Detection method of authentic cordyceps sinensis

A detection method and sample technology, applied in the detection field of genuine Cordyceps sinensis, can solve problems such as difficulty in accurate identification and the like

Inactive Publication Date: 2014-10-01
四川新时康医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the current identification methods of Cordyceps sinensis are mostly identified by shape, size, color and ot...

Method used

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  • Detection method of authentic cordyceps sinensis
  • Detection method of authentic cordyceps sinensis
  • Detection method of authentic cordyceps sinensis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0113] Embodiment 1 Preparation of Cordyceps sinensis specific protein of the present invention

[0114] 1. Medicinal materials

[0115] The medicinal material of Cordyceps sinensis was purchased from Kangding County, Ganzi Prefecture, Sichuan Province, and was identified and confirmed by the chief pharmacist of Sichuan Food and Drug Inspection Institute. The medicinal materials are crushed (passed through a No. 2 sieve), and set aside.

[0116] 2. Preparation method

[0117] (1) Extraction: Take 0.2 g of the coarse powder of Cordyceps sinensis, put it in a pre-cooled mortar, add liquid nitrogen to quickly grind it into a fine powder, add pre-cooled PBS phosphate buffer saline (pH7.3, containing 0.2% mercaptoethanol, 0.5% Tween80) 2ml, ground and leached for 4 hours, transferred to a centrifuge tube, centrifuged for 30 minutes (8000×g, 4°C), scraped off the upper layer of lipids, and took the supernatant to obtain the total protein extract of Cordyceps, which was stored at -...

Embodiment 2

[0145] The detection method of embodiment 2 authentic product Cordyceps sinensis of the present invention

[0146] (1) Extraction: Take 0.2 g of the sample to be tested, put it in a pre-cooled mortar, add liquid nitrogen to quickly grind it into a fine powder, add pre-cooled PBS phosphate buffer (pH7.3, containing 0.2% mercaptoethanol, 0.5% Tween80 ) 2ml, grind and extract for 4 hours, transfer to a centrifuge tube, centrifuge for 30 minutes (8000×g, 4°C), scrape off the upper layer of lipid, and take the supernatant;

[0147] (2) TCA / acetone purification method: Take the supernatant of step (1), add 8 times the volume of -20°C pre-cooled 10% TCA acetone solution (containing 0.07% DTT), let stand at -20°C for 1 hour, centrifuge (8000 ×g, 5min), take the precipitate and wash it twice with pre-cooled acetone, centrifuge (8000×g, 5min) to obtain the precipitate, take the precipitate and add an appropriate amount of 2-DE hydration solution to dissolve, centrifuge at room temperatu...

Embodiment 3

[0156] Embodiment 3 The detection method of genuine product Cordyceps sinensis of the present invention

[0157] (1) Extraction: Take 0.2 g of the sample to be tested, put it in a pre-cooled mortar, add liquid nitrogen to quickly grind it into a fine powder, add pre-cooled PBS phosphate buffer (pH7.3, containing 0.2% mercaptoethanol, 0.5% Tween80 ) 2ml, grind and extract for 4 hours, transfer to a centrifuge tube, centrifuge for 30 minutes (8000×g, 4°C), scrape off the upper layer of lipid, and take the supernatant;

[0158](2) Readyprep2-D cleanup kit purification method:

[0159] ① Take 100 μl of the supernatant from step (1) and place it in a 1.5ml centrifuge tube, add 300 μl of precipitant 1, vortex to mix, and ice-bath for 15 minutes;

[0160] ② Add 300 μl precipitant 2, vortex to mix, high-speed centrifugation (12000×g, 5min), carefully absorb the supernatant, discard, and centrifuge again for 15-30s, try to absorb the supernatant as completely as possible, and discard;...

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Abstract

The invention discloses ten specific proteins of novel authentic cordyceps sinensis and also discloses a detection method of authentic cordyceps sinensis, namely a method for detecting whether a to-be-detected sample contains one or more of the ten proteins or not. The detection method of authentic cordyceps sinensis is high in accuracy, convenient in operation and low in cost; meanwhile, the novel proteins have a function of promoting the proliferation of lymphocyte and can be made into a drug for promoting the proliferation of lymphocyte, thus having good application prospects.

Description

technical field [0001] The invention relates to a detection method of authentic Cordyceps sinensis. Background technique [0002] Cordyceps sinensis is a complex of larva corpses and sporozoites of Cordyceps sinensis (Berk.) Sacc. parasitic on the larvae of Lepidoptera bat moths. It is commonly used clinically to treat lung and kidney diseases such as chronic cough and shortness of breath, impotence and spermatorrhea, hemoptysis due to labor cough, and weakness of the waist and knees. Modern pharmacological studies have proved that Cordyceps sinensis has pharmacological effects such as immune regulation, inhibition of tumor cell growth, protection of liver and kidney functions, promotion of cardiovascular and cerebrovascular circulation, and lowering of blood sugar, and has unique clinical curative effects. [0003] Due to the obvious effect of Cordyceps sinensis, special growth environment, scarce resources, and high price, there are a large number of adulterated and confu...

Claims

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Application Information

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IPC IPC(8): G01N27/447G01N1/28G01N1/34
Inventor 国锦琳陈璐彭成万德光
Owner 四川新时康医疗科技有限公司
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