Title of Invention

A METHJOD FOR THE PRODUCTION OF A LUTEIN-FATTY ACID ESTER CONCENTRATE.

Abstract THE PRESENT INVENTIOON RELATES TO A METHOD FOR THE PRODUCTION OF A LUITEIN FATTY ACID ESTER CONCENTRATE OF HIGH PURITY BY DISSOLVING MATRIGOLD OLEORESIN IN A DETONE SOLVENGT SUCHAS ACETONE, COOLING GHE SOLUTION, AND REMOVING WHEREFROM A KETONE SOLVENT INSOLUBLE COMPONENT TO UNIFROM THE QUALITYU OPFR OLEORESWIN, FOLLOWED BY DISSOLVING THE ACETONE SOLUBLE PORTION OBTAINED AS ABOVE IN BUTANOL, COOLING TGHE SOLUTION, REMOVING A BUTANOL SOLUBLE IMPURITY AFTER ADDING OR WITHOUT ADDING ON3 OR MORE MEMBERWS SELECTEE FROM THE GROU8P CONSISTING OF WATER METHANOL, ETHANOL AND A MIXTURE THEREOF , AND WASHING THE RESULTING BUTANOL INSOLUBLE WITH ETHANOL.
Full Text A METHOD FOR THE PRODUCTION OF A LUTEIN-FATTY ACID ESTER
CONCENTRATE
BACKGROUND OF THE INVENTION
1. FIELD OF THE INVENTION
The present invention relates to a method for the
production of a lutein-fatty acid ester concentrate, more
particularly a method of obtaining a lutein-fatty acid ester
concentrate of high purity by purifying marigold oleoresin.
2. DESCRIPTION OF THE PRIOR ART
Lutein-fatty acid esters are a yellow-orange pigment
contained in fruits such as oranges, peaches, papayas, prunes
and mangos. Lutein-fatty acid esters are also present in many
flowers and vegetables, particularly in petals of marigold
flowers remarkably. Dried and ground marigold flowers have
been used as animal feeds for a long time, and oleoresin which
is obtained by extraction of marigold flowers with a solvent
has also been widely used as food color in recent years in
addition to the above-mentioned application.
The recent scientific researches have reported that
lutein is associated with risk reduction for age-related
macular degeneration (AMD) caused by oxidative damage to
macular area of retina (Journal of the American Medical
Association, 272, 1413-1420, 1994: Investigative
Ophthalmology & Visual Science, 38, 1802-1811, 1997, etc.), and
that lutein is effective for prevention of arteriosclerosis.
prevention of cataract or suppression of carcinogenesis and etc.
(British Medical Journal, 305, 335-339, 1992: The American
Journal of Clinical Nutrition, 70, 517-524, 1992: American
Journal of Epidemiology, 149, 801-809, 1999, etc). As such the
use of lutein as a health food and a dietary supplement is also
expected.
Marigold oleoresin is obtained when dried and ground
marigold flowers are extracted with a hydrocarbon solvent such
as hexane and petroleum ether or with a chlorinated hydrocarbon
solvent such as dichloromethane, and the solvent is removed from
the extract. But the content of lutein-fatty acid ester as an
ester contained in oleoresins produced by the largest producers
of marigold extracts is low, usually 14 to 20 % (cf. U.S. Patent
No. 6,191,293) , and is about 30 to 35 % even in a superior grade
type which is partly sold for food.
Thus , there has been a demand for a lutein-fatty acid ester
concentrate of high purity in order to obtain more vivid color
tones when it is used as a food color and in order to improve
the functional efficiency when it is used as a dietary
supplement.
As to a method of obtaining a lutein-fatty acid ester
concentrate of high purity, "Purification of Lutein-Fatty Acid
Esters From Plant Materials" (U.S. Patent No. 4,048,203) and
"Trans-xanthophyll Ester Concentrates of Enhanced Purity and
Methods of Making Same" (U.S. Patent No. 6,191,293) have been
disclosed.
The former describes a method which comprises
precipitating lutein-fatty acid esters from oleoresin which is
dissolved in hot alkanol containing not more than four carbon
atoms by cooling the solution, removing alkanol wherefrom by
filtration and drying the lutein-fatty acid ester under vacuum.
However, the purity of the lutein-fatty acid ester concentrate
obtained by this method is 51%, which cannot be considered a
high purity.
The latter describes a method of obtaining a purified
trans-xanthophyll ester by contacting coronals (or flowers) of
marigold flowers with a hydrocarbon solvent, removing wherefrom
plant residue after sufficient extraction, evaporating the
extract to obtain oleoresin, admixing the resulting oleoresin
with an alcohol at ambient temperature and removing the alcohol
containing impurity and cis-xanthophyll ester from the mixture.
By this method, however, the purity of lutein-fatty acid
ester in the resulting oleoresin varies widely depending on the
quality of the raw material, i.e. coronals (or flowers) of
marigold flowers, and therefore, the purity of lutein-fatty
acid ester in a concentrate obtained by purifying the said
oleoresin also varies widely. For this reason, control of the
raw material is necessary to obtain a lutein-fatty acid ester
concentrate of high purity by this method, and as such it is
