Rochat Bertrand, Bolay Saskia, Pascual Andres, Calandra Thierry, Marchetti Oscar
Quantitative Mass Spectrometry Facility, Faculty of Biology and Medicine, Centre Hospitalier Universitaire Vaudois, 1011 Lausanne, Switzerland.
J Mass Spectrom. 2007 Apr;42(4):440-9. doi: 10.1002/jms.1171.
Caspofungin [(CASPO) MK-0991] is the first broad-spectrum anti-fungal agent of the echinocandin class approved for clinical use. Measurement of CASPO levels in blood might help monitor therapy in patients who are critically ill, in particular, if high-dose regimens or combinations of CASPO with other anti-fungals are used. The objective of this study was to develop a fast method for the measurement of CASPO levels in clinical blood samples using liquid chromatography coupled to a triple-quadrupole mass spectrometer. Stock solutions were prepared in plasma to avoid CASPO adsorption to glass and plastic surfaces during processing. CASPO and the internal standard (IS) were extracted from 100 microl of plasma using acetonitrile protein precipitation. The supernatant was diluted and directly injected into an analytical column (C8; 2.1 x 30 mm). The total run time was 15 min. CASPO was ionized by electrospray in the positive mode. CASPO and IS [M + 2H]2+ parent ions (m/z 547.3 and 547.8, respectively) and specific product ions (m/z 137.1 and 62.2, respectively) were used for the ion transitions. No carry over or cross-talk was observed on the column. The mean method recovery was 90 +/- 3%. Neither blood from different individuals (n = 6) nor the presence of concomitant drugs (n = 33) in plasma samples interfered with CASPO quantification. Quantification over time of the CASPO levels in plasma and whole blood was investigated at different pre-analysis storage conditions. The calibration curve included the clinically relevant CASPO concentration range from 0.04 to 20 microg/ml. Mean intra- and inter-day accuracy was 96.1 +/- 2.2% and 102.5 +/- 2.4%, respectively. Mean intra- and inter-day precision was 7.9 +/- 3.2% and 6.3 +/- 1.8%, respectively. This simple and robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method may easily be implemented for monitoring CASPO therapy.
卡泊芬净[（CASPO）MK - 0991]是首个被批准用于临床的棘白菌素类广谱抗真菌药物。检测血液中的CASPO水平可能有助于监测重症患者的治疗情况，特别是在使用高剂量方案或CASPO与其他抗真菌药物联合使用时。本研究的目的是开发一种使用液相色谱 - 三重四极杆质谱仪测定临床血液样本中CASPO水平的快速方法。储备液在血浆中制备，以避免在处理过程中CASPO吸附到玻璃和塑料表面。使用乙腈蛋白沉淀法从100微升血浆中提取CASPO和内标（IS）。将上清液稀释后直接注入分析柱（C8；2.1×30 mm）。总运行时间为15分钟。CASPO通过电喷雾在正模式下离子化。CASPO和IS的[M + 2H]2 +母离子（分别为m/z 547.3和547.8）以及特定的产物离子（分别为m/z 137.1和62.2）用于离子跃迁。在柱上未观察到残留或串扰。该方法的平均回收率为90±3%。来自不同个体的血液（n = 6）以及血浆样本中同时存在的药物（n = 33）均未干扰CASPO的定量。在不同的分析前储存条件下，对血浆和全血中CASPO水平随时间的定量进行了研究。校准曲线涵盖了临床相关的CASPO浓度范围，从0.04至20微克/毫升。日内和日间平均准确度分别为96.1±2.2%和102.5±2.4%。日内和日间平均精密度分别为7.9±3.
