed needed needed needed needed Sodium Selenate As As As As
As needed needed needed needed needed Potassium Chloride As As As
As As needed needed needed needed needed
[0398] Male Sprague Dawley rats weighing between 280 and 330 grams
were all fed a normal commercially available Purina Rat Chow for
one week. Rats were then fasted overnight prior to surgery, and
under anesthesia, a laparotomy was performed, and the intestinal
lymph duct was cannulated according to the procedure of Tso et al.,
"The Absorption of Lipid and Lipoprotein Synthesis," Lipid Research
Methodology, Chapter 5: 191-216 (1984) Alan R. Liss, Inc., NY, NY,
hereby incorporated by reference to the extent consistent herewith.
The superior mesenteric artery was isolated, but not occluded. A
silicon infusion tube (1.6 mm OD) was placed in the stomach for
future infusion of a test sample. The fundic incision was closed by
a purse string suture. The rats were allowed to recover for 24
hours before infusion began.
[0399] The rats were randomly assigned to receive one of the five
nutritional compositions being tested. (The number of rats in each
group, e.g. "(n=7)," is shown in the key for FIG. 18.) The animals
were intragastrically infused 24 hours after surgery with 3 ml of
their respective test nutritional compositions.
[0400] Lymph was collected in pre-cooled tubes for 1 hour to
measure fasting levels of triglycerides, palmitic acid, linoleic
acid, and alpha linolenic acid before the nutritional composition
infusions began. Lymph was then collected in pre-cooled tubes
hourly for 6 hours after initiation of infusion. At the end of the
experiment, the rats were sacrificed by exsanguination.
[0401] The lymph lipids were extracted and analyzed for
triglycerides, palmitic acid, linoleic acid, and alpha-linolenic
acid using high-performance liquid chromatography (Craft
Technologies, Wilson, N.C.).
[0402] FIG. 18 shows the lymphatic output of triglycerides over a
period of 6 hours following infusion of the liquid nutritional
compositions. FIG. 19 shows the lymphatic absorption of palmitic
acid over a 6 hour period following infusion of the liquid
nutritional compositions. FIG. 20 shows the lymphatic absorption of
linoleic acid over a 6 hour period following infusion of the liquid
nutritional compositions. FIG. 21 shows the lymphatic absorption of
alpha-linolenic acid over a period of 6 hours following infusion of
the liquid nutritional compositions.
[0403] As shown in FIG. 18, the mean fasting lymphatic triglyceride
output for all groups of rats varied between about 2.0 and 3.5
mg/hr. In all groups, lymphatic triglyceride output increased after
infusion of each nutritional composition began and reached a
maximum output of about 8 to about 17.5 mg/hr two hours after
infusion began. The control group (AET-C) and the group receiving
MDG oil that was not activated (AET-3) showed modest increases in
the lymphatic triglyceride output, with a triglyceride output of
about 8 to about 9 mg/hr two hours after infusion began. However,
the groups receiving either an activated MDG component (AET-2 and
AET-4) or an MDG-protected premix (AET-1) showed greater lymphatic
triglyceride output, with a triglyceride output of about 13 to
about 17.5 mg/hr two hours after infusion of the nutritional
compositions began.
[0404] As shown in FIG. 18, the mean fasting lymphatic triglyceride
output for all groups of rats varied between about 2.0 and 3.5
mg/hr. In all groups, lymphatic triglyceride output increased after
infusion of each nutritional composition began and reached a
maximum output of about 8 to about 17.5 mg/hr two hours after
infusion began. The control group (AET-C) and the group receiving
MDG oil that was not activated (AET-3) showed modest increases in
the lymphatic triglyceride output, with a triglyceride output of
about 8 to about 9 mg/hr two hours after infusion began. However,
the groups receiving either an activated MDG component (AET-2 and
AET-4) or an MDG-protected premix (AET-1) showed greater lymphatic
triglyceride output, with a triglyceride output of about 13 to
about 17.5 mg/hr two hours after infusion of the nutritional
compositions began.
