U.S. patent application number 09/888798 was filed with the patent office on 2001-11-29 for method for treating a subject afflicted with intestinal malabsorption.
Invention is credited to Ayling, June E., Bailey, Steven W..
Application Number | 20010046533 09/888798 |
Document ID | / |
Family ID | 21747940 |
Filed Date | 2001-11-29 |
United States Patent
Application |
20010046533 |
Kind Code |
A1 |
Bailey, Steven W. ; et
al. |
November 29, 2001 |
METHOD FOR TREATING A SUBJECT AFFLICTED WITH INTESTINAL
MALABSORPTION
Abstract
A composition for human or animal consumption for supplying
folate which includes a natural isomer of reduced folate, such as
(6S)-tetrahydrofolic acid, 5-methyl-(6S)-tetrahydrofolic acid,
5-formyl-(6S)-tetrahydrofolic acid, 10-formyl-(6R)-tetrahydrofolic
acid, 5,10-methylene-(6R)-tetrahydro- folic acid,
5,10-methenyl-(6R)-tetrahydrofolic acid,
5-formimino-(6S)-tetrahydrofolic acid, and their polyglutamyl
derivatives is disclosed. Such compositions include multivitamin
preparations (with or without minerals and other nutrients);
breakfast foods such as prepared cereals, toaster pastries and
breakfast bars; infant formulas; dietary supplements and complete
diet and weight-loss formulas and bars; animal feed (for example
pet foods) and animal feed supplements (such as for poultry feed).
The amount of the natural isomer of a reduced folate in a
composition for human consumption can range between about 5% and
about 200% of the daily requirement for folic acid per serving or
dose.
Inventors: |
Bailey, Steven W.; (Mobile,
AL) ; Ayling, June E.; (Mobile, AL) |
Correspondence
Address: |
Braman & Rogalsky, LLP
P.O. Box 352
Canandaigua
NY
14424-0352
US
|
Family ID: |
21747940 |
Appl. No.: |
09/888798 |
Filed: |
June 25, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09888798 |
Jun 25, 2001 |
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09418649 |
Oct 15, 1999 |
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6254904 |
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09418649 |
Oct 15, 1999 |
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09117586 |
Jul 31, 1998 |
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5997915 |
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60010898 |
Jan 31, 1996 |
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Current U.S.
Class: |
426/72 ;
426/2 |
Current CPC
Class: |
A23V 2002/00 20130101;
A61P 3/02 20180101; A23K 50/42 20160501; A23K 50/10 20160501; A23K
50/75 20160501; A23V 2002/00 20130101; Y02A 40/818 20180101; A23V
2002/00 20130101; A23K 50/40 20160501; A23V 2002/00 20130101; A23K
50/80 20160501; Y10S 426/801 20130101; A61P 3/00 20180101; Y10S
426/807 20130101; A23L 33/40 20160801; A23V 2250/705 20130101; A23L
33/15 20160801; A23V 2250/70 20130101; A23V 2250/5482 20130101;
A23V 2250/1886 20130101; A23V 2250/704 20130101; A23V 2250/704
20130101; A23V 2250/708 20130101; A23V 2250/708 20130101; A23V
2250/54246 20130101; A23V 2250/628 20130101; A23V 2250/5118
20130101; A23V 2250/1842 20130101; A23V 2250/628 20130101; A23V
2250/702 20130101; A23V 2250/5488 20130101; A23V 2250/70 20130101;
A23V 2250/5432 20130101; A23V 2250/708 20130101; A23V 2250/702
20130101; A23V 2250/5036 20130101; A23V 2250/5114 20130101; A23V
2250/70 20130101; A23V 2250/156 20130101; A23V 2250/71 20130101;
A23V 2250/71 20130101; A23V 2250/51086 20130101; A23V 2250/304
20130101; A23V 2250/712 20130101; A23V 2250/704 20130101; A23V
2250/156 20130101; A23V 2250/708 20130101; A23V 2250/188 20130101;
A23V 2250/1628 20130101; A23V 2250/218 20130101; A23V 2250/712
20130101; A23V 2250/708 20130101; A23V 2250/5108 20130101; A23V
2250/71 20130101; A23K 20/174 20160501; A23V 2002/00 20130101; A23V
2002/00 20130101 |
Class at
Publication: |
426/72 ;
426/2 |
International
Class: |
A23L 001/302 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 31, 1997 |
US |
PCT/US97/01870 |
Claims
What is claimed:
1. A composition comprising: one or more natural isomers of reduced
folate selected from the group consisting of (6S)-tetrahydrofolic
acid, 5-methyl-(6S)-tetrahydrofolic acid,
5-formyl-(6S)-tetrahydrofolic acid, 10-formyl-(6R)-tetrahydrofolic
acid, 5,10-methylene-(6R)-tetrahydrofolic acid,
5,10-methenyl-(6R)-tetrahydrofolic acid,
5-formimino-(6S)-tetrahydr- ofolic acid, and polyglutamyl
derivatives thereof; a nutritional substance selected from the
group consisting of a food preparation, an essential nutrient
preparation, and combinations thereof; wherein, when the
nutritional substance is a food preparation, the food preparation
comprises two or more food components and each gram of said food
preparation has a natural molar amount, N, of said one or more
natural isomers of reduced folate, wherein N is greater or equal to
zero and wherein each gram of said composition has a total molar
amount, T, of said one or more natural isomers of reduced folate
greater than N; and wherein, when the nutritional substance is an
essential nutrient preparation, the essential nutrient preparation
comprises a vitamin other than ascorbic acid.
2. A composition according to claim 1, wherein the one or more
natural isomers of reduced folate is selected from the group
consisting of 5-methyl-(6S)-tetrahydrofolic acid,
5-formyl-(6S)-tetrahydrofolic acid,
5,10-methenyl-(6R)-tetrahydrofolic acid, and polyglutamyl
derivatives thereof.
3. A composition according to claim 1, wherein the total molar
amount of said one or more natural isomers of reduced folate is
between 5% and 200% of a human daily requirement for folate per
customarily consumed quantity of said composition.
4. A composition according to claim 1, wherein the total molar
amount of said one or more natural isomers of reduced folate is
between 5% and 3000% of an animal daily requirement for folate per
customarily consumed quantity of said composition.
5. A composition according to claim 1, wherein said nutritional
substance is a food preparation.
6. The composition according to claim 5, wherein the nutritional
substance is a food preparation and wherein each gram of said food
preparation further comprises no unnatural isomers of reduced
folate selected from the group consisting of (6R)-tetrahydrofolic
acid, 5-methyl-(6R)-tetrahydrofolic acid,
5-formyl-(6R)-tetrahydrofolic acid, 10-formyl-(6S)-tetrahydrofolic
acid, 5,10-methylene-(6S)-tetrahydrofolic acid,
5,10-methenyl-(6S)-tetrahydrofolic acid,
5-formimino-(6R)-tetrahydr- ofolic acid, and polyglutamyl
derivatives thereof, or one or more of said unnatural isomers of
reduced folate in a molar amount less than T minus N.
7. A composition according to claim 5, wherein the food preparation
is selected from the group consisting of breakfast foods, infant
formulas, dietary supplements, complete diet formulas, and
weight-loss preparations.
8. A composition according to claim 7, wherein the breakfast food
is a prepared cereal, a breakfast drink mix, or a toaster pastry,
and wherein the weight-loss preparations is a weight-loss drink or
a weight-loss bar.
9. A composition according to claim 1, wherein the nutritional
substance is an essential nutrient preparation comprising a vitamin
other than ascorbic acid.
10. A composition according to claim 9, wherein the essential
nutrient preparation further comprises ascorbic acid.
