建立了分子印迹固相萃取（ MISPE）-超高效液相色谱-串联质谱（ UPLC-MS／MS）快速测定猪尿中15种β2-受体激动剂的分析方法。分别优化了上样溶液的 pH值、淋洗和洗脱溶液等 MISPE净化条件及质谱条件。将猪尿样品离心，经 MISPE柱上样，依次用水、乙腈、0.5％（v／v）乙酸乙腈溶液淋洗，10％（v／v）乙酸甲醇溶液洗脱，氮气吹干，0.1％（v／v）甲酸水-乙腈（9∶1，v／v）复溶；采用 BEH C18色谱柱分离，以乙腈-0.1％（v／v）甲酸水溶液为流动相，采用梯度洗脱，在电喷雾正离子多反应监测模式下外标法定量。考察了MISPE对15种β2-受体激动剂的吸附特异性；猪尿中15种β2-受体激动剂在0.1～20μg／L 范围内线性关系良好（ r2≥0.992）；检出限和定量限分别为0.03和0.1μg／L；15种β2-受体激动剂的加标回收率为65.6％～115.0％，批内、批间相对标准偏差分别为0.57％～16.1％和1.11％～16.8％。该方法操作简单、快速、灵敏度高、特异性好。
An ultra performance liquid chromatography-tandem mass spectrometry( UPLC-MS/MS)method coupled with molecularly imprinted solid phase extraction(MISPE)for rapid determination of 15 β2-agonists residues in swine urine has been established. In order to obtain the optimized purification conditions,different pH values of loading,washing and elution sol-vents for the MISPE procedure were optimized. Finally,the swine urine samples( pH 7. 0 ) were performed directly by MISPE column,then the column was washed with water,acetoni-trile and 0. 5%( v/v)acetic acid in acetonitrile,and eluted with 10%( v/v)acetic acid in metha-nol. Meanwhile,the parameters of UPLC-MS/MS have been optimized. The UPLC separation was performed on a BEH C18 column,and eluted with the mobile phase consisting of 0. 1%(v/v) formic acid aqueous solution and acetonitrile with linear gradient program. The analysis was in ESI positive ion scan mode and multiple reaction monitoring( MRM)mode. Under the optimum conditions,the blank swine urine was spiked with different concentrations analytes and taken for analysis to evaluate the established method. To highlight the advantage of MISPE,specificity adsorption for the 15 β2-agonists in swine urine was investigate. The linear ranges were 0. 1-20μg/L for the 15 β2-agonists( r2≥0. 992). The limits of detection( LODs)and the limits of quan-tification( LOQs)were 0. 03 and 0. 1 μg/L,respectively. At the spiked levels of 0. 1,0. 5 and 2. 5 μg/L in blank samples,the mean recoveries were 65. 6%-115. 0% with the relative standard deviations( RSDs,n=6)of 0. 57%-16. 1%. Overall,the proposed method is simple,quick, reliable,sensitive,and can be applied in large scale supervision of illegal usage of the 15 β2-agonists.
参考文献
| [1] | Yan, H.;Xu, D.;Meng, H.;Shi, L.;Li, L..Food poisoning by clenbuterol in China[J].Quality Assurance and Safety of Crops & Foods,20151(1):27-35. |
| [2] | Du, X.-D.;Wu, Y.-L.;Yang, H.-J.;Yang, T..Simultaneous determination of 10 β _2-agonists in swine urine using liquid chromatography-tandem mass spectrometry and multi-walled carbon nanotubes as a reversed dispersive solid phase extraction sorbent[J].Journal of chromatography, A: Including electrophoresis and other separation methods,2012:25-32. |
| [3] | Martín-Esteban, A..Molecularly-imprinted polymers as a versatile, highly selective tool in sample preparation[J].TrAC: Trends in Analytical Chemistry,2013:169-170. |
- 下载量()
- 访问量()
- 您的评分：
-
10%
-
20%
-
30%
-
40%
-
50%