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Method for separating and purifying microcystin by utilizing series-connected solid phase extraction columns

A tandem solid-phase extraction and microcystin technology, which is applied in the preparation methods of peptides, chemical instruments and methods, organic chemistry, etc., can solve the problems of inability to continue, many separation and purification steps, and large loss of toxins, so as to save costs. , low cost, the effect of saving organic solvents

Inactive Publication Date: 2011-02-16
INST OF AQUATIC LIFE ACAD SINICA
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  • Application Information

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Problems solved by technology

However, as mentioned above, the above-mentioned method still has some defects more or less, which are specifically manifested in: (1) the amount of toxin obtained at one time is generally low; Low yield; (3) Pigment and other impurities are still the main factors affecting the toxin separation and purification process, and this problem has not been well resolved

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  • Method for separating and purifying microcystin by utilizing series-connected solid phase extraction columns
  • Method for separating and purifying microcystin by utilizing series-connected solid phase extraction columns
  • Method for separating and purifying microcystin by utilizing series-connected solid phase extraction columns

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Embodiment Construction

[0032] A method for separating and purifying microcystins by means of a tandem solid-phase extraction column, the steps of which are as follows:

[0033] 1. Algae powder material: Harvested wild cyanobacteria or indoor cultured Microcystis aeruginosa ( Microcystis aeruginosa ), freeze-dried to make dry algae powder, and stored at -20 oC.

[0034] 2. Preparation of solid phase extraction (SPE) cartridges: In an SPE column (10 mL, inner diameter 15 mm), load 1 g of C18 filler (15-35 μm, ODS or Sunchrom spherical silica gel, Beijing Jinouya Science and Technology Development Co., Ltd. company), press it with gaskets and save it for later use. The SPE column was activated with 30 mL methanol and 30 mL water before use. The series connection of the SPE column is connected by a stainless steel needle pierced rubber stopper. The ratio of the filler weight (0.5 or 1.0 or 1.5 or 2 g) of the first SPE cartridge to the weight of the algae flour to be treated is 1:10 to 1:40.

[0035]...

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Abstract

The invention discloses a method for separating and purifying microcystin by utilizing series-connected solid phase extraction columns. The method comprises the following steps of: A, preparing an algae powder material, namely harvesting wild bloom-forming cyanobacteria or microcystis aeruginosa cultured indoors, cooling and drying to prepare dried algae powder; B, preparing the solid phase extraction columns, namely putting C18 fillers into Solid Phase Extraction (SPE) columns; C, extracting algae toxin, namely weighing the dried algae powder, adding methanol according to a proportion, putting on a shaking table and oscillating at room temperature; D, removing impurities, namely adding a supernatant and an organic extractant into a separating funnel according to a proportion, mixing and uniformly shaking, standing, and after delamination, removing the organic extractant; E, separating and purifying the algae toxin, namely after pretreatment, enriching sample solution by using pre-activated series-connected SPE columns, and eluting the toxin by using 20 to 50 percent methanol aqueous solution; and F, performing Methyl Cellulose (MC) detection. The method has good separation effect, large loading amount, is easy and convenient to operate and low in cost, can obtain two microcystins with the weight of more than or equal to 10 mg and higher purity each time, and the High Performance Liquid Chromatography-Ultra Violet / Diode Array Detector (HPLC-UV / DAD) detection can reach 85 to 90 percent.

Description

technical field [0001] The invention relates to two methods for separating and purifying microcystins, which belong to the technical field of biochemical separation, and more specifically relate to a method for separating and purifying microcystins by means of tandem solid-phase extraction columns, which is especially suitable for blooming cyanobacteria in the wild or indoors. Purification of microcystins MC-LR and MC-RR from cultured Microcystis algae. Background technique [0002] The eutrophication of water bodies is becoming more and more serious, and the frequent outbreak of harmful cyanobacteria blooms has become a global ecological environmental problem. Cyanotoxins are secondary metabolites produced by cyanobacteria blooms. Studies have shown that cyanotoxins not only cause deterioration of aquatic ecosystems, but may also threaten human health. my country is one of the countries with the highest intensity, highest frequency and longest duration of cyanobacteria blo...

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Application Information

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IPC IPC(8): C07K1/16C07K14/405
Inventor 肖邦定吴幸强李仁辉
Owner INST OF AQUATIC LIFE ACAD SINICA
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