A method for extracting and purifying Solanum nigrum refined polysaccharide with anti-tumor activity

The technology of anti-tumor activity and purification method is applied in the field of extraction and purification of Solanum nigrum refined polysaccharide, which can solve the problems of affecting drug quality and safety, easy residual of toxic chloroform, reducing polysaccharide activity, etc., and achieves low cost, strong anti-tumor activity, The effect of increasing the content of active ingredients

Active Publication Date: 2017-09-29
JIAXING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the traditional method, the Solanum nigrum crude polysaccharide is usually extracted by using water as the solvent; however, the Solanum nigrum crude polysaccharide obtained by using water as the solvent still contains impurities such as protein and pigment, and the existence of these components will affect the quality and safety of the drug
When the commonly used deproteinization method adopts the Sevag method, the efficiency is not high, the operation needs to be repeated many times, the cost is high, the toxic chloroform is easy to remain, and the polysaccharide is easy to be lost; when the hydrochloric acid method is used, the effect is good, but strong acidity will degrade part of the polysaccharide , reduce polysaccharide activity; trifluorotrichloroethane method should not be used in large quantities because of the low boiling point of the solvent and its volatility, and the enzymatic deproteinization will be limited by the specificity of the enzyme

Method used

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  • A method for extracting and purifying Solanum nigrum refined polysaccharide with anti-tumor activity
  • A method for extracting and purifying Solanum nigrum refined polysaccharide with anti-tumor activity
  • A method for extracting and purifying Solanum nigrum refined polysaccharide with anti-tumor activity

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Experimental program
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Effect test

Embodiment 1

[0024] The preparation of embodiment 1 Solanum nigrum refined polysaccharide

[0025] (1) Extraction of Solanum nigrum crude polysaccharide:

[0026] Pretreatment: Weigh 100g Solanum nigrum decoction pieces (purchased from Zhejiang Jiaxin Pharmaceutical Co., Ltd.) and dry them at 60°C for 6-8h, crush them with a grinder and pass through a 40-mesh sieve.

[0027] Degreasing: Add the Solanum nigrum powder in the above step to petroleum ether (400ml) with a solid-to-liquid ratio of 1:4, heat and reflux for extraction in a water bath at 50°C for 2 hours, extract twice in total, and filter the liquid to leave residue. Add 2 times the amount of 80% ethanol (200ml) to the remaining slag, heat and reflux in a water bath at 80°C for 1 hour, twice in total, filter through double-layer gauze, remove the liquid and leave the slag, spread the drug slag flat in a ventilated place to dry .

[0028] Polysaccharide extraction: put the dregs dried in a ventilated place in a flash extractor (J...

Embodiment 2

[0033] The preparation of embodiment 2 Solanum nigrum refined polysaccharide

[0034] (1) Extraction of Solanum nigrum crude polysaccharide:

[0035] Weigh 100g Solanum nigrum decoction slices and dry them at 60°C for 6-8h, pulverize them through a 40-mesh sieve, add petroleum ether with a material-to-liquid ratio of 1:4, heat and reflux in a water bath at 50°C for 2 hours, extract twice in total, filter and discard the liquid For dregs, add 2 times the amount of 80% ethanol, heat and reflux in a water bath at 80°C for 1 hour, twice in total, filter through double-layer gauze, remove the liquid and leave the dregs, spread the dregs flat in a ventilated place to dry. Then put the dregs in the flash extractor, add 30 times the amount of water, and perform flash extraction for 2 minutes with an extraction voltage of 80V, twice in total, filter with 4 layers of gauze, and centrifuge the filtrate at 3000r / min for 10 minutes to obtain the water extract. Concentrate under reduced pr...

Embodiment 3

[0038] The antitumor effect of embodiment 3 Solanum nigrum refined polysaccharide

[0039] (1) In vitro anti-tumor effect of Solanum nigrum refined polysaccharides

[0040] The refined Solanum nigrum polysaccharide prepared in Example 1 was prepared into a stock solution of 10 mg / mL, sterilized by filtration with a 0.22 μm sterile filter membrane, and then stored at 4° C. for later use. Before use, dilute with sterile water for injection to a series concentration of 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6, 1.8, 2.0mg / ml.

[0041] Take H22 mice with liver cancer that have been passaged intraperitoneally for 7 days, and kill the mice by cervical dislocation. Aseptically extract ascites, put it in a centrifuge tube, and centrifuge at 1000r / min for 5min. Carefully absorb the upper layer of cells in the pellet, add PBS to blow and wash twice, Centrifuge at 1000r / min for 5min. Blow and blow with RPMI1640 complete culture solution containing 10% calf serum, dilute and adjust the con...

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Abstract

The invention discloses a novel, relatively safe, high-yield, high-purity and anti-tumor activity of the extraction and purification process route of Solanum nigrum refined polysaccharide. Ethanol degreasing, then flash extraction, concentrated alcohol precipitation, deproteinized with trichloroacetic acid, activated carbon decolorization, secondary alcohol precipitation, and finally dried to obtain Solanum nigrum refined polysaccharide with a yield of 2.554%. The product has inhibitory effect on the growth of H22 hepatoma cells in vitro, with a cell inhibition rate of 35.80-85.4% at a concentration of 1.4-1.8 mg / mL, and has an inhibitory effect on the tumor growth of H22 tumor-bearing mice, with an inhibition rate of 37.73%-42.60% .

Description

technical field [0001] The invention belongs to the technical field of medicine extraction and purification, in particular to an extraction and purification method of Solanum nigrum refined polysaccharide with antitumor activity. Background technique [0002] Solanum nigrum (Solanum nigrum L.) belongs to the Solanaceae plant. It is cold in nature, bitter in taste, slightly sweet, slightly toxic, and returns to the lung and kidney meridians. Solanum nigrum, as a traditional Chinese herbal medicine, is distributed all over my country. It has the effects of clearing away heat and detoxification, reducing swelling and dissipating stagnation, and reducing inflammation and diuresis. Solanum nigrum polysaccharide has obvious growth inhibitory effect on sarcoma S180, gastric cancer MFC, cervical cancer U14, etc., can increase the weight of immune organs spleen and thymus, and can significantly prolong the life cycle of tumor-bearing mice without toxic or side effects Significantly ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/00A61K31/715A61P35/00
Inventor 黄越燕朱琦峰
Owner JIAXING UNIV
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