Identification of Levisticum officinale Adulterated in Angelica sinensis by PCR-RFLP
10.13422/j.cnki.syfjx.20210514
- VernacularTitle:PCR-RFLP鉴别当归药材及饮片中掺混伪品——欧当归的方法
- Author:
Zhong-fei SHI
1
;
Bao-xia TENG
1
;
Jing LAI
1
;
Lin NI
1
;
Ping-shun SONG
1
Author Information
1. NMPA Key Laboratory for Quality Control of Traditional Chinese Medicine and Prepared Slices，Gansu Institute for Drug Control，Gansu Engineering Laboratory for Chinese and Tibetan Medicine Control Technology，Lanzhou 730000，China
- Publication Type:Research Article
- Keywords:
Angelica sinensis;
Levisticum officinale;
polymerase chain reaction -restriction fragment length polymophism （PCR-RFLP）;
admixture identification
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(9):168-175
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a rapid method to identify Levisticum officinale adulterated in Angelica sinensis by polymerase chain reaction -restriction fragment length polymophism（PCR-RFLP）. Method:By comparing sequences restriction sites in ribosomal DNA Internal Transcribed Spacer（ITS） of A. sinensis and L. officinale，the specific restriction site Fnu4HI of L. officinale was selected，and the primers for PCR-RFLP reaction were designed. Different A. sinensis and L. officinale were amplified by PCR. The conditions affecting the PCR-RFLP reaction，such as annealing temperature，primer concentration，cycle number and enzyme digestion reaction time，were optimized，and the accuracy of the method was investigated. The established PCR-RFLP identification method was used to investigate the applicability of L. officinale adulterated in A. sinensis with different aduleration ratios and different origins. Result:A PCR-RFLP method for identifying A. sinensis mixed with L. officinale was established. When the annealing temperature was 62 ℃ and the number of cycles was 30，when the L. officinale adulterated in A. sinensis could be digested by Fnu4H I restriction endonuclease after amplification with specific primers，and the two single DNA bands were detected between 100-500 bp，the A. sinensis were all negative. The minimum detection limit of this method for adulterated L. officinale in A. sinensis was 3%，which could be used for the detection of adulterated L. officinale in A. sinensis. Conclusion:The established PCR-RFLP identification method is sensitive and accurate in detecting whether there is L. officinale in A. sinensis，and it provides inspection reference and basis for the quality control of A. sinensis，with great significance to ensure the safety of its clinical medication.
