Cargando…

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

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 quant...

Descripción completa

Detalles Bibliográficos
Autores principales: Shanmughanandhan, Jeevitha, Shanmughanandhan, Dhivya, Ragupathy, Subramanyam, Henry, Thomas A., Newmaster, Steven G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7900226/
https://www.ncbi.nlm.nih.gov/pubmed/33619286
http://dx.doi.org/10.1038/s41598-020-80465-0
_version_ 1783654180544577536
author Shanmughanandhan, Jeevitha
Shanmughanandhan, Dhivya
Ragupathy, Subramanyam
Henry, Thomas A.
Newmaster, Steven G.
author_facet Shanmughanandhan, Jeevitha
Shanmughanandhan, Dhivya
Ragupathy, Subramanyam
Henry, Thomas A.
Newmaster, Steven G.
author_sort Shanmughanandhan, Jeevitha
collection PubMed
description 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.
format Online
Article
Text
id pubmed-7900226
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-79002262021-02-24 Quantification of Actaea racemosa L. (black cohosh) from some of its potential adulterants using qPCR and dPCR methods Shanmughanandhan, Jeevitha Shanmughanandhan, Dhivya Ragupathy, Subramanyam Henry, Thomas A. Newmaster, Steven G. Sci Rep Article 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. Nature Publishing Group UK 2021-02-22 /pmc/articles/PMC7900226/ /pubmed/33619286 http://dx.doi.org/10.1038/s41598-020-80465-0 Text en © The Author(s) 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Shanmughanandhan, Jeevitha
Shanmughanandhan, Dhivya
Ragupathy, Subramanyam
Henry, Thomas A.
Newmaster, Steven G.
Quantification of Actaea racemosa L. (black cohosh) from some of its potential adulterants using qPCR and dPCR methods
title Quantification of Actaea racemosa L. (black cohosh) from some of its potential adulterants using qPCR and dPCR methods
title_full Quantification of Actaea racemosa L. (black cohosh) from some of its potential adulterants using qPCR and dPCR methods
title_fullStr Quantification of Actaea racemosa L. (black cohosh) from some of its potential adulterants using qPCR and dPCR methods
title_full_unstemmed Quantification of Actaea racemosa L. (black cohosh) from some of its potential adulterants using qPCR and dPCR methods
title_short Quantification of Actaea racemosa L. (black cohosh) from some of its potential adulterants using qPCR and dPCR methods
title_sort quantification of actaea racemosa l. (black cohosh) from some of its potential adulterants using qpcr and dpcr methods
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7900226/
https://www.ncbi.nlm.nih.gov/pubmed/33619286
http://dx.doi.org/10.1038/s41598-020-80465-0
work_keys_str_mv AT shanmughanandhanjeevitha quantificationofactaearacemosalblackcohoshfromsomeofitspotentialadulterantsusingqpcranddpcrmethods
AT shanmughanandhandhivya quantificationofactaearacemosalblackcohoshfromsomeofitspotentialadulterantsusingqpcranddpcrmethods
AT ragupathysubramanyam quantificationofactaearacemosalblackcohoshfromsomeofitspotentialadulterantsusingqpcranddpcrmethods
AT henrythomasa quantificationofactaearacemosalblackcohoshfromsomeofitspotentialadulterantsusingqpcranddpcrmethods
AT newmastersteveng quantificationofactaearacemosalblackcohoshfromsomeofitspotentialadulterantsusingqpcranddpcrmethods