Three-tiered authentication of herbal Traditional Chinese Medicine ingredients used in women's health provides progressive qualitative and quantitative insight - part 1/4

Traditional Chinese Medicine herbal products are increasingly used in Europe but prevalent authentication methods have significant gaps in detection. In this study, three authentication methods were tested in a tiered approach to improve accuracy on a collection of 51 TCM plant ingredients obtained on the European market. We show the relative performance of conventional barcoding, metabarcoding and standardized chromatographic profiling for TCM ingredients used in one of the most diagnosed disease patterns in women, endometriosis. DNA barcoding using marker ITS2 and chromatographic profiling are methods of choice reported by regulatory authorities and relevant national pharmacopeias. HPTLC was shown to be a valuable authentication tool, combined with metabarcoding, which gives an increased resolution on species diversity, despite dealing with highly processed herbal ingredients. Conventional DNA barcoding as a recommended method was shown to be an insufficient tool for authentication of these samples, while DNA metabarcoding yields an insight into biological contaminants. We conclude that a tiered identification strategy can provide progressive qualitative and quantitative insight in an integrative approach for quality control of processed herbal ingredients.

中药（Traditional Chinese Medicine, TCM）草本产品在欧洲的使用量与日俱增，但当前主流的真伪鉴定方法在检测环节存在显著缺陷。本研究针对欧洲市场采集的51份中药植物原料样品，采用分层鉴定策略对三种方法进行测试，以提升鉴定准确率。本次研究针对用于治疗女性最常见确诊病症之一——子宫内膜异位症（endometriosis）的中药原料，对比了传统DNA条形码（DNA barcoding）、宏条形码（metabarcoding）与标准化色谱指纹图谱三种方法的相对检测性能。采用ITS2（Internal Transcribed Spacer 2）作为标记的DNA条形码技术与色谱指纹图谱技术，是监管机构与各国相关药典推荐的首选鉴定方案。研究表明，高效薄层色谱（High Performance Thin Layer Chromatography, HPTLC）与宏条形码技术联用，可提升物种多样性的分辨精度，即便针对高度加工的草本原料依然适用。尽管传统DNA条形码技术为官方推荐方法，但本研究显示其用于此类样品的真伪鉴定时存在明显局限；而DNA宏条形码技术则可揭示样品中的生物性污染物信息。本研究最终得出结论：分层鉴定策略可通过整合性路径，为加工草本原料的质量控制提供逐步递进的定性与定量分析视角。