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.
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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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