Method for extracting and purifying shellfish toxin from toxin-producing algae culture solution

A shellfish toxin and culture medium technology, applied in the biological field, can solve the problems of low shellfish toxin content, unsatisfactory yield, complicated steps, etc., and achieve the effects of saving manpower and material resources, improving yield and purity, and simple extraction and purification steps.

Inactive Publication Date: 2018-03-23
上海泰坦科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the content of shellfish toxin obtained by the current extraction and purification m

Method used

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  • Method for extracting and purifying shellfish toxin from toxin-producing algae culture solution
  • Method for extracting and purifying shellfish toxin from toxin-producing algae culture solution

Examples

Experimental program
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Effect test

Embodiment 1

[0035] A method for extracting and purifying shellfish toxin from toxin-producing algae culture fluid, comprising the steps of:

[0036] (1) Add the toxin-producing algae into 2L natural seawater culture solution according to the wet weight of 50g, and filter the culture solution obtained after 48 hours of cultivation with a filter membrane with a pore size of 0.22μM. volume of ethanol for elution;

[0037] (2) Add the first extractant to the eluted solution and mix and let it stand for 20 minutes, then add the next extractant and mix and let it stand, add n-hexane, chloroform, ethyl acetate and n-butanol in turn to finally obtain the extract ;

[0038] (3) After collecting the extract, perform high-performance liquid chromatography separation and purification after normal phase silica gel column chromatography and reverse phase silica gel column chromatography to obtain shellfish toxin, collect the target chromatographic peak, and freeze-dry to obtain a white solid;

[0039...

Embodiment 2

[0041] A method for extracting and purifying shellfish toxin from toxin-producing algae culture fluid, comprising the steps of:

[0042](1) Add the toxin-producing algae into 2L natural seawater culture fluid according to the wet weight of 50g, and filter the culture fluid obtained after cultivating for 48h with a filter membrane with a pore size of 0.1 μM. times the volume of the filtrate for elution;

[0043] (2) Add the first extractant to the eluted solution and mix and let stand for 20min, then add the next extractant to mix and let stand, add n-chloroform, ethyl acetate and n-butanol in turn to finally obtain the extract;

[0044] (3) After collecting the extract, perform high-performance liquid chromatography separation and purification after normal phase silica gel column chromatography and reverse phase silica gel column chromatography to obtain shellfish toxin, collect the target chromatographic peak, and freeze-dry to obtain a white solid;

[0045] The eluent of th...

Embodiment 3

[0047] A method for extracting and purifying shellfish toxin from toxin-producing algae culture fluid, comprising the steps of:

[0048] (1) Add the toxin-producing algae into 2L of natural seawater culture fluid according to the wet weight of 50g, and filter the culture fluid obtained after cultivating for 48 hours with a filter membrane with a pore size of 0.65 μM. Doubling of the volume of the filtrate in acetone for elution;

[0049] (2) Add the first extractant to the eluted solution and mix and let it stand for 20 minutes, then add the next extractant and mix and let it stand, add n-hexane, chloroform, ethyl acetate and n-butanol in turn to finally obtain the extract ;

[0050] (3) After collecting the extract, perform high-performance liquid chromatography separation and purification after normal phase silica gel column chromatography and reverse phase silica gel column chromatography to obtain shellfish toxin, collect the target chromatographic peak, and freeze-dry to...

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Abstract

The invention relates to a method for extracting and purifying a shellfish toxin from a toxic algae culture solution. The method comprises four steps of filter membrane filtration, adsorption and desalting, extraction and column chromatography. An extraction agent is prepared from n-hexane, chloroform, ethyl acetate and n-butyl alcohol. Normal phase silica gel eluents are prepared from ethyl acetate, n-hexane and chloroform. Reversed phase silica eluents are prepared from water, n-butanol, acetonitrile and methanol. High performance liquid chromatography eluents are prepared from acetonitrileand methanol. By optimizing technology processes, the extraction agent and eluents are reasonably chosen, all steps are interacted collaboratively, product yield and purity are comprehensively improved, yield reaches 2.69g, and purity reaches to 99%. The method is simple and efficient, reduces costs, improves the yield and purity, and is beneficial to large-scale industrial application.

Description

technical field [0001] The invention relates to the field of biotechnology, and mainly relates to a method for extracting and purifying shellfish toxins from toxin-producing algae culture fluid. Background technique [0002] Marine toxins are toxic polymer compounds formed by the toxic algae in the ocean passing through the food chain to algae-eating fish, shrimp and shellfish, and accumulating in their bodies. There are many types of marine toxins, among which shellfish toxins are one of the most harmful. Shellfish toxins include paralytic shellfish toxins, diarrheal shellfish toxins, neurogenic shellfish toxins, and amnestic shellfish toxins. The hazards of shellfish toxins are sudden and widespread, and have the characteristics of strong toxicity and unpredictability. Due to their high toxicity, fast response, and no suitable antidote, many difficulties have been brought to the prevention and control. Shellfish toxins are harmful to the health of consumers. The serious ...

Claims

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Application Information

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IPC IPC(8): C07D493/22C07D487/20
Inventor 谢应波张庆张华徐肖冰罗桂云张维燕
Owner 上海泰坦科技股份有限公司
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