JPH056551B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an improved process for purifying crude defatted phosphatides by chromatographic adsorption onto columns packed with aluminum oxide.

Phosphoratide is obtained from plant sources such as soybeans and peanuts, and is used in the fields of food, cosmetics, and medicine, but high purity is particularly required for use in medicine. Crude phosphatide obtained from plant sources contains large amounts of oils, fats, and polysaccharides, so it is usually defatted with acetone or the like, and the defatted crude phosphatide is purified by extraction with alcohol, particularly ethanol. However, extraction of phosphatide with alcohol has a poor yield and cannot obtain a product of high purity. Unexamined Japanese Patent Publication 1987-
No. 26548 describes a method for extracting oil-containing phosphatide obtained from plant sources with a lower alcohol to obtain defatted crude phosphatide, and purifying the phosphatide by passing the water-containing lower alcohol of the phosphatide through a column packed with aluminum oxide. However, even with this method, the content of phosphatidylcholine is only 92% by weight or less. Furthermore, JP-A No. 59-29694 describes a method of purifying an ethanol extract by treating it with aluminum oxide using a stirring chromatography method at a temperature of 50°C or higher; however, this method can shorten the processing time, but it is not necessary to purify the ethanol extract. No improvement in the purity of the product can be expected.

However, according to the present invention, extremely high purity phosphatylcholine can be purified with a high recovery rate by adsorbing crude defatted phosphatide onto aluminum oxide and eluting with a mixed solvent of a lower alcohol and a low-boiling aliphatic hydrocarbon solvent. I found it. The defatted crude phosphatide purified by the method of the present invention may be defatted by any method. Crude defatted phosphatide contains about 17 to 62% phosphatylcholine, although it varies depending on the raw material plant source and the defatting method. The defatted crude phosphatide treated in the present invention refers to one from which neutral fats have been removed, and may contain free fatty acids. Since free fatty acids are easily removed by the purification method of the present invention, it is not necessary to remove them in advance.

In the present invention, crude defatted phosphatide is adsorbed onto an aluminum oxide packed column. The solvent for adsorbing the defatted crude phosphatide onto the column may be any hydrocarbon solvent such as n-hexane or any solvent capable of dissolving phosphatide such as ethanol. However, it is preferred to use the same solvent from which the adsorbed phosphatide is eluted. The lower alcohol of the mixed solvent used in the present invention is preferably methanol or ethanol, particularly ethanol. In addition, the low boiling point aliphatic hydrocarbon solvent may be one that is mixed with a lower alcohol and has a low boiling point, particularly below 80°C, and specifically, n
-Hexane is preferred. Since n-hexane can be mixed with ethanol in any proportion, an ethanol-n-hexane mixed solvent is advantageous. Methanol and n-hexane are not completely miscible unless one of them is present in a large amount, but the mixed solvent used in the present invention is a mixed solvent containing excess methanol and has a methanol content of 68.4% or more. The mixed solvent of ethanol and n-hexane has an ethanol content of 15 to 80 wt% (n-hexane content of 85 wt%).
~20wt%), preferably 20-50wt%. The boiling points of ethanol and n-hexane are each 78.4
°C and 68.7 °C, but the boiling point of its azeotrope (ethanol content 22%) is 58.7 °C. On the other hand, phosphatidylcholine may decompose at temperatures above about 60°C. However, since the boiling point of the azeotrope is 60° C. or less, decomposition of phosphatidylcholine can be prevented, and it is particularly preferable in terms of solvent recovery and purification after use.

In this way, in the method of the present invention, the crude defatted phosphatide to be treated is dissolved in a suitable solvent, and the solution is poured into a column packed with aluminum oxide in an amount of about 6 times or more than that of the to-be-treated material, and adsorbed. Next, the mixture is developed with the above-mentioned mixed solvent of a lower alcohol and a low-boiling hydrocarbon solvent, and after removing about 5% of the initial portion of the effluent solvent, the entire effluent solvent solution is recovered. The amount of mixed solvent used at this time is about 15 times that of the amount to be treated. If necessary, the recovered solvent solution is removed by distillation under reduced pressure to produce purified phosphatidylcholine with a purity of 92% or more, especially ethanol-n with an ethanol content of about 20 to 50% by weight.
- When using a hexane mixed solvent, the purity is almost
100%, recovery rate over 90%.

Next, examples of the present invention will be shown and specifically explained.

Example 1 Alcohol extract of defatted crude phosphatide (60% phosphatidylcholine, 28% phosphatidylethanolamine, 12% other lysolecithin,
6.5g (oil/fat content: 1% or less) in a mixed solvent of ethanol and n-hexane with an ethanol concentration of 21.8% by weight
It was dissolved in 19.5g. This product was fixed in a column with an inner diameter of 24 mm and a height of 200 mm packed with 39.0 g of aluminum oxide. After fixing, development was performed using 97.5 g of a mixed solvent with the same ethanol concentration. After removing the first 5.0 g of effluent solvent, all the effluent solvent was collected as the main fraction, and the solvent was evaporated under reduced pressure below 50°C to obtain 3.59 g of concentrate (Phosphatidylcholine content 100% by weight, Phosphatidylcholine content 100% by weight, Phosphate). A purification recovery rate of phatidylcholine (90.7% by weight) was obtained.

