NHP Research Alliance, College of Biological Sciences, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada.
Sci Rep. 2021 Feb 22;11(1):4331. doi: 10.1038/s41598-020-80465-0.
The demand for popular natural health products (NHPs) such as Black Cohosh is increasing considerably, which in turn challenges quality assurance (QA) throughout the supply chain. To detect and quantify the target species present in a given NHP, DNA-based molecular techniques such as Real-time quantitative PCR (qPCR) and digital PCR (dPCR) are standard tools in the food and pathogen testing industries. There is a gap in the literature concerning validated quantitative PCR methods for botanicals that can be utilized for QA and good manufacturing practices. The objective of this study is to develop an efficient quantification method using qPCR and dPCR techniques for the detection and quantification of Actaea racemosa (Black cohosh) NHPs from its potential adulterants. These developed methods are validated for applicability on commercial NHPs. Species-specific hydrolysis probe assays were designed to analyze the black cohosh NHPs using qPCR and dPCR techniques. The results confirmed that the developed qPCR and dPCR methods are highly precise for identifying and quantifying black cohosh NHPs, indicating their potential applicability in future routine industrial and laboratory testing. This enables a single qPCR test to determine not only the presence of a specific botanical, but also the amount when mixed with an adulterant.
对黑升麻等热门天然保健品（NHPs）的需求正在大幅增加，这反过来又对整个供应链的质量保证（QA）提出了挑战。为了检测和量化给定 NHP 中存在的目标物种，基于 DNA 的分子技术，如实时定量 PCR（qPCR）和数字 PCR（dPCR），是食品和病原体检测行业的标准工具。在关于可用于 QA 和良好生产规范的植物定量 PCR 方法的文献中存在空白。本研究的目的是开发一种使用 qPCR 和 dPCR 技术的高效定量方法，用于从潜在掺杂物中检测和定量升麻属植物（黑升麻）NHP。这些开发的方法经过验证可适用于商业 NHP。设计了物种特异性水解探针检测法，以便使用 qPCR 和 dPCR 技术分析黑升麻 NHP。结果证实，开发的 qPCR 和 dPCR 方法非常精确，可用于识别和定量黑升麻 NHP，表明它们在未来的常规工业和实验室测试中具有潜在适用性。这使得单次 qPCR 测试不仅可以确定特定植物的存在，还可以确定与掺杂物混合时的数量。
