Development and Validation of a Simple and Selective Chromatographic Method for Quantification of Ceramide-NP in Skin-Simulating Liposome Formulations

Numerous topical products containing ceramide to restore the skin barrier function have been available on the market in recent years. A simple, rapid, and selective analytical method could contribute to the development of innovative formulations such as skin-simulating liposomes, which contain ceramides as stratum corneum components. The similarity of the physicochemical structure of ceramide and lipid excipients in liposome formulation complicates the separative and quantitative analysis. Therefore, this study aims to develop and validate a selective high-pressure liquid chromatography (HPLC) method for the simultaneous quantification of lipids in ceramide-containing skin simulating liposome formulations. The separation was performed with a reversed-phase C18 column (150 mm x 4.6 mm, 5 mu m), the mobile phase system consisted of methanol: acetonitrile (60:40, v/v), the flow rate of 0.5 ml/min, the oven temperature at 45degree celsius, injection volume of 10 mu L, and UV detection at 210 nm. The method was linear (r(2) > 0.99) in the range of concentration from 80 mu g/mL to 480 mu g/mL with acceptable precision, accuracy, and selectivity for the liposome formulations. The limit of detection (LOD) values of ceramide NP (Cer-NP) was 7.90 mu g/mL. The limit of quantification (LOQ) values of Cer-NP was 24.06 mu g/mL. The study indicates that the method is efficient and has accuracy and selectivity for the simultaneous quantification of Cer-NP in skin-simulating liposome formulations.