U.S. patent application number 14/728008 was filed with the patent office on 2015-09-17 for stable and bioavailable compositions of isomers of carotenoids for skin and hair.
The applicant listed for this patent is Nestec S.A.. Invention is credited to Karlheinz Bortlik, Pierre Lambelet, Myriam Richelle.
Application Number | 20150258038 14/728008 |
Document ID | / |
Family ID | 37496603 |
Filed Date | 2015-09-17 |
United States Patent
Application |
20150258038 |
Kind Code |
A1 |
Bortlik; Karlheinz ; et
al. |
September 17, 2015 |
STABLE AND BIOAVAILABLE COMPOSITIONS OF ISOMERS OF CAROTENOIDS FOR
SKIN AND HAIR
Abstract
The present invention relates to compositions that provide
health benefits and methods regarding same are presented. In an
embodiment, the present invention provides a primary composition
comprising at least one carotenoid-containing material, enriched in
Z isomers of the carotenoid compound. For example, the
carotenoid-containing material contains by weight a greater
percentage of an isomer selected from the group consisting of 5-Z,
9-Z and combinations thereof than of 13-Z isomer.
Inventors: |
Bortlik; Karlheinz; (Syens,
CH) ; Lambelet; Pierre; (Saint-Legier, CH) ;
Richelle; Myriam; (Savigny, CH) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Nestec S.A. |
Vevey |
|
CH |
|
|
Family ID: |
37496603 |
Appl. No.: |
14/728008 |
Filed: |
June 2, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12376662 |
May 4, 2009 |
|
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PCT/EP2007/006972 |
Aug 7, 2007 |
|
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14728008 |
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Current U.S.
Class: |
424/59 ;
424/70.1; 514/762 |
Current CPC
Class: |
A23V 2002/00 20130101;
A61K 2800/92 20130101; A61Q 19/00 20130101; A61P 17/00 20180101;
A61Q 5/00 20130101; A61Q 19/08 20130101; A23L 33/155 20160801; A61Q
19/007 20130101; A23V 2250/213 20130101; A61K 31/01 20130101; A61K
8/31 20130101; A61P 17/16 20180101; A61Q 19/005 20130101; A61P 9/00
20180101; A23V 2002/00 20130101; A61K 36/81 20130101; A23L 33/105
20160801; A61K 2800/10 20130101; A61P 17/14 20180101 |
International
Class: |
A61K 31/01 20060101
A61K031/01; A23L 1/30 20060101 A23L001/30; A61Q 19/00 20060101
A61Q019/00; A61Q 5/00 20060101 A61Q005/00; A61K 36/81 20060101
A61K036/81; A61K 8/31 20060101 A61K008/31 |
Foreign Application Data
Date |
Code |
Application Number |
Aug 8, 2006 |
EP |
06118579.9 |
Claims
1. A primary composition comprising: at least one
carotenoid-containing material extracted from a source selected
from the group consisting of tomatoes, carrots, peaches, apricots,
oranges, melons, guavas, papayas, grapefruits, wolfberries,
rosehips, soya, green tea, spices, grapes, cocoa and combinations
thereof, and isomerized in an organic solvent so as to comprise, by
weight, a greater percentage of an isomer selected from the group
consisting of 5-Z, 9-Z and combinations thereof than of a 13-Z
isomer.
2. The primary composition of claim 1, wherein the
carotenoid-containing material contains more than 30% by weight of
the isomer selected from the group consisting of 5-Z, 9-Z and
combinations thereof.
3. The primary composition of claim 1, wherein the
carotenoid-containing material contains less than 10% by weight of
the 13-Z isomer.
4. The primary composition of claim 1, comprising a content in E
isomer that is not greater than 60% of the total carotenoid
content.
5. The primary composition of claim 1, wherein the Z isomers of the
carotenoid compound are present in an amount effective to increase
the bioavailability and/or bioefficacy of the carotenoid
compound.
6. The primary composition of claim 1, wherein the carotenoid
compound is selected from the group consisting of lycopene,
carotenes, zeaxanthine, astaxanthine, .beta.-cryptoxanthin,
capsanthine, canthaxanthine, lutein, phytofluene, phytoene and
combinations thereof.
7. The primary composition of claim 1, wherein the Z/E isomer ratio
of the carotenoid compound is at least 20:80.
8. The primary composition of claim 1, wherein the composition is
in a form selected from the group consisting of liquid, gel and
powder.
9. An oral composition comprising: a primary composition comprising
at least one carotenoid-containing material extracted from a source
selected from the group consisting of tomatoes, carrots, peaches,
apricots, oranges, melons, guavas, papayas, grapefruits,
wolfberries, rosehips, soya, green tea, spices, grapes, cocoa and
combinations thereof, and isomerized in an organic solvent so as to
comprise, by weight, a greater percentage of an isomer selected
from the group consisting of 5-Z, 9-Z and combinations thereof than
of a 13-Z isomer.
10. The oral composition of claim 9, wherein the oral composition
is selected from the group consisting of a foodstuff, a food
supplement, a pet food product and a pharmaceutical
preparation.
11. The oral composition of claim 10, wherein the foodstuff is
selected from the group consisting of a nutritional complete
formula, a dairy product, a chilled or shelf stable beverage, a
mineral water, a liquid drink, a soup, a dietary supplement, a meal
replacement, a nutritional bar, a confectionery product, a milk, a
fermented milk product, a yogurt, a milk based powder, an enteral
nutrition product, an infant formulae, an infant nutritional
product, a cereal product, a fermented cereal based product, an
ice-cream, a chocolate, coffee, a culinary product, a pet food
product and combinations thereof.
12. The oral composition of claim 10, wherein the food supplement
is provided in a form selected from the group consisting of
capsules, gelatin capsules, soft capsules, tablets, sugar-coated
tablets, pills, pastes or pastilles, gums, drinkable solutions or
emulsions, syrups and gels.
13. The oral composition of claim 9 comprising at least one
component selected from the group consisting of a sweetener, a
stabilizer, a flavoring and a colorant.
14. The oral composition of claim 9, wherein the content of the
primary composition is between about 0.001 and 100% by weight.
15. The oral composition of claim 9, wherein the content of the
primary composition is between about 10 and 50% by weight.
16. A cosmetic composition comprising: a primary composition
comprising at least one carotenoid-containing material extracted
from a source selected from the group consisting of tomatoes,
carrots, peaches, apricots, oranges, melons, guavas, papayas,
grapefruits, wolfberries, rosehips, soya, green tea, spices,
grapes, cocoa and combinations thereof, and isomerized in an
organic solvent so as to comprise, by weight, a greater percentage
of an isomer selected from the group consisting of 5-Z, 9-Z and
combinations thereof than of a 13-Z isomer.
17. The cosmetic composition of claim 16, wherein the content of
primary composition is between about 10.sup.-10% and 10% by weight
of the cosmetic composition.
