Development of species-specific PCR for authentication of medicinal plant Bupleuri Radix and its common adulterants

Bupleuri Radix is a widely used herbal plant, and its official source recognized by Chinese pharmacopoeia is the dried roots of Bupleurum chinense DC. or Bupleurum scorzonerifolium Willd. Although two species share core therapeutic functions, differences in the types of chemical components lead to different clinical applications. Currently, two species have not been distinguished in pharmacopeial standards. Therefore, we hypothesize prescribing them separately based on the pharmacological characteristics may provide better clinical efficacy. Additionally, Bupleurum marginatum Wall. ex DC. var stenophyllum (Wolff) Shan et Y. Li, and Bupleurum bicaule Helm were often misused as Bupleuri Radix. To address above issues, we developed species-specific PCR targeting ITS region. The observed fragments of 252bp, 183bp, 272bp and 165bp correspond to B. chinense, B. scorzonerifolium, B. marginatum var. stenophyllum, and B. bicaule respectively. The PCR exhibited strong discriminative capability, even when analyzing artificial adulterate samples. Furthermore, analysis of commercial samples validated the accuracy of the species information. In summary, the species-specific PCR enables accurate authentication of Bupleuri Radix and common adulterants in the market. It provides a practical tool for quality control of Bupleuri Radix and supports standardization of herbal medicine production.