高效液相色谱法测定猪肝组织中3-甲基喹喔啉-2-羧酸

摘要

To develop a high performance liquid chromatography method for determination of 3-methyl-quinoxaline-2-carboxylic acid (MQCA) in porcine liver. MQCA was extracted from porcine liver with 0.3 mol/L hydrochloric acid. The extract was cleaned up on MAX Sep-Pak Silica solid-phase extraction cartridge. Then the eluate was collected and dried under a gentle stream of nitrogen gas and the dissolved residue was analyzed by HPLC. The HPLC separation was performed on a Symmetry Shield reversed phase C18 boned silica column, with mobile phase of acetonitrile-methanol-0.5％ formic acid (15 : 50: 35, V/V/V) and determination wave of 320 nm in an UV detector. The validation was carried out on spiked porcine liver(spiked at 20～250 μg/kg), and the average recoveries of target drug ranged from 72. 35％ to 85.00％, with intra-day relative standard deviations(RSD)(n=5) of 5. 56％ to 9. 49％, and inter-day RSD(n= 3) of 5. 96％ to 9. 63％. The limits of quantification (LOQ) in porcine liver was 20 μg/kg and the limits of detection (LOD) was 6.65 μg/kg. The results demonstrated that the sensitivity , accuracy, and precision of this method meet the requirements of veterinary drug residue analysis. The method is applicable to detect MQCA in porcine liver.%本研究建立了猪肝组织中3-甲基喹喔啉-2羧酸(3-methyl-quinoxaline-2-carboxylic acid,MQCA)的高效液相色谱检测方法.采用0.3 mol/L盐酸提取猪肝组织中药物,经MAX柱净化,以乙腈:甲醇:0.5%甲酸水(15:50:35,V/V/V)为流动相,C18色谱柱进行药物分离,320 nm波长下紫外检测,结果当药物在猪肝组织中添加水平为20～250μg/kg时,平均回收率为72.35%～85.00%,日内测定的相对标准偏差(RSD)(n=5)为5.56%～9.49%,日间RSD(n=3)为5.96%～9.63%.猪肝组织中MQCA的定量限为20μg/kg,检出限为6.65 μg/kg,表明该方法灵敏度、准确度和精密度均符合兽药残留分析技术的要求,适用于猪肝组织中MQCA的检测.