11. A composition according to claim 9, wherein the vitamin is
present in an amount equal to or greater than 25% of the daily
requirement for the vitamin per customarily consumed quantity of
said essential nutrient preparation.
12. A method for increasing the folate content of a nutritional
substance comprising: providing a nutritional substance selected
from the group consisting of a food preparation, an essential
nutrient preparation, and combinations thereof; and incorporating
into the nutritional substance a molar amount of one or more
natural isomers of reduced folate selected from the group
consisting of (6S)-tetrahydrofolic acid,
5-methyl-(6S)-tetrahydrofolic acid, 5-formyl-(6S)-tetrahydrofolic
acid, 10-formyl-(6R)-tetrahydrofolic acid,
5,10-methylene-(6R)-tetrahydrofolic acid,
5,10-methenyl-(6R)-tetrahydrofolic acid,
5-formimino-(6S)-tetrahydr- ofolic acid, and polyglutamyl
derivatives thereof; wherein, when the nutritional substance is a
-food preparation, the food preparation comprises two or more food
components; and wherein, when the nutritional substance is an
essential nutrient preparation, the essential nutrient preparation
comprises a vitamin other than ascorbic acid.
13. A method according to claim 12 further comprising incorporating
into the nutritional substance a molar amount of one or more
unnatural isomers of reduced folate selected from the group
consisting of (6R)-tetrahydrofolic acid,
5-methyl-(6R)-tetrahydrofolic acid, 5-formyl-(6R)-tetrahydrofolic
acid, 10-formyl-(6S)-tetrahydrofolic acid,
5,10-methylene-(6S)-tetrahydrofolic acid,
5,10-methenyl-(6S)-tetrahydrofo- lic acid,
5-formimino-(6R)-tetrahydrofolic acid, and polyglutamyl derivatives
thereof, wherein the molar amount of the one or more unnatural
isomers of reduced folate is less than the molar amount of the one
or more natural isomers of reduced folate.
14. A method according to claim 12, wherein each of the one or more
natural isomers of reduced folate is substantially chirally
pure.
15. A method according to claim 12, wherein the one or more natural
isomers of reduced folate is selected from the group consisting of
5-methyl-(6S)-tetrahydrofolic acid, 5-formyl-(6S)-tetrahydrofolic
acid, 5,10-methenyl-(6R)-tetrahydrofolic acid, and polyglutamyl
derivatives thereof.
16. A method according to claim 12, wherein the nutritional
substance is an essential nutrient preparation comprising a vitamin
other than ascorbic acid.
17. A method according to claim 16, wherein the essential nutient
preparation further comprises ascorbic acid.
18. A method according to claim 12, wherein the nutritional
substance is a food preparation and wherein said method further
comprises: incorporating a vitamin into the food preparation.
19. A method for increasing a subject's dietary intake of folate
comprising: administering a composition according to claim 1 to the
subject.
20. A method according to claim 19, wherein said administering is
carried out by enteral administration.
21. A method according to claim 19, wherein the subject is an
animal.
22. A method according to claim 21, wherein the total molar amount
of said one or more natural isomers of reduced folate is between 5%
and 3000% of the animal's daily requirement for folate per
customarily consumed quantity of said composition.
23. A method according to claim 19, wherein the subject is a
human.
24. A method according to claim 23, wherein the total molar amount
of said one or more natural isomers of reduced folate is between 5%
and 200% of the human's daily requirement for folate per
customarily consumed quantity of said composition.
25. A method according to claim 23, wherein the human is selected
from the group consisting of a pregnant female; a female who has
had a miscarriage; a female who has carried a fetus having a neural
tube defect, a cleft lip defect, or a cleft palate defect; and a
human who suffers vascular disease.
26. A method for treating a subject afflicted with intestinal
malabsorption comprising: administering to the subject an amount of
a composition according to claim 1 effective to increase the
subject's blood folate level.
Description
[0001] This application claims the benefit of U.S. Provisional
Patent Application Ser. No. 60/010,898, filed Jan. 31, 1996.
FIELD OF THE INVENTION
[0002] The present invention relates generally to the field of
nutrition, and more specifically to food and vitamin preparations
containing the natural isomer of reduced folates.
BACKGROUND OF THE INVENTION
[0003] The folates are ubiquitous to nearly all forms of life.
Humans and many other animals lack the capacity to make their own
folate which thus is an essential vitamin, one type of essential
nurient. Anemia especially during pregnancy and in the geriatric
population was an early indication of a dietary requirement for
folate. A major function of folate is to remove one-carbon units
from molecules being metabolized and then deliver them to molecules
being synthesized. As an example, folate participates in the
formation of the nucleic acids. Further, the activity of DNA is
controlled, in part, by methylation, and the primary methylating
agent of the body (S-adenosylmethionine) is made in a metabolic
cycle involving a folate. Many studies have, therefore, focused on
the relationship of folate status to cancer susceptibility,
especially colorectal adenoma.
[0004] The importance of folate to proper growth is clearly evident
in the occurrence of neural tube defects in newborn infants.
Reports from several countries have shown that a majority of such
cases are associated with low folate levels in the mother. The
incidence of these defects as well as of cleft lip/palate is
considerably reduced when women are given folic acid (I) starting
early in pregnancy. Recently, a significant correlation has been
discovered between vitamin deficiency, especially of folate, and
peripheral vascular disease, a major cause of death. A high
percentage of individuals with this affliction have abnormal blood
levels of homocysteine, a precursor to methionine in the folate
dependent step of the S-adenosylmethonine cycle. Folate deficiency
has also been linked to defective maturation of a number of
different cell types, to nervous system disorders, and to decreased
immune response.
[0005] The clear relation of folate intake to health has caused
many governmental agencies around the world (such as the U.S.
National Research Council) to specify a recommended dietary
allowance ("RDA") for folate. In the U.S. these values are used by
the Food and Drug Administration to establish the Reference Daily
Intake ("RDI") that is listed on food labels, currently 0.4 mg for
adults. The highest daily amount of folic acid recommended by a
country is 2.0 mg for healthy adults. Many products are available
that contain RDI or near RDI levels of folic acid (I) including
most daily multiple vitamins. These can be purchased in solid (e.g.
tablet, capsule, or powder) or in liquid formulations, both
over-the-counter and by prescription. In the U.S. folic acid (I) is
also available by itself typically at a dosage of 0.4 mg, but also
up to 0.8 mg in health food stores. Many complete diets, infant
diets, dietary supplements and weight loss products also contain
folic acid (I). In some countries folic acid is added to specific
food types as determined by health officials to provide adequate
folate to the general population without risking excess
consumption. Many breakfast foods, such as cereals, cereal bars,
breakfast drink mixes, breakfast bars and toaster pastries have
folic acid (I) added at a modest fraction of the RDI, typically
10-50% of the adult value per serving. In many of these uses the
folic acid (I) is accompanied by other vitamins, sometimes at RDI
dosages, but also at lower or much higher levels. Frequently,
though not always, essential mineral nutrients are also present.
Further, many products also include compounds hypothesized to have
health related value, but which either have not been officially
recognized as effective, or for which optimal amounts have not been
set. Products such as those described above are meant to fill an
important and wide spread need for folate, especially among those
whose dietary habits would otherwise preclude intake of a
sufficient amount of this vitamin.
[0006] Folic acid (I) is a component of many animal and pet foods.
It is also included in powders or liquids used as animal feed
supplements, often in combination with other nutrients. For
example, the National Research Council (NRC) recommends diets
containing 0.2 mg and 1.0 mg of folic acid (I) per kg of dry diet
(assuming 5 kcal metabolizable energy per gram) for dogs and cats,
respectively. For chicks the NRC has recommended 0.55 mg folic acid
per kg of diet, although recent literature suggests that the
optimal value is about three times higher than this.
