Identification of mixed medicinal powder of Epimedii Folium with different origins based on pyrosequencing technology
10.16438/j.0513-4870.2023-0536
- VernacularTitle:基于焦磷酸测序技术的不同淫羊藿基原混合药材粉末鉴定
- Author:
Zi-yi LIU
1
;
Yi-fei PEI
2
;
Xue FENG
2
;
Gui-ping ZHAO
2
;
Tie-zhu CHEN
3
;
Xi-wen LI
2
Author Information
1. Department of Pharmacy, Southwest Medical University, Luzhou 646000, China; Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China; Sichuan Provincial Key Laboratory of Quality and Innovation Research of Chinese Materia Medica, Sichuan Academy of Traditional Chinese Medicine Sciences, Chengdu 610041, China
2. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
3. Department of Pharmacy, Southwest Medical University, Luzhou 646000, China; Sichuan Provincial Key Laboratory of Quality and Innovation Research of Chinese Materia Medica, Sichuan Academy of Traditional Chinese Medicine Sciences, Chengdu 610041, China
- Publication Type:Research Article
- Keywords:
Epimedii Folium;
single nucleotide polymorphism;
mixed powder;
pyrosequencing;
Sanger sequencing
- From:
Acta Pharmaceutica Sinica
2023;58(8):2532-2540
- CountryChina
- Language:Chinese
-
Abstract:
There are many multi-original medicinal materials in Chinese Pharmacopoeia, and the mixed use of medicinal materials from different sources is common, which has certain influence on the stability of clinical medication. In this study, pyrosequencing technology was used to screen species-specific single nucleotide polymorphisms (SNP) from commonly used DNA barcode sequences, and a rapid and accurate molecular identification method for original species in mixed medicinal powder of Epimedii Folium was established. Multiple sequence alignment analysis showed that the 176th (C/T) mutation and the 196th (A/G) mutation of ITS, the 123rd (C/G) mutation of matK and the 892nd (A/C) mutation of rbcL could be used as the unique SNPs of E. sagittatum, E. koreanum, E. brevicornu and E. pubescens, respectively. In this study, the applicability of pyrosequencing and Sanger sequencing methods in the sequencing of mixture samples was investigated from the perspective of sensitivity and stability. Pyrosequencing method has higher detection sensitivity than Sanger sequencing method for low content samples in the mixed samples. Stability analysis showed that pyrosequencing technology could still obtain effective sequencing results for the amplified products of template DNA after 45 min of 95 ℃ high temperature water bath, while the critical point of Sanger sequencing method was 30 min. In this study, a new identification technology of Epimedii Folium mixed powder primordial species based on pyrosequencing and specific SNP was developed, which can quickly and accurately identify the mixed use of Epimedii Folium with high sensitivity and stability, and can also support the identification of different primordial species and mixed powder primordial herbs, which is conducive to ensuring the consistency and stability of clinical medication.
