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WO1999020587A1 - Procede d'extraction et de purification de la luteine, de la zeaxanthine et de carotenoides rares dans les fleurs et plantes de tagete - Google Patents

Procede d'extraction et de purification de la luteine, de la zeaxanthine et de carotenoides rares dans les fleurs et plantes de tagete Download PDF

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Publication number
WO1999020587A1
WO1999020587A1 PCT/US1998/022229 US9822229W WO9920587A1 WO 1999020587 A1 WO1999020587 A1 WO 1999020587A1 US 9822229 W US9822229 W US 9822229W WO 9920587 A1 WO9920587 A1 WO 9920587A1
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Prior art keywords
lutein
zeaxanthin
extraction
mixture
alcohol
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PCT/US1998/022229
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English (en)
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WO1999020587A8 (fr
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Frederick Khachik
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Office Of Technology Liaison
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Priority to US09/529,815 priority Critical patent/US6262284B1/en
Priority to AU12719/99A priority patent/AU1271999A/en
Publication of WO1999020587A1 publication Critical patent/WO1999020587A1/fr
Publication of WO1999020587A8 publication Critical patent/WO1999020587A8/fr

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    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B67/00—Influencing the physical, e.g. the dyeing or printing properties of dyestuffs without chemical reactions, e.g. by treating with solvents grinding or grinding assistants, coating of pigments or dyes; Process features in the making of dyestuff preparations; Dyestuff preparations of a special physical nature, e.g. tablets, films
    • C09B67/0096—Purification; Precipitation; Filtration
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/40—Colouring or decolouring of foods
    • A23L5/42—Addition of dyes or pigments, e.g. in combination with optical brighteners
    • A23L5/43—Addition of dyes or pigments, e.g. in combination with optical brighteners using naturally occurring organic dyes or pigments, their artificial duplicates or their derivatives
    • A23L5/44—Addition of dyes or pigments, e.g. in combination with optical brighteners using naturally occurring organic dyes or pigments, their artificial duplicates or their derivatives using carotenoids or xanthophylls
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C29/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring
    • C07C29/09—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring by hydrolysis
    • C07C29/095—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring by hydrolysis of esters of organic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C35/00—Compounds having at least one hydroxy or O-metal group bound to a carbon atom of a ring other than a six-membered aromatic ring
    • C07C35/21—Compounds having at least one hydroxy or O-metal group bound to a carbon atom of a ring other than a six-membered aromatic ring polycyclic, at least one hydroxy group bound to a non-condensed ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C403/00—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone
    • C07C403/24—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone having side-chains substituted by six-membered non-aromatic rings, e.g. beta-carotene
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B61/00—Dyes of natural origin prepared from natural sources, e.g. vegetable sources
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/09—Geometrical isomers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
    • C07C2601/16—Systems containing only non-condensed rings with a six-membered ring the ring being unsaturated

Definitions

  • Carotenoids are amongst the most widespread of the naturally occurring groups of pigments and are found in all families of the plant and animal kingdoms. To date, as many as seven hundred carotenoids have been isolated from various sources and their chemical structures have been characterized. Numerous epidemiological studies in various populations have shown that consumption of substantial amounts of fruits and vegetables rich in carotenoids reduces the risk of acquiring several types of cancers. As a result, for nearly two decades, scientists have been focussing most of their attention on investigating the protective effect of beta-carotene in prevention of cancer, cardiovascular and eye diseases. This is despite the fact that beta-carotene is only one of the prominent carotenoids found in fruits and vegetables whose consumption has been associated with health benefits in humans. The reasons for such an intense focus can be attributed to the pro- vitamin A activity of beta-carotene and the lack of commercial availability of other prominent food carotenoids.
  • lutein and zeaxanthin may be obtained from certain fruits and vegetables, the isolation of lutein from extracts of marigold flowers and zeaxanthin from berries ⁇ Lycium Chinese Mill (LCM berries) proves to be most economical.
  • Marigold flowers lutein is the major carotenoid and is normally accompanied by about 3-6% zeaxanthin; in LCM berries zeaxanthin is the major carotenoid and is completely free from lutein.
  • lutein and zeaxanthin are esterified with fatty acids such as lauric, myristic, and palmitic acids.
  • the purification of lutein fatty acid esters from marigold flowers was patented by Philip in 1977 (U.S. Patent No.
