Rathod Dhiraj M, Patel Keyur R, Mistri Hiren N, Jangid Arvind G, Shrivastav Pranav S, Sanyal Mallika
Department of Chemistry, Kadi Sarva Vishwavidyalaya, Gandhinagar, Ahmedabad 382015, India.
Bioanalytical Research Department, Accutest Research Lab, Bodakdev, Ahmedabad 380059, India.
J Pharm Anal. 2017 Feb;7(1):56-62. doi: 10.1016/j.jpha.2016.05.005. Epub 2016 May 26.
The present study describes a simple, reliable and reproducible liquid chromatography-tandem mass spectrometry method (LC-MS/MS) for the simultaneous determination of allopurinol and its active metabolite, oxypurinol in human plasma for a pharmacokinetic/bioequivalence study. After protein precipitation (PPT) of 100 µL plasma sample with 1.0% formic acid in acetonitrile, the recovery of the analytes and allopurinol-d2 as an internal standard ranged from 85.36% to 91.20%. The analytes were separated on Hypersil Gold (150 mm×4.6 mm, 5 µm) column using 0.1% formic acid-acetonitrile (98:2, v/v) as the mobile phase. Quantification was done using electrospray ionization in the positive mode. The calibration concentration range was established from 60.0 to 6000 ng/mL for allopurinol and 80.0-8000 ng/mL for oxypurinol. Matrix effect in human plasma, expressed as IS-normalized matrix factors ranged from 1.003 to 1.030 for both the analytes. The developed method was found suitable for a clinical study with 300 mg allopurinol tablet formulation in healthy subjects.
本研究描述了一种简单、可靠且可重复的液相色谱-串联质谱法（LC-MS/MS），用于在药代动力学/生物等效性研究中同时测定人血浆中的别嘌醇及其活性代谢物氧嘌呤醇。用含1.0%甲酸的乙腈对100 μL血浆样品进行蛋白沉淀（PPT）后，分析物和作为内标的别嘌醇-d2的回收率在85.36%至91.20%之间。使用Hypersil Gold（150 mm×4.6 mm，5 µm）色谱柱，以0.1%甲酸-乙腈（98:2，v/v）作为流动相分离分析物。采用正模式电喷雾电离进行定量。别嘌醇的校准浓度范围为60.0至6000 ng/mL，氧嘌呤醇的校准浓度范围为80.0至8000 ng/mL。人血浆中的基质效应，以IS归一化基质因子表示，两种分析物的范围均为1.003至1.030。所开发的方法被发现适用于在健康受试者中对300 mg别嘌醇片剂制剂进行的临床研究。
