Optimization and Validation of an Ultra-Performance Liquid Chromatography with Quadrupole Detector Mass Spectrometry Quantification Method for the Simultaneous Detection of Tazarotene and Tazarotenic Acid in Porcine Skin: An In Vitro Study.

Exploring tazarotene, a third-generation retinoid for potential hand osteoarthritis treatment, this study presents the development and validation of an ultra-performance liquid chromatography with quadrupole detector mass spectrometry (UPLC-QDa) method for the simultaneous quantification of tazarotene and tazarotenic acid, its active metabolite, in porcine skin. Method development involved a design-of-experiments approach for chromatographic optimization of gradient steepness, organic solvent volume, column temperature, capillary voltage, flow rate, and cone voltage. Central composite orthogonal design was used to optimize peak area, peak width, retention time, and resolution. Validation was performed in accordance with U.S. Food and Drug Administration guidelines. The method was linear over the concentration range of 0.4-18,750 ng/mL for tazarotene and 13.3-12,500 ng/mL for tazarotenic acid, with r values of ≥0.99. Chromatographic analysis demonstrated acceptable accuracy and precision (<15%), and stability tests confirmed the analytes' stability under various conditions. This validated method offers a reliable and accurate approach for the simultaneous analysis of tazarotene and tazarotenic acid, facilitating further research into their therapeutic applications for hand osteoarthritis.