Snyder Marion L, Ritchie James C
Department of Pathology and Laboratory Medicine, Emory University School of Medicine and Emory Healthcare Systems, Atlanta, GA, USA.
Methods Mol Biol. 2010;603:129-36. doi: 10.1007/978-1-60761-459-3_12.
Busulfan is a chemotherapy drug widely used as part of conditioning regimens for patients undergoing bone marrow transplantation (BMT). Challenges of busulfan treatment include a narrow therapeutic window and wide inter- and intra-patient variability. Inappropriately low drug levels lead to relapse and even graft rejection, while higher doses frequently have toxic and sometimes fatal consequences. Maintenance of plasma busulfan concentrations using repeated measurements and proper adjustment of dosage can reduce busulfan-related toxicity and improve treatment outcomes. We describe a rapid (2-minute total analysis time per sample) and simple method for accurate and precise busulfan concentration determination in plasma samples (100 microL) using high performance liquid chromatography combined with electrospray positive ionization tandem mass spectrometry (HPLC-ESI-MS/MS). Busulfan is isolated from plasma after internal standard (busulfan-D(8))-methanol extraction, dilution with mobile phase (ammonium acetate-formic acid-water), and centrifugation. The supernatant plasma is injected onto the HPLC-ESI-MS/MS and quantified using a six-point standard curve. The assay is linear from 0.025 microg/mL (approximately 0.1 micromol/L) to at least 6.2 microg/mL (approximately 25 micromol/L) with precisions of <5% over the entire range.
白消安是一种化疗药物，被广泛用作接受骨髓移植（BMT）患者预处理方案的一部分。白消安治疗面临的挑战包括治疗窗狭窄以及患者间和患者内的差异较大。药物水平过低会导致复发甚至移植物排斥，而较高剂量则常常会产生毒性，有时甚至会导致致命后果。通过重复测量维持血浆白消安浓度并适当调整剂量，可以降低与白消安相关的毒性并改善治疗效果。我们描述了一种快速（每个样品总分析时间为2分钟）且简单的方法，使用高效液相色谱结合电喷雾正离子串联质谱法（HPLC-ESI-MS/MS）准确精确地测定血浆样品（100微升）中的白消安浓度。在加入内标（白消安-D(8)）-甲醇萃取、用流动相（乙酸铵-甲酸-水）稀释并离心后，从血浆中分离出白消安。将上清液血浆注入HPLC-ESI-MS/MS，并使用六点标准曲线进行定量。该测定法在0.025微克/毫升（约0.1微摩尔/升）至至少6.2微克/毫升（约25微摩尔/升）范围内呈线性，在整个范围内精密度<5%。
