Method for separating and purifying DHA ethyl ester from DHA oil fermented by microorganisms

A microbial fermentation, separation and purification technology, which is applied in the field of separation and purification to prepare DHA ethyl ester, can solve the problems of high cost and unsatisfactory purification effect, and achieve the effects of ensuring quality, improving DHA purification degree, and high vacuum degree

Active Publication Date: 2015-11-25
JIANGSU ZHONGBANG PHARMA
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a method for separating and purifying DHA ethyl ester from DHA oil produced by microbial fermentation, which solves the technical problems of high cost and unsatisfactory purification effect of a single application of a certain separation and purification method

Method used

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  • Method for separating and purifying DHA ethyl ester from DHA oil fermented by microorganisms

Examples

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Embodiment 1

[0027](1) Ethyl esterification of DHA oil: Dissolve 4g KOH in 160g ethanol, mix 500g of DHA oil (DHA accounts for 42.23%) produced by the fermentation of Cryptodinium algae with KOH-ethanol solution, under the condition of nitrogen filling, Heated to reflux in a water bath at 60°C for 0.5h. Cool to room temperature under the condition of nitrogen filling, after static layering, take the upper layer of DHA ethyl ester, distill under reduced pressure at 60°C and -0.1Mpa to recover ethanol, then use 40°C warm water to wash repeatedly until the washing water is neutral, after oil-water separation , the oil phase passes through anhydrous Na 2 SO 4 Filter and dry to obtain 464.15 g of ethyl-esterified DHA oil.

[0028] (2) Urea embedding - primary separation of gradient freeze crystallization: Prepare a urea ethanol solution with a mass fraction of 20%, and heat and reflux in a water bath at 65°C until the urea is completely dissolved. After mixing the ethyl-esterified DHA oil an...

Embodiment 2

[0034] (1) Ethylification of DHA oil: Dissolve 7.5g KOH in 500g ethanol, and fully mix 500g of DHA oil (DHA accounts for 42.23%) produced by the fermentation of Cryptodinium algae with KOH-ethanol solution. Heated to reflux in a water bath at 50°C for 4h. Cool to room temperature under the condition of nitrogen filling, after static layering, take the upper layer of DHA ethyl ester, distill under reduced pressure at 60°C and -0.1Mpa to recover ethanol, then use 40°C warm water to wash repeatedly until the washing water is neutral, after oil-water separation , the oil phase was filtered and dried by anhydrous Na2SO4 to obtain 472.1 g of ethyl-esterified DHA grease.

[0035] (2) Urea embedding - primary separation of gradient freeze crystallization: prepare a urea ethanol solution with a mass fraction of 30%, and heat and reflux in a water bath at 70°C until the urea is completely dissolved. Ethyl-esterified DHA oil and 30% urea ethanol were mixed evenly at a mass ratio of 1:10...

Embodiment 3

[0043] (1) Ethyl esterification of DHA oil: Dissolve 5g KOH in 150g ethanol, and fully mix 500g of mixed oil (DHA accounts for 41.24%) produced by Schizochytrium fermentation with KOH-ethanol solution. Heated to reflux in a water bath at 70°C for 0.5h. Cool to room temperature under the condition of nitrogen filling, after standing for stratification, take the fatty acid ethyl ester in the upper layer, distill and recover ethanol under reduced pressure at 60°C and -0.1Mpa, wash with warm water at 40°C repeatedly until the washing water is neutral, and separate the oil and water , the oil phase passes through anhydrous Na 2 SO 4 Filter and dry to obtain 474.8 g of ethyl-esterified DHA oil.

[0044] (2) Urea embedding - primary separation of gradient freeze crystallization: Prepare a urea ethanol solution with a mass fraction of 20%, and heat and reflux in a water bath at 65°C until the urea is completely dissolved. Mix the ethyl-esterified DHA oil and 20% urea-ethanol soluti...

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Abstract

The invention relates to a method for separating and purifying DHA (docosahexaenoic acid) from DHA oil produced by microbial fermentation. The method for separating and purifying DHA from DHA oil produced by microbial fermentation comprises the following steps: (1) taking the DHA oil, and carrying out esterification; (2) carrying out primary separation on esterified DHA oil by adopting a urea embedding-gradient freezing crystallization method; (3) carrying out molecular distillation on primarily separated fatty acid ethyl ester and then separating, so that high-purity DHA ethyl ester is prepared. The method for separating and purifying DHA from DHA oil produced by microbial fermentation is formed on the basis of coupling of multiple separation and purification means, DHA purification is obviously improved, DHA oil produced by crypthecodinium cohnii is taken as a raw material, and DHA ethyl ester with the purity more than 98% can be prepared; DHA oil produced by schizochytrium limacinum is taken as the raw material, and a product with the content of DHA ethyl ester and DPA ethyl ester more than 98% can be obtained.

Description

technical field [0001] The invention belongs to the field of biochemical industry, and relates to a method for separating and purifying DHA ethyl ester from DHA oil produced and fermented by microorganisms. Background technique [0002] Docosahexaenoic Acid (DHA) is an important n-3 long-chain polyunsaturated fatty acid, commonly known as brain gold, which can promote the development of infant brain and vision, prevent and treat cardiovascular and cerebrovascular diseases, Prevention and treatment of senile dementia, inhibition of tumor and other physiological effects. At present, DHA is mainly used in infant formula food and related health products, and pharmaceutical products based on its special physiological effects are under intense research and development. Most of these drugs are derivatives of DHA and combinations of DHA and other drugs. Norwegian Pronova Biopharmaceutical Company has developed more than 26 kinds of DHA derivatives, and has carried out relevant phy...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07C69/587C07C67/08C07C67/48C07C67/54C11C3/04C11C1/10
CPCC07C67/08C07C67/48C07C67/52C07C67/54C11C1/10C11C3/04C07C69/587
Inventor 赵光荣吴剑平邢光才周颖李维思
Owner JIANGSU ZHONGBANG PHARMA
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