Duan Bao-Zhong, Wang Ya-Ping, Fang Hai-Lan, Xiong Chao, Li Xi-Wen, Wang Ping, Chen Shi-Lin
1College of Pharmaceutical Science, Hubei University of Chinese Medicine, Wuhan, 430065 China.
2Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700 China.
Chin Med. 2018 Feb 8;13:8. doi: 10.1186/s13020-018-0162-4. eCollection 2018.
Rhizoma Paridis (Chonglou) is a commonly used and precious traditional Chinese medicine. Smith var. (Franch.) Hand. -Mazz. and Smith var (Franch.) Hara are the two main sources of Chonglou under the monograph of Rhizoma Paridis in Chinese Pharmacopoeia. In the local marketplace, however, this medicine is prone to be accidentally contaminated, deliberately substituted or admixed with other species that are similar to Rhizoma Paridis in shape and color. Consequently, these adulterations might compromise quality control and result in considerable health concerns for consumers. This study aims to develop a rapid and sensitive method for accurate identification of Rhizoma Paridis and its common adulterants.
DNA barcoding coupled with high resolution melting analysis was applied in this research to distinguish Rhizoma Paridis from its adulteration. The internal transcribed spacer 2 (ITS2) barcode was selected for HRM analysis to produce standard melting profile of the selected species. DNA of the tested herbal medicines was isolated and their melting profiles were generated and compared with the standard melting profile of var. .
The results indicate that the ITS2 molecular regions coupled with HRM analysis can effectively differentiate nine herbal species, including two authentic origins of Chonglou and their seven common adulterants. Ten herbal medicines labeled "Chonglou" obtained from a local market were collected and identified with our methods, and their sequence information was analyzed to validate the accuracy of HRM analysis.
DNA barcoding coupled with HRM analysis is a accurate, reliable, rapid, cost-effective and robust tool, which could contribute to the quality control of Rhizoma Paridis in the supply chain of the natural health product industry (NHP).
重楼是一种常用且珍贵的传统中药。七叶一枝花（Paris vietnamensis (Takht.) H. Li）和华重楼（Paris polyphylla Smith var. chinensis (Franch.) H. Li）是《中国药典》中重楼项下的两个主要来源。然而，在当地市场上，这种药材容易受到意外污染、被故意替代或与其他形状和颜色与重楼相似的物种混合。因此，这些掺假行为可能会影响质量控制，并给消费者带来相当大的健康隐患。本研究旨在开发一种快速、灵敏的方法，用于准确鉴定重楼及其常见掺假品。
本研究采用DNA条形码结合高分辨率熔解分析来区分重楼与其掺假品。选择内部转录间隔区2（ITS2）条形码进行高分辨率熔解分析，以生成所选物种的标准熔解曲线。提取受试草药的DNA，生成其熔解曲线，并与七叶一枝花的标准熔解曲线进行比较。
结果表明，ITS2分子区域结合高分辨率熔解分析能够有效区分九种草药物种，包括重楼的两个正品来源及其七种常见掺假品。收集了从当地市场获得的十种标为“重楼”的草药，并采用我们的方法进行鉴定，分析其序列信息以验证高分辨率熔解分析的准确性。
DNA条形码结合高分辨率熔解分析是一种准确、可靠、快速、经济高效且稳健的工具，可有助于天然保健品行业（NHP）供应链中重楼的质量控制。
