Molecular Authentication of Polyherbal Formulation - Triphala Churna

Abstract

Aim: The present study aims to design a molecular authentication protocol for the renowned polyherbal formulation Triphala with the help of recent advances in the DNA barcoding technology. Purpose of Work: In recent decades, ayurvedic polyherbal medications have gained immense popularity worldwide. The quality of some of these herbal products has raised serious concerns regarding their authenticity and safety. The therapeutically potent original herbal species component of the polyherbal formulations are often substituted with cheaper or easily available adulterants or sometimes omitted entirely. In this case, molecular species authentication methods such as DNA barcoding can provide a far safer and efficient way to ensure species authenticity in polyherbal formulations. Methodology: In the present study, ITS DNA barcoding based species authentication was performed for the polyherbal formulation – Triphala. For all the three component species of Triphala i.e. Terminalia bellirica, Terminalia chebula and Phyllanthus emblica, species authenticating primers were designed from their ITS barcoding regions and experimentally validated. The presented method was applied to authenticate all the three herbal species before they are blended in together to generate “Triphala” mixture as well as for quality assurance purpose for the already blended and processed formulation Triphala. This work also addresses the challenges in isolating DNA of satisfactory quality and quantity from highly acidic, polyphenolic, and commercially processed Triphala churna samples after making some critical modifications in the traditional CTAB method; thus enabling their molecular authentication. Utilizing the designed species authenticating primers, a quality assurance inspection was conducted on seven Triphala churna samples from reliable pharmacies that are available in the market. Results: The present study successfully authenticates all the three ingredient species i.e. Terminalia bellirica, Terminalia chebula and Phyllanthus emblica in commercially available Triphala churna samples with the help of ITS DNA barcoding technology.