18. A process of manufacturing a composition, the process
comprising: extracting a carotenoid-containing material from a
source selected from the group consisting of tomatoes, carrots,
peaches, apricots, oranges, melons, guavas, papayas, grapefruits,
wolfberries, rosehips, soya, green tea, spices, grapes, cocoa and
combinations thereof; and isomerizing the carotenoid-containing
material in an organic solvent under conditions sufficient to
increase its content in Z isomers of the carotenoid compound,
wherein the carotenoid-containing material comprises by weight a
greater percentage of an isomer selected from the group consisting
of 5-Z, 9-Z and combinations thereof than of a 13-Z isomer.
19. The process of claim 18, wherein isomerizing the
carotenoid-containing comprises solubilization of Z isomers in the
organic solvent followed by phase separation using centrifugation
or filtration.
20. A method for improving skin health, the method comprising:
administering a composition comprising a primary composition having
at least one carotenoid-containing material extracted from a source
selected from the group consisting of tomatoes, carrots, peaches,
apricots, oranges, melons, guavas, papayas, grapefruits,
wolfberries, rosehips, soya, green tea, spices, grapes, cocoa and
combinations thereof, and isomerized in an organic solvent so as to
comprise, by weight, a greater percentage of an isomer selected
from the group consisting of 5-Z, 9-Z and combinations thereof than
of a 13-Z isomer.
21. The method according to claim 20, wherein the primary
composition is used for a method selected from the group consisting
of protecting the tissues of the skin against aging, prevention or
treatment of sensible, dry and reactive skins, for improving skin
density or firmness, and for increasing skin photoprotection.
22. A method for improving hair and coat quality, the method
comprising: administering a composition comprising a primary
composition having at least one carotenoid-containing material
extracted from a source selected from the group consisting of
tomatoes, carrots, peaches, apricots, oranges, melons, guavas,
papayas, grapefruits, wolfberries, rosehips, soya, green tea,
spices, grapes, cocoa and combinations thereof, and isomerized in
an organic solvent so as to comprise, by weight, a greater
percentage of an isomer selected from the group consisting of 5-Z,
9-Z and combinations thereof than of a 13-Z isomer.
23. The method according to claim 22, wherein the primary
composition is used for a treatment selected from the group
consisting of improving hair or coat density, fiber diameter,
color, oilness, glossiness, and preventing hair or coat loss.
24. A method for preventing or treating cardiovascular disease, the
method comprising: administering a composition comprising a primary
composition having at least one carotenoid-containing material
extracted from a source selected from the group consisting of
tomatoes, carrots, peaches, apricots, oranges, melons, guavas,
papayas, grapefruits, wolfberries, rosehips, soya, green tea,
spices, grapes, cocoa and combinations thereof, and isomerized in
an organic solvent so as to comprise, by weight, a greater
percentage of an isomer selected from the group consisting of 5-Z,
9-Z and combinations thereof than of a 13-Z isomer.
Description
PRIORITY CLAIM
[0001] This application is a continuation of U.S. patent
application Ser. No. 12/376,662, filed on May 4, 2009, which is a
National Stage of International Application No. PCT/EP2007/006972,
filed Aug. 7, 2007, which claims priority to European Application
No. 06118579.9, filed Aug. 8, 2006, the entire contents of which
are being incorporated herein by reference.
FIELD OF TECHNOLOGY
[0002] The present invention relates to a primary composition that
includes at least one carotenoid-containing material enriched in
Z-isomers of the carotenoid compound having an increased stability
and bioavailability, and process of forming the same. It also
relates to an oral composition that contains the primary
composition in a foodstuff, in a food supplement, in a cosmetic
preparation or in a pharmaceutical preparation.
BACKGROUND
[0003] Absorption of carotenoids is a complex process involving
release from the food microstructure matrix, dissolution into mixed
micelles, intestinal uptake, incorporation into chylomicrons,
distribution to the tissues, uptake by liver and re-secretion into
VLDL, which are progressively transformed into LDL.
[0004] Lycopene absorption from food sources is widely documented.
Lycopene bioavailability is quite low from foods such as tomatoes
and tomato juice. Up to now, tomato paste is the best known food
source for bioavailable lycopene. Tomato contains about >90% of
lycopene in its all E configuration.
[0005] Tomato extracts containing a high amount of lycopene are
commercially available in the form of oleoresin but the
bioavailability of the carotenoid in humans is rather limited from
these sources. In concentrated tomato extracts, lycopene is mainly
present in crystalline form, which has been suggested to be one of
the primary factors that reduces its bioavailability.
[0006] To date, most commercially available lycopene sources
display an isomeric profile quite similar to the starting tomatoes
or show only a slight increase in Z-isomers, whether they are
derivatives (such as sauces) or extracts. A number of treatments,
as for instance thermal processing, are known to promote
isomerization. Shi et al., Journal of Food Process Engineering
2003, 25, 485-498, showed that an increase in Z isomers could be
obtained by heating tomato sauces. However certain lycopene isomers
are not stable and prone to retro-isomerization. According to the
literature, 5-Z is the most stable among the predominant lycopene
isomers followed by the all-E, the 9-Z and the 13-Z. Accordingly,
the stability of isomerized lycopene based products depends on
their lycopene isomer profile and thus can be modulated by
technological processing affecting this profile.
[0007] Thermal isomerization of lycopene is known to improve its
bioavailability from food matrices. However, the bioavailability of
individual lycopene isomers has not been investigated yet. As for
stability, it can be assumed that bioavailability of lycopene based
products is dependent on their lycopene isomer profile and thereby
can be modulated by technological means.
[0008] There are already patents that propose technological means
and formulations for improved bioavailability of carotenoids. For
example, WO 2005/075575 provides a primary composition enriched in
Z-isomers, effective to increase the bioavailability of the
carotenoid compound. However, there is still a need for a
carotenoid-containing product that has higher stability and thus
increased bioavailability.
SUMMARY
[0009] It has been found that the stability of individual
Z-lycopene isomers varies from one isomer to another; in particular
the 13-Z lycopene was much less stable than either the 5-Z, or the
9-Z, or the all-E isomers. Consequently, a primary composition
according to the present invention must have a level of 13-Z isomer
as low as possible to exhibit optimal stability. It has also been
shown that some Z isomers (such as 5-Z and 9-Z, for example) of
carotenoids enhance the bioavailability of the composition
containing such carotenoids. The primary composition must therefore
contain mainly the 5-Z isomer, or a combination of 9-Z and 5-Z
isomers to provide an improved bioavailability and bioefficacy.
[0010] Accordingly, it is a first object of the present invention
to provide primary compositions with at least one
carotenoid-containing material enriched in a specific mixture of Z
isomers of the carotenoid compound, the carotenoid-containing
material containing by weight a greater percentage of an isomer
selected from the group consisting of 5-Z, 9-Z and combinations
thereof than of 13-Z isomer.
[0011] In an embodiment, the present invention provides an oral
composition that contains the primary composition in a foodstuff,
in a food supplement, in a cosmetic preparation or in a
pharmaceutical preparation.
[0012] In an embodiment, the present invention provides the primary
composition as an additive in a foodstuff for oral administration,
such as in a nutritional composition, a food supplement, a pet food
product, a cosmetic preparation or a pharmaceutical
preparation.