  • Marigold oleoresin is prepared by extraction of dried marigold petals in boiling n-hexane for extended periods of time. Since lutein/zeaxanthin, and carotenoids in general, are sensitive to heat treatment, this procedure can result in degradation or isomerization of these compounds. Furthermore, according to the Food and Drug administration's (FDA) Federal Register Documents, Code of Federal Regulations & Food, Drug, and Cosmetic act, n-hexane is considered among the solvents whose levels in foods and pharmaceutical products should be limited. The classification of organic solvents by the FDA will be described later in this text.
  • the hydrolysis of lutein and zeaxanthin esters in the marigold oleoresin is conducted in an aqueous solution in the presence of alcohol and propylene glycol in which the fatty acid esters of lutein and zeaxanthin have very low solubility.
  • this process requires high temperatures of up to 70°C and 10 hours to complete the saponification. This can once again result in the degradation and isomerization of lutein and zeaxanthin.
  • the saponified product Due to the high viscosity of propylene glycol, during handling and several purification steps, the saponified product is additionally subjected to high temperatures ranging from 70 to 85°C. This un-necessary exposure to heat in the presence of air can result in oxidative degradation of lutein and zeaxanthin and formation of a number of side products.
  • the process described here provides a convenient and economical route to lutein, zeaxanthin, and several minor carotenoids by employing a simultaneous extraction and saponification procedure at room temperature for only a few hours. Most importantly, this process addresses all the disadvantages and concerns with regard to all of the previously patented procedures described above. As a result, the lutein (from Marigolds) and zeaxanthin (from LCM berries) obtained by this process are in purity of 97% or greater and are therefore suitable for human consumption. This extraction and purification process has also been successfully employed for isolation of lutein and several minor carotenoids from green plants which serve as an alternative source for commercial production of lutein. BRIEF SUMMARY OF THE INVENTION
  • This invention employs a simultaneous extraction and hydrolysis (saponification) procedure for isolation and purification of lutein, zeaxanthin, and several rare carotenoids from Marigold flowers, LCM berries, and green plants.
  • the plants are mixed with tetrahydrofuran (THF) and sufficient amounts of ethanol or methanol (preferably food grade ethanol) with the addition of either 5% or 10% potassium or sodium hydroxide to maintain the pH at 12 and the mixture is homogenized at room temperature for 1 to 2 hours.
  • THF tetrahydrofuran
  • ethanol or methanol preferably food grade ethanol
  • This process not only extracts carotenoids from plants under mild conditions, within a few hours, but it simultaneously hydrolyses the fatty acid esters of major dihydroxycarotenoids such as lutein and zeaxanthin as well as monohydroxycarotenoids such as alpha- cryptoxanthin and beta-cryptoxanthin which are present in minute quantities. As a result, the free and un-esterified forms of these hydroxycarotenoids are regenerated. In the case of green plants, in which lutein is present in its free form, the simultaneous extraction and saponification results in the conversion of chlorophylls to their water soluble derivatives and allows the removal of these compounds from lutein, beta-carotene, and other carotenoids.
  • This treatment removes the salts of fatty acid esters, the base (potassium or sodium hydroxide), and the water soluble plant components.
  • the resulting crystals of lutein or zeaxanthin are approximately 70% pure and contain minor quantities of other carotenoids.
  • lutein or zeaxanthin are dissolved in minimum amount of THF, and water is added until the solution becomes cloudy; at this point the recrystalliztion of these compounds takes place.
  • the mixture is then centrifuged and the solid crystals are washed with water or alcohol to remove the THF. After drying under high vacuum, the crystalline lutein or zeaxanthin are obtained in purities of 97% or greater.
  • 70% lutein or zeaxanthin may be purified by passing a solution of these compounds in THF and water through a column packed with n- silica gel. After collection of the fraction containing lutein or zeaxanthin (greater than 97% purity), the THF is evaporated and the residue is washed with water or alcohol and dried as described above.
  • alpha-cryptoxanthin and beta-cryptoxanthin Rosigolds and LCM berries
  • lutein epoxide from greens
  • all-E and Z isomers of carotenoids refer to all-trans and cis isomers of these compounds.
  • all-trans and cis which have been used with the old nomenclature, are no longer appropriate.
  • lutein or zeaxanthin refer to the most common geometrical forms (all-E or all-trans) of these carotenoids in plants.
  • lutein esters or "zeaxanthin esters” refers to either mono- or di- esters without limitation.
  • the present invention applies a simultaneous extraction and saponification process to three generally different plants sources for isolation of lutein and zeaxanthin; these are: dried petals of Marigolds (Tagete erecta), LCM berries (Lycium Chinese Mill), and several green vegetables such as Kale (Brassica oleracia, var. acephala), Spinach (Spinacia oleracia), and Collard green (Brassica oleracia, var. Champion). These plant sources of carotenoids are described as follows.
