Development and validation of LC-MS/MS method for the determination of UV-filters across human skin in vitro.

A novel High-Performance Liquid Chromatography-Tandem Mass Spectrometry (HPLC-MS/MS) method was developed for the simultaneous determination of the in vitro skin permeation profile of four UV filters. The sunscreen products contained the following components: octocrylene (OC), ethylhexyl methoxycinnamate (EHMC), diethylamino hydroxybenzoyl hexyl benzonate (DHHB) and ethylhexyl salicylate (EHS). The target compounds were analyzed by HPLC-MS/MS method in positive ionization electrospray (ESI) in Multiple Reaction Monitoring (MRM) mode. The proposed method was validated in terms of the detection (LOD) and quantification limits (LOQ), linearity range, intra- and inter- day precision and accuracy of the analysis. The stability of the target compounds in solutions was also studied. All tests provided satisfactory results illustrating acceptable method performance. Samples were analyzed with simple pretreatment procedure, necessary to achieve solvent change and preconcentration. To evaluate matrix effect, the slopes of the standard regression curves with those of the matrix-matched calibration curves were compared using the Student's t-test. Quantitative evaluation of the test samples was performed using external methanolic calibration curves. Accuracy was found within the range 94.37-108.76%. The method was successfully applied to the analysis of UV filters in samples after permeability studies, in Franz's cells, for 24 h, using human skin. Concentration of sunscreens in the acceptor phase at the timescale of 24 h was very low implying the safety of the products.