Vancea Szende, Imre Silvia, Donáth-Nagy Gabriella, Béla Tokés, Nyulas Mária, Muntean Teofil, Borka-Balás Réka
Faculty of Pharmacy, University of Medicine and Pharmacy, Gheorghe Marinescu 38, RO-540139, Târgu Mureş, Romania.
Talanta. 2009 Jul 15;79(2):436-41. doi: 10.1016/j.talanta.2009.04.003. Epub 2009 Apr 10.
A new simple, sensitive and selective liquid chromatography coupled with mass spectrometry (LC/MS) method for quantification of captopril after precolumn derivatization with p-bromo-phenacyl-bromide in human plasma was validated. Plasma samples were analysed on a monolithic column (Cromolith Performance-RP 18e, 100 mm x 4.6 mm I.D., 3 microm) under isocratic conditions using a mobile phase of a 40:60 (v/v) mixture of acetonitrile and 0.1% (v/v) formic acid in water. The flow rate was 1 mL/min at the column temperature of 30 degrees C. In these chromatographic conditions, the retention time was 4.4 min for captopril derivative. The detection of the analyte was in MRM mode using an ion trap mass spectrometer with electrospray positive ionisation. The monitored ions were 216, 253, 255, 268, 270 m/z derived from 415 m/z for derivatized captopril. The sample preparation was very simple and consisted in plasma protein precipitation from 0.2 mL plasma using 0.3 mL methanol after the derivatization reaction was completed. Calibration curves were generated over the range of 10-3000 ng/mL with values for coefficient of correlation greater than 0.993 and by using a weighted (1/y(2)) quadratic regression. The values for precision (CV %) and accuracy (relative error %) at quantification limit were less than 9.9% and 3.9%, for within- and between-run, respectively. The mean recovery of the analyte was 99%. Derivatized samples demonstrated good short-term, long-term, post-preparative and freeze-thaw stability. This is the first reported LC-MS/MS method for analysis of captopril in human plasma that uses protein precipitation as sample processing procedure. The method is very simple and allows obtaining a very good recovery of the analyte. The validated LC-MS/MS method has been applied to a pharmacokinetic study of 50mg captopril tablets on healthy volunteers.
建立了一种新的简单、灵敏且具选择性的液相色谱-质谱联用（LC/MS）方法，用于在人血浆中对卡托普利与对溴苯甲酰溴进行柱前衍生化后进行定量分析，并对该方法进行了验证。血浆样品在整体柱（Cromolith Performance-RP 18e，100 mm×4.6 mm内径，3微米）上，于等度条件下进行分析，流动相为乙腈与0.1%（v/v）甲酸水溶液按40:60（v/v）混合而成的溶液。在柱温30℃时流速为1 mL/min。在这些色谱条件下，卡托普利衍生物的保留时间为4.4分钟。使用带有电喷雾正离子化的离子阱质谱仪，以多反应监测（MRM）模式对分析物进行检测。监测的离子为衍生化卡托普利从415 m/z产生的216、253、255、268、270 m/z。样品制备非常简单，在衍生化反应完成后，用0.3 mL甲醇从0.2 mL血浆中沉淀血浆蛋白。校准曲线在10 - 3000 ng/mL范围内生成，相关系数值大于0.993，采用加权（1/y²）二次回归。定量限处的精密度（CV%）和准确度（相对误差%）值，批内分别小于9.9%，批间分别小于3.9%。分析物的平均回收率为99%。衍生化样品显示出良好的短期、长期、制备后和冻融稳定性。这是首次报道的用于分析人血浆中卡托普利的LC-MS/MS方法，该方法采用蛋白沉淀作为样品处理程序。该方法非常简单，能使分析物获得很好的回收率。经验证的LC-MS/MS方法已应用于50mg卡托普利片剂在健康志愿者身上的药代动力学研究。
