Unveiling Adulteration in Herbal Markets: MassARRAY iPLEX Assay for Accurate Identification of L.

The root of L. is commercially available in herbal markets in both crude and powdered forms. root is a key ingredient in numerous polyherbal formulations. However, has two closely related species, L. and Lam. Since only is traditionally used in Thai polyherbal products, adulteration with other species could potentially compromise the therapeutic efficacy and overall effectiveness of these formulations. To address this issue, a MassARRAY iPLEX assay was developed to accurately identify and differentiate from its closely related species. Five single nucleotide polymorphism (SNP) sites-positions 18, 112, 577, 623, and 652-within the internal transcribed spacer (ITS) region were selected as genetic markers for species identification. The assay demonstrated high accuracy in identifying and was capable of detecting the species at DNA concentrations as low as 0.01 ng/µL. Additionally, the assay successfully identified in commercial crude drug samples, highlighting potential instances of adulteration. Furthermore, it was able to distinguish in mixed samples containing along with either or . The developed MassARRAY iPLEX assay proves to be a reliable and effective molecular tool for authenticating raw materials. Its application holds significant potential for ensuring the integrity of herbal products by preventing misidentification and adulteration.