[0405] FIG. 20 shows the lymphatic absorption of linoleic acid for
6 hours following infusion of the five nutritional compositions.
The lymphatic absorption of linoleic acid increased in all groups
during the first two hours and fell to a steady state from about 4
to 6 hours after infusion of the nutritional compositions began.
However, there was a marked increase in the lymphatic absorption of
linoleic acid two hours after infusion began for the nutritional
compositions containing some form of MDG oil (AET-1 to AET-4), when
compared with the nutritional composition containing the
triglyceride control with no MDG oil (AET-C).
[0406] FIG. 21 shows the lymphatic absorption of alpha-linolenic
acid for 6 hours following infusion of the five nutritional
compositions. The lymphatic absorption of alpha-linolenic acid
increased in all groups during the first two hours and fell to a
steady state from about 4 to 6 hours after infusion of the
nutritional compositions began. However, there was a marked
increase in the lymphatic absorption of alpha-linolenic acid two
hours after infusion began for the nutritional compositions
containing some form of MDG oil (AET-1 to AET-4), when compared
with the nutritional composition containing the triglyceride
control with no MDG oil (AET-C).
Example 55
[0407] In this Example, the nutritional compositions evaluated in
Example 1 were further tested for lutein that is not solubilized by
the fat in the nutritional compositions. The five compositions,
AET-C, AET-1, AET-2, AET-3 and AET-4, were each subjected to high
speed centrifugation (31,000.times.g; 20.degree. C.; 4 h). The
aqueous fraction of each centrifuged nutritional composition was
diluted 2:8 with water, and the diluted aqueous fractions were
filtered through 0.45 .mu.m PTFE membranes. The visible absorbance
of the diluted aqueous fractions was measured at 476 nm (a lutein
absorbance maximum). The absorbance readings (milliabsorbance units
per gram of aqueous fraction (mAU/g)) are reported in Table 3
below, along with a ranking of the in vivo lutein response for the
same compositions tested in Example 1.
TABLE-US-00039 TABLE 38 Relationship between Lutein in the Aqueous
Fraction with in vivo Response Absorbance, Ranking of in vivo
Sample ID mAU/g, 476 nm lutein response AET-C 57.6 1 (lowest
response) AET-1 62.7 4 AET-2 61.3 3 AET-3 59.4 2 AET-4 64.9 5
(highest response)
[0408] As may be seen from the table, the absorbance values
correlate with the in vivo results, in that each of the nutritional
compositions comprising an activated MDG premix (AET-2 and AET-4)
or an MDG-protected premix (AET-1) have higher lutein absorbance
values than do the control nutritional composition (AET-C) or the
nutritional composition that does not comprise an activated premix
(AET-3). More lutein is present in the aqueous fraction of the
nutritional compositions comprising an activated MDG premix or an
MDG-protected premix. Thus, the relative bioavailability of lutein,
as demonstrated by in vivo testing, was accurately predicted by in
vitro testing using this method.
Example 56
[0409] In this Example, the effect on the bioavailability of DHA in
nutritional compositions containing an activated MDG premix or an
MDG-protected premix is evaluated and compared to the
bioavailability of DHA that is in nutritional compositions without
the activated MDG premix or the MDG-protected premix.
[0410] A similar protocol for animal infusion and lymph collection
as described in Example 1 was followed, including the infusion of
the same nutritional compositions (e.g., compositions AET-C, AET-1,
AET-2, AET-3 and AET-4). The lymph lipids were extracted and
analyzed for DHA concentration using high-performance liquid
chromatography. FIG. 22 shows the change in lymphatic output of DHA
over time. There was a significant increase in lymph levels of DHA
from the initial (fasting) levels to the DHA levels in the 6 hrs
following infusion for the nutritional compositions containing an
activated premix (AET-2) or an MDG-protected premix (AET-1). In
comparison, the control nutritional composition (AET-C) and the
nutritional composition that does not comprise an activated premix
(AET-3) showed only a modest increase in the lymph levels of DHA
over time.
