Molecular quantification of herbs(Herb-Q):a pyrosequencingbased approach and its application in Pinellia ternata  

作　　者：PEI Yifei LIU Ziyi YU Dade ZHANG Xiangyu SUN Wei CHEN Xiaofang FENG Xue LI Xiwen 

机构地区：[1]Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China [2]Bijie Institute of Traditional Chinese Medicine,Bijie 551700,China [3]The Key Laboratory for Health Industry of Bijie,Bijie Medical College,Bijie 551700,China

出　　处：《Chinese Journal of Natural Medicines》2024年第7期663-672,共10页中国天然药物（英文版）

基　　金：supported by the Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences(Nos.CI2021A04106 and CI2021A03910);the National Key Research and Development Program of China(No.2019YFC1710601);the Fundamental Research Funds for the Central Public Welfare Research Institutes of China(Nos.ZZ15-YQ-033,ZXKT21026 and ZXKT23004);the Major Special Project of Scientific and Technological Cooperation of Bijie(No.2021-02);the Advantageous Chinese Medicinal Materials R&D Talent Base Project of Bijie,Guizhou Province(No.RCJD2020-21);Bijie Technology Innovation Platform and Talent Team(Bikehe[2023]No.66-BJZDSYS 2024-05).

摘　　要：Variations in herb dosage due to species adulteration and dosing inaccuracies can substantially affect clinical safety and efficacy.Accurate species quantification remains challenging,as current methods often yield inconsistent results.This study introduces a novel pyrosequencing-based technique,termed herb molecular quantification(Herb-Q),designed to precisely quantify herbal products.We evaluated its effectiveness using Pinellia ternata and five of its adulterants.Initially,we assessed commonly used DNA barcodes with sequences from a public database,identifying two candidate regions,Maturase K(matK)and internal transcribed spacer 2(ITS2),for screening specific single nucleotide polymorphism(SNP)loci,allowing for species-specific identification.These loci were validated by amplifying and sequencing genomic material from collected samples.Our validation studies showed that Herb-Q demonstrated excellent linearity,accuracy,repeatability,and detection limits.We established quantitative standard curves with high R^(2)values(>0.99)to enable precise species quantification,which were combined with external standards to provide clear and accurate visual quantification results.The average bias in quantifying the tuber of P.ternata was 2.38%,confirming that Herb-Q can accurately identify and quantify herbal product constituents.Moreover,the entire quantification process took less than 4 h.This study presents a novel,rapid method for accurately quantifying species in herbal products and advances the application of DNA barcoding from species identification to quantitative detection.

关 键 词：Quantification ADULTERANT DNA barcodes Single nucleotide polymorphism 

分 类 号：R917[医药卫生—药物分析学]