Song Ta-Shu, Hsieh Yow-Wen, Peng Ching-Tien, Liu Cheng-Hsiung, Chen Tai-Lin, Hour Mann-Jen
School of Pharmacy, China Medical University, Taichung 404, Taiwan.
Biomed Chromatogr. 2012 Dec;26(12):1575-81. doi: 10.1002/bmc.2734. Epub 2012 Mar 27.
A fast and accurate liquid chromatography/tandem mass spectrometric (LC-MS/MS) assay was first developed and validated for the determination of deferiprone in human plasma. The analytes were extracted with acetonitrile from only 50 μL aliquots of human plasma to achieve the protein precipitation. After extraction, chromatographic separation of analytes in human plasma was performed using a Synergi Fusion-RP 80A column at 30 °C. The mobile phase consisted of methanol and 0.2% formic acid containing 0.2 mM EDTA (60:40, v/v). The flow rate of the mobile phase was 0.8 mL/min. The total run time for each sample analysis was 4 min. Detection was performed using electrospray ionization in positive ion multiple reaction monitoring mode by monitoring the precursor-to-parent ion transitions m/z 140.1 → 53.1 for deferiprone and m/z 143.1 → 98.1 for internal standard. A linear range was established from 0.1 to 20 µg/mL. The limit of detection was determined as 0.05 µg/mL. The validated method was estimated for linearity, recovery, stability, precision and accuracy. Intraday and interday precisions were 4.3-5.5 and 4.6-7.3%, respectively. The recovery of deferiprone was in the range of 80.1-86.8%. The method was successfully applied to a pharmacokinetic study of deferiprone in six thalassemia patients.
首次开发并验证了一种快速准确的液相色谱/串联质谱（LC-MS/MS）分析法，用于测定人血浆中的去铁酮。仅从50μL人血浆等分试样中用乙腈萃取分析物，以实现蛋白质沉淀。萃取后，在30℃下使用Synergi Fusion-RP 80A柱对人血浆中的分析物进行色谱分离。流动相由甲醇和含有0.2 mM EDTA的0.2%甲酸（60:40，v/v）组成。流动相流速为0.8 mL/min。每个样品分析的总运行时间为4分钟。采用电喷雾电离在正离子多反应监测模式下进行检测，通过监测去铁酮的前体离子到母离子的跃迁m/z 140.1→53.1和内标的m/z 143.1→98.1。建立了0.1至20μg/mL的线性范围。检测限确定为0.05μg/mL。对验证后的方法进行了线性、回收率、稳定性、精密度和准确度评估。日内和日间精密度分别为4.3-5.5%和4.6-7.3%。去铁酮的回收率在80.1-86.8%范围内。该方法成功应用于6例地中海贫血患者的去铁酮药代动力学研究。
