A rapid HPLC-MS/MS method for determining busulfan in hemolysis samples from children with hematopoietic stem cell transplantation.

: A rapid and sensitive method for the quantitative detection of busulfan in children's hemolysis samples by HPLC-MS/MS was established. In this study, the sample preparation procedure involved a one-step protein precipitation with acetonitrile solution, and the HPLC-MS/MS method used Hypersil GOLD C18. The mobile phase consisted of 10mM ammonium acetate solution (containing 0.1% formic acid)-acetonitrile with a flow rate of 0.4mL/min. Multiple reaction monitoring modes was used for quantitative analysis and the ion pairs of busulfan and busulfan-d8 were m/z 263.9→150.9 and 272.0→159.0, respectively. Busulfan had a good linearity in the range of 0.01~10 μg·mL-1 . The intra- and inter-day relative error (RE) was between -7.21% and 8.26%, and the coefficient of variation (CV) 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 hemolysis samples could maintain good stability at 4°C, 25°C and -40°C. As a result, this method is particularly suitable for determining busulfan in hemolysis samples from children with HSCT, and this study provides the methodological basis for further research on the pharmacokinetics of busulfan in children with HSCT.