UPLC-MS/MS法检测动物性食品中19种β-受体激动剂残留

Determination of β-agonists in animal derived food by UPLC-MS/MS #$NLYE Ni1, SUN Lei1, YIN Hui1,WANG Yilin1,BI Yanfeng1, WANG Hejia1,XU Shixin1

关键词:  动物性食品  β-受体激动剂  残留  超高效液相色谱-串联质谱

Key words:  animal derived food  β-agonists  residue  UPLC-MS/MS

摘要:

建立了前处理过程较为简便、能同时检测猪、牛和羊的肌肉、肝脏、肾脏和鸡蛋中特布他林、齐帕特罗、沙丁胺醇、西马特罗、西布特罗、克仑塞罗、羟甲基克伦特罗、克仑丙罗、氯丙那林、莱克多巴胺、克仑特罗、妥布特罗、溴布特罗、克仑潘特、班布特罗、马布特罗、马喷特罗、苯乙醇胺A和喷布特罗等19种β-受体激动剂的UPLC-MS/MS方法。样品经乙腈-异丙醇混合溶液（4:1，V/V）提取后，用β-葡萄糖醛苷酶/芳基硫酸酯酶酶解[1]，混合型阳离子交换固相萃取柱净化[2]，然后用UPLC BEH C18色谱柱分离，以0.1%甲酸乙腈溶液和0.1%甲酸水溶液为流动相进行梯度洗脱，基质匹配溶液内标法或外标法定量。结果表明：19种β-受体激动剂在1～50ng/mL基质匹配标准溶液浓度范围内呈现良好线性关系，相关系数R2均大于0.990；在猪、牛和羊的肌肉、肝脏、肾脏和鸡蛋中的检测限均为0.1ng/kg，定量限均为0.5 ng/kg。从0.5、1、2和5 ng/kg四个添加浓度检测结果可以看出，19种药物的回收率范围为60%～120%，批内、批间相对标准偏差均小于20％。该方法具有简便快捷、灵敏度高、定性准确等特点。

Abstract:

A UPLC-MS/MS method was established for the determination of Terbutaline, Zilpaterol, Salbutamol, Cimaterol, Cimbuterol, ,Clenciterol, Hydroxymethyl ClenbuterolS Clenproperol Clorprenaline, Ractopamine, Clenbuterol, Tulobuterol, SBrombuterol , Clenisopenterol , SBambuterol , Mabuterol , Mapenterol, Phenyl ethyl alcohol amine A and Penbutolol residue in animal derived food. After the tissues were extracted by Acetonitrile-2-Methyl-1-propanol, enzymolysis,centrifuged, neutralized, followed by liquid-liquid extraction with ethyl acetate and tert-butyl methyl ether, respectively. The combined extracts were applied to a solid phase extraction MCX cartridge for cleanup. The separation of β-agonists was performed on Waters Acquity UPLC system with the column of BEH C₁₈ and the gradient elutinon solvent of acetonitrile(0.1% formic acid) and water(0.1% formic acid) at a flow rate of 0.3mL/min. The method was quantified by blank tissue spiked standard curves. The calibration curves were good linear between the peak areas and the concentrations of 1～50μg/kg,the correlation coefficient R₂＞0.990. The limits of detection of the 19 β-agonists in animal derived food were 0.1ng/kg , and the limit of quantification was 0.5ng/kg. The average recoveries from spiked animal tissues at three concentrations of 0.5,1,2 and 5ng/kg ranged 60%～120%. The intra- and inter-batch variation coefficients were both less than 20%.