Simultaneous determination of esculetin, quercetin-3-O-β-D-glucuronide, quercetin-3-O-β -D-glucuronopyranside methyl ester and quercetin in effective part of Polygonum Perfoliatum L. using high performace liquid chromatography

Objective: In the present study, a high performance liquid chromatography (HPLC) coupled with photodiode array detection was developed for simultaneous quantitation of esculetin, quercetin-3-O-β-D-glucuronide, quercetin-3-O-β-D- glucuronopyranoside methyl ester and quercetin in Polygonum perfoliatum L. Materials and Methods: The chromatographic separations were performed on a reversed-phase C ₁₈ column using a mobile phase composed of acetonitrile -0.5% aqueous acetic acid with gradient elution. The calibration curves for the analytes demonstrated good linearities within the investigated ranges. The satisfactory intra- and inter-day precision, repeatability and stability of the developed analytical method were shown in the method validation procedure. The recoveries of the established method ranged from 95.76 to 102.10% for all the analytes. Results: This proposed method was successfully applied for simultaneous quantification of the four compounds in effective part of Polygonum perfoliatum L. from different regions. Hierarchical clustering analysis (HCA) and principal components analysis (PCA) were performed to characterize and classify the samples based on the contents of the four compounds in Polygonum perfoliatum L. Conclusion: The established HPLC method combined with chemometric approaches was proven to be useful and efficient for quality control of Polygonum perfoliatum L.