KR830000794B1 - Process for preparing oil-containing high purity phosphatidylcholine - Google Patents

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Description

Process for preparing oil-containing high purity phosphatidylcholine

The present invention relates to a process for producing high purity phosphatidylcholine (lecithin) containing high concentrations of essential fatty acids. Crude lecithin is first extracted with a lower alcohol having 1-3 carbon atoms, the two phases formed are then adversely treated with a conflicting acidic aluminum sorbent containing a large amount of alcohol. The adsorbent is eluted with alcohol (according to the known method) to obtain an oil-containing phosphatidylcholine containing a large amount of essential fatty acids free of cephalin and inositol phosphatide.

German Patent No. 1617679 describes a method of adsorbing phosphatides on aluminum oxide from plant resins and extracting with alcohol to obtain high purity phosphatidylcholine (lecithin) containing large amounts of essential fatty acids.

This process is characterized in that the oil-containing crude phosphide is dissolved in ethyl acetate or a dichlorinated hydrocarbon having 1 to 2 carbon atoms or a mixture of these solvents. The solvent may comprise up to 6% alcohol by volume. The solution which melt | dissolved raw material is processed, stirring and adding aluminum oxide corresponding to the quantity of crude phosphatides at least 5 times. Finally, high purity phosphatidylcholine is liberated with alcohol from the dispersed aluminum oxide. According to Federal Republic of Patent No. 1617680, the oil-containing bath phosphatide solution is contacted (instead of stirring) and the chemically pure phosphatidylcholine is liberated with the alcohol from the aluminum oxide adsorbent. .

Swiss Federal Patent No. 361088 and U.S. Patent No. 2945869 describe a purification method for obtaining a soya-bean phosphatide fraction used as an emulsifier of fatty emulsions intended for intravenous use. According to the above process, the alcoholic solution of crude phosphatide previously deoiled is treated with Al ₂ O ₃ , MgO or activated carbon to reduce the cephalin content of these solutions and lower blood pressure when intravenously administered to cats. Remove all inositol containing phosphatides found to be.

However, the latter method requires a step of pre-oiling commercially useful crude phosphatides before producing an alcoholic solution that reduces the cephalin content, regardless of the use of adsorbents, and also known processes Cannot remove cephalin completely or in large quantities.

German Federal Patent No. 1053299 discloses, among other materials, natural choline phosphate diglycerides from which collagen phosphoric acid diglyceride esters have been removed using column chromatography using aluminum oxide as an adsorbent. Described are methods for preparing esters. This process also uses the method of extracting repeatedly with acetone and deoiling, and extracting the crude phosphatide previously deoiled with alcohol.

For the processes disclosed in Federal Republics Patent Nos. 1617679 and 1617680, the oil-containing crude phosphatide mixture is dissolved in ethyl acetate or chlorinated hydrocarbons without deoiling and the solution is treated with aluminum oxide. do. The deoiling step of crude phosphatides can be selectively separated by treatment of phosphatidylcholine (resin) with alcohol and aluminum oxide, while all phosphatides remain in the adsorbent. According to this process, high purity phosphatidylcholine can be obtained from which oil has been removed.

Oil-containing crude phosphatides do not necessarily have to be treated with ethyl acetate or dihydrocarbon hydrocarbons. The crude phosphatides described above are extracted with lower alcohols having one to three carbon atoms and the extract is extracted. It has been surprisingly found that the adsorbed phosphatides can be separated using alcohols in aluminum oxide, which can be treated directly with aluminum oxide and described in Federal Republics Patent Nos. 1617679 and 1617680.

In contrast, the phosphatidylcholine containing a large amount of essential fatty acids in the molecule can be prepared with crude phosphatides containing vegetable oil by the method described in the present invention. In practice, it can be seen that there is a considerable need to prepare an oil-containing solution of high purity phosphatidylcholine. In particular, there is a need in the pharmaceutical field for oral administration of high purity phosphatidylcholine, for example in the form of gelatin capsules. Absorption studies using phosphatidylcholine labeled as radioactive materials have shown that when orally administered an oily product of phosphatidylcholine with unsaturated polyfatty acid, the absorption rate is multiply higher than the aqueous product.

Natural vegetable oils such as sunflower oil and soybean oil, which always contain phosphatides, mix well with mono and diglycerides made from the same oils. This mixture is very suitable as an oily substance. Thus, liquid phosphatidylcholine fractions that are also useful in other fields such as cosmetics and diet can be obtained directly.

