dos Reis Ederson Oliveira, Vianna-Jorge Rosane, Suarez-Kurtz Guilherme, Lima Edson Luiz da Silva, Azevedo Débora de Almeida
Departamento de Química Orgânica, Instituto de Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
Rapid Commun Mass Spectrom. 2005;19(12):1666-74. doi: 10.1002/rcm.1962.
A sensitive and specific assay for detection of busulfan in human plasma was developed. The assay is based on rapid isolation of busulfan by liquid-liquid extraction with ethyl acetate, and detection by high-performance liquid chromatography with electrospray ionization and tandem mass spectrometry. 1,6-Bis(methanesulfonyloxy)hexane, a synthesized analogue of busulfan, was used as the internal standard (IS). The acquisition was performed in the multiple reaction monitoring mode; busulfan and the IS were detected with no interferences from plasma matrix. The method was linear over the range 5-2500 ng mL(-1), with r2 > 0.99 and a run time of only 3.5 min. The intra- and inter-assay precisions were in the ranges 2.1-11.9% and 3.2-10.1%, respectively, and the intra- and inter-assay accuracies were 92.2-107.6% and 94.7-104.1%, respectively. The absolute recoveries were 82.0% (20 ng mL(-1)), 90.6% (1000 ng mL(-1)) and 80.0% (2000 ng mL(-1)) for busulfan, and 89.1% for the IS (1000 ng mL(-1)). The limits of detection and quantification were 2 and 5 ng mL(-1), respectively. The validated method was successfully applied to analyze plasma samples obtained from six adults receiving doses of 1 mg kg(-1) in a conditioning regimen prior to bone marrow transplantation. A marked intra-patient variation in busulfan concentrations during the steady state was observed, which limits the application of pharmacokinetic modeling and suggests that continuous therapeutic monitoring is necessary for adequate individualized dosing. In this regard, the present assay brings important advantages relative to other methods described in the literature, i.e., it is highly specific and simple to perform, with a rapid chromatographic run time (3.5 min), and the whole procedure can be completed in 4-5 h, which would permit dose corrections after the third dose allowing earlier and better dosing adjustments towards the target level of busulfan.
开发了一种灵敏且特异的检测人血浆中白消安的方法。该方法基于用乙酸乙酯液 - 液萃取快速分离白消安，并通过带有电喷雾电离和串联质谱的高效液相色谱进行检测。1,6 - 双（甲磺酰氧基）己烷，一种合成的白消安类似物，用作内标（IS）。采集在多反应监测模式下进行；白消安和内标检测时不受血浆基质干扰。该方法在5 - 2500 ng mL⁻¹范围内呈线性，r² > 0.99，运行时间仅为3.5分钟。批内和批间精密度分别在2.1 - 11.9%和3.2 - 10.1%范围内，批内和批间准确度分别为92.2 - 107.6%和94.7 - 104.1%。白消安的绝对回收率分别为82.0%（20 ng mL⁻¹）、90.6%（1000 ng mL⁻¹）和80.0%（2000 ng mL⁻¹），内标的绝对回收率为89.1%（1000 ng mL⁻¹）。检测限和定量限分别为2和5 ng mL⁻¹。该经过验证的方法成功应用于分析从六名接受1 mg kg⁻¹剂量进行骨髓移植预处理方案的成年人获得的血浆样本。观察到稳态期间患者体内白消安浓度存在显著差异，这限制了药代动力学模型的应用，并表明持续治疗监测对于适当的个体化给药是必要的。在这方面，本检测方法相对于文献中描述的其他方法具有重要优势，即它高度特异且操作简单，色谱运行时间短（3.5分钟），整个过程可在4 - 5小时内完成，这将允许在第三次给药后进行剂量校正，从而朝着白消安的目标水平更早、更好地调整给药剂量。
