A simple, fast method of high-performance liquid chromatography for the determination and quantification of arbutin and hydroquinone in many different raw materials was developed and validated. The optimum conditions for the separation and detection of these two constituents were achieved on a LiChro-CARD 125-4 Superspher®100 RP-18 column with the water-methanol (gradient elution) mobile phase and recorded at 289 nm. The purities of peaks were verified by PDA analysis of impurities. The results of validation have shown that the HPLC method is stable and accurate for the simultaneous determination of arbutin and hydroquinone in extracts from various plants. The developed method gave a good sensitivity (LOD 1µg/ml for arbutin and 0.49 µg/ml for hydroquinone) with linearity R2 >0.9993 (for both). The relative standard deviation of the method was less than 2.53% for intra-day assays and 3.23% for inter-day assay, the accuracy of the recovery test ranged from 98.96% to 106.4%. This method was used in comparative qualitative analysis of arbutin and hydroquinone in 16 different raw materials from families Lamiaceae, Ericacaeae, Saxifragaceae, Rosaceae. The content of arbutin in B. ciliata, B. cordifolia and Ledum palustre was examined for the first time.
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