Patel Daxesh P, Nair Sneha, Suhagia Bhanubhai N, Patel Bhargav M
Laboratory of Metabolism, Center for Cancer Research, National Cancer Institute, Bethesda, MD 20892, United States.
Department of Chemistry, School of Sciences, Gujarat University, Ahmedabad 380009, India.
J Pharm Biomed Anal. 2016 Nov 30;131:355-363. doi: 10.1016/j.jpba.2016.09.005. Epub 2016 Sep 5.
A specific, rapid, sensitive and selective ultra-performance liquid chromatography - tandem mass spectrometry has been developed for the simultaneous determination of midodrine and desglymidodrine in human plasma. The analytes and its deuterated analogs were quantitatively extracted from 100μL of human plasma by solid phase extraction technique. Separation of analytes was achieved on the Waters Acquity UPLC BEH C18 (50×2.1mm, 1.7μm) column using acetonitrile-4.0mM ammonium formate, pH 2.5(90:10, v/v) as mobile phase. The protonated analytes were quantified by selected reaction monitoring in the positive ionization mode by triple quadrupole mass spectrometer. The calibration plots were linear over the concentration range of 0.050-50.0ng/mL. The intra-batch and inter-batch precision (%CV) across quality control levels was <4.0 and the% mean relative recovery was ≥96%. Various other parameters like stability in different conditions; matrix effect and reproducibility of the method were performed in accordance with the guidelines specified by the USFDA for bioanalytical method development and validation. The developed method was successfully administered to the pharmacokinetics study of 5 mg midodrine tablet in 12 healthy subjects. Reproducibility of assay was proved by reanalysis of 48 incurred samples.
已开发出一种特异性强、快速、灵敏且选择性高的超高效液相色谱-串联质谱法，用于同时测定人血浆中的米多君和去甲米多君。通过固相萃取技术从100μL人血浆中定量提取分析物及其氘代类似物。采用乙腈-4.0mM甲酸铵（pH 2.5，90:10，v/v）作为流动相，在Waters Acquity UPLC BEH C18（50×2.1mm，1.7μm）色谱柱上实现分析物的分离。通过三重四极杆质谱仪在正离子模式下采用选择反应监测对质子化分析物进行定量。校准曲线在0.050 - 50.0ng/mL的浓度范围内呈线性。跨质量控制水平的批内和批间精密度（%CV）<4.0，平均相对回收率（%）≥96%。按照美国食品药品监督管理局（USFDA）生物分析方法开发和验证规定的指南，对该方法在不同条件下的稳定性、基质效应和重现性等各种其他参数进行了测定。所开发的方法成功应用于12名健康受试者口服5mg米多君片的药代动力学研究。通过对48份实际样品的重新分析证明了该测定方法的重现性。
