The Development and Validation of Novel High-Performance Liquid Chromatography Method for Simultaneous Estimation of p-Cymene and Aloe-emodin

The objective of the present investigation was to develop a novel, accurate, precise, and linear High-Performance Liquid Chromatographic (HPLC) method for the simultaneous estimation of p-Cymene and aloe-emodin in the novel topical herbal formulation and validated as per ICH guidelines. In the current study, good chromatographic separation was achieved in isocratic mode using an HPLC C18 column (250mm × 4.6), 5 μm, and a mobile phase consisting of acetonitrile:water in the ratio of 80:20, at a flow rate of 1.0 mL/min and column temperature maintained at 25°C. The response obtained was monitored at 225 nm wavelength with a UV-Visible detector. The retention times of Aloe-emodin and p-Cymene were found to be 4.3 min and 9.0 min respectively. Linearity was established for both p-Cymene and aloe-emodin in the range of 10-90 µg/mL, respectively. For the method, % Recovery was found in the range of 99.67-100.51 % for p-Cymene and 98.68-100.4 % for aloe-emodin respectively. The LOD and LOQ were found to be 0.01 and 0.04 for p-Cymene and 0.12 and 0.36 for aloe-emodin respectively. This method can be successfully employed for simultaneous quantitative analysis of p-Cymene and Aloe-emodin in the novel topical herbal formulation.