RP-HPLC & hptlc analyses and standardization of payapro™ premix-a polyherbal veterinary galactogogue

Objective: The aim of the present study was to standardize Payapro™ Premix by quantitative estimation of apigenin and glycyrrhizin by reverse phase high performance liquid chromatography (RP-HPLC) and high-performance thin layer chromatography (HPTLC) methods. Payapro™ Premix is a polyherbal formulation used as galactogogue containing seven herbs including Cuminum cyminum and Glycyrrhiza glabra. Materials and methods: RP-HPLC and HPTLC methods were developed for the standardization of Payapro™ Premix by quantitative estimation of apigenin and glycyrrhizin, the bio-active constituents of Cuminum cyminum and Glycyrrhiza glabra respectively. The developed methods were validated on various parameters, including linearity, precision, accuracy, LOD, and LOQ as per ICH guidelines. Results: The RP-HPLC and HPTLC analysis methods were selective for the polyherbal formulation. Both the methods had specific linearity range with regression coefficient ≥ 0.995. The apigenin and glycyrrhizin estimation methods were precise (% RSD 0.7 and 0.4), accurate (average recovery values were 96.28 % and 95.83 %), LOD (sensitive) (0.0054 µg/mL & 0.016 µg spot-1), LOQ (reliable) (0.0162 µg/mL & 0.048 µg spot-1), respectively. Methanolic extracts of polyherbal formulation showed the presence of significant amount of apigenin (50 µg/g) and glycyrrhizin (1200 µg/g) using developed methods. The comparison of different batches for these markers was found to be uniform which ensures their quality. Conclusion: The present work emphasized on standardization of the polyherbal formulation by determination of bio-active marker constituents. The developed methods can be used to standardize other samples containing Cuminum cyminum and Glycyrrhiza glabra.