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ES2345529B1 - PROCEDURE FOR THE PURIFICATION OF TRIGLICERIDES CONTAINING GAMMA-LINOLENIC ACID IN POSITION SN-2. - Google Patents

PROCEDURE FOR THE PURIFICATION OF TRIGLICERIDES CONTAINING GAMMA-LINOLENIC ACID IN POSITION SN-2. Download PDF

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ES2345529B1
ES2345529B1 ES200900892A ES200900892A ES2345529B1 ES 2345529 B1 ES2345529 B1 ES 2345529B1 ES 200900892 A ES200900892 A ES 200900892A ES 200900892 A ES200900892 A ES 200900892A ES 2345529 B1 ES2345529 B1 ES 2345529B1
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purification
tgs
procedure
gla
linolenic acid
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ES2345529A1 (en
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Miguel Angel Rincon Cervera
Jose Luis Guil Guerrero
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Universidad de Almeria
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Universidad de Almeria
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
    • B01J20/282—Porous sorbents
    • B01J20/283—Porous sorbents based on silica
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
    • B01D15/32—Bonded phase chromatography
    • B01D15/322—Normal bonded phase
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
    • B01J20/282—Porous sorbents
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B7/00—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B7/00—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils
    • C11B7/0008—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils by differences of solubilities, e.g. by extraction, by separation from a solution by means of anti-solvents

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  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Wood Science & Technology (AREA)
  • Fats And Perfumes (AREA)
  • Edible Oils And Fats (AREA)

Abstract

Procedimiento para la purificación de triglicéridos que
contienen ácido gamma-linolénico en posición sn-2. Con la finalidad de purificar triglicéridos que contienen ácido gamma-linolénico a partir de fuentes naturales, se utiliza una columna cromatográfica gravimétrica en fase normal, trabajando en gradiente de polaridad con solventes biocompatibles. Así se consigue la purificación de triglicéridos que cuentan en su estructura con una o más moléculas de ácido gamma-linolénico, pudiendo ser utilizados con diversos fines. Con esta metodología es posible trabajar a escala industrial, pues es fácilmente escalable, a diferencia de otras técnicas que son aplicables a escala analítica pero presentan serios inconvenientes en cuanto a coste y adiestramiento del personal a la hora de utilizarlas con fines industriales, como por ejemplo la cromatografía líquida de alta resolución (HPLC).Procedure for the purification of triglycerides that
they contain gamma-linolenic acid in sn-2 position. In order to purify triglycerides containing gamma-linolenic acid from natural sources, a normal phase gravimetric chromatographic column is used, working in polarity gradient with biocompatible solvents. Thus, the purification of triglycerides that have in its structure one or more gamma-linolenic acid molecules, can be used for various purposes. With this methodology it is possible to work on an industrial scale, as it is easily scalable, unlike other techniques that are applicable on an analytical scale but have serious inconveniences in terms of cost and training of personnel when using them for industrial purposes, such as high performance liquid chromatography (HPLC).

Description

Procedimiento para la purificación de triglicéridos que contienen ácido gamma-linolénico en posición sn-2.Procedure for purification of triglycerides containing gamma-linolenic acid in sn-2 position.