Example 57
[0411] A study was conducted to compare a commercially
representative nutritional powder with and without MDG. Nutritional
powder batches (control and MDG) were reconstituted (152 g/L),
centrifuged (31,000.times.g/20.degree. C./4 h), and the MDG
concentration in the aqueous fraction of samples was estimated by
HPLC determination of monolinolein and monoolein, and represented
as mg of MDG per kg of aqueous fraction.
TABLE-US-00040 TABLE 39 MDG presence in the Aqueous Fraction after
Nutritional Composition Centrifugation Aug. 15, Sep. 19, Nov. 5,
Feb. 15, Batch 2013 2013 2013 2014 AET-C <70 <40 <20 NT
(no MDG) (not tested) AET-1 144 120 NT NT AET-2 NT 150 NT NT AET-3
NT 90 118 NT AET-4 178 140 115 NT AET4-5 NT NT 127 NT A455 NT NT NT
<10 (no MDG) A456 NT NT NT 30 A457 NT NT NT 40 A458 NT NT NT
40
[0412] A measurable presence of MDG in the aqueous fractions of all
MDG samples was detected, and an absence of MDG in the aqueous
fraction of all control batches, was verified by LC/UV analysis.
The significance of the data is that (a) MDG is present in the
Aqueous Fraction (despite its very low water solubility), and that
(b) the MDG concentration is generally correlated with in vivo
lutein availability.
Example 58
[0413] A study was conducted to investigate the distribution of
lutein and MDG within the nutritional compositions. Sample were
prepared using homogenization at 150/20 bar, and then centrifuged
at 100,000.times.g for 1 hour. The different layers (e.g., cream,
aqueous, and pellet) were then analyzed for the presence of lutein
and MDG.
[0414] The results demonstrate that nutritional compositions
comprising MDG provided MDG and lutein within the aqueous phase,
wherein these molecules have very limited water solubility.
Specifically, it was found that lutein and MDG were present in the
aqueous phase, following centrifugation, at approximately 4 wt %
and approximately 8 wt % of the nutritional composition
respectively. This suggests that the lutein found in the aqueous
phase is associated with water-soluble complexes. It should be
noted that the pellet contained approximately 1% of the lutein,
suggesting that lutein could be associated with a sedimentable
assembly as well.
Example 59
[0415] A study was conducted to investigate the
dispersibility/solubility of MDG (mono- and diglycerides) in
various aqueous preparations as evaluated by HPLC. The MDG was
added to water at 95.degree. C., RT, and then the samples were
subjected to high speed centrifugation at 20.degree. C., and are
reported in the table below.
TABLE-US-00041 TABLE 40 MDG Alone has Limited Solubility in the
Aqueous Fraction Soluble MDG, Soluble MDG, as % MDG Preparation as
mg per kg of total MDG 1.00% (w/w) MDG 1980 19.8% suspension in
95.degree. C. water Room temperature ~32 ~0.32% filtrate of 1.00%
(w/w) MDG suspension Centrifugation ~16 ~0.16% supernatant of room
temperature filtrate
[0416] The MDG present following centrifugation was very limited,
and is in stark contrast to the MDG present within the aqueous
phase following centrifugation of the nutritional composition. This
suggests that the solubility of MDG, along with the lipophilic
compound, is increased in nutritional compositions comprising MDG,
at least one lipophilic compound, and at least one hydrophobic
protein.
Example 60
[0417] A study was conducted to further investigate the
distribution of lutein and MDG within the nutritional composition.
Samples included MDG/lutein/water; MDG/lutein/NFDM/water;
MDG/lutein/WPC/water; and MDG/water.
[0418] The premixes were provided as follows: in samples that
contained lutein, lutein was heated for 2 h at 50.degree. C. with
shaking. In samples that contained MDG (GMO-40) and lutein, lutein
was added to MDG and heated to 50-60.degree. C. and mixed for 10
minutes. In samples that contained protein, non-fat dried milk was
added to water (50-60.degree. C.) with agitation and allowed to
hydrate for approximately 10 min.