difficult to stably obtain a lutein-fatty acid ester
concentrate of high purity when most of the commercially
available oleoresins are used as a starting material.
SUMMARY OF THE INVENTION
An object of the present invention is to provide a
production method by which a lutein-fatty acid ester
concentrate of high purity can be stably obtained even when
oleoresin containing lutein-fatty acid esters of low purity is
used as a starting material, without being affected by variation
of the quality, particularly the content of lutein-fatty acid
esters, of marigold oleoresin.
The present inventors have found that, as a result of
diligent studies to solve the above-mentioned problems, a
lutein-fatty acid ester concentrate of high purity is stably
obtained by dissolving oleoresins in a ketone solvent such as
acetone, cooling the solution, and removing wherefrom an
insoluble component in a ketone solvent such as phospholipid
to uniform the quality of oleoresins, dissolving a ketone
solvent-soluble portion obtained as above in butanol, cooling
the solution, removing a butanol-soluble impurity after adding
or without adding one or more members selected from the group
consisting of water, methanol and ethanol to the solution, and
washing the resulting butanol-insoluble with ethanol. The
present inventors have accomplished the present invention after
further studies based on the finding.
Thus, the present invention is directed to the following
(1) to (9).
(1) A method for the production of a lutein-fatty acid ester
concentrate of high purity from marigold oleoresin, which
comprises:
(a) a step of dissolving marigold oleoresin in a ketone
solvent, cooling the solution and removing wherefrom a ketone
solvent-insoluble impurity,
(b) a step of dissolving a ketone solvent-soluble portion,
obtained by removing a ketone solvent from the ketone solvent
solution, in butanol, cooling the solution and eliminating a
butanol-soluble impurity, and
(c) a step of washing a butanol-insoluble, which is obtained
after the above cooling process, with ethanol to eliminate an
ethanol-soluble impurity, followed by removing a residual
solvent to obtain a lutein-fatty acid ester concentrate of high
purity.
(2) A method for the production of a lutein-fatty acid ester
concentrate of high purity from marigold oleoresin, which
comprises:
(a) a step of dissolving marigold oleoresin in a ketone
solvent, cooling the solution and removing wherefrom a ketone
solvent-insoluble impurity,
(b) a step of dissolving a ketone solvent-soluble portion,
which is obtained by removing a ketone solvent from the ketone
solvent solution, in butanol, cooling the solution, adding
thereto one or more solvents selected from the group consisting
of water, methanol, ethanol and a mixture thereof, and then
eliminating a butanol-soluble impurity, and
(c) a step of washing a butanol-insoluble, which is obtained
upon addition of the said solvent(s) after cooling, with ethanol
to eliminate an ethanol-soluble impurity, followed by removing
a residual solvent to obtain a lutein-fatty acid ester
concentrate of high purity.
(3) A method for the production of a lutein-fatty acid ester
concentrate of high purity, which comprises dissolving marigold
oleoresin in a ketone solvent, cooling the solution and removing
wherefrom a ketone solvent-insoluble impurity.
(4) The method for the production of a lutein-fatty acid ester
concentrate of high purity according to any one of the above
(1) to (3), wherein the ketone solvent is acetone.
(5) A method for the production of a lutein-fatty acid ester
concentrate of high purity, which comprises washing a
butanol-insoluble, which is precipitated from a butanol
solution containing lutein-fatty acid ester obtained by
processing marigold oleoresin, with ethanol.
(6) The method for the production of a lutein-fatty acid ester
concentrate of high purity according to any one of the above
(1) to (5), wherein butanol is n-butanol.
(7) A composition comprising not less than 70wt% of
lutein-fatty acid esters.
(8) A health food or a dietary supplement comprising not less
than 70wt% of lutein-fatty acid esters.
(9) A method of making a health food or a dietary supplement
which comprises incorporating a composition containing not less
than 70wt% of lutein-fatty acid esters into a health food or
a dietary supplement.
DETAILED DESCRIPTION OF THE INVENTION
Marigold oleoresin used in the present invention is a
solid or a viscous liquid having yellow-orange/yellowish-brown
color, which is obtained by drying flowers of marigold which
is a member of the Compositae family (Tagetes erecta WILLD.),
grinding the dried product, optionally converting into pellet,
extracting with an organic solvent, usually hexane, and
removing the solvent from the extract. Its main component is
