Research Institute of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Central South University, 410013 Changsha, Hunan, PR China.
J Pharm Biomed Anal. 2013 Jul-Aug;81-82:8-12. doi: 10.1016/j.jpba.2013.03.015. Epub 2013 Mar 29.
A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of recombinant urate oxidase in human plasma. This assay was based on the determination of enzyme reaction product, (15)N-allantoin, and phenacetin was used as an internal standard (IS). Separation was achieved on a C18 column by the mobile phase of 30% water (containing 0.5% formic acid) and 70% methanol. Quantification was done using multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z 161 → m/z 118 for (15)N-allantoin and m/z 180 → m/z 110.1 for IS at positive ionization mode. The calibration curve was established over the range of 2.077-42.06 U/l and the correlation coefficient was larger than 0.99. The intra-day and inter-day relative standard deviations were less than 10.6%. Accuracy determined at three concentrations ranged between 98.6% and 109.2%. This method was successfully applied to a pharmacokinetic study of intravenous recombinant urate oxidase produced from Escherichia coli in Chinese healthy volunteers.
建立并验证了一种液相色谱-串联质谱（LC-MS/MS）法，用于测定人血浆中的重组尿酸氧化酶。该方法基于酶反应产物（15）N-尿囊素的测定，采用非那西汀作为内标（IS）。采用 C18 柱，以 30%水（含 0.5%甲酸）和 70%甲醇作为流动相进行分离。采用多反应监测（MRM）模式进行定量，监测（15）N-尿囊素的 m/z 161→m/z 118 和 IS 的 m/z 180→m/z 110.1 的前体到产物离子的跃迁。校准曲线的范围为 2.077-42.06 U/l，相关系数大于 0.99。日内和日间相对标准偏差均小于 10.6%。在三个浓度下测定的准确度在 98.6%至 109.2%之间。该方法成功应用于中国健康志愿者中大肠杆菌来源的重组尿酸氧化酶的静脉给药药代动力学研究。