Sector de la técnicaTechnical sector

La presente invención se refiere a un método encuadrado en el sector técnico de la obtención de triglicéridos (TGs) estructurados conteniendo ácido gamma-linolénico (GLA, 18:3n-6) en posición sn-2. Este método se ha adaptado a este fin con el objeto de mejorar la eficacia de los métodos de obtención de los mismos, que son de empleo común en la industria alimentaria y farmacéutica, así como para disminuir los potenciales peligros para la salud de los TGs obtenidos mediante procesos enzimáticos, en los cuales a veces se utilizan solventes potencialmente tóxicos para el ser humano. Por otra parte, mediante este método se obtienen TGs estructurados ricos en GLA en un proceso significativamente más corto, y por tanto con menores probabilidades de alteraciones oxidativas que en los clásticos, en los cuales interviene un largo número de reacciones enzimáticas. Estos TGs estructurados con GLA en posición sn-2 pueden comercializarse como complementos nutricionales.The present invention relates to a method framed in the technical sector of obtaining triglycerides (TGs) structured containing acid gamma-linolenic (GLA, 18: 3n-6) in sn-2 position. This method has been adapted for this purpose. in order to improve the effectiveness of the methods of obtaining the same, which are commonly used in the food industry and pharmaceutical, as well as to diminish the potential dangers for the health of the TGs obtained by enzymatic processes, in the which are sometimes used solvents potentially toxic to the human being. On the other hand, through this method TGs are obtained structured rich in GLA in a significantly more process short, and therefore less likely to be altered oxidative than in the clastic ones, in which a long one intervenes number of enzymatic reactions. These structured TGs with GLA in sn-2 position can be marketed as nutritional supplements 

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Estado de la técnicaState of the art

Los ácidos grasos poliinsaturados, o PUFAs, son moléculas orgánicas consistentes en una cadena hidrocarbonada de 18 o más átomos de carbono con dos o más insaturaciones carbono-carbono en su estructura y un grupo carboxilo al extremo de la misma. La manera más general de proceder a su notación consiste en un número que indica el número de átomos de carbono de su molécula, seguido por dos puntos y el número de insaturaciones que posee. A continuación aparece una "n" en cursiva y un número indicativo de la posición de las insaturaciones en la cadena. En los aceites y grasas naturales de origen vegetal y animal, estos PUFAs se encuentran mayoritariamente esterificados con moléculas de glicerol dando lugar a los triglicéridos. Una vez que los TGs son ingeridos por los seres humanos, son degradados metabólicamente hasta sn-2 monoglicéridos y ácidos grasos libres, siendo de esta forma absorbidos por la mucosa intestinal.Polyunsaturated fatty acids, or PUFAs, are organic molecules consisting of a hydrocarbon chain of 18 or more carbon atoms with two or more carbon-carbon unsaturations in their structure and a carboxyl group at the end thereof. The most general way to proceed with its notation is a number that indicates the number of carbon atoms in your molecule, followed by a colon and the number of unsaturations it has. Below is an "n" in italics and an indicative number of the position of unsaturations in the chain. In natural oils and fats of vegetable and animal origin, these PUFAs are mostly esterified with glycerol molecules giving rise to triglycerides. Once the TGs are ingested by humans, they are metabolically degraded to sn -2 monoglycerides and free fatty acids, thus being absorbed by the intestinal mucosa.

Es ampliamente conocida la acción beneficiosa de una variedad de PUFAs sobre la salud. Entre los más conocidos figuran los ácidos GLA, estearidónico (SDA, 18:4n-3), eicosapentaenoico (EPA, 20:5n-3) y docosahexaenoico (DHA, 22:6n-3). El poder purificar TGs que contengan este tipo de compuestos podría permitir su aplicación en fórmulas farmacológicas y/o alimentarias sin tener que recurrir a incorporar el aceite en su totalidad.The beneficial action of a variety of PUFAs on health is widely known. Among the best known are GLA, stearidonic acids (SDA, 18: 4 n -3), eicosapentaenoic acid (EPA, 20: 5 n -3) and docosahexaenoic acid (DHA, 22: 6 n -3). Being able to purify TGs containing this type of compounds could allow its application in pharmacological and / or food formulas without having to resort to incorporating the oil in its entirety.

