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Method for extracting and purifying microcytis toxin

A technology of microcystin and purification method, which is applied in the preparation methods of peptides, chemical instruments and methods, organic chemistry, etc., can solve the problems of low purity, unsatisfactory, long time consumption, etc., and achieve high purity and high safety. , the effect of high efficiency

Inactive Publication Date: 2003-03-12
INST OF AQUATIC LIFE ACAD SINICA
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Problems solved by technology

[0003] At present, the main way to develop pure toxins is to use combined multi-step purification technology, involving chromatographic technology, preparative or semi-preparative liquid chromatography, and flash chromatography. These two technologies can concentrate and separate and purify microcystins. The disadvantage of these three techniques is that the purity is not high enough to meet the requirements of standard samples; preparative or semi-preparative size exclusion chromatography, ion exchange chromatography, thin layer chromatography, the disadvantage of these three techniques is that they are only suitable for microgram level The preparation of cystic toxins takes a long time and the yield is low

Method used

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Embodiment Construction

[0021] 1. Take 50 grams of Microcystis aeruginosa dry powder, add 1700 grams of 5% acetic acid solution, stir for 40 minutes, centrifuge at 6000 rpm for 10 minutes, and take the supernatant;

[0022] 2. Add 200 grams of 100% methanol to the filter residue, stir for 40 minutes, centrifuge at 6000 rpm for 10 minutes, and take the supernatant;

[0023] 3. Repeat step 2 twice;

[0024] 4. Merge the supernatant extracted three times in step 2 and step 3, and evaporate and concentrate with a rotary evaporator;

[0025] 5. Mix the concentrated solution in step 4 with the supernatant in step 1, and pass through two microfilters of 1 μm and 0.45 μm in turn; then pass the mixed solution through a hollow fiber ultrafilter with a molecular weight cut-off of 6000;

[0026] 6. Rinse C with 10mL of 100% methanol and 10mL of pure water first and then 18 column, pass the mixed solution processed in step 5 through C 18 The column was washed with 20 mL of 20% methanol, and then the toxin was ...

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Abstract

A process for extracting microcystis toxin and purifying it includes immersing microcystis in solution of acetic acid and then in methanol, removing impurities, preparing raw toxin, separating and thin-layer chromatography. Its advantages are high efficiency, low cost, high safety and purity (more than 90%), and no environmental pollution.

Description

Technical field: [0001] The invention relates to a method for extracting and purifying toxin from Microcystis. Background technique: [0002] With the aggravation of water pollution and eutrophication, algal blooms in freshwater lakes and reservoirs in China have occurred frequently and on a large scale, and the situation is becoming more and more serious. Most of the cyanobacteria that form water blooms can release a large amount of algae toxins that pollute water bodies and the environment. Among them, the potential harm of the cyanobacteria Microcystis bloom and the microcystin produced by it to the water environment and human health has attracted widespread attention from all over the world. When microcystin enters the organism, it can inhibit protein phosphatase 1 and 2A. The most common manifestation of its toxic effect is to cause liver poisoning. It has been proved that microcystin is an inducer of liver cancer. The chemical structure of this type of toxin is relat...

Claims

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Application Information

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IPC IPC(8): C07G99/00C07K1/14G01N1/28
Inventor 宋立荣沈强陈伟甘南琴何振荣刘永定雷腊梅
Owner INST OF AQUATIC LIFE ACAD SINICA
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