超高效液相色谱-串联质谱法同时测定牛肉中青霉素类药物及其代谢产物

Simultaneous Determination of 2 Penicillins and Their 5 Major Metabolites in Bovine Muscle by Ultra Performance Liquid Chromatography Tandem Mass Spectrometry

建立了同时检测牛肉中2种青霉素(阿莫西林和青霉素G)及其5种主要代谢产物的超高效液相色谱-串联质谱分析方法。牛肉样品加入青霉素V为内标,经乙腈提取后,在UPLC HSS T3色谱柱(100 mm×2.1 mm,1.8μm)上分离,以乙腈和添加0.15%甲酸与5 mmol/L乙酸铵的水溶液(pH 2.8)作为流动相,采用梯度洗脱程序,正离子电喷雾多反应监测(MRM)模式检测,一次进样分析需8 min,基体匹配标准内标法定量。方法学确证表明:牛肉样品的线性范围为25～2500μg/kg,各被测物质的线性相关系数均大于0.99,加标回收率为91%～106%,相对标准偏差为2%～9%(n=6),样品的检出限为0.05～3.0μg/kg(S/N>3),定量限均为25μg/kg(S/N>10)。本方法灵敏度高,重复性良好,为生物组织样品中青霉素类药物的代谢研究提供了分析基础。

A comprehensive analytical method based on ultra performance liquid chromatography tandem mass spectrometry has been developed for the simultaneous determination of 2 penicillins（amoxicillin and penicillin G） and their 5 major metabolites.Penicillin V was used as the internal standard in this method.Bovine muscle sample was prepared by precipitation of proteins with acetonitrile.The penicillins was analyzed on an ACQUITY UPLC HSS T3 column 100 mm×2.1 mm,1.8 μm using a mixture of 0.15% formic acid in water with 5 mmol/L ammonium acetate（pH 2.8） and acetonitrile as the mobile phase and a gradient program with a cycle time of 8 min,and detected by positive electrospray ionization tandem mass spectrometry in the MRM mode,and quantified by matrix-match internal standard solution.The standard curves were linear in the range of 25-2500 μg/kg for bovine muscle.The average recoveries were 91%-106% with RSDs of 2%-9%（n=6）.The detection limits（S/N3） of the 8 analytes were 0.05-3.0 μg/kg,the limits of quantitation（S/N10） were all 25 μg/kg.The method is sensitive and repeatable,and can be applied to the field of residue analysis about penicillins and their metabolites.