lutein (one of xanthophylls) dipalmitate.
According to the method of the present invention,
marigold oleoresin (also called oleoresin for short) is
dissolved in a ketone solvent. A ketone solvent used here
includes a ketone solvent containing 3 to 9 carbon atoms such
as acetone, methylethylketone and diethylketone, among which
acetone is preferable. The amount of acetone is about 0.5 to
10 parts by weight, preferably about 2 to 3 parts by weight,
relative to 1 part by weight of marigold oleoresin which is the
raw material.
A mixture of oleoresin and acetone is stirred for about
0.5 to 1 hour at about 40 to 55 °C, preferably at about 45 to
50 °C while keeping moderate refluxing. Next the mixture is
slowly cooled to about 10 to 30 °C, preferably about 15 to 25
°C over a period of usually about 2 to 4 hours. The cooled
mixture is filtered through filter paper or filter fabric with
filter aid, if necessary, such as diatomite. The filtrate is
evaporated under reduced pressure and acetone is removed
wherefrom to obtain a solid or a viscous liquid consisting of
acetone-soluble portion in oleoresin.
According to the method of the present invention,
acetone-soluble portion in oleoresin is dissolved in butanol.
Examples of butanol used here are n-butanol, i-butanol.
sec-butanol and tert-butanol. among which n-butanol is
preferable. The amount of butanol is about 1 to 5 parts by
weight, preferably about 2 to 3 parts by weight relative to 1
part by weight of the said soluble portion.
The mixed solution of the said soluble portion and butanol
is stirred for about 0. 5 to 1 hour at about 40 to 80 °C, preferably
at about 45 to 55 °C. Next the mixture is slowly cooled to about
0 to 20 °C, preferably about 3 to 10 °C over a period of usually
about 2 to 5 hours. The cooled mixture is filtered through
filter paper or filter fabric to recover a butanol-insoluble
solid, and the filtrate containing a butanol-soluble impurity
is discarded.
According to the method of the present invention, one or
more solvents selected from the group consisting of water,
methanol, ethanol and a mixture thereof is added, preferably
while stirring, to the above cooled mixture. By adding these
solvent(s), an insoluble solid is precipitated rapidly, by
which operating efficiency can be improved. The amount of water
to be added here is about 0.01 to 1 part by weight, preferably
about 0.05 to 0.5 part by weight relative to 1 part by weight
of the butanol used; the amount of methanol is about 0.01 to
5 parts by weight, preferably about 0.1 to 1 part by weight
relative to 1 part by weight of the butanol used; and the amount
of ethanol is about 0.01 to 10 parts by weight, preferably about
0.5 to 5 parts by weight relative to 1 part by weight of the
butanol used. Although the temperature of solvent(s) to be
added here may be ambient temperature, the same temperature as
the cooled mixture, if possible, is preferable. The mixture
consisting of mixed solvents is filtered through filter paper
or filter fabric to recover a solid which is insoluble in the
mixed solvent, and the filtrate containing a impurity which is
soluble in the mixed solvent is discarded.
According to the method of the present invention, the
butanol-insoluble or the mixed solvent-insoluble wherein
butanol is a main component is washed with ethanol. The amount
of ethanol is about 5 to 80 parts by weight, preferably about
10 to 40 parts by weight relative to 1 part by weight of the
said insoluble. By washing with ethanol, butanol or the mixed
solvent consisting mainly of butanol which is adhered to the
surface of the said insoluble is eliminated, and consequently
the removal of solvent in finishing process becomes easier.
A mixture of the said insoluble and ethanol is stirred
for about 0.5 to 2 hour(s), preferably about 1 to 1.5 hour at
ambient temperature. Next the mixture is filtered through
filter paper or filter fabric to recover a lutein-fatty acid
ester concentrate, and the filtrate containing an
ethanol-soluble impurity is discarded.
Finally, the residual solvent is removed from the
purified lutein-fatty acid ester concentrate by evaporation
using, for example, a vacuum shelf dryer at a temperature of,
for instance, lower than 50 °C preferably in the atmosphere of
nitrogen gas.
The purity of lutein-fatty acid ester contained in the
lutein-fatty acid ester concentrate which is obtained by the
method according to the present invention is about not less than
70%, and that with about more than 80% purity is frequently
obtained. As a result of this, the utilization range as a food
color is broadened due to the vivid color tone, and it becomes
possible to blend a lutein fatty-acid ester concentrate having
high concentration into a dietary supplement by
micro-encapsulation.
WORKING EXAMPLE
The following examples specifically illustrate the
