超高效液相色谱串联质谱法检测水产品中四环素类药物及其差向异构体

Simultaneous Determination of Tetracyclines and Their Epimers in Aquatic Products by Ultra Performance Liquid Chromatography with Tandem Mass Spectrometry

建立了同时检测水产品中四环素母体及其差向异构体四环素、土霉素、金霉素、强力霉素、4-差向四环素、4-差向土霉素、4-差向金霉素共7种药物的超高效液相色谱-串联质谱(UPLC-MS/MS)方法。样品采用0.1mol/LNa2EDTA-Mcllvaine缓冲溶液提取2次,经滤纸过滤和高速离心除去提取液中大颗粒物质,取提取液的一半体积上HLB固相萃取柱净化,应用WatersAcquityUPLCBEHC18柱(100mm×2.1mm,1.7μm)高效分离,以0.1%甲酸水溶液-乙腈为流动相梯度洗脱,电喷雾正离子多反应监测模式监测,质谱分段扫描方式扫描。结果表明,7种四环素药物呈现良好的线性关系(R2>0.995),4种四环素母体的定量限(S/N=10)为2.0μg/kg,3种代谢物的定量限为5.0μg/kg。各分析物的加标回收率为70.3%~90.5%,RSD为5.5%~11%。本方法分析时间短,8min内即完成7种药物的分离,且分离效果好,峰形尖锐。方法的灵敏度、准确度高,适用于水产品中四环素类母体和代谢物的同时测定。

An ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the simultaneous determination of 7 kinds of tetracyclines and their epimers including tetracycline, chlortetracycline, oxytetracycline, doxycycline, 4-epi-tetracycline, 4-epi-chlortetracycline, 4-epi-oxytetracycline in edible parts of aquatic products. The samples were extracted with 0.1 mol/L EDTA-Mcllvaine buffer solution (pH 4.0). The extract was filtered and high-speed centrifuged in order to remove some large particulate matters before further purification. Then Half of the volume was cleaned-up and pre-concentrated using HLB cartridge. All the target compounds were separated on an ACQUITY UPLC BEH C18 column (100 mm×2.1 mm, 1.7 μm) with gradient elution by using acetonitrile and 0.1% formic acid aqueous as eluent, and detected by UPLC-MS/MS under ESI+ ionization, MRM mode and section scanning mode. Under optimized conditions, this method had a good linearity(R2>0.995) and the limits of quantification were in the range of 2.0-5.0 μg/kg (S/N=10). The recoveries of the method for the 7 TCs spiked at three different levels were 70.3%-90.5% with the relative standard deviations (RSDs) of 5.5%-11%. The analysis was rapid and can be achieved in less than 8 min with good separation and sharp peak shapes. The method had the characteristics is of high accuracy and good precision, was suitable for simultaneous determination of tetracycline antibiotics and their metabolites in aquatic products.