Peptide Nucleic Acid Based Molecular Authentication for Identification of Four Medicinal Paeonia Species Using Melting Array Analysis of the Internal Transcribed Spacer 2 Region.

Wook Kim,Byeong Moon,Goya Choi,Sungyu Yang

doi:10.3390/molecules22111922

Wook Kim, Byeong Moon + Show 2 more

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https://doi.org/10.3390/molecules22111922

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Abstract

Accurate taxonomic identification of plant materials in herbal medicines is important for product quality control. The genus Paeonia (Saxifragales) is the source of the herbal preparations Paeoniae Radix (Paeoniae Radix Alba and Paeoniae Radix Rubra) and Moutan Radicis Cotex. However, confusion has arisen regarding their contents due to linguistic and taxonomic ambiguities, similar morphologies and different definitions of Paeoniae Radix in the Korean and Chinese national pharmacopoeias, leading to the distribution of adulterated products. To develop a method for identifying the four Paeonia species used in these medicines, three fluorescently-labeled peptide nucleic acid (PNA) probes were designed against ITS2 sequences containing single nucleotide polymorphisms (SNPs) and used in a real-time PCR melting curve assay. Each of the four Paeonia species was accurately identified using this analysis. The accuracy and analytical stability of the PNA melting curve assay was confirmed using commercially available samples of the four Paeonia species. This assay is a reliable genetic tool to distinguish between different Paeonia-derived herbal medicines and identify the botanical origins of Paeoniae Radix and Moutan Radicis Cortex. This technique may also contribute to quality control and standardization of herbal medicines by providing a reliable authentication tool and preventing the distribution of inauthentic adulterants.

Highlights

Herbal medicine derived from plants has been used in traditional medicine for conditions including injury and acute pain for more than one thousand years [1,2]
ITS2 regions were amplified from the 17 Paeonia samples listed by Kim et al [8] using universal primers
Inter-species distances were 1.84%, 1.87%, 2.00% and 2.25%, respectively (Table 1). These results indicated that P. veitchii and P. suffruticosa had the highest genetic divergences within and between species, respectively

Summary

Introduction

Herbal medicine derived from plants has been used in traditional medicine for conditions including injury and acute pain for more than one thousand years [1,2]. One or more species having equal effects can be used to treat the same disease, leading to plant species with different taxonomic classifications having the same herbal medicinal name. Definitions of botanical origin for numerous herbal remedies differ between pharmacopoeias [6], and some inauthentic species are sometimes included. The use of synonyms and homonyms has led to the inclusion of inauthentic substitutions or adulterants in traditional herbal medicines [7]. These inauthentic substitutions or adulterants, which have different pharmacological efficacy or may have toxic components in some cases, are not an appropriate use as Molecules 2017, 22, 1922; doi:10.3390/molecules22111922 www.mdpi.com/journal/molecules

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