present invention.
A METHOD OF MESURING THE CONTENT OF LUTEIN-FATTY ACID ESTER
About 0.lg of test sample was accurately measured and
dissolved in hexane to make a 100ml solution. It was diluted
with hexane so that the resultant solution had absorbance
ranging from 0.3 to 0.7, and then the maximum absorbance of the
diluted solution around 445nm was measured by a
spectrophotometer. The content of lutein-fatty acid ester was
calculated by the following formula.
Content(%)=
((Absorbance ÷ Weight of Sample)x Dilution Ratio)÷ 1,394 × 100
Note: 1,394 is absorbance coefficient at 445nm
EXAMPLE 1
107g of marigold oleoresin containing 26wt% of
lutein-fatty acid ester was mixed with 214ml of acetone and
dissolved at 50°C under heating. Next the solution was slowly
cooled to 20°C over a period of about 3 hours and the resulting
insoluble was eliminated by filtration, and then the filtrate
was evaporated under reduced pressure to obtain a concentrate
of the acetone-soluble portion. 300g of n-butanol was added
to this concentrate(l00g) , and then the solution was dissolved
at 45°C under heating and slowly cooled to 4°C over a period
of about 4.5 hours.
After further keeping it at 4°C for 16 hours, the resulting
insoluble was filtered through filter paper. The solid which
remained was mixed with 800ml of ethanol, and the mixture was
stirred, dispersed, washed at ambient temperature, and then
filtered again through filter paper. The resulting solid was
dried under vacuum to remove residual solvent, whereby 8.3g of
a concentrate containing 89wt% of lutein-fatty acid ester was
obtained.
COMPARATIVE EXAMPLE 1
107g of marigold oleoresin as used in Example 1 was mixed
with 214ml of acetone and dissolved at 50°C under heating. The
solution was slowly cooled to 20°C over a period of about 3 hours
and the resulting insoluble was eliminated by filtration, and
then the filtrate was evaporated under reduced pressure to
obtain a concentrate of acetone-soluble portion.
Next, according to the method mentioned in Example 1 of
USP 6,191,298, this concentrate( l00g) was stirred for 3 hours
with 200g of isopropanol at ambient temperature to obtain a
suspension. The suspension was filtered through filter paper
and the solid left on the filter paper was dried under vacuum
at ambient temperature to eliminate residual solvent, whereby
34.6g of a solid containing 44wt% of lutein-fatty acid ester
was obtained.
COMPARATIVE EXAMPLE 2
According to the method mentioned in Example 1 of USP
6,191,298, l00g of marigold oleoresin as used in Example 1 was
stirred for 3 hours with 200g of isopropanol at ambient
temperature to obtain a paste-like suspension. The suspension
was filtered through filter paper and the solid left on the
filter paper was dried under vacuum at ambient temperature to
eliminate residual solvent, whereby 35.6g of a concentrate
containing 37wt% of lutein-fatty acid ester was obtained.
EXAMPLE 2
106g of marigold oleoresin containing 22wt% of
lutein-fatty acid ester was mixed with 212ml of acetone and
dissolved at 50°C under heating. Next the solution was slowly
cooled to 20°C over a period of about 3 hours and the resulting
insoluble was eliminated by filtration, and then the filtrate
was evaporated under reduced pressure to obtain a concentrate
of acetone-soluble portion. 400g of n-butanol was added to this
concentrate (l00g), dissolved at 45°C under heating, and slowly
cooled to 4°C over a period of about 4.5 hours.
26.3g of water was added thereto and the resulting
insoluble was filtered through filter paper. The solid
obtained as above was mixed with 800ml of ethanol for washing
by stirring and dispersion at ambient temperature, and then
filtered again through filter paper. The resulting solid was
dried under vacuum to remove residual solvent, whereby 15.9g
of a concentrate containing 82wt% of lutein-fatty acid ester
was obtained.
COMPARATIVE EXAMPLE 3 (No acetone treatment)
300g of n-butanol was added to l00g of marigold oleoresin
as used in Example 2, dissolved at 45°C under heating, and slowly
cooled to 4°C over about 4.5 hours. 32.6g of water was added
thereto and the resulting insoluble was filtered through filter
paper. The solid obtained as above was mixed with 800ml of
ethanol for washing by stirring and dispersion at ambient
temperature, and then filtered again through filter paper. The
resulting solid was dried under vacuum to remove residual
solvent, whereby 36.4g of a concentrate containing 46wt% of
lutein-fatty acid ester was obtained.
COMPARATIVE EXAMPLE 4 (No butanol treatment)
106g of marigold oleoresin as used in Example 2 was mixed
with 212ml of acetone and dissolved at 50°C under heating. Next
the solution was slowly cooled to 20°C over about 3 hours and
the resulting insoluble was eliminated by filtration, and then