[0007] The form of folate currently added to all commercial vitamin
preparations or which is added to foods, folic acid (I) (also known
as pteroyl-L-glutamic acid), is not one of the major forms found in
natural fresh foods. The structure of folic acid (I) differs from
the most abundant natural folate in several aspects. First, the
side-chain of natural folates in almost all fresh foods contains
more than one L-glutamic acid moiety. Frequently, five to seven
(but covering a considerable span of more or fewer) of this amino
acid are linked together into a polyglutamate chain. It is well
known, however, that the primary form by which folates are absorbed
has only a single glutamate residue. Cleavage of the extra
glutamates of dietary folates is usually accomplished by an enzyme
in the digestive tract. In this aspect folic acid (I) is not at a
disadvantage in comparison to naturally occurring folates.
[0008] The second difference between folic acid (I) and natural
folates is that whereas the pteridine ring of the former (I) is
fully oxidized, natural folates in fresh uncooked foods are mostly
present as the tetrahydro forms. Almost all of the known
physiological functions of folate are performed by tetrahydrofolic
acid, (6S)-FH.sub.4 (II), or by a one carbon derivative of it
illustrated as follows: 5-methyl-(6S)-FH.sub.4 (III),
5-formyl-(6S)-FH.sub.4 (IV), 10-formyl-(6R)-FH.sub.4 (V),
5,10-methylene-(6R)-FH.sub.4 (VI), 5,10-methenyl-(6R)-FH.sub.4
(VII), and 5-formimino-(6S)-FH.sub.4 (VIII). The structural formula
for each of these compounds is provided below. 1
[0009] There is no known direct cofactor function for folic acid
(I) itself in humans. Some (6S)-tetrahydrofolic acid polyglutamate
is found in plants or animals, but the majority of folate is
polyglutamate forms of either 5-methyl-,
5-formyl-(6S)-tetrahydrofolic acid, and in some cases
10-formyl-tetrahydrofolic acid. Presumably, most of the folic acid
(I) found in biological food sources results from oxidation,
especially on storage. When folic acid (I) is absorbed by the
digestive tract it is eventually reduced to active
(6S)-tetrahydrofolic acid (II) by the enzyme dihydrofolate
reductase.
[0010] The oral bioavailability of folic acid (I) has been shown to
be widely variable. The literature contains reports of individuals
having poor intestinal uptake of folic acid (I) who respond
normally to intramuscular injection of folic acid (I), or had
normal serum folate status prior to any folic acid challenge.
Several small scale investigations in which the values have been
averaged have concluded that the oral uptake of several of the
reduced folates is similar to folic acid (I). However, there is
reason to believe that a segment of the population possesses
adequate oral response to reduced folates, but not to oral folic
acid (I).
[0011] 5-Formyl-tetrahydrofolic acid (also known as leucovorin or
folinic acid) has long been used in therapeutic doses for several
diseases. Examples include rescue from the toxicity of methotrexate
chemotherapy, and the synergistic combination with fluorouracil for
treatment of various cancers. It is also given to treat acute
anemia not due to B.sub.12 deficiency. 5-Methyl-tetrahydrofolic
acid in high doses (for example, 50 mg/day) has been patented for
treatment of depression (and other neurological disorders)
(EP382019 and EP388827 to Le Grazie 1990, and EP482493 to Le Greca
1992).
[0012] That reduced folates have been overlooked as an improved
source for providing the RDA level is in part due to the
stereochemistry of these compounds. In addition to the single
chiral center of the L-glutamate chain in folic acid (I, the
tetrahydrofolates contain a second stereochemical center at
carbon-6. Chemical reduction of folic acid (I) produces a nearly
racemic mixture of the two isomers at this position. This is in
contrast to the reduced folates found in nature which all consist
of a single diastereoisomer, all having the same L-configuration at
carbon-6. (Compounds II-VIII are shown as the natural isomer). For
many years only the racemic 6(R,S) mixture of
5-formyl-tetrahydrofolic acid (leucovorin) has been used for
therapy of diseases. Recently, however, concern over the possible
effects of the unnatural isomer component has resulted in the
commercial introduction of the pure natural isomer for these high
dose disease treatments by Lederie, although at very high cost.
Most therapeutic regimes utilizing leucovorin last a few weeks or
perhaps months. The effect of a long term exposure to the unnatural
isomer of reduced folates is unknown. For example, although little
5-formyl-(6R)-tetrahydrofolic acid is absorbed, there is
considerable uptake of the unnatural isomer of
5-methyl-tetrahydrofolic acid by the intestinal tract and other
cells of the body which with continuous intake may lead to adverse
consequences.
[0013] Until recently, processes for making the natural isomer of
reduced folates have been limited in scale, or costly, or both.
These include chromatographic separation, enzymatic reduction, and
fractional crystallization. The use of reduced folates as a daily
source of vitamin requires a method that is applicable to large
scale production of the natural isomer having high purity at a cost
that will not place a burden on the average consumer.
SUMMARY OF THE INVENTION
[0014] The present invention relates to a composition which
includes one or more natural isomers of reduced folate and a
nutritional substance. The one or more natural isomers of reduced
folate is selected from the group consisting of
(6S)-tetrahydrofolic acid, 5-methyl-(6S)-tetrahydrofo- lic acid,
5-formyl-(6S)-tetrahydrofolic acid, 10-formyl-(6R)-tetrahydrofol-
ic acid, 5,10-methylene-(6R)-tetrahydrofolic acid,
5,10-methenyl-(6R)-tetr- ahydrofolic acid,
5-formimino-(6S)-tetrahydrofolic acid, and polyglutamyl derivatives
thereof. The nutritional substance is a food preparation, an
essential nutrient preparation, or a combination thereof. When the
nutritional substance is a food preparation, the food preparation
includes two or more food components. Each gram of the food
preparation has a natural molar amount, N, of the one or more
natural isomers of reduced folate, N being greater than or equal to
zero, and each gram of the composition has a total molar amount, T,
of the one or more natural isomers of reduced folate greater than
N. When the nutritional substance is an essential nutrient
preparation, the essential nutrient preparation includes a vitamin
other than ascorbic acid.
[0015] The present invention also relates to method for increasing
the folate content of a nutritional substance. The method includes
providing a nutritional substance selected from the group
consisting of a food preparation, an essential nutrient
preparation, and combinations thereof. The method further includes
incorporating into the nutritional substance a molar amount of one
or more natural isomers of reduced folate selected from the group
consisting of (6S)-tetrahydrofolic acid,
5-methyl-(6S)-tetrahydrofolic acid, 5-formyl-(6S)-tetrahydrofolic
acid, 10-formyl-(6R)-tetrahydrofolic acid,
5,10-methylene-(6R)-tetrahydrofolic acid,
5,10-methenyl-(6R)-tetrahydrofolic acid,
5-formimino-(6S)-tetrahydr- ofolic acid, and polyglutamyl
derivatives thereof. When the nutritional substance is a food
preparation, the food preparation comprises two or more food
components. When the nutritional substance is an essential nutrient
preparation, the essential nutrient preparation comprises a vitamin
other than ascorbic acid.
