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Method for rapidly distinguishing true honeysuckle and false honeysuckle

A honeysuckle, authenticity technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of large component differences and achieve low-cost effects

Inactive Publication Date: 2015-03-18
TIANJIN INSTITUE OF QUALITY STANDARD & TESTING OF AGRICULTUAL PRODS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, from the analysis of chemical components, the content of luteolin, which is the main medicinal ingredient, is relatively high in honeysuckle, while the content in mountain silver flower is very small. Mountain silver flower mainly contains chlorogenic acid, and the composition of the two is quite different.

Method used

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  • Method for rapidly distinguishing true honeysuckle and false honeysuckle
  • Method for rapidly distinguishing true honeysuckle and false honeysuckle
  • Method for rapidly distinguishing true honeysuckle and false honeysuckle

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] (1) The samples include the DNA of genuine and counterfeit honeysuckle, and sample 23# of honeysuckle for testing, which was taken from Shandong.

[0038] (2) The sequence list of PCR primers is as follows:

[0039]

[0040] (3) Sampling and DNA extraction

[0041] Take the complete flower buds of the honeysuckle samples to be tested, freeze and grind them with liquid nitrogen (Retsch MM400 biocrusher), add 500 μL of CTAB lysis buffer (20g / L CTAB, 1.4M NaCl, 0.1M Tris-HCl, 20mM Na2EDTA), and place in a constant temperature water bath at 65°C (Neslab) was lysed for 30 min, and the subsequent DNA extraction and purification were performed according to the resin-type genomic DNA purification kit (Saibaisheng). After DNA extraction, the concentration was uniformly diluted to 50±1ng / μL, Nanodrop ND1000, Therm0l). The DNA extraction method can also be carried out according to a commercially available kit.

[0042] (4) PCR reaction

[0043] Carry out PCR amplification (in...

Embodiment 2

[0047] (1) Samples include DNA of genuine and counterfeit honeysuckle, as well as sample 90# of honeysuckle for testing, sampled from Sichuan.

[0048] (2) The sequence list of PCR primers is as follows:

[0049]

[0050] (3) Sampling and DNA extraction

[0051] Take the complete flower buds of the honeysuckle samples to be tested, freeze and grind them with liquid nitrogen (Retsch MM400 biocrusher), add 500 μL of CTAB lysis buffer (20g / L CTAB, 1.4M NaCl, 0.1M Tris-HCl, 20mM Na2EDTA), and place in a constant temperature water bath at 65°C (Neslab) was lysed for 30 min, and the subsequent DNA extraction and purification were performed according to the resin-type genomic DNA purification kit (Saibaisheng). After DNA extraction, the concentration was uniformly diluted to 50±1ng / μL, Nanodrop ND1000, Thermol). The DNA extraction method can also be carried out according to a commercially available kit.

[0052] (4) PCR reaction

[0053] Carry out PCR amplification (instrument...

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Abstract

The invention discloses a method for distinguishing a true honeysuckle and a common false product lonicerae flos thereof. According to the method, symbolic nucleolide sites of the honeysuckle and the lonicerae flos are analyzed by utilizing a high-resolution solubility curve technology. The distinguishing method disclosed by the invention can be used for true and false evaluation of a honeysuckle medicinal material, can finish the true and false distinguish work of the honeysuckle within 1-2 hours, and has the advantages of being rapid, low in cost, convenient to operate, free of pollution due to detection in a closed tub, and the like.

Description

technical field [0001] The invention belongs to the field of drug analysis, in particular to a method for identifying the authenticity of the traditional Chinese medicinal material honeysuckle, which is a molecular identification method based on high resolution melting curve analysis technology (high resolution melting, HRM). Background technique [0002] Honeysuckle is the dry flower bud or the first flower of Lonicera iaponica Thunb., which has the effects of clearing away heat and detoxifying, cooling wind-heat, and is a commonly used bulk Chinese medicinal material. Modern medical research shows that honeysuckle contains luteolin, inositol, saponins, chlorogenic acid, sugars, proteins, vitamins, various compounds and various trace elements. Some commonly used drugs such as Yinqiao Jiedu pills and Shuanghuanglian Liquid, Qingkailing injection and oral liquid, Yinhuang tablets, compound Daqingye mixture, etc. all use honeysuckle as the main raw material. In recent years, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q2561/101C12Q2563/107
Inventor 兰青阔王永刘征辉赵新朱珠徐石勇陈锐郭永泽
Owner TIANJIN INSTITUE OF QUALITY STANDARD & TESTING OF AGRICULTUAL PRODS