the filtrate was evaporated under reduced pressure to obtain
a concentrate of acetone-soluble portion. This concentrate
(l00g) was mixed with 2,010 ml of ethanol for washing by stirring
and dispersion at ambient temperature, and filtered again
through filter paper. The resulting solid was dried under
vacuum to remove the residual solvent, whereby 35.2g of a
concentrate containing 52wt% of lutein-fatty acid ester was
obtained.
EXAMPLE 3
214g of methanol was used in place of 26.3g of water in
Example 2, and 18.7g of a concentrate containing 73wt% of
lutein-fatty acid ester was obtained.
EXAMPLE 4
l,250g of ethanol was used in place of 26. 3g of water in
Example 2, and 18.4g of a concentrate containing 77wt% of
lutein-fatty acid ester was obtained.
INDUSTRIAL APPLICABILITY
The method of the present invention provides a
lutein-fatty acid ester concentrate of high purity, and as a
result of this, the utilization range as a food color is
broadened due to the vivid color tone of a lutein-fatty acid
ester concentrate, and it becomes possible to blend a
lutein-fatty acid ester concentrate having high concentration
into a dietary supplement by micro-encapsulation.
WE CLAIM:
1. A method for the production of a iutein-fatty acid ester
concentrate having purity of not less than 70 wt.% from
marigold oleoresin, which comprises ;
(a) a step of dissolving marigold oleoresin in a ketone solvent,
cooling the solution and removing wherefrom a ketone
solvent-insoluble impurity,
(b) a step of dissolving a ketone solvent-soluble portion,
obtained by removing a ketone solvent from the ketone
solvent solution, in butanol, cooling the solution and
eliminating a butanol-oluble impurity, and
(c) a step of washing a butanol-insoluble, which is obtained
after the above cooling process, with ethanol to eliminate an
ethanol-soluble impurity, followed by removing a residual
solvent to obtain a Iutein-fatty acid ester concentrate of high
purity.
2. A method for the production of a lutein-fatty acid ester
concentrate of high purity from marigold oleoresin, which
comprises:
(a) a step of dissolving marigold oieoresin in a ketone solvent,
cooling the solution and removing wherefrom a ketone
solvent-insoluble impurity,
(b) a step of dissolving a ketone solvent-soluble portion, which
is obtained by removing a ketone solvent from the ketone
solvent solution, in butanol, cooling the solution, and adding
thereto one or more solvents selected from the group
consisting of water, methanol, ethanol and a mixture thereof,
and then eliminating a butanol-solubte impurity, and
(c) a step of washing a butanol-insoluble, which is obtained
upon addition of the said solvent(s) after cooling, with
ethanol to eliminate an ethanol-soluble impurity, followed by
removing a residual solvent to obtain a lutein-fatty acid ester
concentrate of high purity.
3. A method for the production of a lutein-fatty acid ester
concentrate of high purity, which comprises dissolving
marigold oieoresin a ketone solvent, cooling the solution and
removing wherefrom a ketone solvent-insoluble impurity.
4. The method for the production of a lutein-fatty acid ester
concentrate of high purity as claimed in any one of claims 1 to
3, wherein the ketone solvent is acetone.
5. A method for the production of a lutein-fatty acid ester
concentrate of high purity, which comprises washing a
butanol-insoluble, which is precipitated from a butanol
solution containing lutein-fatty acid ester obtained by
processing marigold oleoresin, with ethanol.
6. The method for the production of a lutein-fatty acid ester
concentrate of high purity as claimed in any one of claims 1
to 5, wherein butanol is n-butanol.
7. A composition comprising lutein-fatty acid ester concentrate
containing not less than 70 wt.% of lutein-fatty acid ester.
8. A health food or a dietary supplement comprising lutein-fatty
acid ester concentrate containing not less than 70 wt.% of
lutein-fatty acid ester.
9. A method of making a health food or a dietary supplement
which comprises incorporating a composition containing
lutein-fatty acid ester concentrate containing not less than 70
wt.% of lutein-fatty acid ester into a health food or a dietary
supplement.
The present invention relates to a method for the
production of a lutein-fatty acid ester concentrate of high
purity by dissolving marigold oleoresin in a ketone solvent such
as acetone, cooling the solution, and removing wherefrom a
ketone solvent-insoluble component to uniform the quality of
oleoresin, followed by dissolving the acetone-soluble portion
obtained as above in butanol, cooling the solution, removing
a butanol-soluble impurity after adding or without adding one
or more members selected from the group consisting of water,
methanol, ethanol and a mixture thereof, and washing the
resulting butanol-insoluble with ethanol.