Previamente, se ha conseguido purificar TGs que contienen PUFAs de interés a partir de sus fuentes naturales mediante los procedimientos de cromatografía líquida de alta resolución (HPLC) y de cromatografía en capa fina (TLC) (Christie, W.W., 1999. Ind. Crop. Prod. 10: 73-83; Redden, P.R., Huang, Y.S., Lin, X. and Horrobin, D.F. 1995. J. Chrom. A 694:381-38 9). Sin embargo, aunque estas metodologías posibilitan la purificación a escala analítica o semipreparativa, no son adecuadas para las necesidades de la industria, que precisa métodos operativos a mayor escala. El procedimiento de purificación de componentes de aceites y grasas por columna cromatográfica gravimétrica con sílice y nitrato de plata ha sido aplicado con éxito para la purificación de ésteres metílicos de ácidos grasos. No obstante, supone además una herramienta útil a la hora de fraccionar TGs, presentando ciertas ventajas como su reducido coste, sus posibilidades de escalado y su facilidad de uso (no siendo necesaria una preparación especifica del operario) frente a otras técnicas anteriormente citadas como HPLC, más costosas y que requieren una preparación previa más exhaustiva por parte del personal encargado de su manejo.Previously, it has been possible to purify TGs containing PUFAs of interest from their natural sources by means of high-performance liquid chromatography (HPLC) and thin layer chromatography (TLC) procedures (Christie, WW, 1999. Ind. Crop. Prod . 10: 73-83; Redden, PR, Huang, YS, Lin, X. and Horrobin, DF 1995. J. Chrom . A 694: 381-38 9). However, although these methodologies enable purification on an analytical or semi-preparative scale, they are not suitable for the needs of the industry, which requires larger-scale operational methods. The process of purification of oil and fat components by gravimetric chromatographic column with silica and silver nitrate has been successfully applied for the purification of fatty acid methyl esters. However, it is also a useful tool when splitting TGs, presenting certain advantages such as its reduced cost, its scalability and ease of use (not requiring a specific operator preparation) compared to other techniques previously mentioned as HPLC , more expensive and that require a more thorough previous preparation by the personnel in charge of their management.

Previamente, la separación de mezclas artificiales de triglicéridos utilizando una fase estacionaria de gel de sílice y nitrato de plata ha sido llevada a cabo exitosamente, pero no fue realizada con solventes biocompatibles ni ha sido ensayada para separar TGs enriquecidos en GLA (Vries, B., 1964. J. Am. Oil Chem. Soc. 41:403-406). Únicamente, este procedimiento anterior ha sido utilizado para separar exitosamente TGs ricos en DHA de otros de TGs (Monsanto Co, 2002. US Patent 6399803). Por otra parte, el uso de este tipo de columna, formada por cationes y gel de sílice, también ha sido utilizada para separar fracciones de TGs con distinto índice de yodo, pero no fue ensayada para separar TGs estructurados conteniendo ácidos grasos específicos (Logan, T.D., 1982. European Patent Aplication 81200401.8).Previously, the separation of artificial triglyceride mixtures using a stationary phase of silica gel and silver nitrate has been carried out successfully, but it was not performed with biocompatible solvents nor has it been tested to separate TGs enriched in GLA (Vries, B. , 1964. J. Am. Oil Chem. Soc . 41: 403-406). Only, this previous procedure has been used to successfully separate TGs rich in DHA from others from TGs (Monsanto Co, 2002. US Patent 6399803). On the other hand, the use of this type of column, formed by cations and silica gel, has also been used to separate fractions of TGs with different iodine levels, but was not tested to separate structured TGs containing specific fatty acids (Logan, TD, 1982. European Patent Application 81200401.8). 