[0013] In an embodiment, the present invention provides a method of
manufacturing the primary compositions or food supplements,
cosmetic preparations or pharmaceutical preparations containing the
same.
[0014] In another embodiment, the present invention provides the
use of the primary composition as described above, for the
preparation of an oral, cosmetic or pharmaceutical composition
intended for improving skin health, in particular for
photoprotection of the skin or for protecting skin tissue against
aging.
[0015] In an alternative embodiment, the present invention provides
the use of the primary compositions for the preparation of an oral,
cosmetic or pharmaceutical composition for preventing or treating
cardiovascular diseases or cancers.
[0016] An advantage of the present invention is to provide
compositions of Z isomers of carotenoids that exhibit a higher
stability, bioavailability and bioefficacy.
[0017] Additional features and advantages are described herein, and
will be apparent from, the following Detailed Description and the
Figure.
BRIEF DESCRIPTION OF THE DRAWING
[0018] FIG. 1 illustrates the area under the curve (AUC) of plasma
lycopene/triglycerides of TRL following the consumption of a
standard meal containing 25 mg total lycopene from either tomato
paste (all-E lycopene) or tomato oleoresin rich in 5-Z lycopene
(5-Z oleoresin) or tomato oleoresin rich in 13-Z lycopene (13-Z
oleoresin) or tomato oleoresin rich in 9 and 13-Z lycopene (9-&
13-Z oleoresin).
DETAILED DESCRIPTION
[0019] The present invention generally relates to compositions that
provide health benefits. More specifically, the present invention
relates to beneficial nutritional compositions that can be used to
improve skin and hair and methods regarding the same.
[0020] The present invention now makes available to the consumer an
improved composition obtained from natural products. The primary
composition provides carotenoids in a particularly highly
bioavailable and/or bioeffective form.
[0021] In a preferred embodiment, the invention provides tomato
extracts or derivatives thereof with an isomer ratio different from
the naturally occurring one in products to date available. In
particular, the invention relates to extracts or derivatives with
an E isomer content not higher than 60% on total lycopene content,
preferably with an E isomer content not higher than 40% on total
lycopene content (by HPLC).
[0022] In an embodiment, the present invention provides a primary
composition containing a specific combination of Z isomers.
Preferably, the ratio of Z/E isomers in the primary compositions of
the present invention should be above 1.
[0023] Moreover, the composition is preferably rich in 5-Z and 9-Z
and poor in 13-Z. In a preferred embodiment, the amount of 5-Z and
9-Z is greater than 30% on total carotenoid content, preferably
greater than 40%, most preferably greater than 50%. Also, the
amount of 13-Z is less than 10% on total carotenoid content,
preferably less than 5%, most preferably less than 3%. By
increasing the specific 5-Z and 9-Z isomers and/or decreasing the
13-Z isomers, for example, a stable form of the primary composition
that is more bioavailable and more bioeffective can be obtained.
Moreover, the extracts or derivatives of the invention are stable
under the usual storage conditions and do not undergo
retro-isomerization. Under ordinary protective conditions (absence
of light and oxygen), the lycopene total content and E isomer
content remains constant. The latter does not increase, even when
keeping the extracts at room temperature.
[0024] Such a profile (i.e. low amount of unstable isomer, such as
13-Z isomer of the carotenoid) may be for example obtained by
isomerizing the carotenoid using catalysis on a solid matrix such
as clays, or by prolonged heating.
[0025] In an embodiment, the carotenoid-containing material can be,
for example, in the form of an extract, a concentrate or an
oleoresin. In the present specification, the term "oleoresin"
should be understood to mean a lipid extract of a
carotenoid-containing material, which includes carotenoids,
triglycerides, phospholipids, tocopherols, tocotrienols,
phytosterols and other less significant compounds. It has been
surprisingly found that retro-isomerization of lycopene in
isomerized tomato oleoresin can be minimized by reducing its
content in 13-Z isomer.
[0026] In an embodiment, the carotenoid-containing material can be
an extract, a concentrate or an oleoresin, which is obtained,
extracted, enriched or purified from a plant or vegetable material,
a microorganism, a yeast or a product of animal origin. It is
further subjected to a treatment to increase its Z isomer content
of carotenoid, as described below.
[0027] If the source of carotenoid is from plant origin, it may be
vegetables, leaves, flowers, fruits and other parts of the plant.
In a preferred embodiment, the source of carotenoids is tomatoes
(i.e., whole tomato, tomato extract, tomato flesh, tomato puree,
tomato skin, with or without the seeds), carrots, peaches,
apricots, oranges, melons, guavas, papayas, grapefruit, wolfberry,
rosehips, soya, green tea, spices such as ginger or others, grapes
and/or cocoa. Suitable plant or vegetable concentrates are
obtainable e.g. by drying or freeze-drying the fresh-cut plants or
vegetables or the respective roots, fruits or seeds thereof and
then optionally grinding or granulating the dried material.
Suitable methods of obtaining extracts of the above-mentioned
plants or vegetables are known in the art. The plant or vegetable
extracts can be obtained, for example, by extracting the fresh-cut
or processed plants or vegetables or the respective roots, fruits
or seeds thereof with water or with one or more food grade solvents
or with a mixture of water and one or more food grade solvents.
Preferably, the extracts and concentrates according to the present
invention may be lipidic or aqueous. Because carotenoids are
liposoluble, extraction with water will remove unwanted
constituents that are water-soluble such as, for example, sugars,
amino acids, soluble proteins and/or organic acids.
[0028] If the carotenoid-containing material is obtained from
microorganism, any microorganism that produces carotenoid may be
used, in particular probiotic microorganism such as, for example,
lactic acid bacterium. Also, the product of animal origin may be
from, for example, salmon, shrimps, krill or a liver extract or a
milk fraction. In the present specification, the term "milk
fraction" should be understood to mean any part of the milk.
[0029] In an alternative embodiment, the carotenoid-containing
material can be an oleoresin. Suitable methods for obtaining
oleoresins from the above-mentioned plants or vegetables are well
known in the art. For example, oleoresins can be obtained by
lipidic extraction using a solvent compatible with the food
business, cosmetics or pharmaceuticals. For example, oleoresins
prepared by conventional methods have a content in carotenoid of
about 0.05% to 50% by weight. Their content of all-E isomer of
carotenoids is usually higher than that of Z-isomers, e.g. the
ratio of Z/E isomers of lycopene in a selected tomato oleoresin is
about 7:93.
[0030] Oleoresins are preferred starting material for obtaining the
primary composition according to the present invention because they
contain other carotenoids or antioxidants such as Vitamin E, which
also stabilize the composition. The bioactivity and stability of
the carotenoid compound in the oleoresin can be improved, in
particular, during the isomerization process and the yield of the Z
lycopene in the primary composition can also be increased.
[0031] The carotenoid-containing material preferably includes
carotenes and xanthophylls such as, for example, lycopene,
zeaxanthin, astaxanthine, .beta.-cryptoxanthin, capsanthine,
canthaxanthine, lutein and derivatives thereof such as esters, for
example. The carotenoid compounds have been subjected to a
treatment to increase the Z isomer fraction in the primary
composition.
