Abstract
There is considerable potential for the use of DNA barcoding methods to authenticate raw medicinal plant materials, but their application to testing commercial products has been controversial. A simple PCR test targeting species-specific sequences within the nuclear ribosomal internal transcribed spacer (ITS) region was adapted to screen commercial products for the presence of Hypericum perforatum L. material. DNA differing widely in amount and extent of fragmentation was detected in a number of product types. Two assays were designed to further analyse this DNA using a curated database of selected Hypericum ITS sequences: A qPCR assay based on a species-specific primer pair spanning the ITS1 and ITS2 regions, using synthetic DNA reference standards for DNA quantitation and a Next Generation Sequencing (NGS) assay separately targeting the ITS1 and ITS2 regions. The ability of the assays to detect H. perforatum DNA sequences in processed medicines was investigated. Out of twenty different matrices tested, both assays detected H. perforatum DNA in five samples with more than 103 ITS copies µL−1 DNA extract, whilst the qPCR assay was also able to detect lower levels of DNA in two further samples. The NGS assay confirmed that H. perforatum was the major species in all five positive samples, though trace contaminants were also detected.
Highlights
Traditional plant-based medicines were once used as the primary source of healthcare, but since the advent of modern pharmaceuticals these preparations have been relegated to the status of ‘complementary’ medicine
Around half of these products were not suitable for either quantitative PCR (qPCR) or Next Generation Sequencing (NGS) analysis due to the low yield and/or poor quality of DNA extracted from these materials
In this situation the anomalies become of interest such as samples 235 and 244 that are labelled as containing a mass of ‘St John’s Wort” (SJW)’, but yield no DNA
Summary
Traditional plant-based medicines were once used as the primary source of healthcare, but since the advent of modern pharmaceuticals these preparations have been relegated to the status of ‘complementary’ medicine. Recent years have seen the resurgence of biologically active botanicals in many different guises: Food supplements, herbal medicines, nutraceuticals, natural health products and herbal remedies. Each of these types of product have been gaining in popularity, with a recent report showing that the sales of dietary supplements in the US are worth more than ever [1]. The booming market for botanicals has been felt in the areas of production; few of the herbals on the market are cultivated and the majority are wild harvested This has had a drastic effect on the natural
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