高效液相色谱-串联质谱法检测猪肝中苯并咪唑类药物及其代谢物残留

Simultaneous determination of benzimidazoles and metabolites residues in Liver by HPLC-MS/MS

关键词:  肝  苯并咪唑  代谢物  残留  高效液相色谱-串联质谱

Key words:  Liver  benzimidazoles  metabolites  residue  HPLC-MS/MS

摘要:

建立了猪肝中阿苯哒唑、阿苯哒唑砜、阿苯哒唑亚砜、胺基阿苯哒唑砜、噻苯咪唑、5-羟基噻苯咪唑、甲苯咪唑、胺基甲苯咪唑、5-羟基甲苯咪唑、芬苯哒唑、芬苯哒唑砜、奥芬哒唑、氟苯咪唑、2-胺基-氟苯咪唑等14种苯并咪唑类药物及其代谢物残留检测的高效液相色谱-串联质谱方法。样品用乙酸乙酯提取后，用MCX固相萃取柱净化。用Waters Xterra C18色谱柱（2.1mm×150 mm，5μm）分离 ，流动相A相为0.1%甲酸乙腈溶液；B相为0.1%甲酸水溶液，梯度洗脱，外标法定量。结果表明：14种苯并咪唑类药物及其代谢物标准溶液在10ng/mL～1000ng/mL的浓度范围内呈现良好的线性关系，R2均大于0.99，方法检测限为5µg/kg，定量限为10µg/kg， 14种苯并咪唑类药物及其代谢物在10µg/kg～200 µg/kg添加浓度范围内回收率均为70%～120%。批内、批间相对标准偏差均小于20%。方法设计独特，灵敏度高、定性、定量准确，完全满足食品安全检测法规的要求。

Abstract:

A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was established for the simultaneous determination of benzimidazoles and metabolites：Albendazole, Albendazole Sulfone, Albendazole Sulfoxide, Albendazole-2-aminosulfone, Thiabendazole, Thiabendazole-5-hydroxy, Mebendazole, Mebendazole-amine, 5-Hydroxymebendazole, Fenbendazole, Fenbendazole Sulfone, Oxfendazole, Flubendazole, Flubendazole-2-aminosulfone residues in animal Liver. The samples were extracted by Ethyl acetate, cleaned up with Waters Oasis MCX solid phase extraction (SPE) column. The separation of benzimidazoles and metabolites was performed on Waters 公司2695-Quattro micro system with a Xterra C18 column ，（2.1mm×150 mm，5μm) and the gradient elution solvent of acetonitrile (containing 0.1% formic acid) and water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min. The method was quantified by external standard method. The calibration curves were good linear between the peak areas and the concentrations of 10-1000 ng/mL with the correlation coefficient r＞0.99. The limit of detection of the 14 benzimidazoles and metabolites was 5 μg/kg, and the limit of quantification was 10 μg/kg. The average recoveries from spiked animal tissues at four concentrations of 10µg/kg～200 µg/kgμg/kg ranged 70%～120%. The relative standard deviations of intra- and inter-batch were both less than 20%.