[0032] In order to obtain a primary composition with such an isomer
profile, a carotenoid-containing material is subjected to a
treatment under conditions sufficient to increase its content in Z
isomers of the carotenoid compound, in particular the
carotenoid-containing material contains by weight a greater
percentage of an isomer selected from the group consisting of 5-Z,
9-Z and combinations thereof than of 13-Z isomer.
[0033] In an embodiment, the carotenoid-containing material which
is in the form of an extract, a concentrate or an oleoresin, is
subjected to an isomerization by using neutral, acidic or basic
solid catalysts (e.g. clays, zeolites, molecular sieves, ion
exchangers) to produce mixtures with high Z/E ratio. The use of
solid catalysts to enrich the carotenoid in Z-isomers is not
polluting and harmful to the food since the catalysts can be
conveniently removed by simple filtration or centrifugation. Also,
combinations of solid catalysts with other common means (e.g. heat,
light and radical initiators) can further enhance the geometrical
isomerization.
[0034] In another embodiment, the extracts or derivatives according
to the invention can be prepared starting from tomatoes, parts of
tomatoes (such as the skin), derivatives (such as sauces and
concentrates) or extracts. Isomerization is carried out by
prolonged heating in a solvent. In particular, when tomatoes or
derivatives thereof are used as starting materials, they can be
treated with a solvent able to extract lycopene. The resulting
extract is then heated; the solvent is removed, thus recovering the
isomerized extract.
[0035] On the other hand, when an extract or derivative is used as
starting material, this is taken up in a solvent, the mixture is
heated for a suitable time, then the solvent is removed, thus
recovering the isomerized extract. Solvents which can be used for
the isomerization step are hydrocarbons, chlorinated hydrocarbons,
esters, ketones, alcohols; particularly C3-C10 aliphatic
hydrocarbons, C1-C3 chlorinated solvents, C3-C6 esters, C3-C8
ketones and C1-C8 alcohols; more particularly hexane, carbon
tetrachloride, ethyl acetate, acetone and butanol. Isomerization in
solvents is carried out at temperatures ranging from 50 to
150.degree. C., preferably at temperatures ranging from 60 to
130.degree. C. Isomerization time ranges from 4 to 240 h,
preferably from 10 to 180 h.
[0036] The Z/E isomer ratio in the primary composition may then be
increased up to at least 20:80, preferably between 20:80 and 95:5,
more preferably from 30:70 to 90:10. In a preferred embodiment, the
(5Z+9Z)/E ratio is above 1, and the 13Z is partly removed.
[0037] In an embodiment, the present invention provides a primary
composition, in the form of a powder, liquid or gel, comprising a
carotenoid compound which has a better bioavailability and/or
bioefficacy than the compound alone. Also, the primary composition
may be in the form of a highly water-dispersible composition, if
the powder form is chosen. In this instance, the powder is
dispersible in water at ambient temperature. The primary
composition also provides carotenoids in a particularly highly
soluble form in lipids and organic solvents, less prone to
crystallisation, and having a lower tendency to aggregate.
[0038] In another embodiment of the present invention, the primary
composition may be used either alone or in association with other
active compounds such as vitamin C, vitamin E (tocopherols and
tocotrienols), carotenoids (carotenes, lycopene, lutein,
zeaxanthine, .beta.-cryptoxanthine, etc.) ubiquinones (e.g. CoQ10),
catechins (e.g. epigallocatechin gallate), coffee extracts
containing polyphenols and/or diterpenes (e.g. kawheol and
cafestol), extracts of chicory, ginkgo biloba extracts, grape or
grape seed extracts rich in proanthocyanidins, spice extracts (e.g.
rosemary), soy extracts containing isoflavones and related
phytoestrogens and other sources of flavonoids with antioxidant
activity, fatty acids (e.g. n-3 fatty acids), phytosterols,
prebiotic fibers, probiotic microorganisms, taurine, resveratrol,
aminoacids, selenium and precursors of gluthathione, or proteins
such as, for example, whey proteins.
[0039] The primary composition can additionally comprises one or
more of emulsifiers, stabilizers and other additives. Emulsifiers
compatible in the food field are, for example, phospholipids,
lecithin, polyoxyethylene sorbitan mono- or tristearate,
monolaurate, monopalmitate, mono- or trioleate; a mono- or
diglyceride. Any type of stabilizer that is known in the food
business, in cosmetics or in pharmaceuticals can be added. Also,
flavorings, colorants and any other suitable additives known in the
food business, in cosmetics or in pharmaceuticals can be added.
These emulsifiers, stabilizers and additives can be added according
to the final uses of the primary compositions.
[0040] In an alternative embodiment, the present invention provides
an oral composition comprising the primary composition described
above in a foodstuff, in a food supplement, in a pet food product,
in a cosmetic preparation or in a pharmaceutical preparation.
[0041] In a preferred embodiment, a food composition for human
consumption can be supplemented by the primary composition. This
food composition may be, for example, a nutritional complete
formula, a dairy product, a chilled or shelf stable beverage, a
mineral water, a liquid drink, a soup, a dietary supplement, a meal
replacement, a nutritional bar, a confectionery, a milk or a
fermented milk product, a yogurt, a milk based powder, an enteral
nutrition product, an infant formulae, an infant nutritional
product, a cereal product or a fermented cereal based product, an
ice-cream, a chocolate, coffee, a culinary product such as
mayonnaise, tomato puree or salad dressings or a pet food.
[0042] For use in food compositions, the primary composition can be
added to the above-mentioned foods or drinks so as to have a daily
intake between about 0.001 and 50 mg of carotenoid contained in the
primary composition, for example, such as lycopene. A daily intake
of about 5 to 20 mg per day is preferably envisaged.
[0043] The nutritional supplement for oral administration may be in
capsules, gelatin capsules, soft capsules, tablets, sugar-coated
tablets, pills, pastes or pastilles, gums, or drinkable solutions
or emulsions, syrups or gels, with a dose of about 0.001% to 100%
of the primary composition, which can then be taken directly with
water or by any other known means. This supplement may also include
a sweetener, a stabiliser, an additive, a flavoring or a colorant.
A supplement for cosmetic purposes can additionally comprise a
compound active with respect to the skin. It should be appreciated
that the supplements can be made by any methods known by those
skilled in the art.
[0044] In another embodiment, a pharmaceutical compositions
containing the primary compositions can be administered for
prophylactic and/or therapeutic treatments, in an amount sufficient
to cure or at least partially arrest the symptoms of the disease
and its complications. In the present specification, an amount
adequate to accomplish this is defined as "a therapeutically
effective dose." Amounts effective for this will depend on the
severity of the disease and the weight and general state of the
patient.
[0045] In prophylactic applications, primary compositions according
to the invention can be administered to a patient susceptible to or
otherwise at risk of a particular disease. Such an amount is
defined to be "a prophylactic effective dose." In this use, the
precise amounts again depend on the patient's state of health and
weight.
