Many members of the genus Artemisia are important for medicinal purposes with multiple pharmacological properties. Often, these herbal plants sold on the markets are in processed forms so it is difficult to authenticate. Routine testing and identification of these herbal materials should be performed to ensure that the raw materials used in pharmaceutical products are suitable for their intended use. In this study, five commonly used Artemisia species included Artemisia argyi, Artemisia annua, Artemisia lavandulaefolia, Artemisia indica, and Artemisia atrovirens were analyzed using high resolution melting (HRM) analysis based on the internal transcribed spacer 2 (ITS2) sequences. The melting profiles of the ITS2 amplicons of the five closely related herbal species are clearly separated so that they can be differentiated by HRM method. The method was further applied to authenticate commercial products in powdered. HRM curves of all the commercial samples tested are similar to the botanical species as labeled. These congeneric medicinal products were also clearly separated using the neighbor-joining (NJ) tree. Therefore, HRM method could provide an efficient and reliable authentication system to distinguish these commonly used Artemisia herbal products on the markets and offer a technical reference for medicines quality control in the drug supply chain.
Psammosilene tunicoides is an important herb used in traditional Chinese medicine. It has been proved to benefit the stomach and treat rheumatism and gout. On the market, this herbal medicine is frequently adulterated by the related species Silene viscidula. Correct identification of P. tunicoides is important to ensure herbal quality, safety, authenticity and health for consumers. However, the identification of the P. tunicoides and S. viscidula species is complicated because of their morphological similarities. Therefore, a reliable method to authenticate these two medicinal plants is needed. In this study, the ITS2 barcode region coupled with high-resolution melting (Bar-HRM) was evaluated as a novel approach for differentiating P. tunicoides from its adulterant S. viscidula. Our findings showed that Bar-HRM not only detected the adulteration, but also quantified the most common admixture. Bar-HRM sensitivity in adulterant detection was assessed by analysing samples mixed with different proportions of S. viscidula and P. tunicoides control. The results are presented as a linear regression with R 2 D 0.9852, which implied the capability of the method to detect adulteration. This study is significant to verify the authenticity for better quality control of this herbal species.
BackgroundThe morphological identification of different Hippophae species (Shaji) was difficult. This study aims to discriminate between medicinal and non-medicinal Hippophae species by DNA barcodes, the ITS2, psbA-trnH, and a combination of ITS2 and psbA-trnH (ITS2 + psbA-trnH).MethodsDNA was extracted from the dried fruit samples. Primer pairs ITS2F/3R for ITS2 and psbAF/trnHR for psbA-trnH were used for PCR amplification. The purified PCR products were bidirectionally sequenced. Genetic distances were calculated according to the Kimura 2 parameter model and phylogenetic tree was constructed based on neighbor-joining (NJ) method, barcoding gap was also analyzed to assess identification efficiency.ResultsAmplification and sequencing efficiencies for both ITS2 and psbA-trnH were 100 %. Sequence data revealed that ITS2 + psbA-trnH was the most suitable candidate barcode at the species and subspecies level. The closely related Hippophae species were effectively differentiated in the NJ tree.ConclusionThe combination of the two loci, ITS2 + psbA-trnH is applicable to the identification of medicinal and non-medicinal Hippophae species.
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