  • Marigold flowers The dried petals of marigold flowers, are harvested and prepared in Central America and are imported into the U.S. Flowers are hand-picked and ensiled to preserve them until they can be economically dried.
  • the marigold blossoms are then placed in a freeze drying apparatus and are dehydrated under controlled conditions. After dehydration, the flowers are put through a series of air separators and mechanical separators where the petals are separated from any other materials and converted to a homogenous Marigold meal.
  • the carotenoids in Marigold meal are esterified with fatty acids such as lauric, myristic, and palmitic acid, however, upon hydrolysis the parent hydroxycarotenoids are regenerated.
  • the major hydroxycarotenoid in Marigold is lutein which is normally accompanied by 3-6% of its isomeric compound, zeaxanthin.
  • LCM berries are normally grown in China and can be obtained from most of the Chinese supermarkets across the U.S. However, a variety of this fruit is also currently grown on a commercial scale by Rehnborg Center for Nutrition in Lakeview, California.
  • the author isolated several grams of zeaxanthin from LCM berries for a human supplementation study and demonstrated that this plant is an excellent source of zeaxanthin (Khachik et al., J. Cellular Biochem. 1995, 22:236-246).
  • Zeaxanthin in LCM berries is mostly esterified with palmitic acid and only trace amounts of this compound is esterified with lauric and myristic acids.
  • LCM berries do not contain any measurable amount of lutein but several other carotenoids such as alpha-cryptoxanthin, beta- cryptoxanthin, and beta-carotene are present in this fruit in minute quantities.
  • carotenoids such as alpha-cryptoxanthin, beta- cryptoxanthin, and beta-carotene are present in this fruit in minute quantities.
  • large quantities of the berries were purchased from a local Chinese supermarket.
  • the draft guideline recommends acceptable amounts of residual solvents in pharmaceuticals for the safety of the patient as well as recommending the use of less toxic solvents in the manufacture of drug substances and dosage forms.
  • solvents are divided into three classes. These are:
  • Class 1 Solvents to be avoided. Known human carcinogens, strongly suspected human carcinogens, and environmental hazards.
  • Class 2 Solvents to be limited. Nongenotoxic animal carcinogens or possible causative agents of other irreversible toxicity such as neurotoxicity or teratogenicity; solvents suspected of other significant but reversible toxi cities.
  • Class 3 Solvents with low toxic potential to man. No health based exposure limit is needed. Class 3 solvents have Permitted Daily Exposure (PDE) of 50 milligrams (mg) or more per day.
  • PDE Permitted Daily Exposure
  • Ethanol and THF employed by the present invention are listed by the FDA in Class 3 and are therefore quite safe for commercial production of carotenoids for human use.
  • the advantage of THF in compare to other organic solvents of Class 3 is due to the strong solubility of carotenoids in this solvent which allows the extraction of these pigments from the matrices of various plants in an efficient and speedy manner.
  • THF is water soluble and therefore can be used in a homogenous phase extraction of carotenoids from plants in which significant amount of water may be present (Kale, Spinach, Collard green).
  • the solubility strength of THF for carotenoids and the homogeneity of THF, ethanol, and water allow for the simultaneous extraction and saponification of carotenoids from plants.
  • Marigold meal 100 g
  • tetrahydrofuran 1000 ml
  • sodium- or potassium hydroxide 25 g
  • food grade ethanol 250 ml
  • Preferred solvents are also selected for their boiling points. Solvents with boiling points of about 75°C to 120°C are preferred. Ethanol, with a
  • the pH of the mixture is monitored and automatically maintained at about pH 12.
  • An additional amount of sodium- or potassium hydroxide (25 g) in 250 ml of ethanol is normally needed to maintain the pH at 12.
  • the extraction and saponification is completed in about 2 hours.
  • the solvents are then evaporated and the residue is washed with 1000 ml of a 1/1 mixture water and alcohol to remove the base and the water soluble components.
  • the lutein crystals are collected by centrifugation, washed with alcohol (100 ml), and dried under high vacuum overnight at room temperature.
  • the lutein crystals (2.0 g) obtained are about 70% pure and can be recrystallized from a mixture of THF (20 ml) and water (30 ml) and dried under high vacuum overnight to give 1.2 g of lutein containing about 5% zeaxanthin in 97% purity.
