title: ITS2 barcoding DNA region combined with high resolution melting (HRM) analysis of Hyoscyami Semen, the mature seed of Hyoscyamus niger.
abstract: Hyoscyami Semen, the mature dried seed of Hyoscyamus niger L., has long been used as a traditional Chinese medicine to treat human diseases. Hyoscyami Semen is found in local markets in China. In markets, sellers and buyers commonly inadvertently mix the seeds of H. niger with the seeds of related species such as Hygrophila salicifolia (Vahl) Nees, Astragalus complanatus R. Br., Cuscuta australis R. Br., Cuscuta chinensis Lam., and Impatiens balsamina L. because of their similar morphologies or similar names. Thus, developing a reliable method for discriminating H. niger seeds from its adulterants is necessary to reduce confusion and ensure the safe use of Hyoscyami Semen. The present study was designed to evaluate the efficiency of high-resolution melting analysis combined with DNA barcoding (Bar-HRM) with internal transcribed spacer 2 to discriminate H. niger. Our results show that Bar-HRM successfully identified the adulterants and detected the proportion of H. niger DNA extract within an admixture. In particular, HRM detected H. niger DNA extract in A. complanatus DNA extract at concentrations as low as 1%. In conclusion, the Bar-HRM method developed in the present study for authenticating H. niger is rapid and cost-effective. It can be used in the future to guarantee the purity of Hyoscyami Semen for the clinical use.
authors.affiliation: College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China; Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Xiong
authors.affiliation: College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China; Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Zhi-Gang
authors.initials: ZG
authors.lastname: Hu
authors.affiliation: College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China; Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China; Traditional Chinese Medicine Resources Research Center, Tianjin Tasly Pharmaceutical Co., Ltd., Tianjin 300402, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Tu
authors.affiliation: College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China.
authors.forename: He-Gang
authors.initials: HG
authors.lastname: Liu
authors.affiliation: College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
authors.affiliation: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Ming-Ming
authors.initials: MM
authors.lastname: Zhao
authors.affiliation: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Yu-Hua
authors.initials: YH
authors.lastname: SHIi
authors.affiliation: College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China; Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Lan
authors.initials: L
authors.lastname: Wu
authors.affiliation: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China. Electronic address: wsun@icmm.ac.cn.
authors.forename: Wei
authors.initials: W
authors.lastname: Sun
authors.affiliation: College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China; Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China. Electronic address: slchen@icmm.ac.cn.
authors.forename: Shi-Lin
authors.initials: SL
authors.lastname: Chen
mesh_terms: D002681:China; D058893:DNA Barcoding, Taxonomic / Q000379:methods*; D021901:DNA, Intergenic / Q000737:chemistry / Q000235:genetics*; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics*; D016002:Discriminant Analysis; D004340:Drug Contamination; D004365:Drugs, Chinese Herbal / Q000737:chemistry*; D032304:Hyoscyamus / Q000235:genetics / Q000254:growth & development*; D012639:Seeds / Q000235:genetics / Q000254:growth & development*; D044366:Transition Temperature
chemical_list: D021901:DNA, Intergenic; D018744:DNA, Plant; D004365:Drugs, Chinese Herbal
keywords: Bar-HRM; Clinical safe medication; Hyoscyami Semen; Identification