Development and Validation of HPLC–Photodiode Array Method for Detecting Steroids in Skin Whitening Products Simultaneously

Abstract

Since steroids are prohibited to be present in cosmetic products, thus identification of these outlawed compounds intentionally incorporated in the skin whitening product is sufficient for market surveillance purposes. Simultaneous methods for detecting prednisone (PRN), triamcinolone acetonide (TCA), hydrocortisone acetate (HCA), clobetasol propionate (CBP), and mometasone furoate (MTF) have not been developed. The goal of this study is to develop and validate the screening of these five steroid compounds in a cosmetic product sing reverse phase (RP) HPLC equipped with a photodiode array (PDA) detector. The sample was prepared by diluting it with methanol. Furthermore, the sample solution was vortexed, sonicated and then centrifuged at 4500 rpm. The filtered solution was then put into a chromatographic system. A validation method was applied including the system suitability test, determination of selectivity and detection limit. In the optimization step, we succeeded in distinguishing and identifying five steroid compounds with different retention times.The retention times of PRN, TCA, HCA, CBP and MTF were detected at 3.267, 4.260, 4.910, 15.360, and 16.707 min, respectively. The method performed good selectivity with a resolution of more than 1.5. While for determination of the limit of detection (LOD) was carried out by calibration curve approach over the range 0.5-5 µg/mL and resulted in LOD values for PRN, TCA, HCA, CBP and MTF were 0.36, 0.44, 0.41, 0.47, and 0.42 µg/mL, respectively. The developed method can be applied for routine analysis of PRN, TCA, HCA, CBP and MTF in various types of cosmetic products.