[0046] In an alternative embodiment, the primary compositions of
the invention can be administered with a pharmaceutical acceptable
carrier, the nature of the carrier differing with the mode of
administration, for example parenteral, intravenous, oral and
topical (including ophthalmic) routes. The desired formulation can
be made using a variety of excipients including, for example,
pharmaceutical grades of mannitol, lactose, starch, magnesium
stearate, sodium saccharin cellulose, magnesium carbonate. The
pharmaceutical compositions may be a tablet, a capsule, a pill, a
solution, a suspension, a syrup, a dried oral supplement, or a wet
oral supplement.
[0047] Preferably, for humans the pharmaceutical compositions
according to the present invention can comprise an amount of the
primary composition as described above, for a daily administration,
so that the carotenoid amount ranges from about 0.01 mg to 100 mg.
When administered daily to pets, the carotenoid amount can range
from about 0.01 mg to 100 mg.
[0048] It will be appreciated that the skilled person will, based
on his own knowledge, select the appropriate components and galenic
form to target the active compound to the tissue of interest, e.g.
the skin, colon, stomach, kidney or liver, taking into account the
route of administration which may be by way of injection, topical
application, intranasal administration, administration by implanted
or transdermal sustained release systems, and the like.
[0049] In another embodiment, the present invention provides a
cosmetic composition comprising the primary composition described
above. It may be formulated in lotions, shampoos, creams,
sun-screens, after-sun creams, anti-aging creams and/or ointments,
for example. Preferably, the content of primary composition can be
between 10.sup.-10% and 10% by weight of the cosmetic compositions.
More preferably, the cosmetic compositions comprise between
10.sup.-8% and 5% by weight of carotenoid compound. The cosmetic
compositions that can be used topically can additionally comprise a
fat or an oil which can be used in cosmetics such as, for example,
those mentioned in the CTFA work, Cosmetic Ingredients Handbook,
Washington.
[0050] The cosmetic compositions of the present invention can also
include any other suitable cosmetically active ingredients. The
composition additionally comprises a structuring agent and an
emulsifier. Other excipients, colorants, fragrances or opacifiers
can also be added to the cosmetic compositions. It will be
appreciated that the present cosmetic products will contain a
mixture of different ingredients known to the skilled person,
ensuring a fast penetration of the objective substance into the
skin and preventing degradation thereof during storage.
[0051] It should also be understood that the concepts of the
present invention may likewise be applied as an adjuvant therapy
assisting in presently used medications. Because the primary
compounds of the present invention may easily be administered
together with food material, special clinical food may be applied
containing a high amount of the primary compositions. It should be
clear that on reading the present specification together with the
appending claims the skilled person will envisage a variety of
different alternatives to the alternative embodiments mentioned
herein.
[0052] The present invention additionally relates to the use of the
primary composition, or the oral composition or the cosmetic
composition described above for the preparation of a product
intended to protect the tissues of the skin against aging, in
particular for inhibiting damage to the skin and/or mucous
membranes by inhibiting collagenases and enhancing the synthesis of
collagen. In fact, the use of the primary composition as described
above, for example, makes it possible to enhance the
bioavailability of the carotenoid compound in the body and to slow
down the aging of the skin. The primary compositions may also be
useful in the prevention or treatment of sensible, dry or reactive
skins, or for improving skin density or firmness, for ameliorating
skin photoprotection, for preventing or treating cardiovascular
diseases or disorders and cancers. They have also particular
benefits on hair and coat of pet animals, such as an improved hair
or coat density, fiber diameter, color, oilness, glossiness and a
help to prevent hair or coat loss.
[0053] The positive effects of the primary compositions of the
present invention on the skin of humans or pets can be measured by
using conventional methods such as, for example, minimal erythemal
dose (MED), colorimetry, transepidermal water loss, DNA repair,
measure of interleukins and proteoglycans production, or
collagenase activity, barrier function or cell renewal or
ultrasonic echography.
EXAMPLES
[0054] By way of example and not limitation, the following examples
are illustrative of various embodiments of the present invention
and further illustrate experimental testing conducted in accordance
with embodiments of the present invention.
Example 1
Study of the Stability of Lycopene Isomers
[0055] The stability of lycopene isomers was evaluated both in an
organic solvent and in a tomato extract.
[0056] Materials
[0057] Lycopene-rich tomato oleoresin has been obtained from Indena
s.p.a. (Milan, Italy). Its total lycopene content amounted to 9.1%,
of which the all-E and the 5-Z isomers represented 93.5% and 6.5%,
respectively. Two isomerized oleoresins were prepared by heating a
suspension of tomato oleoresin in ethyl acetate (1:10 w/w) either
for 1 h or for 48 h. After cooling at room temperature, the
suspensions were centrifuged and ethyl acetate in the recovered
supernatants was removed by distillation under reduced pressure.
Di-t-butyl-hydroxy-toluene (BHT) and N-ethyldiisopropylamine were
from Fluka AG. All solvents were HPLC grade and were used without
purification.
[0058] Isolation of Pure Lycopene Isomers
[0059] Pure 5-Z, 9-Z, 13-Z and all-E lycopene were isolated from
isomerised tomato oleoresin (submitted to 1 hour heating), by
collecting the fractions containing the corresponding peaks after
HPLC separation (see below the experimental conditions). Peaks were
collected during two consecutive HPLC runs and the corresponding
fractions were pooled.
[0060] Lycopene Analysis
[0061] Amount of total lycopene was determined by reverse phase
HPLC on a C.sub.18 precolumn (ODS Hypersil, 5 .mu.m, 20.times.4 mm;
Hewlett Packard, Geneva, Switzerland) and a C.sub.18 column (Nova
pak, 3.9 .mu.m internal diameter.times.300 mm length, Millipore,
Volketswil, Switzerland). The separation was achieved at room
temperature under isocratic conditions with a mobile phase
consisting of acetonitrile/tetrahydrofuran/methanol/ammonium
acetate 1% (533.5:193.6:53.7:28, wt/wt/wt/wt). The mobile phase
flow rate was 1.5 mL/min. Lycopene isomer profiles were determined
by normal phase HPLC according to the method described by Schierle
et al. (1997). Food. Chem. 59: 459. Samples of isomerized
oleoresins were dissolved in n-hexane containing 50 ppm BHT and
spun at maximum speed in an Eppendorf Lab centrifuge. The resulting
supernatants were immediately analyzed by HPLC. The HPLC system
used was a 1100 series Hewlett-Packard model equipped with an
ultraviolet-visible photodiode array detector. Data were
simultaneously acquired at 470 nm, 464 nm, 346 nm and 294 nm.
Samples (10 .mu.l) were separated using a combination of three
Nucleosil 300-5 columns (4 mm internal diameter.times.250 mm
length, Macherey-Nagel). The separation was achieved at room
temperature under isocratic condition with a mobile phase
consisting of n-hexane with 0.15% N-ethyldiisopropylamine. Flow
rate was 0.8 mL/min. Lycopene Z-isomers were identified according
to literature data.
[0062] Amounts of lycopene isomers were calculated based on surface
areas of the HPLC peaks using the same extinction coefficient as
the all-E lycopene. Therefore, the lycopene concentration in
products containing Z-isomers is slightly underestimated since it
is recognized that the extinction coefficients of Z-isomers are
lower than that of the all-E isomer.