[0016] A significant number of people are folate deficient;
especially vulnerable are those whose life style does not include
sufficient fresh food sources of folates. An object of this
invention is nutritional compositions in which the natural isomer
of tetrahydrofolic acid, or a derivative thereof, is substituted
for the usual folic acid (I) for the satisfaction or partial
satisfaction of the dietary requirement for this vitamin. While
some may not be greatly affected by the inclusion of reduced
folates in multivitamin preparations and breakfast foods, still a
substantial number of people, and thus the average health of the
population, will be improved by addressing the needs of those for
whom folic acid (I) bioavailability is poor. Consumer confidence
with regard to consumption of a food or other nutritional product
will be increased with the knowledge that the folate content is
chemically identical to the most abundant natural forms of this
vitamin, except for the advantageous absence of multiple glutamate
residues. A further advantage is that health agencies will be aided
in recommending optimal levels when a more uniformly absorbed form
of folate is widely used.
DETAILED DESCRIPTION OF THE INVENTION
[0017] The present invention relates to a composition which
includes one or more natural isomers of reduced folate and a
nutritional substance. Natural isomers of reduced folate suitable
for use in the present invention include, for example,
(6S)-tetrahydrofolic acid, 5-methyl-(6S)-tetrahydrofolic acid,
5-formyl-(6S)-tetrahydrofolic acid, 10-formyl-(6R)-tetrahydrofolic
acid, 5,10-methylene-(6R)-tetrahydrofolic acid,
5,10-methenyl-(6R)-tetrahydrofolic acid, and
5-formimino-(6S)-tetrahydrofolic acid. Other natural isomers of
reduced folate suitable for use in the present invention include
the polyglutamyl, such as the diglutamyl, triglutamyl,
tetraglutamyl, pentaglutamyl, and hexaglutamyl, derivatives of
(6S)-tetrahydrofolic acid, 5-methyl-(6S)-tetrahydrofolic acid,
5-formyl-(6S)-tetrahydrofolic acid, 10-formyl-(6R)-tetrahydrofolic
acid, 5,10-methylene-(6R)-tetrahydro- folic acid,
5,10-methenyl-(6R)-tetrahydrofolic acid, and
5-formimino-(6S)-tetrahydrofolic acid. Any or all of the natural
isomers of reduced folate can be present in its chirally pure form,
or, alternatively, the composition can optionally contain a molar
amount of one or more unnatural isomers of reduced folate, such as
(6R)-tetrahydrofolic acid, 5-methyl-(6R)-tetrahydrofolic acid,
5-formyl-(6S)-tetrahydrofolic acid, 10-formyl-(6S)-tetrahydrofolic
acid, 5,10-methylene-(6S)-tetrahydrofolic acid,
5,10-methenyl-(6S)-tetrahydrofo- lic acid,
5-formimino-(6R)-tetrahydrofolic acid, and polyglutamyl derivatives
thereof. The molar amount of the natural isomer of reduced folate
can be equal to the molar amount of its corresponding unnatural
isomer (as where the unnatural and natural isomer are present as a
racemic mixture), or, preferably, the natural isomer of reduced
folate can be present in a molar amount greater than the molar
amount of the corresponding unnatural isomer. The total molar
amount of the one or more natural isomers of reduced folate present
in the composition can be between 5% and 200% of a human daily
requirement for folate per a customarily consumed quantity of the
composition. As used herein, the total molar amount of the one or
more natural isomers of reduced folate includes natural isomers of
reduced folates which are naturally present in the nutritional
substance as well as natural isomers of reduced folates which might
have been added to the nutritional substance. The customarily
consumed quantity of various compositions depends, of course, on
the nature of the composition. Where the composition includes a
food preparation, the customarily consumed quantity is the amount
of the food preparation customarily consumed per eating occasion,
for example, as set forth by the U.S. Food and Drug Administration
for the purpose of establishing realistic and consistent serving
sizes for use in food labeling. Examples of customarily consumed
quantities for various food groups can be found in 21 C.F.R.
.sctn.101.12, which is hereby incorporated by reference.
[0018] The human daily requirement for folate varies from person to
person, depending on factors such as body weight, age, health, sex,
and the like. Suitable values for the human daily requirement for
folate include RDI and RDA values, promulgated, respectively, by
the FDA and the National Research Council ("NRC"). Presently, RDI
values, expressend in terms of micrograms of folic acid (nanomoles
of folic acid), are 400 .mu.g (907 nmoles) for adults and 800 .mu.g
(1814 nmoles) for pregnant women. In view of this, the compositions
of the present invention can have between 45 and 1814 nnmoles or
between 91 and 3625 nmoles of folate per customarily consumed
quantity of the nutritional substance. Current RDA values are
published in National Research Council: Recommended Daily
Allowances, 10th ed., Washington, D.C. (1989), which is hereby
incorporated by reference. They are, again expressend in terms of
micrograms of folic acid (nanomoles of folic acid), 25 .mu.g (57
nmoles) for infants 0-6 months of age, 35 .mu.g (79 nmoles) for
infants 6 months to one year of age, 50 .mu.g (113 nmoles) for
infants 1-3 years of age, 100 .mu.g (227 nmoles) for infants 7-10
years of age, 200 .mu.g (454 nmoles) for male adults, 180 .mu.g
(408 nmoles) for female adults other than pregnant adult females,
and 400 .mu.g (907 nmoles) for pregnant females. In view of this,
typical compositions of the present invention can have between 2.8
and 113 nmoles, between 4 and 159 nmoles, between 28 and 227
nmoles, between 11 and 454 nmoles, between 22 and 907 nmoles,
between 20 and 816 nmoles, or between 45 and 1812 nmoles of folate
per customarily consumed quantity of the nutritional substance.
Suitable values for the human daily requirement for folates are
also established by the World Health Organization as 7.03 nmoles/kg
of body weight. For pregnant women the value calculated based upon
body weight should be increased by about 454 to about 680
nmoles.
[0019] The total molar amount of the one or more natural isomers of
reduced folate present in the composition can, alternatively, be
between 5% and 3000% of an animal daily requirement for folate per
a customarily consumed quantity of the nutritional substance. The
animal, whose daily requirement for folate is referred to above,
can be, for example, a dog, a cat, a chicken, a cattle, a domestic
animal, a goat, a horse, a mink, a fox, a sheep, or a swine.
Suitable values for the an animal's daily requirement for folate
are promulgated, for example, by the NRC in Nutrient Requirements
of Domestic Animals (Washington:National Academy Press),
particularly in those publications having the following subtitles:
"Nutrient Requirements of Beef Cattle," Seventh Revised Edition
(1996, ISBN 0-309-05426-5); "Nutrient Requirements of Cats,"
Revised Edition (1986, ISBN 0-309-03682-8); "Nutrient Requirements
of Dairy Cattle," Sixth Revised Edition, Update (1989, ISBN
0-309-03826-X); "Nutrient Requirements of Dogs, Revised (1985, ISBN
0-309-03496-5); "Nutrient Requirements of Fish," (ISBN
0-309-04891-5); "Nutrient Requirements of Goats: Angora, Dairy, and
Meat Goats in Temperate and Tropical Countries," (ISBN
0-309-03185-0); "Nutrient Requirements of Horses," Fifth Revised
Edition, (1989, ISBN 0-309-03989-4); "Nutrient Requirements of
Laboratory Animals," Fourth Revised Edition (1995, ISBN
0-309-05126-6); "Nutrient Requirements of Mink and Foxes," Second
Revised Edition, (1982, ISBN 0-309-03325-X); "Nutrient Requirements
of Poultry," Ninth Revised Edition, (1994, ISBN 0-309-04892-3);
"Nutrient Requirements of Sheep," Sixth Revised Edition, (1985,
ISBN 0-309-03596-1); and "Nutrient Requirements of Swine," Ninth
Revised Edition, (1988, ISBN 0-309-03779-4) (collectively referred
to as "NRC Animal Nutrient Requirements"), which are hereby
incorporated by reference.
