Abstract
Objective: The objective of this study was to develop and validate a novel, simple, rapid, accurate, and precise reverse-phase high-performance liquid chromatographic (RP-HPLC) method for simultaneous estimation of aloe-emodin and piperine in the ayurvedic formulation as per the International Conference on Harmonization guidelines.
 Methods: Chromatographic separation was achieved using a Prontosil C18 (250 × 4.6 mm, 5 μ), with a mobile phase consisting of 0.05% orthophosphoric acid and acetonitrile in the ratio of 50:50, at a flow rate of 1 ml/min and column temperature maintained at 28°C and ultraviolet (UV) detection at 225 nm.
 Results: The retention time of aloe-emodin and piperine was found to be 9.38±0.2 min and 13.45±0.2 min, respectively. The linearity of aloe-emodin and piperine was tested in the range of 1–20 μg/ml. The correlation coefficient for aloe-emodin and piperine was found to be 0.998 and 0.997, respectively. The recovery values (98–102%) indicate a satisfactory accuracy. The percentage relative standard deviation for precision was found to be <2% which indicates that the method is precise.
 Conclusion: A precise method for the simultaneous quantification of aloe-emodin and piperine was developed using high-performance liquid chromatography. Hence, the developed method can be used for quantitative and quality control analysis of formulations containing these phytoconstituents.
Highlights
Ayurveda, the ancient system of Indian medicine, is widely practiced in modern times due to its safety and efficacy [1]
Each ayurvedic formulation is a source of many different phytochemicals, in which individuals may have different medicinal properties [2]
The present study focuses on the standardization of an ayurvedic formulation using high- performance liquid chromatography
Summary
The ancient system of Indian medicine, is widely practiced in modern times due to its safety and efficacy [1]. Ayurvedic medicines are polyherbal formulations and each herb consists of a number of chemical constituents. Each ayurvedic formulation is a source of many different phytochemicals, in which individuals may have different medicinal properties [2]. Standardization and analysis of chemical markers in ayurvedic or polyherbal formulations are important aspects for maintaining and assessing the quality and safety of the polyherbal formulation. Standardization assures safety, efficacy, and quality and minimizes batch-to-batch variation of the polyherbal formulations. An attempt for the standardization of polyherbal formulation has been carried out with respect to the active principles [3]
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