[0063] Conditions for Stability Tests
[0064] Stability of lycopene isomers was investigated both in
n-hexane and in a tomato oleoresin isomerized by 4 hour heating in
ethyl acetate. For this purpose, pure lycopene isomers were stored
for 33 days in n-hexane at room temperature and in the absence of
light, and the isomerized tomato oleoresin was kept for 55 days at
room temperature in the absence of light. Total lycopene
concentration and lycopene isomer profiles were measured at various
time intervals during the storage.
[0065] Results
[0066] Stability of Lycopene Isomers in N-Hexane
[0067] Results of the stability test of pure lycopene isomers
during storage in n-hexane at room temperature in the absence of
light are reported Table 1. All isomers, i.e. included the all-E
isomer, underwent a geometrical isomerization during storage. The
13-Z was the less stable isomer: whereas less than 50% of 5-Z, 9-Z
and all-E lycopene were transformed after 33 day storage, more than
80% of 13-Z lycopene was converted into other isomers during this
period of time. Also, the transformation pathway was different for
the 13-Z lycopene compared to the other Z-isomers: while the 13-Z
isomer was mainly converted into the all-E isomer, the 5-Z and 9-Z
isomers were principally transformed into other Z-isomers during
storage in n-hexane.
TABLE-US-00001 TABLE 1 Stability of pure lycopene isomers in
n-hexane during storage at room temperature. time Concentration (%
of total isomers) (days) all E 13-Z 9-Z 5-Z x-Z all-E 0 97.6 1.4
0.5 0.5 0.1 lycopene 1 86.0 10.1 1.2 1.2 1.5 2 78.4 15.0 1.1 2.6
3.0 5 69.3 19.6 2.2 3.8 5.1 12 67.8 18.2 2.0 6.4 5.6 33 58.7 15.7
3.8 13.4 8.4 5-Z 0 1.1 n.d. n.d. 95.5 3.4 lycopene 1 2.2 n.d. n.d.
84.3 13.5 2 2.4 n.d. n.d. 76.9 20.6 5 3.9 n.d. n.d. 68.4 27.7 12
5.6 1.7 0.7 65.3 26.7 33 10.7 2.8 2.3 53.5 30.7 9-Z 0 4.4 0.6 93.3
1.8 0 lycopene 1 5.8 2.3 87.1 2.1 2.8 2 6.0 3.1 83.5 11.6 5.9 5 5.6
5.2 79.2 1.5 8.6 12 7.0 8.2 66.5 2.3 15.9 33 9.8 10.5 56.0 4.1 19.7
13-Z 0 2.4 96.6 0 0 1.0 lycopene 1 42.6 57.0 0.4 0 0 2 59.8 38.7 0
0 1.5 5 68.9 23.4 1.5 1.9 4.3 12 65.5 20.8 2.6 5.3 5.8 33 57.0 16.9
4.2 11.7 10.2
[0068] Stability of Lycopene Isomers in Tomato Oleoresin
[0069] Results of the stability test of lycopene isomers in a
tomato oleoresin heated for 48 hours in ethyl acetate are reported
in Table 2.
TABLE-US-00002 TABLE 2 Stability of lycopene isomers in isomerized
tomato oleoresin during storage at room temperature (n = 2).
Storage Total time lycopene (days) (mg/g) 13-Z % 9-Z % all-E % 5-Z
% 0 55.6 .+-. 3.0 17.4 .+-. 0.4 32.7 .+-. 0.7 18.7 .+-. 0.6 12.0
.+-. 0.4 3 56.6 .+-. 0.8 12.4 .+-. 0.1 31.4 .+-. 0.3 25.6 .+-. 0.5
13.0 .+-. 0.1 5 58.7 .+-. 0.4 9.5 .+-. 0.0 30.6 .+-. 0.5 30.6 .+-.
0.6 14.0 .+-. 0.4 7 59.2 .+-. 0.1 7.3 .+-. 0.2 30.9 .+-. 0.7 32.9
.+-. 0.3 14.4 .+-. 0.2 11 59.3 .+-. 0.8 5.2 .+-. 0.1 29.4 .+-. 0.0
36.5 .+-. 0.1 15.0 .+-. 0.1 17 60.1 .+-. 0.9 3.3 .+-. 0.4 29.0 .+-.
0.4 38.9 .+-. 0.4 15.4 .+-. 0.4 20 58.8 .+-. 0.9 2.9 .+-. 0.3 29.2
.+-. 0.4 39.9 .+-. 1.3 15.2 .+-. 0.6 34 51.3 .+-. 7.7 2.2 .+-. 0.1
30.0 .+-. 0.1 40.1 .+-. 0.7 14.1 .+-. 1.0 47 53.9 .+-. 1.2 1.9 .+-.
0.6 30.0 .+-. 0.3 41.4 .+-. 0.7 14.6 .+-. 1.3
[0070] Total lycopene content was stable during storage at room
temperature. However, the lycopene isomer profile markedly changed
with a decrease of 13-Z lycopene content and an increase of the
all-E lycopene. The content of 9-Z and 5-Z lycopene remained stable
during the storage period.
[0071] Conclusion
[0072] Both stability tests have shown that the 13-Z lycopene was
much less stable than either the 5-Z, or the 9-Z, or the all-E
isomers. Consequently, an isomerized tomato oleoresin with a low
level of 13-Z lycopene should exhibit a good stability of its
lycopene isomer profile.
Example 2
Isomerized Tomato Oleoresin with Increased Bioavailability
[0073] Objective:
[0074] The objective of the present work was to investigate the
bioavailability of various Z-lycopene isomers in humans. To
elucidate the bioavailability of specific Z-lycopene isomer in
human, tomato oleoresins have been enriched in different Z-lycopene
isomers reaching about 60% of the content of total lycopene i.e.
one rich in 5-Z lycopene, another one rich in 13-Z lycopene and the
last one rich in a mixture of 9-Z lycopene and 13-Z lycopene.
[0075] Material and Method
[0076] Subject
[0077] Thirty healthy men were enrolled in the study. The inclusion
criteria were that the subjects should be nonvegetarians and
nonsmokers and that they have no metabolic disorders such as
diabetes; hypertension; renal, hepatic, or pancreatic disease; or
ulcers. Subjects were normolipidemic, i.e. they had a ratio of
plasma cholesterol to HDL cholesterol <5.0 and plasma
triacylglycerol (TAG) concentrations <1.5 mmol/L. Because of the
large amount of blood that was drawn during the study, subjects
were required to have a blood hemoglobin concentration >13 g/dL.
Subjects were excluded from the study if they used
cholesterol-altering medication or hypolidemic treatment or vitamin
and mineral supplements from 3 months before the start of the study
until the completion of the study or had had major gastrointestinal
surgery; exercised intensively, such as running marathons; and
consumed daily >2 glasses of wine (3 dL), >2 beers (3 dL), or
>1 glass (shot glass) of hard liquor. Twenty-seven of the 30
volunteers completed the 4 postprandial tests. Three volunteers
abandoned the trial before the end for the following reasons:
unavailability, medical treatment related to an eye injury, nausea
related to the consumption of fatty meals. Subjects were 24.+-.1 y
old with a body weight of 70.+-.0.1 kg and body mass index (BMI) of
22.5.+-.0.3 kg/cm.sup.2.