[0020] As indicated above the nutritional substance can be a food
preparation or an essential nutrient preparation. Essential
nutrient preparations are materials which contain one or more
essential nutrients. Where only one essential nutrient is present
in the essential nutrient preparation, that essential nutrient can
be a vitamin other than ascorbic acid. The essential nutrient
preparation can, alternatively, include a vitamin other than
ascorbic acid and, in addition, ascorbic acid. As used herein,
essential nutrients are those nutients which are required to
sustain health but which cannot be effectively produced by one or
more animals or by humans. Examples of essential nutrients are
compiled in a number of published sources, including Modern
Nutrition in Health and Disease, 8th ed., Shils et al., eds.,
Philadelphia:Lea and Febiger (1994), which is hereby incorporated
by reference. Essential nutients are meant to include essential
vitamins and provitamins thereof, essential fats, essential
minerals, such as those minerals for which daily values have been
recommended, and essential amino acids. One example of an essential
nutrient preparation is a formulation which contains a vitamin and
a caloric content of less than 2.5 cal per dry gram, preferably
less than 2 cal per dry gram, most preferably less than 1.8 cal per
dry gram. Essential nutrient preparations also include those
materials which contain at least one vitamin in an amount greater
than 15%, preferably greater than 20%, more preferably greater than
40% of the U.S. adult RDA for that essential nutrient per gram of
essential nutrient preparation. Still other suitable essential
nutrient preparations contain at least two vitamins, each in an
amount greater than 10%, preferably greater than 15%, more
preferably greater than 20% of the U.S. adult RDA for that
essential nutrient per gram of essential nutrient preparation.
Suitable essential nutrient preparations are commonly referred to
as dietary supplements, vitamin supplements, and mineral
supplements, multiple vitamin supplements, and the like, and are
typically available commercially in the form of pills, tablets,
capsules, powders, syrups, and suspensions. Preferably, the
essential nutrient composition contains at least one essential
nutrient in an amount greater than 25%, more preferably greater
than 50%, and most preferably greater than or equal to 100% of the
daily requirement for that essential nutrient per customarily
consumed quantity of the essential nutrient preparation.
[0021] As indicated above, the nutritional substance can also be a
food preparation. Food preparations are materials which contain one
or more amino acid, carbohydrate, or fat, which are suitable for
human or animal consumption, and which are not essential nutrient
preparations. It is preferred that the food preparation be a two or
more component food preparation. For example, a two or more
component food preparation can be a mixture of two or more
one-component foods. One component foods are foods which are
derived substantially from a single natural source. A small
percentage of the one-component food can be derived from a second
natural source, but that percentage, by weight, is preferably less
than 5%, more preferably less than than 1%, more preferably less
than 0.1%. One component foods include, for example, sugar,
unsweetened juice, nectar, or puree from a single species of plant,
such as unsweetened apple juice (including a blend of different
varieties of apple juice), grapefruit juice, orange juice, apple
sauce, apricot nectar, tomato juice, tomato sauce, tomato puree,
and the like. Grain plants of a single species and materials
produced from grain plants of a single species, such as corn syrup,
rye flour, wheat flour, oat bran, and the like are also considered
to be one component foods. Alternatively, the two or more component
food preparation can be a mixture of one or more one component
foods and one or more essential nutrients. Preferably, the amount
of at least one of the one or more essential nutrients present in
the two component food is greater than the amount of the at least
one essential nutrient that is naturally present collectively in
the one or more one component foods. For example, where the
essential nutrients are vitamin X and vitamin Y and where the one
component food is orange juice and where the orange juice naturally
contains vitamin X and vitamin Y in amounts "Nx" and "Ny",
respectively, it is preferred that the composition contain vitamin
X and vitamin Y in amounts "Tx" and "Ty", respectively, so that Tx
is greater than Nx, Ty is greater than Ny, or both.
[0022] Food preparations particularly well suited to the practice
of the present invention include breakfast foods, such as prepared
cereals, toaster pastries, and breakfast drink mixes; infant
formulas; dietary supplements; complete diet formulas; and
weight-loss preparations, such as weight-loss drinks and
weight-loss bars.
[0023] The food preparation can be one which naturally contains no
natural isomer of reduced folate. Alternatively, it can contain a
natural molar amount of a natural isomer of reduced folate. For
purposes of this application, the molar amount of natural isomer of
reduced folate (i.e., collectively, the number of moles of
(6S)-tetrahydrofolic acid, 5-methyl-(6S)-tetrahydrofolic acid,
5-formyl-(6S)-tetrahydrofolic acid, 10-formyl-(6R)-tetrahydrofolic
acid, 5,10-methylene-(6R)-tetrahydrofolic acid,
5,10-methenyl-(6R)-tetrahydrofolic acid,
5-formimino-(6S)-tetrahydr- ofolic acid, and polyglutamyl
derivatives thereof) contained per gram of food is designated "N".
For some foods, the molar amount of natural isomer of reduced
folate naturally present is known. For others, the molar amount of
natural isomer of reduced folate can be determined by a number of
sensitive and specific methods, such as microbial growth
dependence, folate binding protein based assays, high-performance
liquid chromatography ("HPLC") and gas chromatography ("GC") .
Suitable methods are described, for example, in Cossins, E. A.,
"Folates in Biological Materials," in Folates and Pterins, Vol. 1,
Blakley et al., eds., New York:John Wiley & Sons, pp. 1-60
(1984), which is hereby incorporated by reference.
[0024] The molar amount of natural isomer of reduced folate present
in the composition of the present invention is greater that the
molar amount of natural isomer of reduced folate present in the
food preparation. For purposes of this application, the molar
amount of natural isomer of reduced folate (i.e., collectively, the
number of moles of (6S)-tetrahydrofolic acid,
5-methyl-(6S)-tetrahydrofolic acid, 5-formyl-(6S)-tetrahydrofolic
acid, 10-formyl-(6R)-tetrahydrofolic acid,
5,10-methylene-(6R)-tetrahydrofolic acid,
5,10-methenyl-(6R)-tetrahydrofo- lic acid,
5-formimino-(6S)-tetrahydrofolic acid, and polyglutamyl derivatives
thereof) contained per gram of composition is designated "T". Thus,
T necessarily must be greater than N. Preferably, T/N is greater
than 105%; more preferably, it is greater that 110%; and, most
preferably, it is greater than 120%.
[0025] As indicated above; the composition can, optionally, include
one or more unnatural isomers of reduced folate. When present in
the composition, the one or more unnatural isomers of reduced
folate is preferably present in a molar amount which is less than T
minus N.
[0026] The present invention also relates to a method for
increasing the folate content of a nutritional substance. The
nutritional substance can be a food preparation, an essential
nutrient preparation, or a combination of a food preparation and an
essential nutrient preparation. The folate content of the
nutritional substance is increased by incorporating one or more
natural isomers of reduced folate into or with the nutritional
substance. This can be achieved by methods well known in the art of
food and essential nutrient preparation, such as by homegenizing,
coating, spraying, coarsely mixing, tossing, kneading, pilling, and
extruding one or more unnatural isomer of reduced folate, singly or
in combination, onto or with the nutritional substance.
[0027] One or more of the one or more natural isomers of reduced
folate that are added to the nutritional substance in accordance
with the present invention can be substantially chirally pure or
each of the one or more natural isomers of reduced folate can be
chirally pure. Alternatively, one or more of the one or more
natural isomers of reduced folate can be present in a mixture with
one or more unnatural isomers of reduced folate. The molar amount
of the one or more natural isomers of reduced folate and the one or
more unnatural isomers of reduced folate present in the mixture
added to the nutritional substance can be the same, as in the case
where a racemic mixture is added, or they can be different.