  • the above 70% pure lutein can also be purified by passing a solution of this compound in THF/water (1.5/1.0) through a column of n-silica gel and subsequent of washing of the column with this solvent mixture. After collection of the lutein fraction, the THF is evaporated, the crystals of pure lutein are collected by centrifugation, and washed with 20 ml of an alcohol, preferably ethanol, to remove the residual water from the product. Drying under high vacuum gives 1.2 g of 97% pure lutein containing about 5% zeaxanthin. In another fraction from this purification, about 0.3 g of a mixture of lutein, anhydrolutein, alpha- cryptoxanthin, and beta-cryptoxanthin is also obtained.
  • Marigold meal 100 g was vigorously homogenized in a blender with tetrahydrofuran (THF, 1000 ml) and 10% ethanolic KOH (250 ml) at room temperature.
  • the pH of the solution was kept at 12 by monitoring the extraction and hydrolysis with an automatic pH-meter.
  • an additional 250 ml of 10% ethanolic KOH was needed to maintain the pH at 12. It is essential to avoid the addition of excessive amounts of the base to the mixture during the extraction and hydrolysis. This is because at a later purification step, the removal of the excess base will then require additional and un-necessary washing of the residue with large volumes of water which can be time consuming.
  • the lutein crystals (2.0 g, 70% pure) were dissolved in THF (20 ml) and water (30 ml) was added to commence recrystallization. The mixture was stirred at room temperature for 10 minutes and the solids were separated by centrifugation. After removal of the solution, the lutein crystals were washed with ethanol (20 ml), centrifuged, and dried under high vacuum overnight at room temperature. This gave 1.2 g of lutein which was shown by HPLC-UV/Visible photodiode array detection coupled with Mass Spectrometry (MS) to be 97% pure but contained about 4% of all-E-zeaxanthin (Khachik et al. J. Chrom. Biomed. Appl. 582: 153-166, 1992).
  • MS Mass Spectrometry
  • the 70% pure lutein (2.0 g) was dissolved in THF (20 ml) and was passed through a glass column of n-Silica gel (15 cm L X 4 cm I.D.) packed under slight air pressure using THF/water (1.5/1.0). The column was washed with 200 ml of THF/water (1.5/1.0) and fractions 1 (50 ml) and 2 (150 ml) were collected. After evaporation of THF under reduced pressure at 40°C, and removal of water by centrifugation, fractions 1 and 2 were washed with 4 ml and 20 ml of ethanol, respectively, and dried under high vacuum at room temperature overnight.
  • the first fraction (0.3 g) was shown by HPLC-UV/Vis-MS to be a mixture of anhydrolutein, alpha-cryptoxanthin, and beta-cryptoxanthin and the second fraction (1.2 g) was shown to be pure all-E-lutein (97%) but contained approximately 4% zeaxanthin.
  • LCM berries (455.5 g) were rehydrated in a beaker containing 1000 ml of water overnight at room temperature. The water was decanted off and the LCM berries were vigorously homogenized with THF (2000 ml) and 10% ethanolic KOH (250 ml) at room temperature. The pH of the solution was maintained at 12 by monitoring the extraction and hydrolysis with an automatic pH-meter. During the course of the extraction an additional amount of 250 ml of 10% ethanolic KOH was needed to keep the pH at 12.
  • the zeaxanthin crystals (0.48 g, 75% pure) were dissolved in THF (5 ml) and water (about 10 ml) was added to commence recrystallization. The mixture was stirred at room temperature for about 10 minutes and the solids were separated by centrifugation. After removal of the solution, the zeaxanthin crystals were washed with ethanol (about 5 ml), centrifuged, and dried under high vacuum for about 8 hours at room temperature. This gave 0.35 g of zeaxanthin which was shown by HPLC-UV/Visible photodiode array detection coupled with Mass Spectrometry (MS) to be 97% pure but contained about 3.5% of Z-zeaxanthins (Khachik et al. J. Chrom. Biomed. Appl. 582: 153-166, 1992).
  • MS Mass Spectrometry
  • the THF/water solution from the above recrystallization was evaporated to remove the THF and to precipitate small amount of zeaxanthin along with other carotenoids (0.1 g) which were separated by centrifugation.
  • the mixture of carotenoids were washed with small amount of ethanol (1 ml) and dried under high vacuum. HPLC analysis of the dried solid showed that in addition to zeaxanthin, this fraction was enriched in alpha-cryptoxanthin, beta-cryptoxanthin, and beta- carotene.
  • TLC Thin Layer Chromatography
  • HPLC High Performance Liquid Chromatography
  • chlorophylls were completely hydrolyzed to their water soluble derivatives.
  • the mixture was filtered off and the solids were washed with THF (1000 ml).
  • the solvents were evaporated under reduced pressure at approximately 40°C to almost dryness and the solids were stirred at room temperature with a 1/1 mixture of water and food grade ethanol (100 ml) for 10 minutes.