[0419] The samples were prepared as follows: for solution 1, water
was heated to 50-60.degree. C., an lutein in GMO-40 blend and
turrax (30 sec 10000 rpm) was added; for solution 2, NFDM was
placed in water solution heated to 50-60.degree. C., and lutein was
added in GMO-40 blend and turrax (30 sec 10000 rpm); for solution
3, water was heated to 50-60.degree. C., and GMO-40 blend and
turrax (30 sec 10000 rpm) was added. The solutions are listed in
the table below as X.1, wherein 1 is the solution.
TABLE-US-00042 TABLE 41 Batch Details for Example 60 sample MDG +
MDG + MDG + lutein + lutein + lutein NFDM WPC MDG 3.1 3.2 3.2-1
3.2-2 3.3 lutein 0.0695 0.0601 0.0601 0.0601 0 GMO-40 9.18 7.94
7.94 7.94 9.18 dried milk 0 107.98 107.98 0 0 (nonfat dried milk,
NFDM) Whey protein 0 0 0 107.98 0 concentrate 80 (WPC) water 790.75
684.02 684.02 684.02 790.82 TOTAL [g] 800 800 800 800 800
[0420] Samples were formulated as detailed above, and then
characterized by ultrafiltration with various membranes, as well as
ultracentrifuged at various times and speed (e.g., 100,000.times.g,
4,500.times.g, and 1,000.times.g). The details of the different
centrifugation/filtration method are listed below.
[0421] Speed of centrifugation: 1000.times.g, Centrifugation time:
15 min, Centrifugation temperature: 20.degree. C., Centrifuge
tubes: Cellstar tube 50 ml from Greiner bio-one, Tube diameter: 23
mm, Amount of product in tube: 45 ml, Centrifuge: type 5810 R from
Eppendorf, aqueous phase isolation by syringe through the cream
layer.
[0422] Speed of centrifugation: 4500.times.g, Centrifugation time:
15 min, Centrifugation temperature: 20.degree. C., Centrifuge
tubes: Cellstar tube 50 ml from Greiner bio-one, Tube diameter: 23
mm, Amount of product in tube: 45 ml, Centrifuge: type 5810 R from
Eppendorf, aqueous phase by syringe through the cream layer.
[0423] Speed of centrifugation: 100,000.times.g, Centrifugation
time: 1 h, Centrifugation temperature: 20.degree. C., Centrifuge
tubes: Ultra Clear Centrifuge tubes from Beckmann Coulter, Tube
size: 14.times.89 mm, Amount of product in tube: 11.3 g,
Centrifuge: type L-90K from Beckmann Coulter, aqueous phase
isolation by freezing the tube with content in liquid nitrogen and
cutting the part with the serum phase including the tube wall
followed by thawing the content of this part of the tube.
[0424] Membrane: Centrifugal filter units from Amicon Ultra; 100
kDa and 10 kDa, Centrifugation speed: 4000 g, Centrifugation time:
30 min, Centrifugation temperature: 20.degree. C., Centrifuge
tubes: Cellstar tube 50 ml from Greiner bio-one, Tube diameter: 23
mm, Amount of product in tube: 45 ml, Centrifuge: type 5810 R from
Eppendorf, Serum isolation by removing the filter form the tube and
decanting the filtrate.
[0425] Centrifugation speed (Lumifuge): 1000 g, Centrifugation
time: 255 min, Light factor: 1, Centrifugation temperature:
20.degree. C., Lumifuge: type Centrifuge lumiview 110 from LUM
Gmbh.
[0426] Following centrifugation and/or ultrafiltration, samples
were analyzed for the presence of lutein, and MDG within the
different phases.
