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Preparation method for separating and purifying EPA and DHA

A separation, purification and semi-preparation technology, applied in the field of separation and purification of EPA and DHA, can solve the problems of high cost, complicated operation, low purity, etc., and achieve the effect of low production cost and simple post-treatment

Inactive Publication Date: 2014-05-14
JIANGSU HANBON SCI & TECH CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a new method for preparing high-purity DHA and EPA monomers by using a semi-preparative SFC separation system for the problems of high cost, complicated operation and low purity in the process of separating and purifying DHA and EPA. , the method includes the following steps:

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] 1. Take the crude fish oil extract containing a mixture of EPA and DHA, dissolve it in methanol, the sample solubility is 5mg / ml, put it in a filter device, and filter to remove solid particles;

[0011] 2. Inject the sample solution into the semi-preparative SFC separation system, the chromatographic column size is Φ10×250mm, C18 packing, the particle size is 5~45um, the sample volume is 10mg, CO 2 The flow rate is 0.8~3.0ml / min, methanol is used as modifier, the flow rate is 0~0.5ml / min, the temperature is 40°C, and the back pressure is 9~15MPa. The detection wavelength of the ultraviolet photometric detector used is 210nm, and the fractions with retention times of 5-27min (corresponding to EPA) and 6-30min (corresponding to DHA) are collected, and the purity is ≥99.8% by HPLC analysis.

Embodiment 2

[0013] 1. Take the crude fish oil extract containing a mixture of EPA and DHA, dissolve it in methanol, the sample solubility is 10mg / ml, put it in a filter device, and filter to remove solid particles;

[0014] 2. Inject the sample solution into the semi-preparative SFC separation system, the chromatographic column size is Φ10×250mm, C18 packing, the particle size is 5~45um, the sample volume is 20mg, CO 2 The flow rate is 1.0~5.0ml / min, methanol is used as modifier, the flow rate is 0~1.0ml / min, the temperature is 40°C, and the back pressure is 9~15MPa. The detection wavelength of the ultraviolet photometric detector used is 210nm, and the fractions with retention times of 5-25min (corresponding to EPA) and 6-30min (corresponding to DHA) are collected, and the purity is ≥99.5% by HPLC analysis.

Embodiment 3

[0016] 1. Take the crude fish oil extract containing a mixture of EPA and DHA, dissolve it in methanol, the sample solubility is 10mg / ml, put it in a filter device, and filter to remove solid particles;

[0017] 2. Inject the sample solution into the semi-preparative SFC separation system, the chromatographic column size is Φ25×250mm, C18 filler, the particle size is 5~45um, the sample volume is 50mg, CO 2 The flow rate is 1.0~10.0ml / min, methanol is used as modifier, the flow rate is 0~2.0ml / min, the temperature is 40°C, and the back pressure is 10~18MPa. The detection wavelength of the ultraviolet photometric detector used is 210nm, and the fractions with retention times of 5-25min (corresponding to EPA) and 6-30min (corresponding to DHA) are collected, and the purity is ≥99.7% by HPLC analysis.

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Abstract

The invention relates to a preparation method for separating and purifying EPA and DHA. The method comprises the following steps: employing a fish oil extract product crude product containing a mixture of EPA and DHA as a raw material, acquiring an appropriate mobile phase and a chromatographic column separating condition through a SFC optimization condition, and is amplified to a semi-preparation SFC separating system for further to obtain a EPA and DHA monomer. The preparation method for separating and purifying EPA and DHA has the advantages that the EPA and DHA mixture crude product extracted in the fish oil directly use the semi-preparation SFC separating system to obtain the EPA and DHA monomer, so that good separating and purifying effects are reached, the operation is simple, the period is short, the efficiency is high, the purity can reach more than 99.2%, and individual impurity can be controlled below 0.2%.

Description

technical field [0001] The invention relates to the technical field of extraction and separation of biomolecules such as fat and oil in marine organisms, in particular to a preparation method for separating and purifying EPA and DHA. Background technique [0002] In 1978, Dyerberg et al. reported for the first time that the low incidence of cardiovascular disease in Eskimos in Greenland was related to eating fish, and EPA and DHA in fish oil attracted people's attention. A large number of studies have confirmed that EPA and DHA have the functions of lowering blood fat, anti-platelet aggregation, alleviating thrombosis, and protecting cerebrovascular. In particular, DHA can also increase brain function, improve discrimination, and enhance memory and attention. In the brain and retina, DHA can also act on the structure and function of nerve cell nuclei and retinal photoreceptors through the activity of annexin, which helps the growth and development of fetal and infant brain v...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C57/03C07C51/47
CPCC07C51/47C07C57/03Y02P20/54
Inventor 王冰冰刘根水张大兵
Owner JIANGSU HANBON SCI & TECH CO
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