NHP Research Alliance, College of Biological Sciences, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
J AOAC Int. 2021 Jun 12;104(3):836-846. doi: 10.1093/jaoacint/qsaa167.
Actaea racemosa (black cohosh) herbal dietary supplements are commonly used to treat menopausal symptoms in women. However, there is a considerable risk of contamination of A. racemosa herbal products in the natural health product (NHP) industry, impacting potential efficacy. Authentication of A. racemosa products is challenging because of the standard, multi-part analytical chemistry methods that may be too costly and not appropriate for both raw and finished products.
In this paper, we discuss developing and validating quick alternative biotechnology methods to authenticate A. racemosa herbal dietary supplements, based on the use of a species-specific hydrolysis PCR probe assay.
A qPCR-based species-specific hydrolysis probe assay was designed, validated, and optimized for precisely identifying the species of interest using the following analytical validation criteria: (1) specificity (accuracy) in determining the target species ingredient, while not identifying other non-target species; (2) sensitivity in detecting the smallest amount of the target material; and (3) reliability (repeatability and reproducibility) in detecting the target species in raw materials on a real-time PCR platform.
The results show that the species-specific hydrolysis probe assay was successfully developed for the raw materials and powders of A. racemosa. The specificity of the test was 100% to the target species. The efficiency of the assay was observed to be 99%, and the reliability of the assay was 100% for the raw/starting and powder materials.
The method developed in this study can be used to authenticate and perform qualitative analysis of A. racemosa supplements.
白头翁（黑升麻）草本膳食补充剂常用于治疗女性更年期症状。然而，在天然健康产品（NHP）行业中，白头翁草本产品存在相当大的污染风险，这影响了其潜在疗效。由于标准的多部分分析化学方法可能成本过高，且不适合原材料和成品，因此白头翁产品的鉴定具有挑战性。
在本文中，我们讨论了开发和验证快速替代生物技术方法，以鉴定白头翁草本膳食补充剂，方法是基于使用物种特异性水解 PCR 探针检测。
设计、验证和优化了基于 qPCR 的物种特异性水解探针检测方法，使用以下分析验证标准来精确鉴定目标物种：（1）确定目标物种成分的特异性（准确性），同时不识别其他非目标物种；（2）检测目标材料最小量的灵敏度；以及（3）在实时 PCR 平台上检测原材料中目标物种的可靠性（重复性和再现性）。
结果表明，成功开发了用于白头翁原材料和粉末的物种特异性水解探针检测方法。该测试对目标物种的特异性为 100%。该检测方法的效率观察到为 99%，且对原材料和粉末的检测可靠性为 100%。
本研究中开发的方法可用于鉴定和进行白头翁补充剂的定性分析。
