Determination of Capsaicin and Dihydrocapsaicin in Capsicum Fruits by Liquid Chromatography-Electrospray/Time-of-Flight Mass Spectrometry
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A simple, highly selective, sensitive, and reproducible liquid chromatography-electrospray ionization/time-of-flight mass spectrometry method has been developed for the direct and simultaneousdetermination of capsaicin and dihydrocapsaicin in Capsicum fruit extracts. Capsaicin and dihydrocapsaicin are the two major members of the so-called capsaicinoid family, which includes other minoranalogues, and usually account for at least 90% of the pungency trait in Capsicum fruits.Chromatographic separation of capsaicin and dihydrocapsaicin was achieved with a reversed-phasechromatography column, using a gradient of methanol and water. Quantification was done using asan internal standard (4,5-dimethoxybenzyl)-4-methyloctamide, a synthetic capsaicin analogue notfound in nature. Analytes were base-peak resolved in less than 16 min, and limits of detection were20 pmol for capsaicin and 4 pmol for dihydrocapsaicin. The intraday repeatability values were lowerthan 0.5 and 12% for retention time and peak area, respectively, whereas the interday repeatabilityvalues were lower than 0.6 and 14% for retention time and peak area, respectively. Analyte recoveriesfound were 86 and 93% for capsaicin and dihydrocapsaicin, respectively. The method developedhas been applied to the identification and quantification of capsaicin and dihydrocapsaicin in fruitextracts from different Capsicum genotypes, and concentrations found ranged from 2 to 6639 mgkg-1.

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