  • the present invention has accomplished the isolation of lutein and zeaxanthin as well as a mixture of several rare carotenoids from plants in substantially pure form under controlled and mild condition to preserve the integrity of these essential dietary compounds.
  • This economically viable process employs solvents which are not toxic and as a result, the carotenoids purified by this procedure can be safely used as nutritional supplements or food coloring additives.

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  • Chemical Kinetics & Catalysis (AREA)
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  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

L'invention concerne un procédé qui permet, simultanément, d'extraire, de saponifier et d'isoler la lutéine et la zéaxanthine, ainsi qu'une combinaison de plusieurs caroténoïdes rares, avec une pureté élevée, dans des plantes, sans utiliser de solvants organiques nocifs. On a recueilli des cristaux de lutéine renfermant 5 % de zéaxanthine dans des pétales séchés de fleurs de tagète (Tagete erecta), la zéaxanthine étant isolée et purifiée à partir de baies de Lycium chinese Mill (LCM). On a utilisé le même procédé pour isoler et purifier un mélange de lutéine, de bêta-carotène, de néoxanthine, de violaxanthine et d'époxyde de lutéine à partir de végétaux verts (de préférence, chou fourrager, chou rosette et épinards). Dans une moindre mesure, ces végétaux ont également été la source de plusieurs caroténoïdes rares comme l'alpha-cryptoxanthine (tagète) et la bêta-cryptoxanthine (baies de LCM). Le procédé fait appel à une combinaison de tétrahydrofurane (THF), de solution alcoolique de potassium ou d'hydroxyde de sodium, à température ambiante et en conditions douces, pour déclencher quantitativement dans un délai de deux heures l'hydrolyse en formes libres des esters d'acides gras de la lutéine et de la zéaxanthine dans les fleurs de tagète et les baies de LCM. S'agissant des végétaux verts, qui contiennent la lutéine sous forme libre, l'opération simultanée d'extraction et d'hydrolyse permet d'éliminer rapidement les chlorophylles indésirables. Après filtration, on laisse évaporer le THF et l'alcool, et on soumet le résidu à un lavage séquentiel à l'eau et à l'alcool de manière à éliminer le THF, la base, et dans le cas de végétaux verts, les chlorophylles et leurs dérivés. Les cristaux de lutéine et de zéaxanthine résultants, issus de leurs sources végétales respectives, sont purs à environ 70 %, et on peut les purifier plus avant, soit par recristallisation, soit par injection de leur mise en solution de THF/eau dans une colonne à gel de silice N. On peut obtenir comme sous-produits des dernières étapes de purification mentionnées plusieurs caroténoïdes rares comme l'anhydrolutéine et l'alpha-cryptoxanthine (à partir des tagètes) et la bêta-cryptoxanthine (à partir de baies de LCM). Les caroténoïdes purifiés isolés selon le procédé décrit sont dépourvus d'impuretés et constituent une source de suppléments nutritifs sans risques pour la consommation humaine, fournissant par ailleurs des colorants pour les aliments destinés à la consommation humaine.
PCT/US1998/022229 1997-10-21 1998-10-21 Procede d'extraction et de purification de la luteine, de la zeaxanthine et de carotenoides rares dans les fleurs et plantes de tagete WO1999020587A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US09/529,815 US6262284B1 (en) 1998-10-21 1998-10-21 Process for extraction and purification of lutein, zeaxanthin and rare carotenoids from marigold flowers and plants
AU12719/99A AU1271999A (en) 1997-10-21 1998-10-21 Process for extraction and purification of lutein, zeaxanthin and rare carotenoids from marigold flowers and plants