The fluid oily solution of high purity phosphatidylcholine used to fill gelatin capsules is a natural vegetable oil or other edible oil or monoglyceride or free fatty acid produced during the purification process of the oil-free phosphatidylcholine obtained by the method described in German Patent No. 1617679. Prepared by mixing with a mixture again. However, the aforementioned method requires the step of removing the oil adhering to the phosphatide mixture during or before the adsorption process on Al ₂ O ₃ to remove cephalin and inositol phosphatide.

The prior art processes and the processes described in German Patent No. 1053299 or related patents are quite uneconomical and have a number of disadvantages. First, when acetone is contained as a commercially available crude phosphatide, a small amount of the metal oxide produced by chemically assembling with two acetone molecules is always produced. Due to its toxicity, character and peculiar smell, the mesityin group has a very detrimental effect even when present in very small amounts. Second, when the oil and phospholipids are separated by chromatography, a solvent mixture is formed, which must always be fractionated before reuse.

As an example, the present invention is directed to extracting an oil-containing phosphatide extract from an oil-containing crude phosphatide with a lower alcohol and directly contacting the oily phosphatide extract with an aluminum oxide adsorbent. Next, there is provided a method for preparing a highly purified oil-containing phosphatidylcholine, consisting of recovering the adsorbed phosphatidylcholine. The present invention also provides the product of the process.

The process described in the present invention first proceeds by extracting commercially useful crude lecithin containing 20-40% of oil at a temperature of about 50 ° C. with methanol, ethanol or propanol. The two phases formed during the cooling of the mixture are separated, for example by decantation or centrifugation. The alcohol-rich upper layer is treated with aluminum oxide by stirring with aluminum oxide or through a column of chromatography filled with adsorbent. Elution with alcohol yields an oil-containing phosphatidylcholine that does not contain cephalin and inositol phosphatide and contains a large amount of essential fatty acids.

In contrast to the methods according to the prior art in the art, when the oil-containing alcoholic phosphatide solution is separated by chromatography on aluminum oxide, no pretreatment step is necessary, and the solution containing a large amount of oil It does not interfere with the adsorption step of acid phosphatide at all. This is particularly surprising in that small amounts of oil dissolve in alcohol, which, during contact with aluminum oxide, separates the oil on the phosphatide and interferes with the active center of the adsorbent. It can also be seen that the amount of adsorption of phosphatidylcholine from the alcoholic solution does not occur in a sufficient amount as in the prior known case of using this solvent as an element.

The oil-containing alcoholic extract used in the process of the present invention is prepared by one or several treatments of the phosphatide raw material with alcohol at the boiling point of the solvent, most preferably at a temperature of 20 to 50 ° C. The mixture thus obtained can be separated by treatment by decantation or centrifugation. The alcoholic top layer is stirred and treated with aluminum oxide, and then the adsorbent separated from the solution is stirred with ethanol for several minutes to separate with a decanter. Rinse repeatedly with a small amount of ethanol and de-aluminum the aluminum oxide containing unnecessary material or introduce it into the next process. When using a chromatography column to separate acidic phospholipids. The column is eluted with alcohol, preferably ethyl alcohol, to give an oil-containing phosphatidylcholine concentrate. If necessary, the eluate from the column can be separated to reduce the oil content of the phosphatidylcholine fraction.

According to this process, the alcoholic solution obtained by stirring with aluminum oxide or by treatment with column chromatography is separated from the solvent under vacuum and under the protection of an inert gas. If necessary, the solution may be filtered.

Small amounts of filter aids, such as diatomaceous earth, sold under the trademark celite may be used. The product is an oil-free fraction of high purity natural phosphatidylcholine containing little cephalin and containing at least 66-70% essential fatty acids.

Suitable alcohols for the extraction of crude oil-containing phosphatides and the chromatographic process of the present invention are lower alcohols (preferably ethyl alcohol) having one to three carbon atoms. The alcohol can be used in a mixture containing 85 to 96% by weight of alcohol (ie, a solution containing 4 to 15% by weight of water) and a small amount of water, the latter being particularly advantageous as it is commercially useful. .

Basic and neutral aluminum oxides, preferably basic aluminum oxides of activity 1-3 graded by Brockmann can be used as adsorbents.

The process proposed in the present invention is an oil-containing phosphatide in cephalin and inositol phosphatide by treating with aluminum oxide or stirring with aluminum oxide in a chromatography column while maintaining an oily phase. The fractions are separated and prepared into oil-containing phosphatidylcholine fractions containing large amounts of essential fatty acids using a single process.

The products according to the process of the present invention are atherosclerosis, diabetes mellitus, and hyperlipidemia due to the high phosphatidylcholine content in excess of 90% of the phosphatide fraction and the inclusion of more than 70% essential fatty acids relative to all fatty acids present It has a useful therapeutic effect on various conditions, and even a very small amount of 2 grams per day has the effect of reducing the pathologically suggested blood glucose level.

