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Molecular biology authentication method of bulbus fritillariae cirrhosae in Chinese patent medicine containing fritillariae

An identification method and technology of Chinese patent medicine, which is applied in the field of molecular biology identification of Fritillaria sichuanensis, can solve the problems of high requirements and identification, and achieve the effects of less sampling, automation, and optimization of PCR amplification reaction system

Inactive Publication Date: 2013-04-10
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have higher requirements for the test materials, and can only identify fresh bulbs and leaves, but cannot accurately identify raw materials in various Chinese patent medicines containing Fritillaria, such as tablets, powders, pills, etc.

Method used

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  • Molecular biology authentication method of bulbus fritillariae cirrhosae in Chinese patent medicine containing fritillariae
  • Molecular biology authentication method of bulbus fritillariae cirrhosae in Chinese patent medicine containing fritillariae
  • Molecular biology authentication method of bulbus fritillariae cirrhosae in Chinese patent medicine containing fritillariae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] In this example, 12 dried bulbs collected from the six base sources of Fritillaria fritillaria and one dried bulb of each of the other four types of Fritillaria chinensis and Snake Gallbladder were used to prepare each Snake Gallbladder Fritillaria Powder according to the composition ratio (6:1) of Snake Gallbladder Fritillaria As the test material (see Table 1), genomic DNA was extracted, RAPD molecular marker analysis was performed with specified primers, and standard fingerprints were established with agarose gel electrophoresis bands for the identification of components of the Chinese patent medicine Chuanbei.

[0029] The specific operation steps are as follows:

[0030] 1), after each Snake Gallbladder Fritillaria powder is crushed into very fine powder, then through TNE solution [200 μmol / L tris-hydrochloric acid buffer solution, pH 8.0; 50 μmol / L ethylenediaminetetraacetic acid, pH 8.0 and 250 μmol / L NaCl] pretreatment, and the genomic DNA was extracted by the ...

Embodiment 2

[0046] The test materials and operation steps in this example are the same as in Example 1, wherein the random amplification polymorphic primer is S62GTGAGGCGTC, and a standard fingerprint is established for the identification of different sources of Chinese patent medicine Chuanbei.

[0047] figure 2 The electropherogram of primer S62 (GTGAGGCGTC) on the RAPD marker analysis of sixteen snake gallbladder powders, the marker and figure 1 same. It can be seen that the primer can identify the source of each Fritillaria in Shedan Chuanbei Powder (the arrows indicate the specific bands of each Shedan Chuanbei Powder).

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Abstract

The invention belongs to the technical field of authentication of traditional Chinese medicines, in particular discloses a molecular biology authentication method of bulbus fritillariae cirrhosae in a Chinese patent medicine containing fritillariae, which is characterized by comprising the following steps of: on the basis of RAPD (Random Amplified Polymorphism DNA) analysis of genomes, establishing a random amplified polymorphism DNA standard atlas of the bulbus fritillariae cirrhosae and a random amplified polymorphism DNA standard atlas of the Chinese patent medicine to be authenticated, comparing the two random amplified polymorphism DNA standard atlases to find a stripe corresponding to the standard atlas in the random amplified polymorphism DNA standard atlas of the Chinese patent medicine to be authenticated, and then judging that the fritillariae component in the Chinese patent medicine to be authenticated is the bulbus fritillariae cirrhosae, and vice versa. The invention overcomes the shortages of characters, microscopy, physicochemical authentication and the like of the conventional authentication method.

Description

technical field [0001] The invention belongs to the technical field of identification of traditional Chinese medicines, and specifically relates to a molecular biological identification method for various Chinese patent medicines containing Fritillaria fritillaria Zhongchuan Fritillaria based on genome RAPD analysis. Background technique [0002] Fritillary is the dry bulb of many plants of the genus Fritillaria L. in the family Liliaceae. It is one of the commonly used traditional Chinese medicines. The 2010 edition of "Pharmacopia of the People's Republic of China" records that Fritillaria is divided into five categories: Chuan Fritillaria, Hubei Fritillaria, Flat Fritillaria, Yi Fritillaria, and Zhejiang Fritillaria, among which Chuan Fritillaria is F.Cirrhosa D.Don, dark purple Fritillaria F.unibracteata Hsiaoet K.C.Hsia, Gansu Fritillaria F.przewalskii Maxim. Dried bulbs of ex Batal, F.delavayi Franch, F.taipaiensis P.Y.Li and F.wabuensis S.Y.Tang et S.C.Yueh. [0003]...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 李会军李萍徐传林
Owner CHINA PHARM UNIV