[0427] Lutein analysis as done by RP-HPLC-DAD, and MDG analysis was
done by RP-HPLC-UV-ELSD. In MDG analysis, samples were prepared as
follows: samples were weighed at 0.15 g in a 20 mL tell bottle; the
solution was stirred on a magnetic stirrer in the fume hood; 15.0
mL or 13.5 mL, 80% acetone+20% chloroform were added with a
measuring cylinder.
[0428] Stir the solution for 10 minutes on a magnetic stirrer in a
water bath at .+-.50.degree. C. (warm water from the tap); the
solution was sonicated for 10 minutes at room temperature; 1.5 mL
of sample was centrifuged in a 1.5 mL centrifuge tube, 10 min at
20.000.times.g; 20 uL of the supernatant was injected on the HPLC
column. UV spectroscopy of the samples was performed with
wavelength scans (200-900 nm at 5 nm intervals) at 20.degree. C.
using a Cary 4000 UV-Vis Spectrophotometer (Varian BV). All
measurements were carried out in quartz cuvettes with a 1 mm path
length.
[0429] Qualitatively, samples without protein resulted in more
appreciable amounts of precipitated lutein along the side of the
sample flasks. This suggests that there is an optimal interaction
between lutein, hydrophobic protein and MDG to achieve the water
soluble assembly.
[0430] In the ultrafiltration analysis, no lutein or MDG passed
through the 100 kD filter, this was across all samples (FIG.
23).
[0431] The results also demonstrated that the presence of protein
resulted in more lutein and MDG in the aqueous phase. Furthermore,
when comparing WPC vs. NFDM, WPC resulted in both more lutein and
MDG in the aqueous phase. There may be different particle
characteristics (e.g., size) of for the WPC batch, that results in
greater distribution of lutein and MDG in the aqueous phase,
relative to NFDM.
[0432] Centrifugation at 100,000.times.g: lutein was only found in
the aqueous phase of samples containing protein; negligible lutein
was found in aqueous phases of samples containing only lutein and
MDG.
[0433] Centrifugation at 4,500.times.g: similar amounts of lutein
were found in the aqueous phase of samples containing NFDM and WPC.
More MDG was found in the aqueous phase of samples containing WPC
than NFDM. Additionally, concentrations of lutein and MDG in the
aqueous were considerably higher than in those of protein-free
systems centrifuged at 1,000.times.g.
[0434] Centrifugation at 1,000.times.g: resulted in considerably
higher amounts of lutein in the aqueous phase of samples with NFDM,
than without. Specifically, .about.20% lutein was found in the
aqueous phase at this centrifugation speed. Overall, the data
suggest that there is an optimal interaction between lutein,
hydrophobic protein and MDG to achieve the water soluble
assembly.
TABLE-US-00043 TABLE 42 Centrifugation/Ultrafiltration Analysis of
Variable MDG Batches sample 3.1 3.2 3.2-1 3.2-2 3.3 Lutein MDG
Lutein MDG Lutein MDG Lutein MDG Lutein MDG Ultrafiltration n.a.
n.a. n.a. n.a. n.d. n.d. n.d. n.d. n.a. n.a. Centrifugation 0.1
0.047 3.3 3.572 n.d. n.d. n.d. n.d. n.a 0.234 method A (1000 g)
Centrifugation n.d. n.d. n.d. n.d. 1.6 1.967 1.6 3.01 n.d. n.d.
method B (4500 g) Centrifugation n.a. n.a 0.2 1.59 n.d. n.d. n.d.
n.a. n.a method C (100,000 g) Overall 13.1 7.33 15.4 8.44
sample
Example 61
[0435] A study was conducted to investigate the effect of
shelf-life on MGD batches. Different MDG bathes included, 18 month
old batch, 24 month old batch (AET-1), and the control. The
different MDG batches were examined in the lymphatic model as
described in Example 20.
The results demonstrate that the older batch of bounty performed at
similar levels as the newer batch in the bioavailability of lutein
(FIG. 24). This suggests that the assembly of the nutritional
composition is stable for at least 24 months.
* * * * *