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Extraction method of pheretima thrombolysis component

An extraction method, the technology of Ceruleus cylindrica, is applied in blood diseases, extracellular fluid diseases, medical raw materials derived from leeches/worms, etc., to achieve high thrombolytic activity and good clinical application prospects

Active Publication Date: 2017-05-10
高努
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing extraction process is still completely effective in extracting the thrombolytic active ingredients of C.

Method used

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  • Extraction method of pheretima thrombolysis component
  • Extraction method of pheretima thrombolysis component
  • Extraction method of pheretima thrombolysis component

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 extraction method of the present invention

[0032] method of extraction:

[0033] (1) Quick-frozen fresh green worms, chopped with a medicine cutter until the length is 1-2cm, add 1-2 times of water (w / w), and add sodium benzoate 1-1.5‰ (w / w), at 38 Stir for 9 hours at ℃, the stirring speed is 120-180rmp, centrifuge for 10min, the centrifugation speed is 2500rpm, discard the precipitate, and collect the supernatant for later use;

[0034] (2) Removal of heavy metals and decolorization: take the supernatant, add 0.1-0.3% of polyaluminum chloride or quartz sand, stir for 10 minutes at a speed of 120-180 rpm, centrifuge for 10 minutes at a speed of 2500 rpm, discard the sediment, and collect the centrifugate ;

[0035] (3) Alcohol precipitation: add ethanol to the centrifugate to make the ethanol concentration of the mixed solution 75%, stir for 10 minutes to fully mix the solution, stir at a rotating speed of 120-180rmp, centrifuge for 10min at a centrifug...

Embodiment 2

[0072] The screening test of embodiment 2 extraction method of the present invention

[0073] 1. Extraction method

[0074] ① Stirring temperature and time in step (1)

[0075] Take quick-frozen fresh green worms, chop them with a medicine cutter, put the same amount into 6 insulated containers with stirring, add 2 times of water (w / w), add 1.5‰ of sodium benzoate (w / w), and store them at room temperature , 36, 37, 38, 39, 40°C temperature insulation, stirring, centrifugation for 10min at a speed of 2500rpm. The measurement was started from the first hour, and at the same time, 20 microliters of liquid were drawn from 6 heat preservation containers, and the thrombolytic activity was measured. Then, samples were taken every hour until the 11th hour.

[0076] ②Concentration of ethanol in step (2)

[0077] Take the supernatant obtained in step (1) of Example 1, divide it into 5 parts on average, add ethanol at room temperature to concentrations of 65, 70, 73, 75, and 80%, sti...

Embodiment 3

[0104] Embodiment 3 extraction method of the present invention

[0105] 1. Extraction method:

[0106] (1) Take the worm worm (purchased from the medicinal material market), chop it with a medicine cutter, add water 1-2 times (w / w), and add sodium benzoate 1-1.5‰ (w / w) at the same time, at 38 ° C Stir for 9 hours at a speed of 120-180rpm, centrifuge for 10min at a speed of 2500rpm, discard the precipitate, and collect the supernatant for later use;

[0107] (2) Remove heavy metals and decolorize: take the supernatant, add polyaluminum chloride or quartz sand 0.1-0.3%, stir at 120-180rmp, centrifuge for 10min at a speed of 2500rpm, discard the precipitate, and collect the centrifugate;

[0108] (3) Alcohol precipitation: add ethanol to the centrifugate to make the ethanol concentration of the mixture 75%, stir for 10 minutes to fully mix the solution, rotate at a speed of 120-180 rpm, centrifuge for 10 minutes at a speed of 2500 rpm, and collect the precipitate;

[0109] (4) ...

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Abstract

The invention discloses an extraction method of a pheretima thrombolysis component, comprising the following steps: (1) pretreatment: fetching a fresh pheretima product or quick frozen product, cutting or mincing, adding 1-2 times by weight of water, stirring at 36-40 DEG C for 7-10 h, centrifuging, removing sediment, and collecting a supernatant; and (2) ethanol precipitation: adding ethanol until concentration is 73-80%, uniformly mixing, centrifuging, collecting sediment, and drying. The invention also discloses an extract prepared by the above method. By the extraction method, the pheretima thrombolysis component can be effectively extracted. The obtained extract has higher thrombolysis activity than extracts prepared by existing extraction method. The product has a good clinical application prospect.

Description

technical field [0001] The invention relates to a method for extracting a thrombolytic component of Ectropis circatus. Background technique [0002] Animals of the genus Cyclochaetus have a body length of 230-245 mm and a body width of 7-12 mm. The dorsal foramen starts between the 12th and 13th segments. The back is purple-red or purple-gray. Annulus occupies 3 segments, located at segments 14-16, without setae. There are ring-shaped setae on the body, including Pheretima aspergillum (E.Perrier), Pheretima vulgaris Chen, Pheretimaguillelmi (Michaelsen), Pectinifera Michaelsen and Hubei Ringworm Pheretima hupeienis Michaelsen1895 and so on. [0003] Lumbritin, Lumbrofebrin (a derivative of tyrosine), Terrestro-lumbrolysim, Xanthine, Choline, Cholesterol, Fatty Acid Classes, lipid compounds, nucleic acid derivatives, various amino acids, vitamins and inorganic salts and other substances have thrombolytic effect. [0004] At present, the thrombolytic products prepared fr...

Claims

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Application Information

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IPC IPC(8): A61K35/62A61P7/02
CPCA61K35/62
Inventor 高努高让
Owner 高努
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