Documents:

00654-cal-2002-abstract.pdf

00654-cal-2002-claims.pdf

00654-cal-2002-correspondence.pdf

00654-cal-2002-description (complete).pdf

00654-cal-2002-form 1.pdf

00654-cal-2002-form 18.pdf

00654-cal-2002-form 2.pdf

00654-cal-2002-form 26.pdf

00654-cal-2002-form 3.pdf

00654-cal-2002-form 5.pdf

00654-cal-2002-letter patent.pdf

00654-cal-2002-priority document others.pdf

00654-cal-2002-priority document.pdf

00654-cal-2002-reply f.e.r.pdf


Patent Number 211007
Indian Patent Application Number 00654/CAL/2002
PG Journal Number 42/2007
Publication Date 19-Oct-2007
Grant Date 16-Oct-2007
Date of Filing 25-Nov-2002
Name of Patentee RIKEN VITAMIN CO., LTD.
Applicant Address TOKYYO 101-8370 JAPAN
Inventors:
# Inventor's Name Inventor's Address
1 HARADA KOICHI C/O. ROLEM VITAMIN CO. LTD. KYOTO 621-0007 JAPAN.
2 SDADANO SHIN C/O RIKEN VITAMIN CO. LTD 102, DIGASHIKAKIUCHI JAPAN.
PCT International Classification Number C07C 403/00
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 NA