高分子印记固相萃取-液相色谱质谱法测定水产品中孔雀石绿、结晶紫、亮绿及其代谢产物  

Determination of multi-residues for malachite green,crystal violet,brilliant green and their metabolites in fish by using molecularly imprinted solid-phase extraction followed by high performance liquid chromatography-tandem mass spectrometry

作　　者：李晓丽[1,2] 李娜[1] 李鹏[1,3] 刘力[2] 苗虹[1] 

机构地区：[1]中国疾病预防控制中心营养与食品安全所化学污染与健康安全重点实验室,北京100021 [2]西南大学动物科技学院,重庆400715 [3]中国农业大学动物医学院,北京100193

出　　处：《中国食品卫生杂志》2012年第3期209-214,共6页Chinese Journal of Food Hygiene

基　　金：北京市自然科学基金(7082065);卫生行业科研专项项目(200902009)

摘　　要：目的建立一种高分子印记固相萃取-液相色谱质谱联用(MISPE-HPLC/MS2)测定水产品中孔雀石绿、结晶紫、亮绿及其代谢产物的检测方法。方法样品经乙腈提取后,经中性氧化铝柱和高分子印记固相萃取柱净化,Waters Atlantis T3色谱柱(2.1 mm×150 mm,3μm)分离,乙腈和0.1%甲酸水等度洗脱,采用选择反应监测(SRM)模式进行正离子扫描,内标法定量。结果待测化合物在0.2～10μg/L范围内有很好的相关性,相关系数大于0.99,加标水平为1、2、4μg/kg,孔雀石绿、结晶紫和亮绿的平均回收率分别为93.2%～105.9%、92.7%～107.5%和60.6%～94.4%,相对标准偏差均小于12.5%,孔雀石绿、结晶紫和亮绿的检出限分别为0.02、0.03、0.03μg/kg,定量限分别为0.07、0.10、0.10μg/kg。结论本方法前处理净化效果更好、灵敏度更高,同时采用稳定性同位素稀释技术及基质匹配标准曲线,将基质抑制效应降低到最低,适用于大批量鱼类产品中孔雀石绿、结晶紫和亮绿的检测。Objective To establish a method based on molecularly imprinted solid-phase extraction and liquid chromatography-mass spectrometry （MISPE-HPLC/MS2） for the determination of malachite green, crystal violet, brilliant green and their metabolites in aquatic. Methods The aquatic samples were extracted with acetonitrile and the extract were cleaned up by Alumina-N and molecularly imprinted solid-phase extraction catridge （MISPE）. The samples were separated on a Waters Atlantis T3 column （2.1 mm × 150 ram, 3 μm） , acetonitrile and 0. 1% formic acid-water were used as mobile phases for isocratic elution, ESI positive ion scan mode was used with selected reaction monitoring （SRM, MS2） , and MG-D5 and CV-D6 were used as internal standards for quantification. Results The leaner relation for the malachite green, crystal violet and brilliant green were in the range of 0.2 - 10 μg/L with a correlation coefficient not less than 0. 99. The mean recoveries of spiked samples at the level of 1.0, 2.0 and 4. 0 μg/kg were in the range of 93.2% - 105.9% , 92.7% -107.5% and 60.6% -94.4% for malachite green, crystal violet and brilliant green, respectively, the relative standard deviations all less than 12.5%. The detection limits of malachite green, crystal violet and brilliant green were 0.02, 0.03 and 0.03μg/kg, and the quantification limits were 0. 07, 0. 10 and 0. 10 μg/kg, respectively. Conclusion This method used the stable isotope dilution technique and matrix-matched calibration was validated to be high extraction selectivity, sensitivity and better purification for the determination of malachite green, crystal violet, brilliant green and their metabolites in a large of real aquatic samples.

关 键 词：孔雀石绿 结晶紫 亮绿 高分子印记固相萃取 液相色谱-质谱法 水产品 兽药残留 

分 类 号：O657.72[理学—分析化学]