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Descripción de la invenciónDescription of the invention

El procedimiento consiste en hacer pasar los TGs disueltos en hexano a través de una columna cromatográfica gravimétrica cuya fase estacionaria consiste en una mezcla de sílice y nitrato de plata. Esta fase estacionaria, una vez activada a una temperatura apropiada, es introducida en la columna y acondicionada con varios volúmenes de disolventes biocompatibles. Para un desarrollo óptimo del proceso se carga por la parte superior de la columna una cantidad de aceite (mezcla de TGs) equivalente al 2-20% en masa de la fase estacionaria. La cantidad de fase estacionaria utilizada y por tanto la cantidad de aceite que se puede procesar es variable y dependerá de las necesidades especificas de cada aplicación, adoptándose un tamaño de columna mayor o menor en función de las mismas.The procedure consists in passing the TGs dissolved in hexane through a chromatographic column gravimetric whose stationary phase consists of a mixture of silica and silver nitrate. This stationary phase, once activated at a appropriate temperature, is introduced into the column and conditioned with several volumes of biocompatible solvents. For a optimal development of the process is loaded by the top of the column an amount of oil (mixture of TGs) equivalent to 2-20% by mass of the stationary phase. The amount of stationary phase used and therefore the amount of oil that It can be processed is variable and will depend on the needs specific to each application, adopting a column size major or minor depending on them.

Las diferentes fracciones son extraídas y purificadas haciendo pasar una fase móvil en gradiente creciente de polaridad consistente en mezclas de hexano, acetona y etanol en proporción variable en función de la composición de la mezcla inicial de TGs. Los TGs que contienen los PUFAs son así purificados en fracciones de polaridad variable en función de la naturaleza de dichos TGs. Alícuotas de dichas fracciones son analizadas posteriormente por HPLC y GLC para comprobar sus perfiles de TGs y ácidos grasos respectivamente (figura 1) . Los análisis demuestran que se consiguen fracciones enriquecidas en GLA hasta de un 500% con respecto a la riqueza original de GLA en el aceite.The different fractions are extracted and purified by passing a mobile phase in increasing gradient of polarity consisting of mixtures of hexane, acetone and ethanol in variable proportion depending on the composition of the mixture initial of TGs. TGs containing PUFAs are thus purified in fractions of variable polarity depending on the nature of said TGs. Aliquots of these fractions are analyzed subsequently by HPLC and GLC to check their TG profiles and fatty acids respectively (figure 1). The analyzes show that fractions enriched in GLA of up to 500% are achieved with regarding the original richness of GLA in the oil.

A continuación se presenta un ejemplo que ilustra el método de la invención y que en ningún caso debe considerarse limitativo del alcance de la presente invención.Below is an example that illustrates the method of the invention and that in no case should be considered as limiting the scope of the present invention. 

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Descripción de una realización modeloDescription of a model embodiment

Se utiliza esta técnica para purificar TGs que contienen GLA a partir de aceite de semillas de onagra (Oenothera biennis L.). Para ello se procede al rellenado de una columna hueca de vidrio con una fase estacionaria formada por gel de sílice y nitrato de plata. A continuación se adicionan por la parte superior de la columna aceite de onagra disuelto en hexano. Seguidamente, se hacen pasar una serie de volúmenes de eluyente en polaridad creciente que son recogidos por separado. Los eluyentes utilizados son biocompatibles, en proporciones variables.This technique is used to purify TGs containing GLA from evening primrose oil ( Oenothera biennis L.). This is done by filling a hollow glass column with a stationary phase formed by silica gel and silver nitrate. Then, evening primrose oil dissolved in hexane is added through the top of the column. Next, a series of eluent volumes in increasing polarity are passed through and collected separately. The eluents used are biocompatible, in varying proportions.

Una vez recogidos todos los eluyentes se analiza una alícuota de cada uno por HPLC en fase reversa para comprobar su perfil de TGs. Mediante este procedimiento se consiguen purificar los picos de TGs con mayor contenido en GLA (figura 2) . De dichas fracciones se separa una alícuota que es metilada para comprobar su perfil de ácidos grasos. De este análisis se obtiene una composición de GLA sobre el total de ácidos grasos que forman cada pico.Once collected all the eluents are analyzed an aliquot of each by reverse phase HPLC to check its TG profile. Through this procedure they are able to purify the peaks of TGs with the highest GLA content (figure 2). Of said fractions an aliquot that is methylated is separated to check its fatty acid profile. From this analysis a composition is obtained of GLA over the total fatty acids that form each peak. 

