SU1289440A1 - Method of producing phospholipids - Google Patents

Abstract

The invention relates to the food pharmaceutical industry and can be used in the preparation of phospholipids from animal raw materials, which are used in the manufacture of medicinal and cosmetic products, feed and food additives, as well as reagents for scientific research. The aim of the invention is to increase the yield of phospholipids. To obtain phospholipids, before extraction, mineral or organic acid is introduced into the buttermilk with a ratio of buttermilk and acid (1: 0.001 - 1: 0.0023). Extraction of lipids from the precipitate is carried out with a mixture of chloroform and methanol in the ratio (I: 1.5) - (1: 2) twice in the cold (0-4 ° C), followed by release of phospholipids from the extract: when the ratio of extract and acetone

Description

11289440

The invention relates to the food and pharmaceutical industry and can be used in the preparation of phospholipids from animal raw materials, which are used in the manufacture of pharmaceutical and cosmetic products, feed and food additives, as well as reagents for scientific research. ABOUT

The purpose of the invention is to increase the yield of phospholipids.

The method consists in the use of buttermilk as the raw material of animal origin, in which mineral or organic acid is introduced before extraction with a ratio of buttermilk and acid (1: 0.001) - (1: 0.0023). Extraction of lipids from the precipitate is carried out with a mixture of chloroform 20 and methanol in a ratio of (1: 1.5) (1: 2), and the release of phospholipids from the extract is carried out at a ratio of extract and acetone (1: 2) - (1: 3) or - „. „,

N at h / final fraction. The final fraction

by chromatography on alumina oxide 4, „„,. „„ „„ „. ,,„ „„ „„ „l,

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The use of acid as a coagulant in ratios less than 1: 0.001 causes incomplete coagulation. At ratios above 1: 0.0023, oxidation of lipids is observed (the studies were carried out using microtonoel chromatography), which degrades the quality of the product and unreasonable coagulant overrun

The lipid extraction of fat globule membranes is carried out with chloroform: methanol in a ratio of (1: 1.5) - (1: 2) twice in the cold (), followed by separation of the extracted lipids from proteins by centrifuging at 1000 rpm for 5 minutes, concentration of lipids, evaporation of solvents on a rotary evaporator.

As can be seen from the table. 3, the optimum ratio of a mixture of chloroform: methanol is 1: (1.5–2, ensuring a maximum yield of

phospholipids are isolated from the extract by precipitation with acetone at an extract / volume ratio of acetone 1: (2-3).

mini mixture of chloroform and methanol in the ratio (3: 1) - (3: 1.5), while 35-36% hydrochloric or 98-100% acetic acid are used as a mineral or organic acid.

Treatment of buttermilk with acid in the ratio of buttermilk and acid (1: 0.001) - (1-0.0023) provides coagulation of the fatty buttermilk membranes, which are then separated from the aqueous phase, lipids are extracted from the precipitate and phospholipids are precipitated from the extract using selected organic solvents at specific ratios. The use of a new source of raw materials under optimal conditions for the distribution of phospholipids provides a yield of phospholipids up to 90-95% while reducing the cost of the final product.

Mineral or organic acids can be used as coagulants. The most accessible of these are salt and acetic acid. Due to coagulation of the membranes of the fat globules, the yield of dry buttermilk mass increases by 78- 90 times, which makes it possible to significantly speed up and simplify the mode of its separation.

As a result of numerous experiments, optimal amounts of used reagents were determined.

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The use of acid as a coagulant in ratios less than 1: 0.001 causes incomplete coagulation. At ratios above 1: 0.0023, an oxidation of lipids is observed (the studies were performed by micro-thin layer chromatography), which degrades the quality of the product and there is an unreasonable excess of coagulants.

The lipid extraction of fat globule membranes is carried out with a mixture of chloroform: methanol in a ratio of (1: 1.5) - (1: 2) twice in the cold (), followed by separation of the extracted lipids from proteins, by centrifuging at 1000 rpm for 5 minutes , concentration of lipids, evaporation of solvents on a rotary evaporator.

As can be seen from the table. 3, the optimum ratio of a mixture of chloroform: methanol is 1: (1.5-2), ensuring maximum yield

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phospholipids are isolated from the extract by precipitation with acetone at an extract / volume ratio of acetone 1: (2-3).

As can be seen from the table. 4, when precipitating phospholipids with acetone in an extract: acetone ratio less than 1: 2, the yield of the final product decreases due to incomplete precipitation, and with more acetone at a ratio of 1: 4 to 6, part of the phospholipids goes into an acetone solution. The yield of phospholipids is at a ratio of extract: acetone 1: (2-3) 60-65.7%, at a ratio of 1: (0.8-1), 50-51%, at a ratio of 1: (4-6 ) 33.37%.

Column chromatography makes it possible to isolate phospholipids in almost pure form, but the use of chromatographic methods is accompanied by loss of substance.

When phospholipids are selected by column chromatography,

the output of phosphatidylcholine (lecithin) 89-91%.

