固相萃取-超高效液相色谱-串联质谱法测定蜂产品中10种头孢类药物

Determination of ten cephalosporins in bee products by solid phase extraction-ultra-performance liquid chromatography-tandem mass spectrometry

建立了蜂产品中10种头孢类药物(头孢喹肟、头孢噻肟、头孢洛宁、头孢哌酮、头孢匹林、头孢氨苄、头孢乙腈、头孢拉定、去乙酰基头孢匹林、头孢唑林)含量的液相色谱-串联质谱测定方法。蜂产品样品中头孢类药物用乙腈-水(80∶20, v/v)溶液提取,离心,上清液经Oasis PRIME HLB固相萃取柱净化,氮吹后复溶,进行液相色谱-串联质谱分析。采用Acquity BEH C18色谱柱,以0.1%(v/v)甲酸水溶液-甲醇体系作为流动相进行梯度洗脱,ESI源正离子模式电离,多反应离子监测模式(MRM)检测,基质校准外标法定量。结果表明,10种头孢类药物在一定浓度范围内峰面积与质量浓度的相关系数(r^2)大于0.999,线性关系良好;检出限为0.15~1.5μg/kg,定量限为0.50~5.0μg/kg;在阴性蜂产品样品中的加标回收率为75.0%~89.8%,相对标准偏差(RSD)为1.4%~4.6%(n=5)。该方法检测周期短,准确度和精密度高,能满足多种蜂产品样品中头孢类药物的检测需要。

An ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) method for the simultaneous quantitative determination of ten cephalosporins in bee products, including honey, royal jelly, and lyophilized royal jelly powder, has been established. The samples were extracted with an acetonitrile-water(80∶20, v/v) solution. After purification by solid phase extraction using an Oasis PRIME HLB cartridge, the extract was blown to dryness under a stream of nitrogen gas and then re-dissolved in 1 mL 0.1%(v/v) formic acid solution and methanol(95∶5, v/v). The samples were separated on an Acquity UPLC BEH C18 column, using a mixture of 0.1%(v/v) formic acid solution and methanol as the mobile phase under gradient elution. The analysis was carried out using a positive electrospray ion source in the multiple reaction monitoring mode. The matrix-matched external standard method was applied to quantitative analysis. Good linear relationships were obtained for the ten cephalosporins in certain concentration ranges, and the correlation coefficients were more than 0.999. The limits of detection and limits of quantification for the ten cephalosporins were in the ranges 0.15-1.5 μg/kg and 0.5-5 μg/kg, respectively. The recoveries for the analytes in the bee products were in the range of 75.0%to 89.8%, with relative standard deviations of 1.4% to 4.6%. This method is characterized by a short analysis time and is suitable for the determination of cephalosporins in different bee products by virtue of its simplicity and reliability.