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Base del funcionamientoPerformance Base

En estas columnas los ácidos grasos se separan en función del número y configuración de los dobles enlaces debido a que los iones de plata interaccionan reversiblemente con los dobles enlaces para formar complejos polares doble enlace-Ag^{+}. Por tanto a mayor número de dobles enlaces más fuertemente es retenido el ácido y mayor es el tiempo de elución. (Ryu, S., Lee, J., Jeong, B and Hur, H. 1997. US Patent 5.672.726).In these columns the fatty acids are separated depending on the number and configuration of the double links due to that silver ions interact reversibly with doubles links to form double polar complexes link-Ag +. Therefore a greater number of doubles bonds more strongly the acid is retained and the longer is the time of elution (Ryu, S., Lee, J., Jeong, B and Hur, H. 1997. US Patent 5,672,726). 

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Utilidad de la invenciónUtility of the invention

La presente invención se refiere a un método para obtención de TGs estructurados conteniendo GLA en posición sn-2. Estos TGs son biodiponibles en el organismo humano, ya que las lipasas actúan preferentemente en posición sn-1 y sn-3, con lo que los sn-2 monoglicéridos conteniendo GLA remanentes en el aparato digestivo, pueden ser fácilmente absorbidos por la mucosa intestinal, incorporándose al sistema circulatorio y realizando un amplio número de funciones fisiológicas beneficiosas.The present invention relates to a method to obtain structured TGs containing GLA in position sn-2 These TGs are biodiponible in the body human, since lipases preferably act in position sn-1 and sn-3, so that sn-2 monoglycerides containing GLA remnants in the digestive system, can be easily absorbed by the mucosa intestinal, joining the circulatory system and performing a large number of beneficial physiological functions. 

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Descripción de los dibujosDescription of the drawings

Figura 1 - Representa el esquema del proceso de purificación de TGs en columna cromatográfica gravimétrica con sílice y nitrato de plata como fase estacionaria.Figure 1 - Represents the process scheme of purification of TGs in gravimetric chromatographic column with silica and silver nitrate as stationary phase.

Figura 2 - Muestra el perfil de TGs del aceite de onagra, obtenido por HPLC en fase reversa.Figure 2 - Shows the oil TGs profile of evening primrose, obtained by reverse phase HPLC.

Figura 3 - Muestra cromatogramas que corresponden a las fracciones de TGs enriquecidas en GLA. Los números 1 y 2 indican los picos que son purificados por este procedimiento.Figure 3 - Shows chromatograms that correspond to the fractions of TGs enriched in GLA. The numbers 1 and 2 indicate the peaks that are purified by this process.

Claims (7)

