Tandem Mass Spectrometry Method for the Quantification of Serum Busulfan
Tandem Mass Spectrometry Method for the Quantification of Serum Busulfan
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DOI：
10.1097/01.ftd.0000173372.04945.7b
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发表时间：
2005-10
影响因子：
2.5
通讯作者：
Mark D. Kellogg;T. Law;M. Sakamoto;N. Rifai
中科院分区：
文献类型：
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作者：
Mark D. Kellogg;T. Law;M. Sakamoto;N. Rifai
布磺胺是一种烷基化剂，是骨髓移植术前治疗方案中最常用的成分。患者间和患者内药代动力学的显著变化需要根据时间-浓度曲线下的面积来个性化剂量。及时报告结果对于调整剂量以降低与方案相关的发病率和死亡率至关重要。作者建立了一种快速、准确、灵敏的血清丁硫丹直接注射串联质谱定量方法。血浆样品（50 μL）用含1,6-二-（甲磺酰氧基）己烷为内标的1 mL甲醇提取。上清液在氮气（40℃，30分钟）下干燥，然后溶解在200 μL甲醇中，转移到适合LC/MS/MS分析的干净玻璃小瓶中。样品通过高效液相色谱泵输送，每分钟输送0.2 mL甲醇，20 μL样品注入配备涡轮离子喷雾的串联质谱仪。总分析时间为5分钟。在内部标准中，对第一季度/第三季度的丁硫丹（BU）转换在269/55和297.1/55.1进行监控。该方法对10 μmol/L呈线性，对至少0.5 μmol/L敏感。在1、2.2和4.4 μmol/L浓度下，测定结果重复性分别为4.2%、5.6%和6.3%。3种不同对照样品的运行内精密度分别为3.9%、3.9%和6.9%。4种不同BU浓度加入10种不同BU游离血浆样品的平均回收率为98%。与建立的HPLC-UV方法的相关斜率为0.98，截距为0.1,r2 = 0.95 （n = 48）。没有发现明显的干扰物质或离子抑制。该方法对现有的HPLC-UV法进行了改进。该方法准确度高，重复性好，需要较少的样品、样品制备和分析时间。
Busulfan, an alkylating agent, is most commonly used as a component of bone marrow transplantation preoperative regimens. Significant interpatient and intrapatient variations in pharmacokinetics require individualizing the dosage based on area under the time-versus-concentration curve. Timely result reporting is critical to dose adjustment to reduce morbidity and mortality associated with the regimen. The authors developed a rapid, accurate, and sensitive method for the quantification of serum busulfan using direct inject tandem mass spectrometry. Plasma samples (50 μL) are extracted in 1 mL of methanol containing 1,6-bis-(methanesulfonyloxy)hexane as an internal standard. The supernatant is dried under nitrogen (40°C, 30 minutes) and then dissolved in 200 μL methanol and transferred into a clean glass vial suitable for LC/MS/MS analysis. The sample is delivered using an HPLC pump that delivers 0.2 mL of methanol per minute, and 20 μL of sample is injected into a turbo ion spray-equipped tandem mass spectrometer. Total analysis time is 5 minutes. The Q1/Q3 transition for busulfan (BU) is monitored at 269/55 and 297.1/55.1 for the internal standard. The assay is linear to 10 μmol/L and sensitive to at least 0.5 μmol/L. The interassay reproducibility at 1, 2.2, and 4.4 μmol/L were 4.2%, 5.6%, and 6.3%, respectively. Within-run precision using 3 different control samples was 3.9%, 3.9%, and 6.9%. Mean recovery of 4 different BU concentrations spiked into 10 different BU free plasma samples was 98%. Correlation with an established HPLC-UV method revealed a slope of 0.98, an intercept of 0.1, and r2 = 0.95 (n = 48). No significant interfering substances or ion suppression was identified. This method is a significant improvement over the existing HPLC-UV method for BU determination. The method is highly accurate, reproducible, and requires less specimen, sample preparation, and analysis time.