Next, the above method is repeated by changing the ethanol concentration of the mixed solvent of ethanol and n-hexane,
The content of phosphatidylcholine in the obtained purified product and the recovery rate of phosphatidylcholine were measured, and the relationship with the composition of the mixed solvent was investigated. The results are shown in FIGS. 1 and 2. According to that, if the ethanol concentration is 20-80%, the recovery rate is 90%.
It will be understood that from the above, phosphatidylcholine with a purity of 93% or more can be obtained.

Example 2 6.5 g of defatted crude phosphatide (18% phosphatidylcholine content, 18% phosphatidylethanolamine, 15% inositol, 49% serine, lysolecithin, etc. (fat and oil content 1% or less)) was added to an ethanol concentration of 40.0 It was dissolved in 19.5 g of a mixed solvent of ethanol and n-hexane (wt%). This product is filled with 39.0g of aluminum oxide, has an inner diameter of 24mm, and a height of
It was fixed on a 200 mm column. After fixing, development was performed using 97.5 g of a mixed solvent with the same ethanol concentration.
After removing the first 5.0 g of effluent solvent, all the effluent solvent was collected as the main fraction, and the solvent was evaporated under reduced pressure below 50 °C to obtain 1.04 g of concentrate (Phosphatidylcholine content 100% by weight, Phosphatidylcholine content 100% by weight, A purification recovery rate of phatidylcholine (90.4% by weight) was obtained.

Reference Example 1 6.5 g of the same alcoholic extract of defatted crude phosphatide used in Example 1 was dissolved in 19.5 g of a mixed solvent of ethanol and n-hexane with an ethanol concentration of 10.0% by weight. This item is filled with 39.0g of aluminum oxide, has an inner diameter of 24mm, and a height of 200mm.
It was fixed on a mm column. After fixing, development was performed using 97.5 g of a mixed solvent with the same ethanol concentration. After removing the first 5.0 g of effluent solvent, all the effluent solvent was taken as the main fraction, and the solvent was evaporated under reduced pressure below 50°C to obtain 2.54 g of concentrate (Phosphatidylcholine content 85.8% by weight, Phosphatidylcholine content 85.8% by weight, A phatidylcholine purification recovery rate of 55.1% by weight was obtained.

Example 3 6.5 g of the same alcoholic extract of defatted crude phosphatide as in Example 1 was added to a methanol concentration of 75.0% by weight.
It was dissolved in 19.5 g of a mixture of methanol and n-hexane. This product was fixed in a column with an inner diameter of 24 mm and a height of 200 mm packed with 39.0 g of aluminum oxide. After fixing, use a mixed solvent with the same methanol concentration.
It was developed using 97.5g. Initial effluent solvent 5.0g
After removing the solvent, all the effluent solvent was collected as the main fraction, and the solvent was evaporated under reduced pressure below 50°C to obtain 3.98 g of concentrate.
(Phosphatidylcholine content 90% by weight, purification recovery rate 90.5% by weight).

Reference Example 2 6.5 g of the same alcoholic extract of defatted crude phosphatide used in Example 3 was dissolved in 19.5 g of a mixture of methanol and n-hexane with a methanol concentration of 3.0% by weight. This product was fixed in a column with an inner diameter of 24 mm and a height of 200 mm packed with 39.0 g of aluminum oxide. After fixing, development was performed using 97.5 g of a mixed solvent with the same methanol concentration. After removing the first 5.0 g of effluent solvent, all the effluent solvent was collected as the main fraction, and the solvent was evaporated under reduced pressure below 50 °C to obtain 2.38 g of concentrate (phosphatidylcholine content 83.5
Weight%, phosphatidylcholine purification recovery rate 50.2
% by weight) was obtained.

[Brief explanation of drawings]

Figures 1 and 2 show the relationship between the ethanol concentration of the ethanol-n-hexane mixed solvent on the horizontal axis and the content and recovery rate (wt%) of phosphatidylcholine on the vertical axis. be.

Claims (1)

[Scope of Claims] 1. Purification of phosphatidylcholine, characterized by adsorbing crude defatted phosphatide onto an aluminum oxide-packed column and then eluting it with a mixed solvent of a lower alcohol and a low-boiling aliphatic hydrocarbon solvent. Method. 2. The method according to claim 1, wherein the lower alcohol is methanol or ethanol, and the low-boiling aliphatic hydrocarbon solvent is n-hexane. 3. The method according to claim 1, which uses an ethanol-n-hexane mixed solvent having an ethanol content of 15 to 80% by weight.