高效液相色谱-串联质谱法研究猪毛发中克仑特罗和莱克多巴胺的残留及代谢规律

Studies on the Determination of Clenbuterol and Ractopamine inSwine Hairs Using HPLC-MS/MS and Its Residue Elimination

关键词:  高效液相色谱-串联质谱  猪毛发  克伦特罗  莱克多巴胺  代谢

Key words:  HPLC-MS/MS  swine hairs  clenbuterol  ractopamine  residue elimination

摘要:

建立了高效液相色谱-串联质谱（HPLC-MS/MS）同时检测猪毛发中克仑特罗和莱克多巴胺残留的方法。猪毛发经1 mol/L氢氧化钠溶液水解、乙酸乙酯萃取、MCX固相萃取柱净化，流动相溶解后用HPLC- MS/MS进行检测。克仑特罗在1.9～463.2 ng/mL浓度范围内线性关系良好（R2=0.9990）；回收率为83.3 %～86.6 %，相对标准偏差为6.7 %～9.2 %；检出限为0.3 μg/kg，定量限为0.8 μg/kg。莱克多巴胺在2.8～562.9 ng/mL浓度范围内线性关系良好（R2=0.9992）；回收率为83.4 %～88.4 %，相对标准偏差为7.2 %～9.6 %；检出限为0.4 μg/kg、定量限为1.2 μg/kg。应用该方法研究了克仑特罗、莱克多巴胺在猪毛发中的代谢规律并进行了猪毛发样品检测。结果喂药7 d至停药21 d猪毛发中均检出克仑特罗、莱克多巴胺残留；检测猪毛发样品25份，1份样品检出克仑特罗，残留量为58.68 μg/kg。

Abstract:

A method for simultaneous determination of clenbuterol and ractopamine residues in swine hairs was developed using high performance liquid chromatography coupled with tandem mass spectrometry.After hydrolyzed by 1 mol/L Sodium hydroxide aqueous solution,extracted by Ethyl aetate,purified by MCX solid phase extraction cartridge,the sample was dissolved in mobile phase for analysis by HPLC-MS/MS.The calibration curve of clenbuterol was linear in the range of 1.9~463.2 ng/mL（R2=0.9990）,the recoveries of different quantities of clenbuterol in swine hairs were from 83.3 % to 86.6 % with the RSD from 6.7 % to 9.2 %,The limit of determination and limit of quantitative of clenbuterol were 0.4 μg/kg and 0.8 μg/kg,respectively.The calibration curve of ractopamine was linear in the range of 2.8~562.9 ng/mL（R2=0.9992）,the recoveries of different quantities of ractopamine in swine hairs were from 83.4 % to 88.4 % with the RSD from 7.2 % to 9.6 %,The limit of determination and limit of quantitative of ractopamine were 0.4 μg/kg and 1.2 μg/kg, respectively.The result indicated that the residues of clenbuterol and ractopamine were found in swine hairs from the seventh day that the swines were fed with these drugs to the 21st day that the swines had not been fed;25 copy of swine hair samples were determined by the method,the residue of clenbuterol was found in one of these samples,its content was 58.68 μg/kg.