Preferably the molar amount of the natural isomer exceeds the molar
amount of the unnatural isomer. Additionally or alternatively,
unnatural isomer of reduced folate can be incorporated in a
separate step subsequent to or prior to incorporating the one or
more natural isomers of reduced folate into the nutritional
substance. It is preferred that the collective molar amount of
unnatural isomer of reduced folate added before, during, and/or
after the addition of natural isomer of reduced folate be less than
the collective molar amount of natural isomer added.
[0028] Natural isomers of reduced folates that are substantially
chirally pure can be prepared by any suitable method, including,
for example, by the method described in U.S. Pat. No. 5,350,851 to
Bailey et al., which is hereby incorporated by reference.
[0029] When the nutritional substance is a food preparation, in
addition to incorporating a natural isomer of reduced folate, one
or more essential nutrients, optionally, can be incorporated into
the food preparation. The essential nutrients can, for example, be
added to the food preparation before, during, or after addition of
the natural isomer of reduced folate.
[0030] The compositions of the present invention can be used to
increase a subject's dietary intake of folate by administering the
composition to the subject. The subject can be an animal, such as a
dog or a cat; alternatively, the subject can be a human. Certain
classes of individuals are viewed to be especially benefitted by
increasing dietary intake of folate. These include pregnant
females; females who have had a miscarriage; females who have
carried a fetus having a neural tube defect, a cleft lip defect, or
a cleft palate defect; and humans who suffer vascular disease.
[0031] The compositions can also be used to treat a subject
afflicted with intestinal malabsorption, especially folate
malabsorption. When treating a subject afflicted with intestinal
malabsorption, the amount of composition administered is preferably
effective to cause an increase in the subject's blood folate level.
More preferably, the amount administered is effective to produce
blood folate levels in an normal range, as determined by
conventional blood-folate analysis methods, such as with the Quanta
Phase II assay from BioRad Laboratories, Hercules, Calif.
[0032] The compositions can be administered enterally, such as
orally, intragastricly, or transpyloricly. Many factors that may
modify the action of the composition can be taken into account by
those skilled in the art; e.g., body weight, sex, diet, time of
administration, route of administration, rate of excretion,
condition of the subject, and reaction sensitivities and
severities. Administration can be carried out continuously or
periodically, such as once daily, or once with every meal.
[0033] Compositions containing the natural isomer of reduced
folates are preferably for enteral consumption (enteral including
oral, intragastric, or transpyloric), and are of any, food
preparation, food supplement, essential nutrient preparation, or
vitamin preparation. Typical examples of such food or vitamin
preparations are those to which folic acid (I) is currently added
for use by either humans or other animals. Natural isomer means a
tetrahydrofolate having the natural configuration at both the
glutamate a- and the pteridine 6-carbons. More specifically, these
compositions are, but are not limited to, multivitamin (with or
without minerals and other nutrients) preparations (such
preparations can be in solid, liquid or suspension forms);
breakfast foods such as prepared cereals, breakfast drink mixes,
toaster pastries and breakfast bars; infant formulas; dietary
supplements and complete diet and weight-loss formulas and bars;
animal feed or animal feed supplements (for example, for poultry),
and pet foods. The natural isomer of reduced folates can also be
used in products which health officials might in the future deem an
appropriate vehicles for satisfaction of the daily folate
requirement. The composition of the present invention can include a
natural isomer of a tetrahydrofolate, such as II-VIII, preferably
the monoglutamate form, in a vitamin formulation preferably
containing at least one other vitamin (for example another vitamin
such as a B vitamin) other than ascorbic acid (vitamin C), although
ascorbic acid can be an additional component. Such nutrients or
vitamins can be those intended for either human or animal use.
Other ingredients may also be present, such as fillers, binding
agents, stabilizers, sweeteners, including nutitive sweeteners
(e.g. sucrose, sorbitol and other polyols) and non-nutritive
sweeteners (e.g. saccharin, aspartame, and acesulfame K),
colorants, flavors, buffers, salts, coatings, and the like that are
known to those skilled in the art of vitamin formulation.
[0034] For many applications of the described reduced folates (II
through VIII) 5-methyl-(6S)-tetrahydrofolic acid (III),
5-formyl-(6S)-tetrahydrof- olic acid (IV), and
5,10-methenyl-(6R)-tetrahydrofolic acid (VII) are preferred. All of
the reduced folates are to differing extent subject to oxidation by
air. Several factors influence this susceptibility, foremost being
derivatization of the 5-nitrogen which considerably slows
oxidation. The 5-methyl- and 5-formyl derivatives are the most
abundant forms of folate found in most tissues of the body. The
predominate folate in human blood is 5-methyl-(6S)-tetrahydrofolic
acid (III).
[0035] In using tetrahydrofolates for increasing the folate in a
nutritional composition, several factors should be considered. Many
nutrients are unstable to processing, including several of the
vitamins. For example, vitamins A, B.sub.1 (thiamin), and ascorbic
acid are especially labile under some conditions. Many procedures
are known for enhancing the stability of the various nutrients such
as pH and moisture control of the composition. Components which are
to a degree incompatible with each other, for example ascorbic acid
with the more oxidized forms of iron or copper, can be made to be
present heterogeneously in the composition. Nutrients which are
unstable to heat are often added after baking steps; for example,
vitamins are often sprayed onto breakfast cereals subsequent to
toasting. Nutrients which are unstable to air can be packaged in a
reduced oxygen condition, and/or in containers that have low or no
permeability to oxygen. These and other procedures known to those
skilled in the art are useful for maintaining folates in their
natural reduced form.
[0036] The rate of oxidation of a reduced folate in the presence of
air is increased in water solutions. The shelf life of liquid
preparations will be greatly extended if they are preferably kept
in air-tight containers. Replacing air with an inert gas such as
nitrogen or argon also retards loss. Reduced folates can also be
protected from oxidation by a number of reducing agents and
antioxidants, the most relevant of these being other vitamins that
are often included in multivitamin mixtures or nutritional
compositions. Ascorbic acid has been used for protection of reduced
folates in biochemical experiments and procedures for the
laboratory analysis of biological samples. Such protection need not
be limited to use of ascorbic acid or other vitamins; other agents
suitable for human or animal consumption are useful, for example
iso-ascorbic acid and certain thiols, such as glutathione. Further,
known packaging and formulation technologies which increase the
stability of compounds such as ascorbic acid or other air labile
materials (for example, coated forms, blister packaging, and use of
reduced metals or metal complexes) are useful for the maintenance
of reduced folates. The salt form of a reduced folate also somewhat
affects stability and solubility, and this can be optimized for the
needs of each product. The pH of the final composition can also be
optimized according to the stability properties of the particular
reduced folate derivative used and of the other components present,
as is well understood in the arts of processing nutrients and of
folate compounds. For example, in the presence of moisture
5,10-methenyl-(6R)-FH.sub.4 (VII) can be transformed into
10-formyl-(6R)-FH.sub.4 and 5-formyl-(6S)-FH.sub.4 (IV) (the latter
also a preferred compound) in a pH dependent manner. Compostions
containing 5,10-methenyl-(6R)-FH.sub.4 (VII) are most stable to
oxidation when either substantially dry and/or have an acidity less
than about pH 4. With proper attention to the above factors, the
lability of reduced folates need not limit the life of a product,
especially with III and IV which are more resistant than ascorbic
acid to many oxidation reactions.
