Fast Determination of Chloramphenicol, Florfenicol, Thiamphenicol in Eggs by Ultra Performance Liquid Chromatography-tandem Mass Spectrometry Combined with Library Search
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摘要: 采用超高效液相色谱-三重四级杆/复合线性离子阱质谱(QTrap UPLC-MS/MS)技术结合谱库检索分析,建立了鸡蛋中氯霉素、氟苯尼考和甲砜霉素的快速检测确证方法。样品加入氯霉素-D5后,经乙腈提取,浓缩至干,3.00 mL水复溶,正己烷去脂净化,以乙腈-水为流动相,经Phenomenex Kinetex F5色谱柱(100 mm×3.0 mm,2.6 μm)分离,用QTrap UPLC-MS/MS进行多反应监测(MRM)、信息依赖性扫描(IDA)、增强子离子扫描(EPI)和谱库检索分析。氯霉素内标法定量,氟苯尼考、甲砜霉素外标法定量。结果表明,氯霉素和氟苯尼考在0.03~50.0 μg/L,甲砜霉素在0.06~50.0 μg/L的线性范围内线性关系良好,相关系数r>0.999;氯霉素、氟苯尼考和甲砜霉素在3个加标水平下,平均回收率为90.1%~120.0%,精密度为4.2%~11.9%,检出限为0.05~0.10 μg/kg。该方法高效、简便、准确,可用于鸡蛋中酰胺醇类抗生素定性和定量分析。Abstract: A new method had been developed for the fast determination of chloramphenicol, florfenicol and thiamphenicol in eggs by ultra performance liquid chromatography-triple quadrupole/linear ion trap combined with library search. The egg extract was concentrated up to dryness after extracted with acetonitrile and then dissolved in water with n-hexa purified. After gradient elution separation on Phenomenex Kinetex F5 LC column (100 mm×3.0 mm, 2.6 μm) using acetonitrile and water as the mobile phases, a multiple reaction monitoring (MRM) scan as survey scan and an enhanced product ion (EPI) scan as dependent scan were performed in an information dependent acquisition (IDA) experiment. Finally, the identification of the drugs were carried out by library search with a newly developed MS/MS library. Chloramphenicol was determined by internal standard method, as florfenicol and methamphenicol by external standard method. The linear ranges of chloramphenicol and florfenicol were good in the range of 0.03~50.0 μg/L, and of thiamphenicol was good in the range of 0.06~50.0 μg/L. The correlation coefficient (R) of them were all over 0.999.The average recoveries of chloramphenicol, florfenicol and thiamphenicol in eggs varied from 90.1% to 120.0%, and the relative standard deviations were between 4.2% and 11.9% at three spiked levels. The limits of detection (LODs) ranged from 0.05 to 0.10 μg/kg. The method is simple, effective, and can be applied to real samples.
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