Analytical method development and validation for simultaneous determination of triamcinolone acetate, betamethasone valerate and clobetasol propionate in cream formulation by RP-HPLC

Abstract

Topical steroids are usually used for the treatment of inflammation and irritation of the skin; however, their inappropriate use results in serious side effects. Thus, the Thai Food and Drug Administration regulation forbade the use of corticosteroids in cosmetics. In this study, a method of high-performance liquid chromatography with a photodiode array detector was developed and validated for the simultaneous determination of three corticosteroids in cream formulation: triamcinolone acetonide (TA), betamethasone valerate (BV), and clobetasol propionate (CP). Chromatographic separation was performed on ACE® C18 (4.6 mm × 150 mm, 5 μm particle size) using a mobile phase consisting of water and acetonitrile in gradient elution. The detection wavelength was set at 254 and 240 nm. The validation results revealed the specificity and linearity of the proposed method with r≥0.999 within the concentration range of 2.5–50 μg/mL. The relative standard deviation (%) of the repeatability and intermediate precision of all corticosteroid samples at three different concentration levels were in the range of 0.04%–2.04% and 0.41%–4.93%, respectively, and the recovery (%) was 98.72 %–105.41%. The detection limits of TA, BV, and CP were 0.69, 0.46, and 0.48 μg/mL, respectively, and their quantitative limits were 2.08, 1.39, and 1.47 μg/mL, respectively. The validated method can be applied to quantify the amount of active compounds in commercial formulations and to determine the corticosteroid adulterants present in cream samples from online shops.