[0037] The substitution of a reduced folate for folic acid (I)
should take into account the differences in molecular weights of
the various forms. For example, the current U.S. Reference Daily
Intake of 0.4 mg of folic acid (I) corresponds to 0.91 micromole
using an anhydrous molecular weight of 441.4. The effective
molecular weight of reduced folates depends upon the derivative
employed (i.e. II-VIII), the salt form, and water content. For
example, 0.91 micromole of 5-formyl-(6S)-tetrahydrofol- ic acid
(IV) calcium salt-pentahydrate would weigh 0.545 mg, and 0.91
micromole of 5-methyl-(6S)-tetrahydrofolic acid (III) disodium salt
would weigh 0.456 mg. Several salt forms of the reduced folates are
described in the literature, such as hydrochloride, sodium,
potassium, magnesium, calcium, and others and having various water
content. For each of these forms a similar calculation can be made.
The amount required to achieve the mole equivalent to a desired
fraction of the RDI would then be that fraction of this new weight.
As an example, 25% RDI of 5-methyl-(6S)-tetrahydrofolic acid (III)
disodium salt would be 0.25.times.0.456=0.114 mg, the mole
equivalent of 0.10 mg of folic acid (I). Previous investigations of
groups of individuals having a normal uptake of folic acid (I) have
shown that the bioavailability of the reduced folates is similar on
a mole basis.
[0038] As mentioned above, loss of nutrients during processing,
especially of foods is well known to those skilled in this art. An
often practiced procedure is the addition of an initial excess, an
"overage", of a particular nutrient or nutrients, such that the
final post-processing amount is at the desired level. Many highly
sensitive and specific methods are known (such as microbial growth
dependence, folate binding protein based assays, HPLC and GC) for
the analysis of folates, in both the reduced and oxidized forms as
well as for their various derivatives. These assays permit
adjustment of the added amount of the natural isomer of a reduced
folate so as to yield the desired final amount subsequent to
processing and packaging. The range of the natural isomer of a
reduced folate in the composition of this invention is preferably
that fulfilling between about 5% and about 200% of the RDI of
humans for folate, and should be taken to encompass both the
situation where allowance is made for processing loss, and also
where no such allowance is made. Separate RDA dosages are specified
for different groups of people, for example pregnant and
non-pregnant women. Further, the RDI level although relying on RDA
values, can be different from RDA values. The above range of
"between about 5% and about 200% of the RDI for folate" should be
taken to operate independently on each of these separate RDA and
RDI specifications, or their foreign equivalents, as presently
stated or as modified in the future. For the purpose of this
invention these several specifications shall be referred to as the
daily requirement for folate. Unless the recommended dietary
allowance for folate in humans is increased, the maximum final
amount of the natural isomer of a reduced folate in composition for
human use in satisfying the daily requirement for folate preferably
should not exceed about 4.5 micromole per dose or customarily
consumed serving. However, for individuals afflicted with
intestinal malabsorption, such as celiac disease or tropical sprue,
compositions containing higher amounts of the natural isomer of a
reduced folate will be useful.
[0039] For the purpose of this invention an essential nutrient
composition can be a dietary supplement or the like, the
substantial folate component of which is derived from substantially
pure tetrahydrofolic acid or derivative thereof, such as compounds
II through VIII. Essential nutrient compositions encompassed by
this invention comprise the natural isomer of a reduced folate
preferably within the above described range along with other
vitamins and/or other nutrients which are preferably each present
in an amount that is considered to be safe. In formulating
compositions for animal consumption manufacturers often
considerably exceed the dosage recommended by the NRC for folate
(by 10-fold, 20-fold, or more in some cases), not only to overcome
losses during processing, but also to cover occasions of possible
increased need for folate, such as during antibiotic treatment.
Other vitamin and nutrient components can be present in amounts
that vary considerably from NRC recommendations. The following
examples are given to further illustrate the invention, and are not
intended to limit its scope in any way.
EXAMPLES
[0040] 1) A typical ready to eat breakfast cereal: corn (and/or
other grains), sugar, salt, malt flavoring, such that a 30 g
serving provides about 2 g of protein, 26 g total carbohydrate, and
330 mg of sodium, also containing per serving size vitamin A
palmitate (15% of RDI), ascorbic acid (25% of RDI), reduced iron
(45% of RDI), vitamin D (10% of RDI), thiamin hydrochloride (25% of
RDI), riboflavin (25% of RDI), niacinamide (25% of RDI), pyridoxine
hydrochloride (25% of RDI), and 0.114 mg of
5-methyl-6(S)-tetrahydrofolic acid (III) disodium salt (the mole
equivalent of 0.1 mg folic acid, 25% of RDI).
[0041] 2) A typical daily multivitamin tablet: calcium carbonate,
ascorbic acid (60 mg, 100% RDI), gelatin, vitamin E acetate (30
I.U., 100% RDI), starch, niacinamide (20 mg, 100% RDI),
hydroxypropyl-methylcellulose, calcium pantothenate (10 mg, 100%
RDI), calcium silicate, hydroxypropylcellulose, pyridoxine
hydrochloride (2 mg, 100% RDI), riboflavin (1.7 mg, 100% RDI),
thiamin mononitrate (1.5 mg, 100% RDI), beta carotene & vitamin
A acetate (5000 I.U., 100% RDI), sodium hexametaphosphate,
magnesium stearate, vitamin D (400 I.U., 100% RDI), vitamin
B.sub.12 (6 .mu.g, 100% RDI), lecithin, and 0.437 mg of
5-methyl-6(S)-tetrahydrofolic acid (III) magnesium salt (the mole
equivalent of 0.4 mg folic acid, 100% of RDI).
[0042] 3) A typical daily multivitamin and minerals tablet: calcium
phosphate (130 mg of elemental calcium), magnesium hydroxide &
stearate (100 mg, 25% RDI), cellulose, potassium chloride, ascorbic
acid (60 mg, 100% RDI), gelatin, ferrous fumarate (18 mg elemental
iron, 100% RDI), zinc sulfate (15 mg, 100% RDI), modified cellulose
gum, vitamin E acetate (30 I.U., 100% RDI), citric acid,
niacinamide (20 mg, 100% RDI), magnesium stearate,
hydroxypropyl-methylcellulose, calcium pantothenate (10 mg, 100%
RDI), selenium yeast, polyvinylpyrrolidone, hydroxypropylcellulose,
manganese sulfate, silica, copper oxide (2 mg, 100% RDI), chromium
yeast, molybdenum yeast, pyridoxine hydrochloride (2 mg, 100% RDI),
riboflavin (1.7 mg, 100% RDI), thiamin mononitrate (1.5 mg, 100%
RDI), beta carotene & vitamin A acetate (5000 I.U., 100% RDI),
potassium iodide (150 .mu.g, 100% RDI), sodium hexametaphosphate,
biotin (30 .mu.g, 10% RDI), vitamin D (400 I.U., 100% RDI), vitamin
B.sub.12 (6 .mu.g, 100% RDI), lecithin, and 0.545 mg
5-formyl-(6S)-tetrahydrofolic acid (IV) calcium salt-pentahydrate
(the mole equivalent of 0.4 mg of folic acid, 100% RDI).
