高效液相色谱法测定辣椒中的辣椒碱和二氢辣椒碱

摘要

A method to determine capsaicin and dihydrocapsaicin in pepper products simultaneously by high performance liquid chromatography (HPLC) was developed. The effects of mobile phase composition, proportion, column temperature, velocity of flow and injection volume on separation were studied. The best chromatographic conditions were determined: a ZORBAX SB C18 column (250 mm x 4.6 mm,5 (μm) ; a DAD detector ; the optimal detection wavelengths of capsaicin and dihydrocapsaicin were 280 ran ; the column temperature was 20 °C . The mobile phase was water-methanol(Volume ratio was 35 : 65) with flow rate of 1.0 mL / min ; the injection volume was 5 μL. Under the optimal conditions, calibration curves of each substitutes were established, and the related coefficients were 0.999 9. The recoveries of capsaicin and dihydrocapsaicin in samples were 94.1%-96.8%, 92.6%-95.0% respectively. The RSD was less than 2%(n=11). The method has achieved the simultaneous determination on capsaicin and dihydrocapsaicin in 25 minutes.%建立了高效液相色谱法(HPLC)测定辣椒中辣椒碱、二氢辣椒碱的检测方法.实验考察了仪器条件、不同流动相体系、流动相配比、柱温、流速等因素对分离的影响.确定了最佳色谱条件为ZORBAX SB-C18色谱柱(250mm×4.6 mm,5μm)；二极管阵列检测器,辣椒碱、二氢辣椒碱的最佳检测波长为280 nm；色谱柱温度为20℃;最佳流动相配比为水-甲醇(体积比为35:65),流速为1.0 mL/min；进样量为5 μL.在最佳色谱条件下,建立了辣椒碱的标准曲线,相关系数r=0.9999.辣椒碱、二氢辣椒碱的加标回收率分别为94.1％～96.8％、92.6％～95.0％,测定结果的相对标准偏差小于2.0％(n=11).该法实现了25 min内对辣椒碱、二氢辣椒碱的同时测定.