超高效液相色谱-串联质谱法分析鸡蛋中利巴韦林及其代谢物残留

Analysis of Ribavirin and Its Metabolites in Fresh Eggs by Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry

建立了超高效液相色谱-串联质谱法同时快速测定鸡蛋中利巴韦林及其两种主要代谢物TCONH2和RTCOOH的分析检测方法。样品采用乙腈-水（9∶1,V/V）提取,乙腈饱和正己烷除脂,C18结合GCB进行固相分散萃取除杂,Agilent ZORBAX SB-Aq色谱柱（100 mm×3.0 mm,1.8μm）分离,超高效液相色谱-串联质谱测定。结果表明：利巴韦林、TCONH_2和RTCOOH分别在2.0~200μg/L,0.5~200μg/L,5.0~200μg/L浓度范围内,线性良好,相关系数R2〉0.99,检出限分别为0.54,0.09和1.54μg/L,定量限分别为1.79,0.31和5.13μg/L。在5.0,10.0和50.0μg/L加标水平下,利巴韦林和RTCOOH回收率分别为96.1%~99.6%和42.9%~58.3%;在0.5,2.0和5.0μg/L加标水平下,TCONH2的回收率为75.9%~106.7%,相对标准偏差均为4.2%~12.7%。实际样品测定结果表明,本方法操作简单、快速、准确,能够满足鸡蛋中利巴韦林及其两种主要代谢物的分析检测。

A rapid and convenient ultra high performance liquid chromatography-tandem mass spectrometry （UPLC-MS / MS） method for analysis of ribavirin and its two main kinds of metabolites, 1,2,4-triazole-3-carboxamide （TCONH2 ） and 1-β-D-ribofuranosyl-1,2,4-triazole-3-carboxylic acid （RTCOOH）, in fresh eggs has been developed and validated. The sample was extracted with acetonitrile water （9 ∶ 1, V/ V）, added N-hexane to move liposoluble substances, after centrifugation, the upper solvent was cleaned-up by C18 and GCB, then determined by UPLC-MS / MS coupled with Agilent ZORBAX SB-Aq column （100 mm×3. 0 mm, 1. 8 μm）. Over the concentration range of 2. 0-200. 0 μg / L （0. 5-200. 0 μg / L and 5. 0-200. 0 μg / L） for ribavirin （TCONH2 and RTCOOH） in fresh egg matrix, the correlation coefficients （R2 ） of the calibration curves were above 0. 99, and the limits of detection （LODs） were 0. 54 μg / L （0. 09 and 1. 54 μg / L）. The limits of quantitation （LOQs） were 1. 79, 0. 31 and 5. 13 μg / L, respectively. At three spiked concentration levels of 5. 0, 10. 0 and 50. 0 μg / L, the recoveries of ribavirin and RTCOOH ranged from 96. 1% to 99. 6%and 42. 9% to 58. 3% . And at three spiked concentration levels of 0. 5, 2. 0 and 5. 0 μg / L, the recoveries of TCONH2 ranged from 75. 9% to 106. 7% . The results of the determination in various real samples showed that the method is simple, accurate, rapid and suitable determination of ribavirin and its two main kinds of metabolites in fresh eggs.