[0043] 4) A typical daily multivitamin and minerals tablet for
older adults: calcium carbonate, calcium phosphate (200 mg Ca, 20%
RDI; 48 mg phosphorous, 5% RDI), magnesium oxide, magnesium
stearate (100 mg, 25% RDI), potassium chloride (80 mg, 2% RDI),
microrystalline cellulose, ascorbic acid (60 mg, 100% RDI),
gelatin, d'l-alfa-tocopheryl acetate (45 I.U., 150% RDI), modified
food starch, maltodextrin, crospovidone, reduced iron (4 mg, 22
RDI), hydroxypropyl methylcellulose, niacinamide (20 mg, 100% RDI),
zinc oxide (15 mg, 100% RDI), calcium pantothenate, manganese
sulfate (3.5 mg), vitamin D (400 I.U., 100% RDI), titanium dioxide,
vitamin A and .beta.-carotene (5000 I.U., 100% RDI), stearic acid,
pyridoxine hyrochloride (3 mg, 150% RDI), riboflavin (1.7 mg, 100%
RDI), silicon dioxide, copper oxide (2 mg, 100% RDI), dextrose,
thiamin mononitrate (1.5 mg, 100% RDI), triethyl citrate,
polysorbate 80, chhromium chloride (130 .mu.g),, artificial colors,
potassium iodide ((150 .mu.g, 100% RDI), sodium metasilicate (2
mg), sodium molybdate (160 .mu.g), borates, sodium selenate (20
.mu.g), biotin (30 .mu.g, 10% RDI), sodium metavanadate (10 .mu.g),
cyanocobalamin (25 .mu.g, 417% RDI), nickelous sulfate (5 .mu.g),
and phytonadione, and 5,10-methenyl-(6R)-tetrahydrofolic acid
hydrochloride (VII)(0.44 mg, the mole equivalent of 0.4 mg of folic
acid, 100% RDI).
[0044] 5) A typical complete diet drink: water, sugar, calcium and
sodium caseinates, maltodextrin, high-oleic safflower oil, soy
protein, soy oil, canola oil, cocoa, sodium and potassium citrates,
calcium carbonate and phosphate (250 mg Ca, 25% RDI), magnesium
chloride and phosphate (100 mg Mg, 25% RDI), sodium chloride, soy
lecithin, choline chloride, flavor, ascorbic acid (30 mg, 50% RDI),
carrageenan, zinc sulfate (5.6 mg, 37% RDI), ferrous sulfate (4.5
mg Fe, 25% RDI), alfa-tocopheryl acetate (11.3 I.U., 37.7% RDI),
niacinamide (5 mg, 25% RDI), calcium pantothenate (2.5 mg, 25%
RDI), manganese sulfate (1.3 mg), copper salt (25% RDI), vitamin A
palmitate (1250 I.U., 25% RDI), thiamin hydrochloride (0.375 mg,
25% RDI), pyridoxine hydrochloride (0.5 mg, 25% RDI), riboflavin
(0.425 mg, 25% RDI), biotin (75 .mu.g, 25% RDI), sodium molybdate
(38 .mu.g), chromium chloride (25 .mu.g), potassium iodide (37.5
.mu.g, 25% RDI), sodium selenate (18 .mu.g), phylloquinone (vitamin
K.sub.1), cyanocobalamin (1.5 .mu.g, 25% RDI), vitamin D.sub.3 (100
I.U., 25% RDI), and 0.136 mg 5-formyl-(6S)-tetrahydrofolic acid
(IV) calcium salt-pentahydrate (the mole equivalent of 0.1 mg of
folic acid, 25% RDI), packaged in an air-tight container, and
supplying about 225 calories.
[0045] 6) A typical enhanced B-vitamin/tetrahydrofolate tablet:
dibasic calcium phosphate, pyridoxine hydrochloride (50 mg, 2,500%
RDI), cellulose, stearic acid, magnesium stearate, and 0.912 mg of
5-methyl-6(S)-tetrahydrofolic acid (III) disodium salt (the mole
equivalent of 0.8 mg folic acid, 200% of RDI for adults, 100% RDA
for pregnant women).
[0046] 7) A typical poultry feed vitamin supplement: (amounts per
kg of diet) vitamin A (trans retinyl acetate, 5500 I.U.), vitamin E
(11 I.U.), menadione sodium bisulfite (1.1 mg), vitamin D.sub.3
(1100 I.U.), riboflavin (4.4 mg), vitamin B.sub.12 (10 .mu.g),
vitamin B.sub.6 (3.0 mg), thiamin mononitrate (2.2 mg), biotin (0.3
mg), ethoxyquin (125 mg), and 2.0 mg 5-formyl-(6S)-tetrahydrofolic
acid (IV) calcium salt-pentahydrate (the mole equivalent of 1.45 mg
of folic acid).
[0047] 8) A typical dry cat food: ground yellow corn, corn gluten
meal, soybean meal, poultry by-product meal, animal fat, fish meal,
meat and bone meal, ground wheat, phosphoric acid calcium
carbonate, dried animal digest, salt, brewers dried yeast,
potassium chloride, dried whey solubles, choline chloride, dried
skimmed milk, taurine, L-lysine, zinc oxide, ferrous sulfate,
niacin, vitamin A, vitamin D.sub.3, vitamin B.sub.12, calcium
pantothenate, citric acid, manganese sulfate, riboflavin
supplement, biotin, copper salt, thiamine mononitrate, pyridoxine
hydrochloride, menadione sodium bisulfate complex, such that the
crude protein is not less than 31%, crude fat is not less than 8%,
crude fiber is not more than 4.5%, moisture is not more than 12%,
calcium is not less than 1.2%, phosphorous is not less than 1.0%,
sodium chloride is not more than 1.5%, the metabolizable energy is
about 3,600 kcal/kg, taurine, iron, vitamins A, D.sub.3, B.sub.12,
and E are at least 100% of levels recommended by the Association of
American Feed Control Officials, and containing not less than 0.97
mg/kg diet 5-methyl-6(S)-tetrahydrofoli- c acid (III) calcium salt
dihydrate (the mole equivalent of 0.8 mg/kg diet of folic
acid).
[0048] 9) A typical soy based infant formula: 75.5% water; 13%
sucrose; 6.6% oleo oil: coconut, high oleic (safflower or
sunflower), and soybean oils; 3.8% soy protein isolate; (protein
2.7 g, fat 5.3 g, carbohydrate 10.2, linoleic acid 500 mg);
potassium citrate and bicarbonate (potassium 105 mg); monobasic
potassium and dibasic calcium phosphates (phosphorous 63 mg); soy
lecithin; taurine; calcium carrageenan; calcium hydroxide, chloride
and citrate (calcium 90 mg); sodium chloride (sodium 30 mg);
L-methionine; zinc (Zn 0.8 mg), ferrous (Fe 1.8 mg), and manganese
(Mn 30 .mu.g) sulfates; copper salt (Cu 70 .mu.g); taurine;
L-carnitine; potassium iodide (I 9 .mu.g); ascorbic acid (8.3 mg);
choline chloride; alpha-tocopheryl acetate (1.4 I.U.); niacinamide
(750 .mu.g); vitamin A palmitate and betacarotene (300 I.U.);
calcium pantothenate (450 .mu.g); thiamin hydrochloride (100
.mu.g); riboflavin (150 .mu.g); pyridoxine hydrochloride (62.5
.mu.g); vitamin K, (15 .mu.g); biotiri (5.5 .mu.g); vitamin D.sub.3
(60 I.U.); cyanocobalamin (0.3 .mu.g); and 9.1 .mu.g of
5-methyl-6(S)-tetrahydrofolic acid (III) calcium salt dihydrate
(the mole equivalent of 7.5 .mu.g of folic acid), packaged in an
air-tight container (amounts are per 150 ml of 1:1 diluted
formula).
[0049] One skilled in the art will readily appreciate that the
present invention is well adapted to carry out the objects and
obtain the ends and advantages mentioned. While the above
description contains many specificities, these should not be
construed as limitations on the scope of the invention, but rather
as an exemplification of preferred embodiments thereof. Changes
therein and other uses will occur to those skilled in the art which
are encompassed within the spirit of the invention as defined by
the scope of the claims and their legal equivalents.
* * * * *