Abstract
There is an unmet need for herbal medicine identification using a fast, sensitive, and easy-to-use method that does not require complex infrastructure and well-trained technicians. For instance, the detection of adulterants in Lobelia chinensis herbal product has been challenging, since current detection technologies are not effective due to their own limits. High Resolution Melting (HRM) has emerged as a powerful new technology for clinical diagnosis, research in the food industry and in plant molecular biology, and this method has already highlighted the complexity of species identification. In this study, we developed a method of species specific detection of L. chinensis using HRM analysis combined with internal transcribed spacer 2. We then applied this method to commercial products purporting to contain L. chinensis. Our results demonstrated that HRM can differentiate L. chinensis from six common adulterants. HRM was proven to be a fast and accurate technique for testing the authenticity of L. chinensis in herbal products. Based on these results, a HRM approach for herbal authentication is provided.
Highlights
Lobelia chinensis Lour. belongs to the family Campanulaceae, and is distributed widely in East Asian countries including China, Korea, and Japan (Tada et al, 1995)
In order to develop a fast method combining DNA barcoding and High Resolution Melting (HRM) analysis for discrimination of L. chinensis and its adulterant species, as well as to authenticate herbal products containing this species, HRM was performed on amplicons produced from primers for the nuclear internal transcribed spacer 2 (ITS2) region
By assigning L. chinensis as a genotype and using its melting curve as the baseline, by subtracting the melting curves of the other species, we were able generate difference data to estimate the similarity between the melt profiles of the ITS2 amplicons of L. chinensis and the five adulterant species tested in this study
Summary
Lobelia chinensis Lour. belongs to the family Campanulaceae, and is distributed widely in East Asian countries including China, Korea, and Japan (Tada et al, 1995). Various parts of the plant have been used for the treatment of snakebite, edema, diarrhea, and jaundice in Chinese folk medicine (Yang et al, 2014). Many active chemical compounds have been identified in the plant, including: piperidine alkaloids, such as lobeline, norlobelanine, and lobelanine; coumarins, such as 6,7-dimethoxycoumarin, 5-hydroxy-7-methoxycoumarin, and 5,7dimethoxy-6-hydroxy-coumarin; and terpenoids, including phytol, phytenal, cycloeucalenol, and 24-methylene-cycloartanol (Ishimaru et al, 1992; Shibano et al, 2001; Yang et al, 2014). Bar-HRM for Lobelia chinensis Products herbal health products has become more frequent (Han et al, 2016). Fraudulent labeling of this herbal product has become commonplace, giving rise to health concerns. Fraud control is desirable as a method of supporting fair trade and safeguarding consumer health
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