柱前衍生-高效液相色谱-串联质谱法测定鲜茶叶中的春雷霉素

Determination of kasugamycin in fresh tea using high performance liquid chromatography-tandem mass spectrometry after pre-column derivation

中文关键词:  春雷霉素  液相色谱串联质谱仪  鲜茶叶  衍生化

英文关键词:kasugamycin  high performance liquid chromatography-tandem mass spectrometry  fresh tea  derivation

中文摘要:

      目的 建立柱前衍生高效液相色谱-串联质谱法(high performance liquid chromatography-tandem mass spectrometry, HPLC-MS/MS)测定鲜茶叶中春雷霉素的检测方法。方法 样品经甲醇/水溶液提取, MCX SPE阳离子交换柱(mixed-mode cation exchanger SPE column)净化, 9-氯甲酸芴甲酯[9-fluorenylmethyloxycarbonyl (Fmoc) chloride, Fmoc-Cl]衍生后测定, 内标法定量。结果 目标化合物在0.31~20.00 μg/L范围内, 线性关系良好, 相关系数大于0.998 (r2)。鲜茶叶中春雷霉素的检出限(limit of detection, LOD)、定量限(limit of quantification, LOQ)分别为2.0 μg/kg和5.0 μg/kg。在LOQ、2×LOQ和10×LOQ水平上加标, 平均回收率为95.2%~104.0%, 相对标准偏差为7.4%~13.3%。结论 基于Fmoc-Cl衍生化的测定法, 能有效提高检测的灵敏度、降低基质的干扰; 将西马特罗作为内标使用, 可以提供更为准确的检测结果, 该方法满足鲜茶叶中春雷霉素的监测。

英文摘要:

      Objective To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the determination of kasugamycin in fresh tea. Methods After methanol/water extraction and MCX SPE column purification, the kasugamycin-derivant derived with Fmoc-Cl was measured with HPLC-MS/MS aided by internal standard quantification. Results The calibration curve showed good linearity in the certain ranges (0.31～20.00 μg/L) with the correlation coefficient greater than 0.998 (r2). The limit of quantification (LOQ) and detection (LOD) for fresh tea was 5.0 μg/kg and 2.0 μg/kg respectively. The average recoveries varied from 95.2%-104.0% with the relative standard deviations of 7.4%-13.3% at three spiked levels (LOQ, 2×LOQ, and 10×LOQ). Conclusion This method depended on derivation with Fmoc-Cl can effectively increase analytical sensitivity and decrease sample matrix interference. Taking cimaterol as internal standard will offer more accuracy determination result. This method is conformity with the monitoring requirement for kasugamycin in fresh tea.