[0078] The protocol was approved by the ethical committee of
Marseille (Marseille, France). Subjects received information on the
background and design of the study and gave written informed
consent before participation. They were free to withdraw from the
study at any time.
[0079] Study Design
[0080] This was a double-blind, randomized, 4-periods, 4-treatments
cross-over clinical trial with a washout period of 3 weeks minimum.
After an overnight fast, subjects arrived at the Clinical
Pharmacology and Therapeutic Trial Center of University of
Marseille and consumed a standard meal consisting of 25 mg lycopene
incorporated in 40 g peanut oil that was mixed with 70 g wheat
semolina (cooked with 200 mL tap water). In addition, they consumed
40 g bread, 60 g cooked egg whites, a 125 g yoghurt containing 5 g
of white sugar and drank 330 mL of water (Aquarel, Nestle). This
standard meal provided 842 kcal (3520 kJ) with the following
nutrient composition: protein (11.7%), carbohydrates (39.3%) and
lipids (49.0%). This meal was consumed within 15 min. No other food
was allowed over the subsequent 6 h, but subjects were allowed to
drink up to a bottled water (330 ml) during the last 3 h
post-absorption (Aquarel, Nestle).
[0081] Lycopene Supplements
[0082] Four different tomato products were tested providing each
one 25 mg of total lycopene. They consisted of:
[0083] Tomato paste (THOMY, Switzerland) containing lycopene mostly
in all-E configuration;
[0084] Tomato oleoresin enriched in 5-Z lycopene; and
[0085] Tomato oleoresin enriched with 13-Z lycopene Tomato
oleoresin enriched with a mixture of 9-Z and 13-Z lycopene.
[0086] Table 3 presents the lycopene content as well as the
lycopene isomer profile of these four tomato products.
TABLE-US-00003 All-E 5-Z 9-Z 13-Z X*-Z (% of total (% of total (%
of total (% of total (% of total lycopene) lycopene) lycopene)
lycopene) lycopene) Totato paste 94.9 4.1 nd 0.1 nd 5-Z 33.4 65.3
1.3 nd nd 13-Z 29.3 7.6 9.6 41.5 12.0 9- & 13-Z 27.7 7.7 30.8
23.5 10.2 *unidentified lycopene isomers is a pool of unknown
lycopene isomers calculated from the corresponding peak areas in
the HPLC chromatogram.
[0087] Collection of Blood Samples
[0088] Fasting blood was drawn from an anticubital vein by
venipuncture into an evacuated tube containing potassium
EDTA/K.sub.3 that was immediately placed in an ice-water bath and
covered with an aluminium foil to avoid light exposure. Fasting
blood samples were collected before i.e. 20 minutes and 5 minutes
before consumption of the standard meal as well as 2 h, 3 h, 4 h, 5
h, 6 h post-absorption. The tube containing the blood was protected
from light, stored at 4.degree. C. and then centrifuged within 2 h
(10 min, 4.degree. C., 2.800 rpm) to separate the plasma. A
cocktail of inhibitors (10 .mu.L/mL) was added (Cardin et al.,
Degradation of apolipoprotein B-100 of human plasma low density
lipoproteins by tissue and plasma kallikreins, Biol Chem 1984;
259:8522-8.).
[0089] Isolation of Plasma Triglyceride-Rich Lipoproteins (TRL)
[0090] After consumption of a fatty meal, dietary lipophilic
molecules are incorporated into chylomicrons, which are secreted
into blood. Lipoproteins are separated by ultracentrifugation
methodology based on their density. Due to the quite similar
density of chylomicrons (0.95 g/ml) and VLDL (1.006 g/ml), it is
not possible to separate one from the other and they are collected
altogether in a fraction called triglyceride-rich lipoproteins
(TRL). However, in the postprandial state, this plasma TRL fraction
contains mainly chylomicrons secreted from the intestine, which is
a good assessment of the intestinal bioavailability.
[0091] Triglyceride-rich lipoproteins (TRL) containing mainly
chylomicrons with little amount of VLDL were immediately isolated
by ultracentrifugation as follows: 6 mL of plasma were overlaid
with a 0.9% NaCl solution and ultracentrifuged for 28 min at 32,000
rpm, at 10.degree. C. in a SW41TI rotor (Beckman), in a L7
ultracentrifuge (Beckman). Immediately after centrifugation, the
TRL were aliquoted and stored at -80.degree. C. before analytical
determinations. Lycopene analyses were performed within 10 days,
and triacylglycerol analyses within 30 days.
[0092] Analytical Determination
[0093] Triglycerides were assayed by an enzymatic and colorimetric
method using a commercial kit (Kit Bio-Merieux). Total lycopene and
lycopene isomer profiles were determined by reverse phase and
normal phase HPLC method, respectively (M. Richelle, K. Bortlik, S.
Liardet, C. Hager, P. Lambelet, L. A. Applegate, E. A. Offord, J.
Nutr. (2002) 132, 404-408.). Total lycopene content was calculated
as the sum of the 5-Z, 9-Z, 13-Z, x-Z and all-E-lycopene isomers.
Lycopene isomer was quantified using the extinction coefficient of
all-E lycopene since the exact value for all individual Z-lycopene
is still unknown. Profile of lycopene isomers is determined by the
ratio of individual lycopene isomer to total lycopene expressed in
percentage.
[0094] Statistical Analysis
[0095] Lycopene bioavailability was assessed by measuring the area
under the lycopene concentration in TRL-time curve (AUC). This area
was calculated over the 0-6 hour period using the trapezoidal
method (AUC(0-6 h)). Data are presented as mean.+-.SEM. The
baseline concentration was the average of the concentrations
measured in the two plasma samples collected before consumption of
the standard meal containing 25 mg lycopene from the tomato matrix.
For each subject and each lycopene treatment, calculation of the
AUC(0-6 h) was performed by subtracting the baseline concentration
from the concentration value measured at each time point
post-absorption. If this value was negative, it was considered as
zero.
[0096] For each treatment, if the distribution of the
AUC.sub.(0-6h) was normal (Skewness and Kurtosis tests) with or
without logarithmic transformation, comparison was performed by
using a linear mixed model with treatment as fixed effect and
subject as random effect. All statistical analyses were done with
SAS software (version 8.2; SAS Institute, Cary, N.C.). The
rejection level in statistical tests was equal to 5%.
[0097] Results
[0098] Lycopene Bioavailability
[0099] Because the four tomato treatments induced a variation of
the extent in triglyceride secretion, lycopene bioavailability has
been normalized using triglyceride absorption (AUC.sub.(0-6h)).
Normalized lycopene bioavailability was markedly different between
the four tomato treatments (FIG. 1).