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US6257297P 1997-10-21 1997-10-21
US60/062,572 1997-10-21
US7836098P 1998-03-18 1998-03-18
US60/078,360 1998-03-18

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WO1999020587A1 true WO1999020587A1 (fr) 1999-04-29
WO1999020587A8 WO1999020587A8 (fr) 1999-08-26

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Cited By (35)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6191293B1 (en) 1998-04-20 2001-02-20 Inexa, Industria Extractora C.A. Trans-xanthophyll ester concentrates of enhanced purity and methods of making same
WO2001055100A1 (fr) * 2000-01-27 2001-08-02 Dsm N.V. Isolation de cristaux de carotenoide
US6329557B1 (en) 2000-06-09 2001-12-11 Prodemex, S.A. De C.V. Purification of xanthophylls from marigold extracts that contain high levels of chlorophylls
WO2002004415A3 (fr) * 2000-07-12 2002-05-10 Univ Maryland Procede de purification et de cristallisation de carotenoides d'huile palmitique
WO2002060864A1 (fr) * 2001-01-30 2002-08-08 Sabinsa Corporation Procede permettant d'obtenir des compositions de luteine stable et des derives de luteine
EP1361281A1 (fr) * 2001-01-26 2003-11-12 Fuji Chemical Industry Co., Ltd. Procede de purification de xanthophylles brutes
EP1371641A1 (fr) * 2002-06-10 2003-12-17 Adisseo France S.A.S. Procédé de préparation et isolement de cristaux caroténoides
WO2004018417A1 (fr) * 2002-08-26 2004-03-04 Kancor Flavours And Extracts Ltd Procede ameliore pour la preparation de cristaux de xanthophylle
EP1408737A2 (fr) * 2001-06-29 2004-04-21 Ball Horticultural Company Oeillets d'inde tagetes erecta renfermant des compositions a base de carotenoides et des teneurs en carotenoides modifiees
US6737535B2 (en) 2002-06-05 2004-05-18 Kancor Flavours And Extracts Limited Trans-lutein enriched xanthophyll ester concentrate and a process for its preparation
US6818798B1 (en) 2000-05-03 2004-11-16 University Of Maryland, College Park Process for making a (3R,3′R)-zeaxanthin precursor
EP1531665A2 (fr) * 2002-06-26 2005-05-25 Ball Horticultural Company Soucis tagetes erecta a compositions carotenoides et teneurs en carotenoides modifiees
US6911564B2 (en) 2000-07-27 2005-06-28 University Of Maryland, College Park Method for production of rare carotenoids from commercially available lutein
US7115786B2 (en) 2002-02-06 2006-10-03 University Of Maryland, College Park Method for production of β-cryptoxanthin and α-cryptoxanthin from commercially available lutein
US7173145B2 (en) 2001-11-29 2007-02-06 University Of Maryland, College Park Process for extraction and purification of lutein, zeaxanthin and rare carotenoids from marigold flowers and plants
WO2007087815A2 (fr) 2004-12-17 2007-08-09 Metanomics Gmbh Procédé de contrôle de production de produits chimiques fins
EP1857441A1 (fr) * 2006-05-17 2007-11-21 ZheJiang Medicine Co., Ltd. Xinchang Pharmaceutical Factory Procédé de séparation et de purification d'esters d'acide gras de xanthophyll provenant d'oléorésine de soucis
US7351424B2 (en) 2004-07-22 2008-04-01 Bio Lut S.A. De C.V. Enhanced purity trans-lutein-ester compositions and methods of making same
CN101402597A (zh) * 2008-10-31 2009-04-08 南京泛成生物化工有限公司 叶黄素提取工艺
CN102174005A (zh) * 2011-01-25 2011-09-07 石河子大学医学院第一附属医院 一种从万寿菊干花中提取叶黄素的方法
JP4848420B2 (ja) * 2005-04-25 2011-12-28 カトラ フィトケム（インディア） プライヴェイト リミテッド マリーゴールド由来カロテノイドの単離及び精製
CN102558008A (zh) * 2011-09-08 2012-07-11 黑龙江省嘉宝生物技术开发有限公司 一种叶黄素的制备方法
CN103091446A (zh) * 2013-01-31 2013-05-08 甘肃省农垦农业研究院 叶黄素酯皂化程度快速测定方法
CN103275088A (zh) * 2013-05-30 2013-09-04 湖南炎帝生物工程有限公司 从念珠藻中分离叶绿素a与类胡萝卜素的方法
CN103992256A (zh) * 2014-05-17 2014-08-20 昆明涞章医药科技有限公司 一种从金盏菊中制备高纯度叶黄素酯的方法
