Simultaneous quantification of gallic acid and ellagic acid in Triphala using reverse-phase highperformance liquid chromatography (RP-HPLC)
Runjhun Pallavi, Shivesh Jha, Swadesh Pattanik

An Ayurvedic formulation, Triphala consists of three dried fruits of Phyllanthus emblica, Terminalia belerica, and Terminalia chebula in equal proportion was found to be potentially effective and used as an antioxidant, antiinflammatory, antibacterial, and antimutagenic, antineoplastic, and incorporated in prevention and treatment of many Gastrointestinal diseases like constipation, chronic ulcer, jaundice anaemia, asthma. The polyphenols present in the formulation is responsible for their therapeutic efficacy. The proposed method of reverse-phase high-performance liquid chromatography RP-HPLC method was found to be simple, sensitive, precise, and accurate and validation of the method was performed to demonstrate its selectivity, linearity, precision, accuracy, and robustness. The chromatographic conditions used for the separation were mobile phase comprised of O- phosphoric acid (0.3%) OPA in Water: Acetonitrile (80:20% v/v) and pH was maintained at 4.5. The flow rate was 1.0mL/min with detection at 270nm. The linearity was found to be in the range of 20-100µg/mL and the correlation coefficient of Gallic acid (GA) and Ellagic acid (EA) was 0.998 and 0.995 respectively. The retention time was found to be 5.40 and 10.01 for GA and EA respectively in the in-inhouse preparation of Triphala (Ti) and 5.42 and 10.04 in Triphala marketed (Tm). The result indicates that Triphala contains several markers that are responsible for its therapeutic activity. The developed method of RP-HPLC assists in the standardization of Triphala as the marker contents of in-house Triphala were found to have higher than that of marketed Triphala which may be due to the use of inferior quality material used in the preparation of Triphala.

Keywords: triphala, gallic acid, ellagic acid, reverse-phase-HPLC