基于萃取和超高效液相色谱-串联质谱法的贝类抗生素快速测定方法

建立一种简单、快速的超声波萃取–固相萃取净化–超高效液相色谱串联质谱分析方法, 可以同时测定淡水贝类软组织中磺胺类、喹诺酮类、大环内酯类、β-内酰胺类和氨基糖苷类中15种抗生素。采取超声波萃取法, 对贝类体内的抗生素进行萃取, 用固相萃取方法净化提取液, 最后用超高效液相色谱–三重四级杆质谱对目标物进行分析检测。主要比较两种固相萃取柱Oasis HLB和Oasis PRiME HLB对污染物的净化效率, 后者展现出较好的结果。15种抗生素加标回收率实验结果表明: 当加标浓度为50 ng/g时, 回收率为64%~121%, 相对标准偏差为0.5%~19% (n=3); 添加高浓度(100 ng/g)样品时, 回收率为67%~117%, 相对标准偏差为1%~9%(n=3)。方法检测限(LOD)为0.004~0.5 ng/g (干重), 定量限(LOQ)为0.013~1.67 ng/g (干重)。该方法具有回收率高、检测限低的特点。使用该方法对鄱阳湖3个采样点三角帆蚌中的抗生素进行分析, 结果显示, 在15种抗生素中, 有9种均不同程度地检出, 检出频率和浓度最高的是甲氧苄胺嘧啶, 最高浓度达到78.8 ng/g, 其次是奇霉素(41.2 ng/g)和环丙沙星(39.8 ng/g)。

Fast Determination of Multiple Antibiotics in Mussels Based on Extraction and UPLC-MS/MS

A fast and simple method for simultaneous determination of fifteen antibiotics (sulfadiazine, ciprofloxacin, penicillin, spechromycin and roxithromycin) in mussels using ultrasound extraction, solid phase extraction purification and ultra performance liquid chromatography-tandem mass spectrometric was established. The mussel samples were extracted by ultrasonic extraction, followed by solid-phase extraction (SPE) as the cleanup procedure. Ultra performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS) was used for identification and quantification of antibiotics. The performance of two kinds of SPE cartridges, Oasis HLB and Oasis PRiME HLB, was compared, with the latter showing better results. Recoveries ranged from 64% to 121% at spiking levels of 50 ng/g, with RSD being 0.5%–19%, and from 67% to 117% at spiking levels of 100 ng/g, with RSD being 1%–9%. Limits of detection (LODs) ranged from 0.004 to 0.5 ng/g (dry weight) and limits of quantification (LOQs) ranged from 0.013 to 1.67 ng/g (dry weight). The recoveries were reasonable and the detection limit was low. Finally, the method was successfully applied to the determination of the target antibiotics in mussel samples (Hyriopsis cumingii) collected from Poyang Lake. 9 out of 15 selected antibiotics were detected in the collected mussel tissues. The concentration and detection frequency of trimethoprim in mussels were the highest. The concentration of trimethoprim was 78.8 ng/g, followed by spectinmycin (41.2 ng/g) and ciprofloxacin (39.8 ng/g).