1. Procedimiento para la purificación de triglicéridos que contienen GLA en posición sn-2 mediante cromatografía en columna gravimétrica en fase normal (CGFN), que incorpora:1. Procedure for the purification of triglycerides containing GLA in position sn -2 by normal phase gravimetric column chromatography (CGFN), which incorporates: - Una fase estacionaria (FE) y una fase móvil (FM)- A stationary phase (FE) and a mobile phase (FM) - FE formada por una mezcla de gel de sílice y nitrato de plata- FE formed by a mixture of silica gel and silver nitrate - FM constituida por una serie de disolventes biocompatibles- FM consisting of a series of solvents biocompatible - Un proceso de purificación mediante CGFN usando la FE y FM descrita para obtener TGs enriquecidos en GLA en posición sn-2.- A purification process using CGFN using the FE and FM described to obtain TGs enriched in GLA in position sn -2. 
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2. Procedimiento para la purificación de TGs que contienen GLA en posición sn-2 según la reivindicación 1 caracterizado porque la CGFN se usa para la purificación de TGs obtenidos de fuentes naturales.2. Procedure for the purification of TGs containing GLA in position sn -2 according to claim 1 characterized in that the CGFN is used for the purification of TGs obtained from natural sources. 3. Procedimiento para la purificación de TGs que contienen GLA en posición sn-2 según cualquiera de las reivindicaciones anteriores, y caracterizado por una fase estacionaria que contenga cualquier tipo de metales que pudieran formar complejos con los dobles enlaces contenidos en los PUFAs y así retardar su elución.3. Procedure for the purification of TGs containing GLA in position sn -2 according to any of the preceding claims, and characterized by a stationary phase containing any type of metals that could form complexes with the double bonds contained in the PUFAs and thus delay Your elution. 4. Procedimiento para la purificación de TGs que contienen GLA en posición sn-2 según reivindicación 2, y caracterizado por el uso de la CGFN para aplicar a cualquier tipo de aceite de procedencia microbiológica, vegetal o animal.4. Procedure for the purification of TGs containing GLA in position sn -2 according to claim 2, and characterized by the use of the CGFN to apply to any type of oil of microbiological, vegetable or animal origin. 5. Procedimiento para la purificación de TGs que contienen GLA en posición sn-2 según reivindicación 4 y caracterizado por el uso de disolventes biocompatibles (FM) en gradiente de polaridad.5. Procedure for the purification of TGs containing GLA in position sn -2 according to claim 4 and characterized by the use of biocompatible solvents (FM) in polarity gradient. 6. Uso del procedimiento para la purificación de TGs que contienen GLA en posición sn-2 según las reivindicaciones anteriores, en la industria alimentaria y farmacéutica.6. Use of the method for the purification of TGs containing GLA in position sn -2 according to the preceding claims, in the food and pharmaceutical industry. 7. Uso del procedimiento para la purificación de TGs que contienen GLA en posición sn-2 según la reivindicación 6, en la formulación de complementos nutricionales.7. Use of the method for the purification of TGs containing GLA in position sn -2 according to claim 6, in the formulation of nutritional supplements.
ES200900892A 2009-03-13 2009-03-13 PROCEDURE FOR THE PURIFICATION OF TRIGLICERIDES CONTAINING GAMMA-LINOLENIC ACID IN POSITION SN-2. Active ES2345529B1 (en)

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ES200900892A ES2345529B1 (en) 2009-03-13 2009-03-13 PROCEDURE FOR THE PURIFICATION OF TRIGLICERIDES CONTAINING GAMMA-LINOLENIC ACID IN POSITION SN-2.
PCT/ES2010/000112 WO2010103146A1 (en) 2009-03-13 2010-03-08 Procedure for the purification of triglycerides containing gamma-linolenic acid in position sn-2.

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Publication number Priority date Publication date Assignee Title
US4961881A (en) * 1988-02-17 1990-10-09 Uop Process for separating triglycerides and regenerating absorbent used in said separation process
ATE273373T1 (en) * 1999-02-26 2004-08-15 Martek Biosciences Corp METHOD FOR SEPARATING A TRIGLYERIDE CONTAINING DOCOSAHEXAENIC ACID FROM A TRIGLYCERIDE MIXTURE

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CHAKRABARTY, M. M. et al. "{}Detection of rearrangement reaction of natural glycerides by chromatography"{} Journal of Chromatography, 1966, Volumen 22, páginas 84-89. Ver página 84, párrafo 2; página 85, párrafo 1. *
CHRISTIE, W. W. "{}Silver ion and chiral chromatography in the analysis of triacylglycerols"{}. Progress in Lipid research, 1994, Volumen 33, Número 172, páginas 9-18. Ver página 11, párrafo 2; página 12, párrafo 1. *
VRIES, B. "{}Separation of Triglycerides by Column Chromatography on Silica Impregnated with Silver Nitrate"{}. The Journal of the American Oil Chemists´Society, 1964, Volumen 41, páginas 403-406. Ver página 403, resumen. *

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