Development and validation of a RP-HPLC method for the simultaneous determination of Mangiferin, Ellagic acid and Hydroxycitric acid in polyherbal formulation

Abstract:

The US patented polyherbal formulation for the prevention and management of Type II diabetes and its vascular complications was used for the present study. The formulation consists of roots of Salacia species, leaves of Lagestroemia parviflora and fruit rind of Garcinia indica. The use of reversed phase C₁₈ HPLC column was used and eluted with isocratic mobile phase of acetonitrile and phosphoric acid buffer solution enabled the efficient separation of chemical markers within 20min. Validation of the method was performed in order to demonstrate its selectivity, accuracy, precision, repeatability and recovery. All calibration curve shows good linear correlation coefficients (r²>0.995) within tested ranges. Three markers in this polyherbal formulation were quantified were Mangiferin (1.53% w/w), Ellagic acid (0.9655 w/w), Hydroxycitric acid (5.3% w/w). Intra and inter day RSDs of retention times and peak areas were less than 3%. The recoveries were between 95% and 102.5%. In conclusion a method has been developed for the simultaneous quantification of three markers in this polyherbal formulation. The established RP-HPLC method was simple, precise and accurate and can be used for the quality control of the raw materials as well as formulations.

Key words: Polyherbal formulation, Mangiferin, Ellagic acid, Hydroxycitric acid, RP-HPLC.