[0100] Surprisingly, lycopene was better bioavailable, by about two
times, from tomato oleoresin rich in 5-Z lycopene than from the
other three treatments, i.e tomato paste, tomato oleoresin rich in
13-Z lycopene as well as tomato oleoresin rich in a mixture of 13-Z
and 9-Z lycopene (p<0.0001) (FIG. 1). While lycopene was
similarly bioavailable from tomato paste as from the mixture of
13-Z and 9-Z tomato oleoresin. Lycopene present in 13-Z tomato
oleoresin exhibited a slight but significant lower bioavailability
(p<0.03) compared to tomato paste.
[0101] Conclusion
[0102] These results indicate that the configuration of the
lycopene molecule affects markedly the trafficking of lycopene
within the gastrointestinal tract and in consequence the amount of
lycopene that is absorbed. Lycopene bioavailability from tomato
extract rich in 5-Z lycopene is about double than that from tomato
paste. In contrast, lycopene present in tomato extract rich in a
mixture of 9-Z and 13-Z lycopene is similarly bioavailable to that
present in tomato paste while tomato oleoresin rich in 13-Z
lycopene presents a slightly less bioavailable lycopene. Several
authors have already pointed out that the presence of Z-lycopene in
a tomato product is associated with an increase of lycopene
bioavailability. This is the first study demonstrating that the
enhancement of lycopene bioavailability is specifically related to
lycopene configuration, i.e. 5-Z lycopene>9-Z lycopene>13-Z
lycopene.
Example 3
Extraction and Isomerization in Ethyl Acetate
[0103] 52 kg of fresh tomatoes containing 100 ppm of lycopene are
chopped and homogenized. Part of the water is distilled off under
reduced pressure to obtain 18 kg of tomato concentrate. This is
extracted with 36 l of water saturated ethyl acetate; during
extraction, the mixture is kept at room temperature shielded from
light and under stirring for 2 hours. The extract is then separated
from the tomato concentrate. The above described procedure is
repeated twice on such tomato concentrate, totally using 108 l of
solvent. The combined extracts are washed in a separate funnel with
27 l of water. The aqueous phase is then discarded while the
organic phase is concentrated under reduced pressure to obtain a
suspension with 10% w/v dry residue; the dry residue has a total
lycopene content of 9.1% w/w and a Z isomer content of 0.46% w/w.
This mixture is refluxed (76.degree. C.) under stirring for 7 days
before being concentrated to dryness under reduced pressure.
[0104] 46.8 g of final extract with a total lycopene content of 9%
w/w and a Z isomer content of 5.59% w/w are obtained; in
particular, the E isomer content is 3.41% w/w and the 13-Z isomer
content is 0.16% w/w.
Example 4
Extraction and Isomerization in Hexane
[0105] 10 kg of fresh tomatoes containing 140 ppm of lycopene are
chopped and homogenized. Part of the water is distilled off under
reduced pressure to obtain 2.5 kg of tomato concentrate, which is
extracted with 12.5 l of hexane. During extraction, the mixture is
kept at room temperature shielded from light and under stirring for
2 hours. The extract is then separated from the tomato concentrate.
The above described procedure is repeated once on such tomato
concentrate, totally using 25 l of solvent. The extracts are
combined and concentrated under reduced pressure to obtain a
solution with 10% w/v dry residue; the dry residue has a total
lycopene content of 9.1% w/w and a Z isomer content of 0.46% w/w.
This mixture is refluxed (69.degree. C.) under stirring for 6 days
before being concentrated to dryness under reduced pressure. 16.5 g
of final extract with total lycopene content of 9.1% w/w and Z
isomer content of 5.62% w/w are obtained; in particular, the E
isomer content is 3.38% w/w and the 13-Z isomer content is 0.18%
w/w.
Example 5
Isomerization in Butanol
[0106] 10 kg of fresh tomatoes containing 90 ppm of lycopene are
chopped and homogenized. Part of the water is distilled off under
reduced pressure to obtain 3.4 kg of tomato concentrate, which is
extracted with 7 l of water-saturated ethyl acetate. During
extraction, the mixture is kept at room temperature shielded from
light and under stirring for 2 hours. The extract is then separated
from the tomato concentrate. The above described procedure is
repeated twice on such tomato concentrate, totally using 21 l of
solvent. The combined extracts are washed in a separate funnel with
5.3 l of water. The aqueous phase is then discarded while the
organic phase is concentrated to dryness under reduced pressure.
The dry residue (9.8 g), which has a total lycopene content of 7.8%
w/w and a Z-isomer content of 0.40% w/w, is suspended in 98 ml of
n-butanol. The mixture is kept at 130.degree. C. under stirring for
4 hours before being concentrated to dryness under reduced
pressure. 9.8 g of final extract with a total lycopene content of
6.35% w/w and a Z-isomer content of 4.50% w/w are obtained; in
particular, the E-isomer content is 1.85% w/w and the 13-Z isomer
content is 0.47% w/w.
Example 6
Isomerization on Solid Catalysts
[0107] Materials
[0108] Lycopene-rich tomato oleoresin has been obtained from Indena
s.p.a. (Milan, Italy). Its total lycopene content amounted to 9.1%,
of which the all-E and the 5-Z isomers represented 93.5% and 6.5%,
respectively.
[0109] Methods
[0110] A suspension of tomato oleoresin in acetyl acetate (1:100
w/w) was filtered and incubated with 5% of solid catalyst under
constant stirring at room temperature for 2 h. The mixture was
centrifuged at maximum speed in an Eppendorf Lab centrifuge and an
aliquot of supernatant evaporated under N.sub.2 and re-suspended in
n-hexane/BHT.
[0111] Lycopene Analysis
[0112] Amount of total lycopene and lycopene isomer profiles were
determined by reverse phase and normal phase HPLC, respectively,
under the analytical conditions described in example 1.
[0113] Results
[0114] Lycopene isomer profiles measured in tomato oleoresin
isomerized for 2 h at room temperature using solid catalysts are
reported in Table 4.
TABLE-US-00004 TABLE 4 Lycopene isomer profiles in tomato
oleoresins isomerized using solid catalysts Isomer concentration (%
of total isomers) Catalyst All-E 13-Z 9-Z 5-Z x-Z* Control 83.3 3.0
0.9 8.6 4.1 Tonsil Optimum 31.3 7.0 13.4 23.8 24.5 Amberlyst 15
34.5 4.8 11.2 19.4 30.1 *unknown lycopene isomers
[0115] Lycopene was efficiently isomerized during 2 h reaction in
ethyl acetate at room temperature in the presence of either Tonsil
Optimum or Amberlyst 15. With both catalysts a large fraction of
lycopene all-E isomer was converted into Z-isomers. Among the
identified lycopene isomers, the 5-Z was formed in majority,
followed by the 9-Z and the 13-Z, respectively; thus, concentration
of the 13-Z isomer was, thus, below 10% in the isomerized tomato
oleoresins.
[0116] It should be understood that various changes and
modifications to the presently preferred embodiments described
herein will be apparent to those skilled in the art. Such changes
and modifications can be made without departing from the spirit and
scope of the present subject matter and without diminishing its
intended advantages. It is therefore intended that such changes and
modifications be covered by the appended claims.
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