摘要
目的:开发HPLC-ESI-MS/MS方法测定大鼠组织对氨基水杨酸浓度以改善以往方法的复杂前处理过程及低灵敏度的问题,并应用于研究其在大鼠体内的分布。方法:采用一步蛋白沉淀的前处理,选择菲诺门Synergi 4u Polar-RP 80A(150 mm×4.6 mm;4μm)色谱柱以及梯度法进行色谱分离。内标为氯唑沙宗。采用API4000液相色谱串联质谱(LC-MS/MS)法测定。扫描方式采用多重反应监测(MRM),负离子模式。对氨基水杨酸及内标(氯唑沙宗)监测离子对为m/z152.0→108.1,m/z 167.9→131.9。结果:总分析运行时间为6.5 min。对氨基水杨酸在0.05~50μg/m L内线性关系良好,定量下限低至0.05μg/m L。本实验方法学所有验证结果都符合FDA生物样品分析指导原则要求。结论:本分析方法准确可靠,并成功应用于临床前组织分布研究。
AIM：To develop a novel HPLC-ESI-MS/MS method to determine P-aminosalicylic acid （PAS） in different rat tissues to overcome the weaknesses of the conventional complicated pretreatment process and its low sensitivity.METHODS：The assay was performed with a one-step pre-column process of protein precipitation for extraction and the use of a Phenomenex Synergi 4u Polar-RP 80A （150 mm × 4.6 mm;4 μm） column with gradient elution for separation.Chlorzoxazone was selected as an internal standard （IS）.LC-MS/MS （API4000） was applied for determination.PAS and IS were detected by tandem mass spectrometer using multiple reaction monitoring mode （MRM） with negative electrospray ionization.The ion transitions recorded in multiple reaction monitoring mode were m/z 152.0→108.1 for PAS and m/z 167.9→ 131.9 for the internal standard.RESULTS：The total analytical run time was short of 6.5 min.Calibration curve was linear over the range of 0.05-50 μg/mL and the lower limit of quantification （LLOQ） was low up to 0.05 μg/ mL.All validation criteria were fulfilled in compliance with the guidelines of the United States Food and Drug Administration （FDA）.CONCLUSION：This HPLC-MS/MS method is accurate and reliable.The described assay method has been successfully applied for the measurement of PAS in preclinical distribution study.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2017年第3期272-280,共9页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
国家自然科学基金资助项目(81460570)
