Quantification of Actaea racemosa L. (black cohosh) from some of its potential adulterants using qPCR and dPCR methods

Abstract

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.

Strategies for molecular authentication of herbal products: from experimental design to data analysis

“…It would be straightforward to develop similar techniques for the identification of herbs. Recent researches demonstrated that quantification and semi-quantification of target species are possible by different molecular techniques, such as qPCR [ 12 , 13 ], dPCR [ 14 ], vector control quantitative analysis [ 15 ] and double peak detection in nucleotide signature [ 16 , 17 ], further expanding the scope of potential applications of this approach. However, species-specific assays cannot be used to identify unknown samples with no intended target species.…”

Utilizing Big Data to Identify Tiny Toxic Components: Digitalis

“…The world of eukaryotic DNA-based species identification in food and dietary supplements products has undergone a revolution in the past 10 years, and while progress has been swift it has not been without challenges [8][9][10]. Contemporary methods of DNAbased identification include DNA fingerprinting techniques like restriction fragment length polymorphisms [11], microarrays [12], a full suite of polymerase chain reaction (PCR) methods, [13][14][15][16][17], in addition to traditional DNA barcoding [9,18] and a variety of next generation sequencing (NGS) approaches [19][20][21].…”