Abstract
The wide distribution of Valeriana officinalis as a herbal remedy as well as the considerably higher concentration of putative mutagenic valepotriate metabolites in other drug-delivering valerian species like Valeriana procera Kunth and Valeriana jatamansi Jones ex Roxb. illustrate the necessity of secure authentication of roots of Valeriana officinalis s.l., especially as the morphologically similar roots of the acutely toxic Veratrum album can be mistaken for those of Valeriana officinalis. We developed two DNA-based systems, a multiplex amplification refractory mutation system (MARMS), and a high-resolution melting curve analysis (HRMA) assay, both based on a sequence mutation within the atpB-rbcL region. With both methods, identification of Valeriana officinalis s.l. was possible. With the HRMA, the characteristic melting curve of 33 samples of Valeriana officinalis s.l. and of two commercial samples of Valerianae radix was distinct from the melting curves of all other Valeriana species (60 accessions), and from the closely related genera Centranthus and Valerianella. Since adulteration of Valeriana with toxic Veratrum species was reported previously, Veratrum primers were included in a multiplex PCR-HRM analysis. This system allowed the detection of a Veratrum admixture down to the level of 0.01 %. Although the advantages, in terms of sensitivity, specificity and practicality of the HRM for analysis of degraded plant material were superior to the MARMS assay, both methods are suitable for routine analysis. The results demonstrated the general ability of HRMA to detect specific (toxic) adulterations in drugs in a semiquantitative way.
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Acknowledgements
This work has been carried out with financial support from Aboca (Sansepolcro, Italy) and FP7 grant No. 245199 (Plantlibra), which are gratefully acknowledged.
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This study has received funding from the European Community’s Seventh Framework Programme (FP7/2007–2013) under grant agreement No. 245199 (Plantlibra). This report does not necessarily reflect the Commission’s views or its future policy in this area.
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The corresponding author has received research grants from Aboca (Sansepolcro, Italy).
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Ruzicka, J., Schmiderer, C. & Novak, J. DNA-Based Identification of Valeriana officinalis s.l.: a Multiplexed Amplification Refractory Mutation System (MARMS) and High Resolution Melting Curve Analysis (HRMA). Plant Mol Biol Rep 34, 909–922 (2016). https://doi.org/10.1007/s11105-015-0967-6
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DOI: https://doi.org/10.1007/s11105-015-0967-6