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Extracting and purifying process for scorpion toxins

A technology of scorpion toxin and process, which is applied in the field of deep processing of pharmaceutical raw materials, can solve the problem of low survival rate of small scorpions, achieve high purity and avoid waste

Inactive Publication Date: 2018-01-12
NANJING XIANCAOTANG BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Pregnant female scorpions can still give birth to young scorpions, but the survival rate of young scorpions is much lower than that of unvenomed scorpions

Method used

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  • Extracting and purifying process for scorpion toxins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: A kind of extraction and purification process of scorpion toxin

[0027] A process for extracting and purifying scorpion toxin, said process comprising the following steps:

[0028] (1) Collect the adult scorpions bred to adults in March every year, and anesthetize the adult scorpions with ethanol at 4°C, then cut off the palps, put them into a centrifuge tube, and freeze them at -16°C 1 hour; anesthesia treatment refers to immersing the adult scorpion in ethanol or ether, or spraying the adult scorpion with ethanol or ether.

[0029] (2) Take the frozen sample in step (1), add 1 times the mass of ethanol with a volume fraction of 75% to the sample, grind it into a slurry, centrifuge at 3000r / min for 5min, and take the supernatant.

[0030] (3) Add 1 volume of absolute ethanol to the supernatant obtained in step (2), mix well up and down, then centrifuge at 12000r / min for 15min, and take the supernatant protein solution.

[0031] (4) Add proteinase K to ...

Embodiment 2

[0034] Embodiment 2: A kind of extraction and purification process of scorpion toxin

[0035] A process for extracting and purifying scorpion toxin, said process comprising the following steps:

[0036] (1) Collect the adult scorpions bred to adulthood in June every year, anesthetize the adult scorpion with ether at 4°C, then cut off the limbs, put them into a centrifuge tube, and freeze them at -25°C 4 hours; the anesthesia treatment refers to immersing the adult scorpion in ethanol or ether, or spraying the adult scorpion with ethanol or ether.

[0037] (2) Take the frozen sample in step (1), add 2.5 times the mass of ethanol with a volume fraction of 80% to the sample, grind it into a slurry, centrifuge at 4000r / min for 8min, and take the supernatant.

[0038] (3) Add 1.5 times the volume of absolute ethanol to the supernatant obtained in step (2), mix well up and down, then centrifuge at 13000r / min for 20min, and take the supernatant protein solution.

[0039] (4) Add pr...

Embodiment 3

[0042] Embodiment 3: A kind of extraction and purification process of scorpion toxin

[0043] A process for extracting and purifying scorpion toxin, said process comprising the following steps:

[0044] (1) Collect the adult scorpions bred to adults in August every year, anesthetize the adult scorpions with ethanol at 6°C, then cut off each palp, put them into a centrifuge tube, and freeze them at -20°C 3 hours; anesthesia treatment refers to immersing the adult scorpion in ethanol or ether, or spraying the adult scorpion with ethanol or ether.

[0045] (2) Take the frozen sample in step (1), add 2.5 times the mass of ethanol with a volume fraction of 75% to the sample, grind it into a slurry, centrifuge at 3500r / min for 8min, and take the supernatant.

[0046] (3) Add 1.5 times the volume of absolute ethanol to the supernatant obtained in step (2), mix well up and down, then centrifuge at 13,500 r / min for 15 minutes, and take the supernatant protein solution.

[0047] (4) Add...

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Abstract

The invention discloses an extracting and purifying process for scorpion toxins. The extracting and purifying process comprises the following steps: performing anesthesia disposal on scorpions throughalcohol or ethyl ether, freezing to take a sample, adding alcohol into the sample, grinding the sample into slurry, and centrifuging the slurry to obtain liquid supernatant; adding absolute ethyl alcohol into the liquid supernatant, uniformly mixing, and centrifuging to take upper-layer protein fluid; adding protease K into the upper-layer protein fluid, adding absolute ethyl alcohol of equal volume and a statured sodium chloride solution after treatment is ended, and centrifuging to take an upper-layer solution after uniformly mixing; adding a sodium sulfate aqueous solution into the upper-layer solution, adding absolute ethyl alcohol after uniformly mixing, and centrifuging to collect the upper-layer solution after uniformly mixing; and freeze-drying the upper-layer solution to preparefreeze-dried powder. The extracting and purifying process adopts analytical chemical means to directly extract scorpion toxins from pedipalps, and can avoid waste caused by incomplete extracting of the existing method.

Description

technical field [0001] The invention relates to an extraction and purification process of active ingredients, in particular to an extraction and purification process of scorpion toxin, and belongs to the technical field of deep processing of pharmaceutical raw materials. Background technique [0002] Scorpion is a commonly used traditional Chinese medicine, which has the effect of quenching wind, relieving spasm and attacking poison. Modern pharmacological studies have proved that: scorpion has antihypertensive and anticonvulsant effects; the polypeptide contained in it has antiepileptic effects. Scorpion venom can cause respiratory paralysis, increase the excitability of mice, and make them shed tears, excessive salivation, and then convulsions; the alcohol extract still has the effect of inhibiting cysticercosis of pigs. Studies on antifungal effects have not been reported. Clinically, try to make scorpion ointment and tincture to treat skin ringworm and beriberi, and ha...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/30
Inventor 史月龙曾宋君吴英亮刘文斌吴坤林汤静
Owner NANJING XIANCAOTANG BIOLOGICAL TECH CO LTD
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