TWI466709B (zh) * 2011-07-20 2015-01-01 Nat Univ Chung Hsing From the original berry Wolfberry a liquid - liquid freezing method for separation of high purity theaflavins palmitate Maize
CN104447467A (zh) * 2014-10-30 2015-03-25 蒋小春 一种提取枸杞中类胡萝卜素的方法
WO2015109657A1 (fr) * 2014-01-27 2015-07-30 中国科学院大连化学物理研究所 Procédé de fabrication de zéaxanthine naturelle au moyen d'une concentration in situ
CN105017116A (zh) * 2015-07-31 2015-11-04 江西恩泉油脂有限公司 一种叶黄素浸膏的皂化工艺
CN105503681A (zh) * 2014-09-25 2016-04-20 赵军 高纯度枸杞玉米黄质棕榈酸酯的制备方法
CN107082756A (zh) * 2017-03-17 2017-08-22 晨光生物科技集团股份有限公司 一种叶黄素晶体瞬时高效的工业制备方法
CN110563625A (zh) * 2019-09-29 2019-12-13 山东天音生物科技有限公司 一种由万寿菊油树脂分离纯化玉米黄质的方法
CN110835315A (zh) * 2019-10-28 2020-02-25 百瑞源枸杞股份有限公司 一种枸杞红素的制备方法
KR102161387B1 (ko) * 2019-12-09 2020-09-29 대한켐텍 주식회사 루테인 포함 피부건강용 조성물 및 이를 포함하는 피부건강기능식품
WO2021019275A1 (fr) * 2019-07-26 2021-02-04 Universita' Degli Studi Di Cagliari Isolement de caroténoïdes chromoplastidials à partir de fruits

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US4048203A (en) * 1976-12-06 1977-09-13 Thomas Philip Purification of lutein-fatty acid esters from plant materials
US5382714A (en) * 1994-03-17 1995-01-17 The Catholic University Of America Process for isolation, purification, and recrystallization of lutein from saponified marigold oleoresin and uses thereof
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US6191293B1 (en) 1998-04-20 2001-02-20 Inexa, Industria Extractora C.A. Trans-xanthophyll ester concentrates of enhanced purity and methods of making same
WO2001055100A1 (fr) * 2000-01-27 2001-08-02 Dsm N.V. Isolation de cristaux de carotenoide
US7015014B2 (en) 2000-01-27 2006-03-21 Dsm Ip Assets B.V. Isolation of carotenoid crystals
US6818798B1 (en) 2000-05-03 2004-11-16 University Of Maryland, College Park Process for making a (3R,3′R)-zeaxanthin precursor
US6329557B1 (en) 2000-06-09 2001-12-11 Prodemex, S.A. De C.V. Purification of xanthophylls from marigold extracts that contain high levels of chlorophylls
WO2002004415A3 (fr) * 2000-07-12 2002-05-10 Univ Maryland Procede de purification et de cristallisation de carotenoides d'huile palmitique
US7119238B2 (en) 2000-07-12 2006-10-10 University Of Maryland Process for purification and crystallization of palm oil carotenoids
US6911564B2 (en) 2000-07-27 2005-06-28 University Of Maryland, College Park Method for production of rare carotenoids from commercially available lutein
EP1361281A1 (fr) * 2001-01-26 2003-11-12 Fuji Chemical Industry Co., Ltd. Procede de purification de xanthophylles brutes
EP1361281A4 (fr) * 2001-01-26 2004-02-04 Fuji Chem Ind Co Ltd Procede de purification de xanthophylles brutes
WO2002060864A1 (fr) * 2001-01-30 2002-08-08 Sabinsa Corporation Procede permettant d'obtenir des compositions de luteine stable et des derives de luteine
EP1408737A2 (fr) * 2001-06-29 2004-04-21 Ball Horticultural Company Oeillets d'inde tagetes erecta renfermant des compositions a base de carotenoides et des teneurs en carotenoides modifiees
EP1408737A4 (fr) * 2001-06-29 2004-12-22 Ball Horticultural Co Oeillets d'inde tagetes erecta renfermant des compositions a base de carotenoides et des teneurs en carotenoides modifiees
US7173145B2 (en) 2001-11-29 2007-02-06 University Of Maryland, College Park Process for extraction and purification of lutein, zeaxanthin and rare carotenoids from marigold flowers and plants
US7115786B2 (en) 2002-02-06 2006-10-03 University Of Maryland, College Park Method for production of β-cryptoxanthin and α-cryptoxanthin from commercially available lutein
US6737535B2 (en) 2002-06-05 2004-05-18 Kancor Flavours And Extracts Limited Trans-lutein enriched xanthophyll ester concentrate and a process for its preparation
