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Method for preparing microcystin-LR

A microcystin, semi-preparation technology, applied in the direction of algae/bryopeptide, peptide source, etc., can solve the problem of immature pure isomers

Inactive Publication Date: 2011-01-26
UNIV OF JINAN
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, methods for the large-scale preparation of pure isomers are not yet mature

Method used

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  • Method for preparing microcystin-LR
  • Method for preparing microcystin-LR

Examples

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Effect test

Embodiment 1

[0013] In a 150 mL Erlenmeyer flask, add 2 g of dry cyanobacteria powder collected from Dianchi Lake and 40 mL of 65% (v / v) acetonitrile / water solution, stir magnetically for 60 min at room temperature, oscillate ultrasonically for 20 min, and freeze in liquid nitrogen for 10 min. After thawing, centrifuge at 10,000 rpm for 10 min, and remove the supernatant. The residue was extracted twice using the above steps, and the supernatants were combined. Filtrate with a 0.45 μm microfiltration membrane to obtain crude microcystin. Preliminary purification of the crude product by solid-phase extraction: activate the solid-phase extraction column with 10mL 100% methanol and 10mL ultrapure water successively, and flow the crude extract through the solid-phase extraction column at a flow rate of 1mL / min for concentration and enrichment, 5mL 40 % (v / v) methanol / water solution was rinsed to remove impurities, 10mL 70% (v / v) acetonitrile / water solution was eluted and sample components wer...

Embodiment 2

[0015] In a 150 mL Erlenmeyer flask, add 4 g of dry cyanobacteria powder collected from Dianchi Lake and 100 mL of 65% (v / v) acetonitrile / water solution, stir magnetically for 90 min at room temperature, oscillate ultrasonically for 20 min, and freeze in liquid nitrogen for 10 min. After thawing, centrifuge at 10,000 rpm for 10 min, and remove the supernatant. The residue was extracted twice using the above steps, and the supernatants were combined. Filtrate with a 0.45 μm microfiltration membrane to obtain crude microcystin. Preliminary purification of the crude product by solid-phase extraction: sequentially activate the solid-phase extraction column with 10mL 100% methanol and 10mL ultrapure water, and flow the crude extract through the solid-phase extraction column at a flow rate of 1mL / min for concentration and enrichment, 5mL40% (v / v) methanol / water solution was rinsed to remove impurities, 10mL 80% (v / v) acetonitrile / water solution was eluted and sample components were...

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Abstract

The invention relates to a method for preparing microcystin-LR. In the method, blue-green algae obtained from Dianchi in Yunnan are taken as raw materials. The method comprises the following steps of: performing extraction by using 50 to 90 percent (v / v) aqueous solution of acetonitrile to obtain crude microcystin, and removing impurities by membrane filtration; performing primary purification and enrichment by adopting solid phase extraction; and performing separation and purification by semi-preparative high performance liquid chromatography. In the pre-treatment step of the method, most of the impurities are removed, so the purification difficulty of the high performance liquid chromatography is reduced; the method has the advantages of simplicity, easy operation and relatively less microcystin loss; and the purity of the finally obtained microcystin-LR is 91.35 percent which is relatively higher.

Description

technical field [0001] The invention relates to a method for preparing microcystin-LR from cyanobacteria. The method is to extract a microcystin isomer mixture from natural cyanobacteria, and obtain the microcystin isomer-LR after separation and purification. Background technique [0002] The aggravation of eutrophication in the water body causes the massive reproduction of algae. Many freshwater lakes in my country, such as Dianchi Lake in Yunnan, Taihu Lake in Jiangsu and Chaohu Lake in Anhui, have experienced varying degrees of cyanobacteria "bloom" phenomenon. It has become one of the major environmental issues of global concern. With the deepening of research on microcystins, the demand for pure isomers of microcystins is also increasing. [0003] At present, there is no mature technology for the preparation of pure microcystin isomers in my country, and the required pure microcystin isomers are all imported from abroad. With the international inclusion of microcystin...

Claims

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Application Information

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IPC IPC(8): C07K14/405C07K1/36C07K1/20C07K1/14
Inventor 刘思全王立国贾瑞宝陈永芳
Owner UNIV OF JINAN
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