Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for identifying traditional Chinese medicinal material rhizome pinelliae and easily-confused products thereof

A technology of Chinese medicinal material Pinellia, which is applied in the field of identification of Chinese medicinal material Pinellia and its easily mixed products, can solve problems such as difficult to effectively and accurately identify Pinellia and its easily mixed products, and achieve shortened analysis time and cost, accurate Identify and reduce the effects of sequencing

Pending Publication Date: 2021-02-26
河北省中药材质量检验检测研究中心有限公司
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The present invention aims to solve the problem that it is difficult to effectively and accurately identify pinellia and its easily mixed products by traditional methods, and provides multiple site-specific PCR primer pairs and a method for identifying pinellia, tiger's palm and whip eaves.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for identifying traditional Chinese medicinal material rhizome pinelliae and easily-confused products thereof
  • Method for identifying traditional Chinese medicinal material rhizome pinelliae and easily-confused products thereof
  • Method for identifying traditional Chinese medicinal material rhizome pinelliae and easily-confused products thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] Embodiment 1: the primer pair of pinellia and easy-to-mix product is first PCR amplification

[0094] 1. Sample source

[0095] The authentic pinellia used in this example (fresh or dry products can be used, fresh products need to be dried and crushed at 45°C), common easily mixed products such as tiger palm, whip eaves plowshare tip, other easily mixed products Aracea, unicorn lotus From Hebei, Hunan, Anhui and Guangzhou.

[0096] 2. DNA extraction

[0097] DNA extraction was performed on the sample, and the extraction method was as follows:

[0098] 1) Pulverization: sterilize the surface of the sample with 75% ethanol, cut the sample into small pieces, and grind it into powder with a high-throughput tissue grinder;

[0099] 2) Removing starch and polysaccharides: Take 100-150 mg of the pulverized sample, mix with 1 mL of nuclear separation solution, centrifuge at a low speed of 3 500 r / min for 5 minutes, repeat the operation of adding liquid and centrifugation for...

Embodiment 2

[0116] Example 2: Amplification of Multiplex Site-Specific PCR Primer Pairs

[0117] 1. Design multiple site-specific PCR primer pairs

[0118] Download the ITS sequences of Pinellia genus Pinellia, Tiger Palm, Dripdrop, Stone Spider and Pinellia scutellaria from Genbank (accession numbers: KU551864.1, AF469040.1, AF469039.1, AF469033.1, AF469034.1), The ITS sequences of the plants of the genus Aracea, the eaves of the eaves, the unicorn, and the unicorn (accession numbers: KM236658.1, AY863058.1, KM236662.1), the ITS sequences of the plants of the genus Aracea, Aracea heterophyllum, and Aracea dongbei ITS sequence (accession numbers: KT634023.1, MF095999.1, KT634014.1).

[0119] The multiple sequence alignment analysis of the ITS sequences of the above plants was carried out using Mega X software. The ITS sequence of pinellia is A at 77bp, the others are T, the ITS sequence at 662bp is T, and the others are G; the ITS sequence of Tiger palm is T at 84bp, the others are G, C...

experiment example 1

[0141] Experimental Example 1: Process Screening

[0142] 1. Download the ITS sequences of Pinellia, Pinellia and Pinellia scutellaria from Genbank (accession numbers: KU551864.1, AF469040.1, AF469039.1, AF469033.1, AF469034.1) The ITS sequences of the plants Whipcornia, Plowshare and Unicorn (accession numbers: KM236658.1, AY863058.1, KM236662.1), the ITS sequences of Araceae plants Aracea, Heterophyllum and Northeast Aracea (accession numbers No.: KT634023.1, MF095999.1, KT634014.1).

[0143] The multiple sequence alignment analysis of the ITS sequences of the above plants was carried out using Mega X software. The designed primers are listed in Table 2.

[0144] Table 2 Pairs of specific PCR primers for pinellia chinensis, tiger palm and whip eaves

[0145]

[0146]

[0147] Based on the fact that "the specific primers designed for a variety only have specific bands for this variety, and other easily mixed products have no amplified bands", after PCR amplification ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a method for identifying traditional Chinese medicinal material rhizome pinelliae and an easily-confused product thereof. The method comprises the following steps: (1) DNA ofa to-be-identified sample is extracted; (2) primary PCR amplification is conducted, specifically, the DNA extracted in the step (1) is taken as a template, and PCR amplification is conducted by usingan ITS 4 / 5 universal primer pair; (3) secondary PCR amplification is conducted, specifically, a PCR product obtained in the step (2) is taken as a template, PCR amplification is conducted by using a multi-site specific PCR primer pair; and (4) electrophoresis is conducted on a PCR amplification product obtained in the step (3), the to-be-identified sample with the detected 620bp band is identifiedas rhizome pinelliae, the to-be-identified sample with the detected 346bp band is identified as pinellia pedatisecta schott, the to-be-identified sample with the detected 167bp band is identified aswhipformed typhonium rhizome, and other easily-confused products are identified that no amplification band exists.

Description

Technical field: [0001] The invention relates to the field of identification of traditional Chinese medicinal materials, in particular to an identification method of the traditional Chinese medicinal material pinellia chinensis and its easily mixed products. Background technique: [0002] Pinellia ternate is the tuber of the Araceae plant. It has the effects of drying dampness and reducing phlegm, reducing adverse flow and relieving vomiting, and dispelling mass. Pinellia is a bulk medicinal material that is commonly used blindly. It is not only used in clinical formulas, but also as a raw material for various Chinese patent medicines. The standardization and standardization of the planting process is not enough, and the output continues to decrease, which cannot meet the market demand for medicinal materials, and the price is high. Therefore, there are many mixed products of the same family or genus in the Pinellia market, which cause serious problems in terms of planting,...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/6895C12Q1/686C12Q2600/16C12Q2537/143C12Q2565/125
Inventor 张兰兰章飚冯红李细芬
Owner 河北省中药材质量检验检测研究中心有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com