Simultaneous Quantification of Phenolic Compounds and Capsaicinoids in Chili Peppers Using Ultra‐High Performance Liquid Chromatography‐Tandem Mass Spectrometry

ABSTRACT Chili peppers are abundant in bioactive constituents, particularly rich in phenolic compounds and capsaicinoids, which exhibit potent antioxidant capacity. In this study, an ultra‐high performance liquid chromatography‐tandem mass spectrometry (UHPLC‐MS/MS) method was developed for the simultaneous quantification of both phenolic compounds and capsaicinoids in chili peppers. Key parameters, including sample extraction protocols, chromatographic separation conditions, and mass spectrometry parameters were systematically optimized. The validated method demonstrated that all analytes exhibited satisfactory linearity with coefficients of determination ( R 2 ) ≥ 0.9967. The limits of quantification ranged from 0.2 to 100 μg/L, and the recovery values spanned 61.4%–124.7% with intra/inter‐day precisions below 16% relative standard deviation. The proposed method was finally successfully applied to quantify 19 phenolic compounds and four capsaicinoids in six chili pepper samples collected from China. The results demonstrated that this methodology represented a promising analytical approach for simultaneous, rapid, sensitive and accurate detection of targeted phenolic compounds and capsaicinoids in chili peppers.