Determination of capsaicinoids in Capsicum species using ultra performance liquid chromatography‐mass spectrometry

In the present work, a rapid and sensitive ultra performance liquid chromatography‐mass spectrometry method has been proposed for the analysis of capsaicinoids (nordihydrocapsaicin, capsaicin, dihydrocapsaicin, homocapsaicin, and homodihydrocapsaicin) present in different Capsicum samples. Extraction of capsaicinoids was carried out by liquid–liquid extraction using ethanol as an extracting solvent, while the chromatographic separation was achieved by reversed phase C₁₈ column with gradient mobile phase (solvent A: acetonitrile and solvent B: water with 0.1% formic acid). Under the optimum experimental conditions, the linear ranges were 0.5–50 μg/g with correlation coefficient (r²) >0.999 for each capsaicinoids and detection limits were 0.15, 0.05, 0.06, 0.2, and 0.1 μg/g for nordihydrocapsaicin, capsaicin, dihydrocapsaicin, homocapsaicin, and homodihydrocapsaicin, respectively. Run‐to‐run and day‐to‐day precisions of the method with relative standard deviations <1.5% were achieved for all analyzed capsaicinoids. The robustness of the method was determined by utilizing different injection volumes of the extracts. Furthermore, to validate the system robustness, a run of high number of capsaicinoids present in different varieties of Capsicum samples was performed in this study. All the capsaicinoids were separated in a time of less than 9 min by employing the proposed method.