Phenolic Profile of Potentilla anserina L. (Rosaceae) Herb of Siberian Origin and Development of a Rapid Method for Simultaneous Determination of Major Phenolics in P. anserina Pharmaceutical Products by Microcolumn RP-HPLC-UV

摘要

A chemical study of Potentilla anserina L. herb (Rosaceae) of Siberian origin led to the isolation of 17 compounds. Three ellagitannins—potentillin, agrimonic acid A and B—are reported for the first time in this species. With a view to rapid quantitative analysis, a new method was developed for simultaneous determination of major phenolic compounds in P. anserina, including caffeic acid, myricetin-3-O-glucuronide, agrimoniin, ellagic acid, miquelianin, isorhamnetin-3-O-glucuronide, and kaempferol-3-O-rhamnoside. The quantitative determination was conducted by microcolumn reversed phase high-performance liquid chromatography with UV detection. Separation was performed using a ProntoSIL-120-5-C18 AQ column (60 mm × 1 mm × 5 μm) with six-step gradient elution of aqueous 0.2 М LiClO4 in 0.006 M HClO4 and acetonitrile as mobile phases. The components were quantified by HPLC-UV at 270 nm. All calibration curves showed good linearity (r² > 0.999) within test ranges. The reproducibility was evaluated by intra- and inter-day assays, and RSD values were less than 2.8%. The recoveries were between 97.15 and 102.38%. The limits of detection ranged from 0.21 to 1.94 μg/mL, and limits of quantification ranged from 0.65 to 5.88 μg/mL, respectively. Various solvents, extraction methods, temperatures, and times were evaluated to obtain the best extraction efficiency. The developed method was successfully applied for the analysis of selected pharmaceutical products: 12 batches of P. anserina herb collected from three Siberian regions (Yakutia, Buryatia, Irkutsk), two commercial samples of P. anserina herb, and some preparations (liquid extract, tincture, decoction, infusion, and dry extract).