Example 1. K1 l of buttermilk added 35% HCI in the ratio of buttermilk: acid 1: 0.0023 (until the

signs of coagulation). Then the precipitate is isolated by centrifuging in a low-speed refrigeration centrifuge at 1000 rpm for 5 minutes at 4 ° C. The supernatant is removed and k

2.1 l of a mixture of chloroform: methanol in a ratio of 1: 1.5 is added to the precipitate. The mixture is kept cold at 0 ° C for 2 hours with occasional stirring, again centrifuged as before. The first extract is poured, and the precipitate is again treated with 2.1 liters of a mixture of chloroform - methanol in a ratio of 1: 1.5 for 3 hours.

The precipitate was removed by centrifugation in the same way, and the extracts were combined, 2 L of distilled water was added. After separation of the phases, the upper aqueous-methanol phase is decanted, and the lower (chloroform lipid extract) is evaporated on a rotary evaporator under reduced pressure to a volume of 7 ml. To the resulting concentrated extract, cold acetone is added at 4 ° C. The extract – acetone ratio is 1: 3, kept cold at 4 ° C for 2 hours, the phospholipid precipitate is filtered off and dissolved in chloroform. The yield of total lipids is 7.3 g / L, and that of phospholipids, 4.7 g / L or 64%.

The results of the preparation of phospholipids in examples 2-7 are presented in table.

Thus, the proposed method allows to obtain a high yield of the phospholipid complex of 57-65% from easily accessible and cheap natural raw material — buttermilk — a minor product of oil production. The resulting phospholipids can be used.

Table 1

Comparative yield of the main classes of lipids (g / 1 l).

Total Lipid0, 1-0.3

Glycerides 0.07-0, .18

Phospholipids 0,025-0,04

Sterols Sterides

0.004-0.009 1.2-3.0

as in medicine, veterinary medicine, and in the pharmaceutical, perfumery and food industry.

The reduction in the cost of the final product obtained according to the proposed method relative to the complex of phospholipids obtained from egg yolks is due to the low cost of buttermilk.

Claims (1)

Invention Formula A method of producing phospholipids, which involves extracting lipids from animal raw materials with an organic solvent, separating phospholipids from the extract, adding acetone to the latter, characterized in that, in order to increase the yield of phospholipids, buttermilk is used as animal raw material or an organic acid in the ratio of buttermilk: acid 1: 0.001-1: 0.0023, the extraction of lipids from the precipitate is carried out with a mixture of chloroform: methanol in a ratio of 1: 1.5-1: 2: 2, and the release of phos folipids from the extract are carried out at an extract: acetone ratio of 1: 2-1: 3 or by column chromatography on alumina with a mixture of chloroform: methanol in a ratio of 3: 1-3: 1.5, while 35-36 are used as a mineral or organic acid % hydrochloric or 98-100% acetic acid. 24-26 15-20 16.8-19.6 9.7-13.0 5.2-6.0 3.0-4.0 0.75-1.0 12894406 table 2 The dependence of the yield of dry mass and lipid buttermilk from coagulation conditions Coagul nty 35% HC1 98% CHjCOOH 1: 0.0009 18 7  1: 0.001 35 15 1: 0.0023 40 20 1: 0.003 20 20 Table 3 The dependence of the yield of lipids on the conditions of extraction Classes are- Ratio of chloroform: methanol pid, g / l t-1j- | - : 0 I 1: 1 1: 1.5 1: 2 1: 3 Total lipids 18 21 24 26 20 Glycerides 13.5 13.7 6.8 19.6 15.1 Phospholipids 3.0 3.9 5.2 6.0 4.0  Table4 Dependence of lipid yield on precipitation conditions acetone The ratio of extract: acetone in the main lipid composition Components .1: (0.8-1) 1: (2-3). 1: (4-6). g / l% g / l% g / l j% Total lipids 3.5-5.0 51-53 7,: 0-7.6 49-50 4.5-5.6 55-57 Glycerides 1.5-2.0 21-22 2.6-3, 0 18-19 1.8-2.5 23-26 Phospholipids 1.8-2.5 26-26.54.2-4.6 30-31 1.5-2.1 19-20 Sterols, sterides, 0.1-0.2 3-0.5 0.1-0.5 0.6 : Output dry masses buttermilk g / l The output of lipids, g / l 20 45 47 47 9  24 26 26 71289440 8 Table 5 The chemical composition of buttermilk lipids in comparison with the composition of lipids of egg yolk (according to the prototype) Glycerides 65-70 Type and concentration of coagu-, lnta The ratio of buttermilk: acid Sludge separation The ratio of chloroform - methanol in the extraction Phospholipid absorption 36% on 35% on 98% on 100% on 36% on 35% on HC1 HC1 acetic acetic HC1 HC1 1: 0.0018 1: 0.0022 1: 0.0023 1: 0.001 1: 0.0018 1: 0.0022 Centrifuge - Filtro Centrifuge - Filtro Filtro-Centrifugation 1: 1.5 12 1: 2 1: 2 1: 1.5 1: 2 Deposition Deposition- Deposition Deposition-Acetone Acetone Acetone Acetonone Kolonoch- Column Chroma Chromatograficaografy fi 12 1: 3 65 63 31-42 3-5 40-60 1: 2 1: 2 1: 1.5 1: 2 Kolonoch- Column Chroma Chromatograficaografy fi 1: 2 1: 3 3: 1 3: 1, 5 55 57 88 91