Simultaneous determination by UHPLC-PDA of major capsaicinoids and capsinoids contents in peppers

• First UHPLC-PDA method for simultaneous separation of capsaicinoids and capsinoids. • An analysis time of only less than 2 min is required for a complete separation. • The method showed excellent validation results. Capsaicinoids and capsinoids compounds have been a focus of special attention for their health benefits. An effective and rapid Ultra-High-Performance Liquid Chromatography (UHPLC-PDA) method has been developed and validated for the simultaneous separation and quantitative determination of the major capsaicinoids and capsinoids present in peppers. The separation of all the compounds of interest was achieved in less than 2 min by means of an ACQUITY UPLC BEH rp-C18 column (100 mm × 2.1 mm i.d., 1.7 µm particle size). The variables that have been optimized are the mobile phase (water as solvent A and acetonitrile as solvent B, both acidified by adding 0.1% acetic acid), separation gradient, column temperature (35–70 °C), flow rate (0.6–0.95 mL min −1 ), and injection volume (2.5–3.5 µL). The evaluation of the chromatographic performance revealed excellent resolution, retention factor, and selectivity. The method was satisfactorily validated in terms of linearity, detection and quantification limits, precision, and robustness.