A rapid and sensitive method for the quantitative detection of busulfan (BU) in children's hemolytic samples by HPLC–tandem mass spectrometry (MS/MS) was established. In this study, the sample preparation procedure involved a one‐step protein precipitation with acetonitrile (ACN) solution, and the HPLC–MS/MS method used Hypersil GOLD C₁₈. The mobile phase consisted of 10 mM ammonium acetate solution (containing 0.1% formic acid) and ACN with a flow rate of 0.4 mL/min. Multiple reaction monitoring modes were used for quantitative analysis and the ion pairs of BU and BU‐d8 were m/z 263.9 → 150.9 and 272.0 → 159.0, respectively. BU had a good linearity in the range of 0.01–10 μg mL⁻¹. The intra‐ and inter‐day relative error was between –7.21% and 8.26%, and the coefficient of variation was less than 12.64%. The average extraction recovery rate in plasma samples was 99.76% ± 6.53%, and the matrix in normal plasma and hemolyzed plasma had no significant effect on the detection results. Normal and hemolytic samples could maintain good stability at 4, 25 and –40°C. As a result, this method is particularly suitable for determining BU in hemolytic samples from children with hematopoietic stem cell transplantation (HSCT), and this study provides the methodological basis for further research on the pharmacokinetics of BU in children with HSCT.

中文翻译：

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快速HPLC-MS / MS法测定造血干细胞移植患儿溶血样品中的白消安。

建立了一种快速灵敏的HPLC-串联质谱（MS / MS）定量检测儿童溶血样品中白消安（BU）的方法。在这项研究中，样品制备过程涉及用乙腈（ACN）溶液一步沉淀蛋白质，而HPLC–MS / MS方法使用Hypersil GOLD C ₁₈。流动相由10 m M醋酸铵溶液（含0.1％甲酸）和ACN组成，流速为0.4 mL / min。多种反应监测模式用于定量分析，BU和BU-d8的离子对分别为m / z 263.9→150.9和272.0→159.0。BU在0.01–10μgmL ^-1范围内具有良好的线性。日内和日间相对误差在–7.21％和8.26％之间，并且变异系数小于12.64％。血浆样品的平均提取回收率为99.76％±6.53％，正常血浆和溶血血浆中的基质对检测结果无明显影响。正常和溶血样品可以在4、25和–40°C下保持良好的稳定性。因此，该方法特别适用于测定造血干细胞移植（HSCT）儿童溶血样品中的BU，该研究为进一步研究HSCT儿童的BU的药代动力学提供了方法学依据。