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PCR identification method for kangfuxin solution

A technology of Kangfuxin liquid and identification method, which is applied in the field of rapid identification of Kangfuxin liquid medicines, can solve the problems of destroying organic components, no reports, difficulty in identification of Kangfuxin liquid, etc., and achieves the effect of qualitative control.

Pending Publication Date: 2020-07-21
SICHUAN GOODDOCTOR PANXI PHARMA
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0006] At present, although there have been reports on the identification of Periplaneta americana medicinal materials by PCR method (invention patents CN201710426033.6 and CN201710991800.8), the samples of Periplaneta americana extract (water extract and alcohol extract) and extreme conditions treatment and rehabilitation There is no report on the PCR identification method of the new liquid drug
Especially in the production process of Kangfuxin liquid, there are steps such as drying, crushing and mixing of Periplaneta americana medicinal materials, alcohol decoction extraction process, and even high-temperature and high-pressure treatment of the finished product. These conditions will either turn the Periplaneta americana powder into a mixture , or the Periplaneta americana will destroy a large amount of organic components due to the extraction process, making the identification of Kangfuxin solution difficult

Method used

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  • PCR identification method for kangfuxin solution
  • PCR identification method for kangfuxin solution
  • PCR identification method for kangfuxin solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The primers in embodiment 1 in the present invention are to the PCR amplification of Kangfuxin solution Periplaneta americana COI gene

[0034]1. Kangfuxin solution (15 batches of Kangfuxin solution from Sichuan Good Doctor Panxi Pharmaceutical Co., Ltd. Batch numbers: B171207, B171208, B171209, B171210, B180121, B180122, B180123, 171103, 180301, 180101, B171216, 180201, B17121 , 180302, 180107); the Periplaneta americana reference drug (ID: 5RTB-TBMS) was purchased from China National Institutes for Food and Drug Control.

[0035] 2. Sample pre-treatment: Take an appropriate amount of Kangfuxin liquid sample and evaporate it to dryness in a water bath for concentration treatment before use.

[0036] 3. Sample DNA extraction: Take a certain volume of concentrated Kangfuxin liquid into a centrifuge tube, add 275 μl of digestion solution (cell lysate 200 μl; 0.5M EDTA (pH 8.0) 50 μl; proteinase K, 20 mg / mL 20 μl; RNase A solution 5μl) to ensure that the sample is complet...

Embodiment 2

[0046] Example 2 Based on the primers in the present invention, the general primers of Blatella insects and the specific primers of Periplaneta americana, the PCR amplification of Kangfuxin solution and the adult COI gene of Periplaneta americana

[0047] The materials of this embodiment mainly include: Kangfuxin liquid (batch number: 180107), American cockroach adults (lot number: CP180701) from Sichuan Good Doctor Panxi Pharmaceutical Co., Ltd., American cockroach reference drug (ID: 5RTB-TBMS ) were purchased from China National Institutes for Food and Drug Control.

[0048] 1. Primer synthesis: The primers used in this example are as follows:

[0049] Primers in this invention: (JWF1 and JWR1)

[0050] Forward primer JWF1: 5′-ATTCTGATTATTACCACCTTCA-3′

[0051] Reverse primer JWR1: 5′-AAATTGCTAGATCAACAGATGC-3′

[0052] Periplaneta americana COI primers: (LCO1490 and HCO2198)

[0053] Forward primer LCO1490: 5′-TAAACTCCAGGGTGACCAAAAAAATCA-3’

[0054] Reverse primer HCO2...

Embodiment 3

[0059] Example 3 is based on the primers in the present invention to the PCR amplification of Periplaneta americana water extract, alcohol extract and extreme (high temperature, high pressure, long time) condition processing sample

[0060] The materials of this embodiment mainly include: Periplaneta americana adults (batch number: CP180701), Periplaneta americana water extract samples and samples processed under extreme (high temperature, high pressure, long time) conditions, Periplaneta americana alcohol extract samples and extreme ( The samples processed under the conditions of high temperature, high pressure, and long time) were from Sichuan Good Doctor Panxi Pharmaceutical Co., Ltd., and the Periplaneta americana reference material (ID: 5RTB-TBMS) was purchased from China National Institutes for Food and Drug Control.

[0061] Extract DNA according to the DNA extraction method in Example 1, and amplify according to the PCR reaction conditions, primers and reaction procedur...

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Abstract

The invention discloses a PCR identification method for a kangfuxin solution. The method is used for a PCR to amplify a periplaneta Americana L COI gene sequence (140 bp), a PCR product is verified through agarose gel and then sent to a sequencing company for sequencing, a sequencing result is manually proofread, subjected to sequence assembly and compared with a public sequence, and if the consistency with a gene sequence SEQ ID NO.1 is 99% or above, it can be judged that samples are periplaneta Americana L sources. A reliable PCR identification technology is adopted, the kangfuxin solution is quickly and accurately identified, accuracy and reliability are enhanced, and the safety of the kangfuxin solution used as a medicine by adopting the periplaneta Americana L as a medicinal materialis guaranteed.

Description

technical field [0001] The invention relates to the technical field of traditional Chinese medicine, Chinese medicinal materials, and Chinese medicinal extract identification, in particular to a sample capable of treating Periplaneta americana medicinal materials, Periplaneta americana extract (water extract and ethanol extract) and extreme conditions and a new rehabilitation solution Drugs are quickly identified. Background technique [0002] Periplaneta Americana (Periplaneta Americana L) is an insect of the genus Periplaneta in the family Blattidae. Its use as medicine was first recorded in "Shen Nong's Materia Medica", and it was listed as a middle grade; The throat is closed, and the internal cold has no children", "Tongli blood vessels", "Qi under food". Its "[nature, flavor and meridian] salty, flat; return liver, kidney, spleen meridian. [function and indications] promoting blood circulation and removing blood stasis, clearing away heat and detoxification, eliminati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6869G16B30/10G16B10/00
CPCC12Q1/6888C12Q1/6869G16B30/10G16B10/00C12Q2531/113C12Q2565/125
Inventor 耿越飞姜顺日耿福能王杉杉刘彬吴桃清唐艳萍
Owner SICHUAN GOODDOCTOR PANXI PHARMA
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