摘要
目的建立测定人血浆中非布司他及其活性代谢物67M-4质量浓度的LC-MS/MS方法。方法采用乙腈沉淀蛋白法处理血浆样品,采用不同的LC-MS/MS条件分别测定血浆中极性差异较大的非布司他和67M-4的质量浓度。非布司他的测定采用Platisil ODS（150 mm×4.6 mm,5μm）色谱柱和体积分数0.1%甲酸溶液-乙腈（体积比30∶70）为流动相;67M-4的测定采用Zorbax SB-C18（50 mm×4.6 mm,5μm）色谱柱和体积分数0.2%甲酸溶液-乙腈为流动相,梯度洗脱。质谱检测采用多反应离子选择监测（MRM）方式。结果血浆中非布司他和67M-4的质量浓度分别在0.010 00～15.06 mg·L-1和0.505 0～202.0μg·L-1内线性关系良好,提取回收率分别为88.0%～93.5%和88.7%～96.2%,方法的专属性,日内、日间精密度及基质效应均符合测定要求。结论该方法灵敏、可靠,适用于非布司他在人体内的药代动力学研究。
Objective To develop a rapid and sensitive LC-MS / MS method for the determination of febuxostat and its active carboxylic acid metabolite 67M-4 in human plasma. Methods Plasma samples were pretreated using protein precipitation with acetonitrile. Due to the great polarity differences between the two analytes,different LC-M S / M S conditions were employed to determine febuxostat and 67M-4 in human plasma separately. Febuxostat was analysed on a Platisil ODS（ 150 mm × 4. 6 mm,5 μm） column with a mobile phase consisting of acetonitrile and 0. 2% formic acid aqueous solution（ V ∶ V = 70∶ 30）. The analysis of 67M-4was conducted on a Zorbax SB-C18（ 50 mm ×4. 6 mm,5 μm） column using gradient elution with acetonitrile and 0. 1% formic acid aqueous solution as the mobile phase. M ass spectrometry detection was performed in multiple selected reaction monitoring（ M RM） mode. Results The assay was linear over the concentration ranges of 0. 010 00- 15. 06 mg·L- 1for febuxostat and 0. 505 0- 202. 0 μg·L- 1for 67M-4,respectively. The extraction recoveries at low,middle,high level were 88. 0%- 93. 5% and 88. 7%- 96. 2%,respectively. The specificity,inter-and intra-run precisions and matrix effects met the requirements. Conclusions The method is accurate,sensitive,simple,and can be applied to pharmacokinetic study of febuxostat in human.
出处
《沈阳药科大学学报》
CAS
CSCD
北大核心
2015年第9期715-721,共7页
Journal of Shenyang Pharmaceutical University