Oil-containing high-purity phosphatidylcholine fractions obtained from certain crude phosphatides according to the process described above generally have the following composition.

Iodine Number 98-105 Choline 9.5-10.8%

Oil content 15-30% Phosphoric acid: molar ratio of choline 0.996%

Total amount of phosphorus 2.4-2.75%

Gas chromatography analysis of fatty acids shows the following average values.

a) milk-containing phosphatidylcholine fraction according to the invention b) deoiled phosphatidylcholine fraction

The invention will be better explained in the following examples which illustrate the invention. However, the present invention is not limited to the specific description of the embodiment.

Example 1

a) Preparation of oil-containing ethanolic sunflower-phosphatide solution.

1 kg of sunflower phosphatide containing 35% oil is stirred with 5 meters of ethanol under a nitrogen atmosphere of 40 ° C. The solution is allowed to stand at room temperature for 1 hour, and a light weight upper layer rich in ethanol is separated from the viscous lower layer in a sorting funnel.

b) treatment with Al ₂ O ₃

1.5 liter of the above-mentioned upper layer containing 75 grams of solid material and 30 grams of sunflower oil are stirred vigorously with ₃₀₀ grams of Al ₂ O _{3 for} 2 hours and 30 minutes under an N ₂ protective atmosphere at room temperature. When the adsorbent is settled, the ethanol lava is decanted, and Al ₂ O ₃ is washed twice with 150 ml of ethanol. As a filter aid, a mixture of 1 g of celite is added, then the two ethanol solutions are filtered off and the solvent is upstreamed in vacuo under a nitrogen atmosphere with a bath temperature of 40 ° C.

Yield:

Viscous oil-containing phosphatidylcholine fraction containing no cephalin and having the following composition: 45

Oil content 30%

Phosphatidylcholine content 64%

Example 2

According to the method of Example 1 (a), 1 liter of upper layer (containing 53 grams of solid content and 27% of soybean oil) obtained by extracting with ethanol from soybean phosphatide raw material was 280 grams of Al ₂ O ₃ in an inert gas atmosphere. Stir vigorously for 4 hours with. When the adsorbent is settled, the ethanol solution is separated, and each of Al ₂ O ₃ is washed twice with 150 ml of ethanol, and then the mixed ethanol extract is filtered through a celite mixture. The solvent is evaporated in vacuo under nitrogen protection to obtain 33 grams of a flexible, cephalin-free, yellow phosphatidyl fraction that can be changed into a fluid form by a mixture of oil or partial glycerides with at least 50% unsaturated fatty acids. The fraction to be recovered has the following composition.

Soybean oil 25%

Phosphatidylcholine Content 68%

Example 3

1.3 liters of 4.1% ethanol-based soy-phosphatide solution containing 28% solids obtained by extracting the ethanol-containing soy phosphatide raw material with ethanol or the upper layer at 25 ° C. according to the process of Example 1 Under N ₂ protection is introduced into a 31 mm diameter, 400 mm high chromatography column filled with 280 grams of Al ₂ O ₃ suspension dissolved in ethanol. After passing through the solution, the adsorbent is washed with 350 ml of alcohol. The mixed ethanol extract is separated from the solvent under vacuum under inert gas protection with a bath temperature of 40 ° C.

Yield: 35 grams (68%) of high-purity phosphatidylcholine fraction containing no cephalin at all and an oil content of 29% phosphatidylcholine

Example 4

400 grams of soy phosphatide raw material is extracted twice for 30 minutes using 1,000 ml of methanol at 40 ° C., respectively.

The mixed methanol extract cooled to room temperature contains 9% solids, 12% oil and 33% phosphatidylcholine.

One liter of the methanol solution described above is introduced into a chromatography column filled with 500 grams of an aluminum oxide suspension under the protection of an inert gas. After the solution has passed, the adsorbents of the column are washed with 700 ml of methanol. The mixed methanol eluate is concentrated under the protection of an inert gas at 40 ° C. in vacuo.

Yield: 55 g of yellow calcined product containing little cephalin, 30% oil and 66% phosphatidylcholine, a mixture of mono-glycerides containing at least 50% unsaturated fatty acids. Phosphatidylcholine concentrate is obtained.

Claims (1)

A lower alcoholol having 1-3 carbon atoms and an 85-96% aqueous solution thereof, extracting an oil-containing phosphatide extract from an oil-containing tank phosphatide, wherein the oil The oil-containing crude phosphatide, comprising a step of directly contacting the containing phosphatide extract with an aluminum oxide adsorbent and recovering the adsorbed phosphatidylcholine therefrom. Method for preparing high purity phosphatidylcholine.