Zhang Yantu, Zhang Zhujun, Sun Yonghua, Wei Yue
College of Chemistry and Materials Science, Shaanxi Normal University, Xi'an 710062, China.
J Agric Food Chem. 2007 Jun 27;55(13):4949-56. doi: 10.1021/jf070144y. Epub 2007 May 31.
A novel method was developed for the simultaneous determination of beta2-agonist residues such as terbutaline, salbutamol, and clenbuterol by high-performance liquid chromatography (HPLC) coupled with chemiluminescence (CL) detection. The procedure was based on the enhancement effect of beta2-agonists on the CL reaction between luminol and the complex of trivalent copper and periodate ([Cu(HIO6)2]5-), which was on-line electrogenerated by constant current electrolysis. The HPLC separation used a Nucleosil RP-C18 column (250 mm x 4.6 mm i.d., 5 microm; pore size, 100 A) with a mobile phase consisting of 90% acetonitrile and 10% aqueous ammonium acetate (20 mmol L-1, pH 4.0) at a flow rate of 1.0 mL min-1. The effects of several parameters on the HPLC resolution and CL emission were studied systematically. Liver samples were hydrolyzed with beta-glucuronidase followed by a solid-phase extraction procedure using Waters OasisMCX cartridges. Under optimum conditions, the limits of detection at a signal-to-noise ratio of 3 ranged from 0.007 to 0.01 ng g-1 and the limits of quantification at a signal-to-noise ratio of 10 ranged from 0.023 to 0.033 ng g-1 for three beta2-agonists. The relative standard deviations (RSDs) of intra- and interday precision were below 4.5%. The average recoveries for beta2-agonists (spiked at the levels of 0.05-5.0 ng g-1) in pig liver ranged from 84 to 110%, and the RSDs of the quantitative results were from 1.6 to 7.2%. The proposed method was successfully applied to the determination of beta2-agonist residues in pig liver samples.
开发了一种通过高效液相色谱（HPLC）与化学发光（CL）检测联用同时测定β2 - 激动剂残留（如特布他林、沙丁胺醇和克伦特罗）的新方法。该方法基于β2 - 激动剂对鲁米诺与三价铜和高碘酸盐络合物（[Cu(HIO6)2]5 - ）之间化学发光反应的增强作用，该络合物通过恒流电解在线电生成。HPLC分离使用Nucleosil RP - C18柱（250 mm×4.6 mm内径，5微米；孔径，100 Å），流动相由90%乙腈和10%乙酸铵水溶液（20 mmol L - 1，pH 4.0）组成，流速为1.0 mL min - 1。系统研究了几个参数对HPLC分离度和CL发射的影响。肝脏样品先用β - 葡萄糖醛酸酶水解，然后使用Waters OasisMCX柱进行固相萃取。在最佳条件下，三种β2 - 激动剂在信噪比为3时的检测限为0.007至0.01 ng g - 1，在信噪比为10时的定量限为0.023至0.033 ng g - 1。日内和日间精密度的相对标准偏差（RSD）低于4.5%。猪肝中β2 - 激动剂（加标水平为0.05 - 5.0 ng g - 1）的平均回收率为84%至110%，定量结果的RSD为1.6%至7.2%。所提出的方法成功应用于猪肝样品中β2 - 激动剂残留的测定。
