亲水相互作用色谱-串联质谱法同时测定动物组织中金刚烷胺与利巴韦林

Determination of Amantadine and Ribavirin in Animal Tissues by Hydrophilic Interaction Chromatography-Tandem Mass Spectrometry

采用亲水相互作用色谱-串联质谱技术,建立了同时测定动物组织中金刚烷胺和利巴韦林的方法。样品加入同位素内标,采用20 g/L三氯乙酸提取,经磷酸酯酶酶解,PBA固相萃取小柱净化后,采用HILIC色谱柱（100 mm×2.1 mm,3μm）分离,以5 mmol/L乙酸铵水溶液（含0.1%甲酸）-乙腈为流动相梯度洗脱,采用电喷雾离子源串联质谱,在正离子扫描方式下以多反应监测（MRM）模式检测,内标法定量。在优化条件下,金刚烷胺和利巴韦林在0.5~5.0μg/L质量浓度范围内线性关系良好,定量下限分别为1.0μg/kg和2.0μg/kg。在鸡肉和鸡肝样品中,金刚烷胺和利巴韦林在3个加标水平（分别为1.0、2.0、5.0μg/kg和2.0、5.0、10.0μg/kg）下的平均回收率为89.2%~109%,相对标准偏差为2.0%~9.3%。该方法准确可靠、方便快捷,适用于动物组织中金刚烷胺和利巴韦林的同时定量分析。

A hydrophilic interaction chromatography- tandem mass spectrometric（ HILIC- MS /MS） method was developed for the simultaneous determination of amantadine and ribavirin in animal tissues. After added isotopically labeled standards as the internal standards,the sample was extracted with 20 g / L trichloroacetic acid and enzymed with acid phosphatase,then purified with PBA cartridge. The targeted compounds were separated on a HILIC column（ 100 mm × 2. 1 mm,3 μm） by gradient elution with 5 mmol / L ammoniumacetate（ containing 0. 1% formic acid） and acetonitrile as mobile phases,and detected by MS / MS in positive electrospray ionization and multiple reaction monitoring（ MRM） mode. The quantification was performed by the internal standard method. The calibration curves showed good linearity in the range of 0. 5- 5. 0 μg / L. The limits of quantitation for amantadine and ribavirin were 1. 0 μg / kg and 2. 0 μg / kg,respectively. The recoveries for chicken musle and liver samples at three spiked levels were in the range of 89. 2%- 109% with relative standard deviations of 2. 0%- 9. 3%. This method was accurate,reliable and simple,and was suitable for the analysis of amantadine and ribavirin in animal tissues.