超高效液相色谱-电喷雾串联四极杆质谱法测定尿液中9种β_2-受体激动剂

Simultaneous determination of nine β_2-agonists residues in urines by ultra performance liquid chromatography-electrospraytandem mass spectrometry

目的研究尿液中9种β2-受体激动剂残留测定方法。方法样品在醋酸钠缓冲条件下（pH5.2）进行酶解,以高氯酸溶液沉淀蛋白,超声提取,调整提取溶液pH4.0,用HLB、MCX固相萃取柱浓缩和净化,通过Waters ACQUITY UPLCTM BEH C18色谱柱分离,以甲醇和含0.1%甲酸的水溶液为流动相梯度洗脱,采用超高效液相色谱-电喷雾串连四极杆质谱,正离子多反应监测（MRM）模式进行测定。结果检出限0.002-0.025ng/ml,最低定量限为0.007-0.08ng/ml。添加水平为0.1、0.5和2ng/ml时,9种β2-受体激动剂的加标回收率为77.4%-101.7%,相对标准偏差为3.4%-18%。结论该法测定尿液中9种β2-受体激动剂残留物方法准确可靠,灵敏度高。

Objective To study the determination method of nine β2-agonists residues in urine. Methods Urine samples were duconjugated with β2-glucuronidase/arylsulfatase enzyme in acetate buffer and deproteinized by perchloric acid, and then adjusted pH4.0. Sample concentration and purification were performed by Oasis HLB, Oasis MCX. The separation was performed on a Waters ACQUITY UPLCTM BEH C18 column （100mm × 2.1mm i.d., 1.7μm） with gradient elution using methanol and water （containing 0.1% formic acid） at a flow rate of 0.3ml/min. Results The limits of detection （LOD） of the method were from 0.002 to 0.025ng/ml and the limits of quantification （LOQ） ranged from 0.007 to 0.08ng/ml. Average recoveries for nine β2-agonists at the spiking levels of 0.1, 0.5 and 2ng/ml ranged from 77.4% to 101.7 % with relative standard deviations between 3.4 % and 18 %. Conclusion The method can be used to determine the residues of nine β2-agonists in urines.