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High performance liquid chromatography for rapidly identifying cordyceps sinensis and cordceps militaris

A technology of Cordyceps sinensis and high-performance liquid phase is applied in the field of rapid identification of Cordyceps sinensis and Cordyceps militaris by high-performance liquid phase method, which can solve the problems of high cost, complicated operation, incomplete technical means and the like, and achieves the effects of rapid and accurate identification and easy operation.

Inactive Publication Date: 2012-01-25
YUNNAN UNIV CHINESE HERBAL MEDICINE BIOLOGICAL RESOURCE INST
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(2) There are differences in the chemical composition of Cordyceps militaris and Cordyceps sinensis. From the existing reports, Cordyceps militaris contains some components of Cordyceps sinensis. Due to the current equipment conditions, analysis methods and other reasons, it cannot be said that the composition is completely clear. For example, many Trace chemical components have not been fully clarified with current technical means and research (Lin Qunying et al., Microbiology Communications, 2006, 33(4): 154-157)
Related patent applications include "Method for Identifying Cordyceps" (Chinese Invention Patent Application No.: 99106135.7) and "Gene Sequence and Method for Identifying Cordyceps" (Chinese Invention Patent Application No.: 00109084.4) invented by Xu Ruixiang. In order to distinguish Cordyceps sinensis from other Cordyceps by means of molecular biology, the operation is complicated, but it takes a long time and the cost is high, so it is not easy to popularize and apply

Method used

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  • High performance liquid chromatography for rapidly identifying cordyceps sinensis and cordceps militaris
  • High performance liquid chromatography for rapidly identifying cordyceps sinensis and cordceps militaris
  • High performance liquid chromatography for rapidly identifying cordyceps sinensis and cordceps militaris

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0031] Example 1 The distinction between Cordyceps sinensis and Cordyceps militaris in Yushu County, Qinghai

[0032] 1 Preparation of Cordyceps sinensis and Cordyceps militaris samples

[0033] Collected from Cordyceps sinensis in Yushu County, Qinghai, identified as authentic Cordyceps sinensis by Mr. Zang Mu, Kunming Institute of Botany, Chinese Academy of Sciences, crushed through a 60-mesh sieve, and set aside. The fruiting body of Cordyceps militaris purchased from Yiyuan Cordyceps cultivation field in Xinhui District, Jiangmen City, was crushed through a 60-mesh sieve, and set aside.

[0034] 2 Preparation of sample solution

[0035] 2.1 Preparation of Cordyceps militaris test solution: Weigh 0.2000g of Cordyceps militaris sample into a 5ml centrifuge tube, add 3ml of 20% methanol solution, ultrasonicate for 1h, centrifuge at 10000 rpm for 5min, take the supernatant into a 10ml volumetric flask, Repeat 2 more times. Combine the extracts and dilute to 10ml.

[0036] ...

example 2

[0041] Example 2 Distinguishing between Cordyceps sinensis and Cordyceps militaris in Baqing County, Tibet

[0042] 1 Preparation of Cordyceps sinensis and Cordyceps militaris samples

[0043] Collected from Cordyceps sinensis in Baqing County, Tibet, identified as authentic Cordyceps sinensis by Mr. Zang Mu, Kunming Institute of Botany, Chinese Academy of Sciences, crushed through a 60-mesh sieve, and set aside. The fruiting body of Cordyceps militaris purchased from Yiyuan Cordyceps cultivation field in Xinhui District, Jiangmen City, was crushed through a 60-mesh sieve, and set aside.

[0044] 2 Preparation of sample solution

[0045] 2.1 Preparation of Cordyceps militaris test solution: Weigh 0.2000g of Cordyceps militaris sample into a 5ml centrifuge tube, add 3ml of 20% methanol solution, ultrasonicate for 1h, centrifuge at 10000 rpm for 5min, take the supernatant into a 10ml volumetric flask, Repeat 2 more times. Combine the extracts and dilute to 10ml.

[0046] 2.2...

example 3

[0051] Example 3 Distinguishing Cordyceps sinensis cordycepin and Cordyceps militaris cordycepin in Yulong Snow Mountain, Lijiang City, Yunnan

[0052] 1 Preparation of Cordyceps sinensis and Cordyceps militaris samples

[0053] Collected from Yulong Snow Mountain Cordyceps sinensis in Lijiang City, Yunnan Province, identified as authentic Cordyceps sinensis by Mr. Zang Mu, Kunming Institute of Botany, Chinese Academy of Sciences, crushed through a 60-mesh sieve, and set aside. The fruiting body of Cordyceps militaris purchased from Yiyuan Cordyceps cultivation field in Xinhui District, Jiangmen City, was crushed through a 60-mesh sieve, and set aside.

[0054] 2 Preparation of sample solution

[0055] 2.1 Preparation of Cordyceps militaris test solution: Weigh 0.2000g of Cordyceps militaris sample into a 5ml centrifuge tube, add 3ml of 20% methanol solution, ultrasonicate for 1h, centrifuge at 10000 rpm for 5min, take the supernatant into a 10ml volumetric flask, Repeat 2 m...

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Abstract

The invention relates to a method for rapidly identifying cordyceps sinensis and cordceps militaris by adopting high performance liquid chromatography. Cordycepins from different sources (cordyceps sinensis and cordceps militaris) can be separated under a certain chromatograph conditions, the cordyceps sinensis as quality goods is selected as a standard product used for identifying whether unknown samples are cordyceps sinensis or cordceps militaris, and the identification other types of worm grass is not related. The method comprises the steps of: taking 0.1ml of tested solution and 0.4ml of cordyceps sinensis extraction solution, uniformly stirring, feeding samples and detecting, wherein the chromatograph conditions include a C18 column, methanol-0.1 percent phosphoric acid water solution as a flowing phase (14.8:85.2), a flowing velocity of 1.0mL / min, a detection wavelength of 260nm and a column temperature of 30 DEG C. The separation degree of cordycepin peaks of two different kinds (cordyceps sinensis and cordceps militaris) reaches above 90 percent, and cordycepin peaks of a mixed sample of cordyceps sinensis from different geographical provenances are not separated, therefore, the cordyceps sinensis and the cordceps militaris can be rapidly and accurately identified.

Description

technical field [0001] The invention belongs to the field of chemical component detection, and relates to a method for quickly identifying Cordyceps sinensis and Cordyceps militaris by using a high-performance liquid phase method. Background technique [0002] Cordyceps sinensis (Berk.) Sacc. is formed by the parasitic fungus Trichosporum sinensis on the larvae of the genus Hirsutella Pat of the Lepidoptera bat moth family. Cordyceps sinensis is a rare edible and medicinal treasure, mainly distributed in alpine forests and subalpine meadows in Qinghai, Sichuan, Tibet, Gansu and Yunnan. Due to the special habitat in the high cold, its host bat moth needs more than three years to complete a generation, resulting in very limited resources of Cordyceps, and the output is not large. Coupled with artificial over-excavation and the destruction of the ecological environment of the Cordyceps origin, my country's Cordyceps is on the verge of exhaustion. In this context, people use C...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/068G01N30/02G01N30/36
Inventor 虞泓曾文波杨俊媛陈自宏
Owner YUNNAN UNIV CHINESE HERBAL MEDICINE BIOLOGICAL RESOURCE INST
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