Zhou Shilin, Zhou Ting, Zhang Yanmei, Yang Bingqian, Niu Junmei, Li Weixian, Weitu Yiyi, Long Faming, Liu Zhenwen, Zhou Jing
School of Pharmaceutical Science and Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University, Kunming, China.
Yunnan Academy of Forestry and Grassland, Kunming, China.
Front Pharmacol. 2025 Apr 28;16:1584065. doi: 10.3389/fphar.2025.1584065. eCollection 2025.
Ensuring quality and authenticity of traditional medicines is crucial, particularly for multi-ingredient formulations like commercial Chinese polyherbal preparations (CCPPs). This study aims to authenticate Renshen Jianpi Wan (RSJPW), a classical CCPP composed of 11 prescribed botanical drugs, using DNA metabarcoding to overcome challenges in species-level identification of processed biological ingredients.
We analyzed 56 commercial RSJPW products from different manufacturers and production batches, alongside eight laboratory-prepared reference samples serving as authentic controls. A dual-marker protocol combining ITS2 and psbA-trnH regions was employed, with optimized DNA extraction and PCR protocols to mitigate degradation issues.
Detection rates varied across samples, with the highest detection being 10 out of 11 prescribed ingredients in a single sample. The key fungal ingredient () was consistently undetectable, likely due to DNA degradation during processing and challenges in extracting fungal DNA from complex matrices. Multiple high-abundance non-prescribed species from Fabaceae, Apiaceae, Brassicaceae, and other families were frequently detected as potential contaminants.
This study establishes a systematic framework for molecular authentication of complex herbal formulations, providing technical support for reliable identification of botanical drugs. While DNA metabarcoding offers valuable insights into CCPP composition, authentication of heavily processed ingredients remains a significant technical limitation. The integration with complementary analytical methods such as metabolomics could provide more comprehensive quality assessment in future studies, demonstrating the necessity of multi-analytical approaches in ensuring the authenticity of traditional medicine.
确保传统药物的质量和真实性至关重要，特别是对于像商业用中药复方制剂（CCPPs）这样的多成分配方。本研究旨在利用DNA宏条形码技术对由11种法定植物药组成的经典CCPP——人参健脾丸（RSJPW）进行鉴定，以克服加工后生物成分物种水平鉴定方面的挑战。
我们分析了来自不同制造商和生产批次的56种市售RSJPW产品，以及8个实验室制备的作为真实对照的参考样品。采用了结合ITS2和psbA - trnH区域的双标记方案，并优化了DNA提取和PCR方案以减轻降解问题。
不同样品的检出率各不相同，单个样品中11种法定成分的最高检出数为10种。关键真菌成分（此处原文缺失具体成分）始终未被检测到，可能是由于加工过程中的DNA降解以及从复杂基质中提取真菌DNA存在挑战。来自豆科、伞形科、十字花科和其他科的多种高丰度非法定物种经常被检测为潜在污染物。
本研究建立了复杂草药配方分子鉴定的系统框架，为可靠鉴定植物药提供了技术支持。虽然DNA宏条形码技术为CCPP组成提供了有价值的见解，但对深度加工成分的鉴定仍然是一个重大技术限制。与代谢组学等互补分析方法相结合，可能在未来研究中提供更全面的质量评估，表明多分析方法对于确保传统药物真实性的必要性。
