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Method for preparing high-purity dilinoyl phosphatidylcholine through combination of enzymatic modification and reversed-phase column chromatography separation and product of high-purity dilinoyl phosphatidylcholine

A technology of dilinoleoylphosphatidylcholine and phosphatidylcholine, which is applied in the field of preparation of high-purity dilinoleoylphosphatidylcholine by enzymatic modification combined with reversed-phase column chromatography, can solve complex production process and product Low purity, long reaction time and other problems, to achieve the effect of good repeatability, simple and controllable operation, and low equipment cost

Active Publication Date: 2021-07-27
WUHAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It solves the problems of complex production process, long reaction time, low product purity and high cost in the prior art

Method used

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  • Method for preparing high-purity dilinoyl phosphatidylcholine through combination of enzymatic modification and reversed-phase column chromatography separation and product of high-purity dilinoyl phosphatidylcholine
  • Method for preparing high-purity dilinoyl phosphatidylcholine through combination of enzymatic modification and reversed-phase column chromatography separation and product of high-purity dilinoyl phosphatidylcholine
  • Method for preparing high-purity dilinoyl phosphatidylcholine through combination of enzymatic modification and reversed-phase column chromatography separation and product of high-purity dilinoyl phosphatidylcholine

Examples

Experimental program
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Effect test

Embodiment 1

[0037] This example provides a method for preparing high-purity dilinoleoylphosphatidylcholine and its product through enzymatic modification combined with reversed-phase column chromatographic separation.

[0038] figure 1 Show the flow chart of the preparation DLPC of an embodiment of the present invention, see figure 1 , add 3.5% (moisture: high-purity phosphatidylcholine + linoleic acid, m / m) to the mixed solution of 2g high-purity phosphatidylcholine, 4g linoleic acid and 10mL n-hexane, add 5% TL IM fat Enzyme (lipase: high-purity phosphatidylcholine + linoleic acid, m / m), the above mixture was premixed under the ultrasonic continuous flow cell disruptor, the premixing power was 200W, ultrasonic 5s / intermittent 5s, repeated 5 times , total ultrasound 45s. The above mixture was stirred and reacted at 50° C. for 1 h to obtain a modified mixture of phosphatidylcholine and fatty acid. After the phosphatidylcholine mixture was dried at low temperature to remove n-hexane, it...

Embodiment 2

[0041] This example provides a method for preparing high-purity dilinoleoylphosphatidylcholine and its product through enzymatic modification combined with reversed-phase column chromatographic separation.

[0042] Add 3.5% (moisture: high-purity phosphatidylcholine + linoleic acid, m / m) to the mixed solution of 2g high-purity phosphatidylcholine, 4g linoleic acid and 10mL n-hexane, add 5% TL IM lipase (Lipase: high-purity phosphatidylcholine+linoleic acid, m / m), the above mixture is premixed under an ultrasonic continuous flow cell disruptor, the premixing power is 500W, ultrasonic 5s / intermittent 5s, repeat 3 times, A total of 15s of ultrasound. The above mixture was stirred and reacted at 50° C. for 1 h to obtain a modified mixture of phosphatidylcholine and fatty acid. After the phosphatidylcholine mixture was dried at low temperature to remove n-hexane, it was eluted three times with acetone, and the acetone insoluble matter was collected, wherein the content of dilinole...

Embodiment 3

[0045] This example provides a method for preparing high-purity dilinoleoylphosphatidylcholine and its product through enzymatic modification combined with reversed-phase column chromatographic separation.

[0046] Add 3.5% (moisture: high-purity phosphatidylcholine + linoleic acid, m / m) to 20g high-purity phosphatidylcholine, 40g linoleic acid and 100mL n-hexane mixed solution, add 5% TL IM lipase (Lipase: high-purity phosphatidylcholine+linoleic acid, m / m), the above mixture is premixed under an ultrasonic continuous flow cell disruptor, the premixing power is 800W, ultrasonic 5s / intermittent 5s, repeat 10 times, A total of 95s of ultrasound. The above mixture was stirred and reacted at 50° C. for 1 h to obtain a modified mixture of phosphatidylcholine and fatty acid. After the phosphatidylcholine mixture was dried at low temperature to remove n-hexane, it was eluted with acetone three times to collect the acetone insoluble matter, wherein the content of dilinoleoylphosphat...

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Abstract

The invention belongs to the field of phospholipid processing, and particularly relates to a method for preparing high-purity dilinoyl phosphatidylcholine through combination of enzymatic modification and reversed-phase column chromatography separation and a product of the high-purity dilinoyl phosphatidylcholine. According to the method, high-purity phosphatidylcholine (PC) and linoleic acid are used as raw materials, an ultrasonic continuous flow cell crusher is adopted to pre-mix a substrate and lipase before lipase reaction, the reaction product of the system is subjected to acetone desolvation to obtain an acetone insoluble substance rich in dilinoyl phosphatidylcholine, the acetone insoluble substance is added to a chromatographic column with C18 as a stationary phase, and eluting is carried out with low-carbon alcohol or a low-carbon alcohol-water solution to obtain a DLPC product with the purity of more than 95% and the yield of more than 65%. The preparation method disclosed by the invention is mild in condition, efficient, simple and controllable in operation and easy to realize industrial production, and the dilinoyl phosphatidylcholine obtained by the method is high in purity and high in yield and can be an important source of high-purity dilinoyl phosphatidylcholine in the medicine and health care industry.

Description

technical field [0001] The invention belongs to the field of phospholipid processing, and more specifically relates to a method for preparing high-purity dilinoleoylphosphatidylcholine and its product through enzymatic modification combined with reverse-phase column chromatography separation. Background technique [0002] Dilinoleoylphosphatidylcholine (DLPC) belongs to polyene phospholipids and is the main active ingredient of soybean lecithin (PC). DLPC is the basic component of the human cell membrane, and it is of great significance to maintain the cell activity of the biofilm and the normal metabolism of the substrate. DLPC has the functions of treating liver cirrhosis, preventing fibrosis, protecting and treating liver damage, etc. It also helps to reduce blood and liver total cholesterol and low-density lipoprotein cholesterol levels, and increase high-density lipoprotein to improve its lipid metabolism. [0003] As a lipase, Lipozyme TL IM is widely used in lipoenzy...

Claims

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Application Information

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IPC IPC(8): C12P7/64C12P13/00C07F9/10
CPCC12P7/6481C12P7/6454C12P13/001C07F9/103
Inventor 从艳霞张维农齐玉堂刘思敏张海龙
Owner WUHAN POLYTECHNIC UNIVERSITY
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