Development and validation of stability indicating HPLC–PDA method for the simultaneous determination of benzoyl peroxide and tretinoin in topical dosage form

Abstract

Abstract Background Acne vulgaris is a very dangerous skin disease leads to psychological disorders. Benzoyl peroxide and tretinoin in combination with topical dosage are used to improve the appearance of acne-prone skin. The study aimed to develop and validate an HPLC–PDA method for the simultaneous estimation of both drugs. Intentional modifications were implemented in the analytical method to get better optimum conditions. The final method was chosen for the reverse phase chromatographic separation by using the C18 column as a stationary phase and 0.01 M phosphate buffer adjusted pH 2.5 mixed with acetonitrile (25:75 v/v) as a mobile phase. The optimized conditions were 1.5 mL/min flow rate, 30 °C column temperature, 5 °C autosampler temperature, and 20 µL injection volume. The wavelength was chosen for detection of Benzoyl peroxide at 272 nm and Tretinoin at 353 nm by utilizing a PDA detector. All standard and sample solutions were made in methanol. Results The developed method exhibited peak retention times of 2.94 min for Benzoyl peroxide and 11.34 min for Tretinoin. This analytical method was proven to be robust, linear in calibration curve, accurate, precise, and specific. The forced degradation studies results showed that a high degree of specificity was obtained by separating both analytes from produced impurities completely. Conclusions The developed analytical method is fast, economic and stability indicating. It is useful for routine pharmaceutical analysis where the combination of benzoyl peroxide and tretinoin is formulated for their quality and safety.

Keywords

• Benzoyl peroxide
• Tretinoin
• HPLC
• Method validation
• Stress studies