Development of a novel RP-HPLC method for simultaneous estimation of aloe emodin, rhein, and piperine in a polyherbal formulation

The present manuscript describes a rapid, novel, simple, precise, accurate, and robust HPLC method development and validation for simultaneous estimation of aloe emodin, rhein and piperine in a polyherbal formulation. The proposed chromatographic method was carried out isocratically, in a short run time of 10 minutes using Prontosil C18 (250 × 4.6 mm) SH 5.0 µm, usingmobile phase acetonitrile and water (0.05% orthophosphoric acid) in the ratio 55:45. Flow rate was set at 1 mL/min, and the UV detection was at 257 nm. Retention time of aloe emodin, rhein and piperinewas found to be 7.064 ± 0.2 min, 7.580 ± 0.2 min and 8.800 ± 0.2 min respectively. Calibration curves were linear over the concentration range of 1 to 30µg/mL.Regression coefficient were 0.998, 0.996, 0.999 for aloe emodin, rhein and piperinerespectively. This and can be used to for routine analysis of aloe emodin, rhein and piperine in polyherbal formulations, thus conforming to the need of ensuring safety and quality of herbal formulations.