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Species Quantification in Complex Herbal Formulas—Vector Control Quantitative Analysis as a New Method
Product mislabeling and/or species fraud in Traditional Chinese Medicine (TCM) not only decrease TCM quality, but also pose a potential health issue to the end user. Up to now, methods to control TCM quality have been developed to detect specific metabolites or identify the original species. However...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7725679/ https://www.ncbi.nlm.nih.gov/pubmed/33324200 http://dx.doi.org/10.3389/fphar.2020.488193 |
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author | Zhao, Bo Xiong, Chao Li, Jingjian Zhang, Deng Shi, Yancai Sun, Wei Duan, Xiaoqun |
author_facet | Zhao, Bo Xiong, Chao Li, Jingjian Zhang, Deng Shi, Yancai Sun, Wei Duan, Xiaoqun |
author_sort | Zhao, Bo |
collection | PubMed |
description | Product mislabeling and/or species fraud in Traditional Chinese Medicine (TCM) not only decrease TCM quality, but also pose a potential health issue to the end user. Up to now, methods to control TCM quality have been developed to detect specific metabolites or identify the original species. However, species quantification in complex herbal formulas is rarely concerned. Here, we reported a simple Vector Control Quantitative Analysis (VCQA) method for flexible and accurate multiplex species quantification in traditional Chinese herbal formulas. We developed PCR-based strategy to quickly generate the integrated DNA fragments from multiple targeted species, which can be assembled into the quantitative vector in one round of cloning by Golden Gate ligation and Gateway recombination technique. With this method, we recruited the nuclear ribosomal DNA Internal Transcribed Spacer (ITS) region for the quantification of Ligusticum sinense “Chuanxiong,” Angelica dahurica (Hoffm.) Benth. & Hook.f. ex Franch. & Sav., Notopterygium incisum K. C. Ting ex H. T. Chang, Asarum sieboldii Miq., Saposhnikovia divaricata (Turcz.) Schischk., Nepeta cataria L., Mentha canadensis L., and Glycyrrhiza uralensis Fisch. ex DC. in ChuanXiong ChaTiao Wan, a classic Chinese herbal formula with very long historical background. We found that, firstly, VCQA method could eliminate the factors affecting such as the variations in DNA extracts when in combination with the use of universal and species-specific primers. Secondly, this method detected the limit of quantification of A. sieboldii Miq. in formula products down to 1%. Thirdly, the stability of quality of ChuanXiong ChaTiao Wan formula varies significantly among different manufacturers. In conclusion, VCQA method has the potential power and can be used as an alternative method for species quantification of complex TCM formulas. |
format | Online Article Text |
id | pubmed-7725679 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-77256792020-12-14 Species Quantification in Complex Herbal Formulas—Vector Control Quantitative Analysis as a New Method Zhao, Bo Xiong, Chao Li, Jingjian Zhang, Deng Shi, Yancai Sun, Wei Duan, Xiaoqun Front Pharmacol Pharmacology Product mislabeling and/or species fraud in Traditional Chinese Medicine (TCM) not only decrease TCM quality, but also pose a potential health issue to the end user. Up to now, methods to control TCM quality have been developed to detect specific metabolites or identify the original species. However, species quantification in complex herbal formulas is rarely concerned. Here, we reported a simple Vector Control Quantitative Analysis (VCQA) method for flexible and accurate multiplex species quantification in traditional Chinese herbal formulas. We developed PCR-based strategy to quickly generate the integrated DNA fragments from multiple targeted species, which can be assembled into the quantitative vector in one round of cloning by Golden Gate ligation and Gateway recombination technique. With this method, we recruited the nuclear ribosomal DNA Internal Transcribed Spacer (ITS) region for the quantification of Ligusticum sinense “Chuanxiong,” Angelica dahurica (Hoffm.) Benth. & Hook.f. ex Franch. & Sav., Notopterygium incisum K. C. Ting ex H. T. Chang, Asarum sieboldii Miq., Saposhnikovia divaricata (Turcz.) Schischk., Nepeta cataria L., Mentha canadensis L., and Glycyrrhiza uralensis Fisch. ex DC. in ChuanXiong ChaTiao Wan, a classic Chinese herbal formula with very long historical background. We found that, firstly, VCQA method could eliminate the factors affecting such as the variations in DNA extracts when in combination with the use of universal and species-specific primers. Secondly, this method detected the limit of quantification of A. sieboldii Miq. in formula products down to 1%. Thirdly, the stability of quality of ChuanXiong ChaTiao Wan formula varies significantly among different manufacturers. In conclusion, VCQA method has the potential power and can be used as an alternative method for species quantification of complex TCM formulas. Frontiers Media S.A. 2020-11-26 /pmc/articles/PMC7725679/ /pubmed/33324200 http://dx.doi.org/10.3389/fphar.2020.488193 Text en Copyright © 2020 Zhao, Xiong, Li, Zhang, Shi, Sun and Duan http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Pharmacology Zhao, Bo Xiong, Chao Li, Jingjian Zhang, Deng Shi, Yancai Sun, Wei Duan, Xiaoqun Species Quantification in Complex Herbal Formulas—Vector Control Quantitative Analysis as a New Method |
title | Species Quantification in Complex Herbal Formulas—Vector Control Quantitative Analysis as a New Method |
title_full | Species Quantification in Complex Herbal Formulas—Vector Control Quantitative Analysis as a New Method |
title_fullStr | Species Quantification in Complex Herbal Formulas—Vector Control Quantitative Analysis as a New Method |
title_full_unstemmed | Species Quantification in Complex Herbal Formulas—Vector Control Quantitative Analysis as a New Method |
title_short | Species Quantification in Complex Herbal Formulas—Vector Control Quantitative Analysis as a New Method |
title_sort | species quantification in complex herbal formulas—vector control quantitative analysis as a new method |
topic | Pharmacology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7725679/ https://www.ncbi.nlm.nih.gov/pubmed/33324200 http://dx.doi.org/10.3389/fphar.2020.488193 |
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