WO2003104190A1 (fr) * 2002-06-10 2003-12-18 Adisseo France S.A.S. Procede de formation et d'isolation de carotenoides cristallises
EP1371641A1 (fr) * 2002-06-10 2003-12-17 Adisseo France S.A.S. Procédé de préparation et isolement de cristaux caroténoides
AU2003232424B2 (en) * 2002-06-26 2008-04-24 Ball Horticultural Company Tagetes erecta marigolds with altered carotenoid compositions and ratios
EP1531665A2 (fr) * 2002-06-26 2005-05-25 Ball Horticultural Company Soucis tagetes erecta a compositions carotenoides et teneurs en carotenoides modifiees
EP1531665A4 (fr) * 2002-06-26 2006-03-22 Ball Horticultural Co Soucis tagetes erecta a compositions carotenoides et teneurs en carotenoides modifiees
AU2002347590B2 (en) * 2002-08-26 2008-01-03 Omniactive Health Technologies Limited An improved process for the preparation of xanthophyll crystals
US6743953B2 (en) 2002-08-26 2004-06-01 Kancor Flavours & Extracts Ltd. Process for the preparation of xanthophyll crystals
AU2002347590B8 (en) * 2002-08-26 2004-03-11 Omniactive Health Technologies Limited An improved process for the preparation of xanthophyll crystals
WO2004018417A1 (fr) * 2002-08-26 2004-03-04 Kancor Flavours And Extracts Ltd Procede ameliore pour la preparation de cristaux de xanthophylle
EP2292598A3 (fr) * 2002-08-26 2012-04-25 Omniactive Health Technologies Limited Procédé amélioré pour la préparation de crystaux de xanthophyll
US7351424B2 (en) 2004-07-22 2008-04-01 Bio Lut S.A. De C.V. Enhanced purity trans-lutein-ester compositions and methods of making same
WO2007087815A2 (fr) 2004-12-17 2007-08-09 Metanomics Gmbh Procédé de contrôle de production de produits chimiques fins
JP4848420B2 (ja) * 2005-04-25 2011-12-28 カトラ フィトケム（インディア） プライヴェイト リミテッド マリーゴールド由来カロテノイドの単離及び精製
EP1857441A1 (fr) * 2006-05-17 2007-11-21 ZheJiang Medicine Co., Ltd. Xinchang Pharmaceutical Factory Procédé de séparation et de purification d'esters d'acide gras de xanthophyll provenant d'oléorésine de soucis
CN101402597A (zh) * 2008-10-31 2009-04-08 南京泛成生物化工有限公司 叶黄素提取工艺
CN102174005A (zh) * 2011-01-25 2011-09-07 石河子大学医学院第一附属医院 一种从万寿菊干花中提取叶黄素的方法
TWI466709B (zh) * 2011-07-20 2015-01-01 Nat Univ Chung Hsing From the original berry Wolfberry a liquid - liquid freezing method for separation of high purity theaflavins palmitate Maize
CN102558008A (zh) * 2011-09-08 2012-07-11 黑龙江省嘉宝生物技术开发有限公司 一种叶黄素的制备方法
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WO2015109657A1 (fr) * 2014-01-27 2015-07-30 中国科学院大连化学物理研究所 Procédé de fabrication de zéaxanthine naturelle au moyen d'une concentration in situ
CN103992256A (zh) * 2014-05-17 2014-08-20 昆明涞章医药科技有限公司 一种从金盏菊中制备高纯度叶黄素酯的方法
CN103992256B (zh) * 2014-05-17 2016-09-07 昆明涞章医药科技有限公司 一种从金盏菊中制备高纯度叶黄素酯的方法
CN105503681A (zh) * 2014-09-25 2016-04-20 赵军 高纯度枸杞玉米黄质棕榈酸酯的制备方法
CN104447467A (zh) * 2014-10-30 2015-03-25 蒋小春 一种提取枸杞中类胡萝卜素的方法
CN105017116A (zh) * 2015-07-31 2015-11-04 江西恩泉油脂有限公司 一种叶黄素浸膏的皂化工艺
CN107082756A (zh) * 2017-03-17 2017-08-22 晨光生物科技集团股份有限公司 一种叶黄素晶体瞬时高效的工业制备方法
CN107082756B (zh) * 2017-03-17 2018-11-23 晨光生物科技集团股份有限公司 一种叶黄素晶体的工业制备方法
WO2021019275A1 (fr) * 2019-07-26 2021-02-04 Universita' Degli Studi Di Cagliari Isolement de caroténoïdes chromoplastidials à partir de fruits
CN114401775A (zh) * 2019-07-26 2022-04-26 卡利亚里大学 从果实中分离有色体类胡萝卜素
CN114401775B (zh) * 2019-07-26 2023-11-07 卡利亚里大学 从果实中分离有色体类胡萝卜素
US12162827B2 (en) 2019-07-26 2024-12-10 Universita' Degli Studi Di Cagliari Isolation of chromoplastid carotenoids from fruits
CN110563625A (zh) * 2019-09-29 2019-12-13 山东天音生物科技有限公司 一种由万寿菊油树脂分离纯化玉米黄质的方法
CN110835315A (zh) * 2019-10-28 2020-02-25 百瑞源枸杞股份有限公司 一种枸杞红素的制备方法
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