论文部分内容阅读
目的建立准确、灵敏、可靠的LC/MS/MS方法,定量检测家兔血浆和房水中咪哒酚衍生物浓度,并用于家兔给予咪哒酚衍生物滴眼后血浆及房水中药代动力学研究。方法血浆及房水样品经蛋白沉淀前处理,内标选用Amms,液相色谱柱为TOSOHTSKgel Amide,流动相为含0.1%甲酸的甲醇∶水(80∶20,体积比),仪器选用Finni-gan公司TSQ Quantum型液相色谱-质谱联用仪,采用电喷雾离子化电离源(ESI),选择反应离子监测(SRM)方式进行正离子检测,用于定量分析的离子反应分别为m/z 381→308(咪哒酚衍生物)和m/z 285→138(Amms)。结果在本实验条件下,咪哒酚衍生物与内标Amms分离良好,保留时间分别为1.48和1.55 min,血浆和房水中药物浓度分别在5~2000和10~10 000 ng/ml范围内线性关系良好,相关系数(r2)分别为0.9971和0.9999,定量下限分别为5和10 ng/ml,回收率均大于80%,日内、日间精密度均小于15%(n=5)。结论该方法快速、准确、灵敏度高,专属性好,可用于咪哒酚衍生物在家兔体内的药物代谢动力学研究。
Objective To establish an accurate, sensitive and reliable LC / MS / MS method for the quantitative determination of microdrophenol derivatives in rabbit plasma and aqueous humor and to provide pharmacokinetics of plasma and aqueous humor in rabbits after hypnophenol derivatisation Study. Methods The plasma and aqueous humor samples were pretreated by protein precipitation. The internal standard was Amms, the column was TOSOHTSKgel Amide and the mobile phase was methanol: water (80:20, volume ratio) with 0.1% formic acid. The instrument was Finni-gan The company TSQ Quantum liquid chromatography-mass spectrometer, electrospray ionization source (ESI), the choice of reactive ion monitoring (SRM) positive ion detection for the quantitative analysis of the ion reaction were m / z 381 → 308 (picol derivative) and m / z 285 → 138 (Amms). Results Under the conditions of this experiment, the microdrophenol derivatives were separated from the internal standard Amms for a retention time of 1.48 and 1.55 min, respectively. The drug concentrations in plasma and aqueous humor ranged from 5 to 2000 and from 10 to 10 000 ng / ml, respectively The correlation coefficients (r2) were 0.9971 and 0.9999, respectively. The lower limits of quantification were 5 and 10 ng / ml respectively. The recoveries were both higher than 80%. The intra- and inter-day precision was less than 15% (n = 5). Conclusion The method is rapid, accurate, sensitive and specific. It can be used to study the pharmacokinetics of imidacloprid derivatives in rabbits.