Lee Eun Jin, Park Nuri, Lee Sang Hee, Lee Woochang, Kim Hyun Soo, Chun Sail, Min Won-Ki
Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Republic of Korea.
Department of Laboratory Medicine, Hallym University College of Medicine, Dongtan Sacred Heart Hospital, Hwaseong-Si, Republic of Korea.
Ann Clin Lab Sci. 2019 Mar;49(2):212-217.
Busulfan, frequently used as a conditioning regimen for hematopoietic stem cell transplantation, has a narrow therapeutic range and wide intra-and interpatient variabilities. Therefore, therapeutic drug monitoring of busulfan is necessary to ensure that the drug concentrations of patients are within a targeted therapeutic range. In this study, we developed a simple and accurate method for measuring busulfan concentrations using liquid chromatography tandem mass spectrometry (LC-MS/MS).
Separation and detection of busulfan was performed using T3 column equipped with LC-MS/MS. Busulfan was isolated from 50 μL human plasma after mixing with busulfanH (internal standard) solution, calibrator, and quality-control material. The sample was eluted and gradated with a mobile phase composed of ammonium acetate, formic acid, and water or methanol. The busulfan concentration was quantified using a six-point standard curve. Busulfan and busulfanH were detected in positive-ion multiple-reaction-monitoring mode. According to the Clinical and Laboratory Standards Institute guideline, we verified the precision, linearity, limit of detection (LOD), limit of quantification (LOQ), and carryover.
Busulfan and busulfanH were detected at m/z 264.1>151.1 and 272.2>159.1. The total run time was 3 min. Both intra-and inter-assay coefficients of variation were <3%. The calibration curve was linear at 25-5,000 ng/mL. The LOD and LOQ were 2.5 ng/mL and 25 ng/mL, respectively. The recoveries ranged from 92.0-104.8% and the carryover was-0.02%.
Our method for busulfan reduces total run time and has excellent analytical performance. It will be a useful method for therapeutic drug monitoring of busulfan in clinical laboratories.
白消安常用于造血干细胞移植的预处理方案，其治疗窗窄，患者内及患者间差异大。因此，对白消安进行治疗药物监测以确保患者的药物浓度处于目标治疗范围内很有必要。在本研究中，我们开发了一种使用液相色谱串联质谱法（LC-MS/MS）测定白消安浓度的简单准确方法。
使用配备LC-MS/MS的T3柱对白消安进行分离和检测。将白消安与人血浆50μL混合，加入白消安-d8（内标）溶液、校准品和质量控制物质后，分离出白消安。样品用由醋酸铵、甲酸和水或甲醇组成的流动相洗脱并梯度洗脱。使用六点标准曲线对白消安浓度进行定量。以正离子多反应监测模式检测白消安和白消安-d8。根据临床和实验室标准协会指南，我们验证了精密度、线性、检测限（LOD）、定量限（LOQ）和残留。
在m/z 264.1>151.1和272.2>159.1处检测到白消安和白消安-d8。总运行时间为3分钟。批内和批间变异系数均<3%。校准曲线在25 - 5000 ng/mL范围内呈线性。LOD和LOQ分别为2.5 ng/mL和25 ng/mL。回收率在92.0 - 104.8%之间，残留为-0.02%。
我们的白消安检测方法缩短了总运行时间，具有出色的分析性能。它将成为临床实验室对白消安进行治疗药物监测的有用方法。
