A novel method for quantification of sulfolane (a metabolite of busulfan) in plasma by gas chromatography-tandem mass spectrometry

The role of busulfan (Bu) metabolites in the adverse events seen during hematopoietic stem cell transplantation and in drug interactions is not explored. Lack of availability of established analytical methods limits our understanding in this area. The present work describes a novel gas chromatography-tandem mass spectrometric assay for the analysis of sulfolane (Su) in plasma of patients receiving high-dose Bu. Su and Bu were extracted from a single 100 μL plasma sample by liquid-liquid extraction. Bu was separately derivatized with 2,3,5,6-tetrafluorothiophenolfluorinated agent. Mass spectrometric detection of the analytes was performed in the selected reaction monitoring mode on a triple quadrupole instrument after electronic impact ionization. Bu and Su were analyzed with separate chromatographic programs, lasting 5min each. The assay for Su was found to be linear in the concentration range of 20-400ng/mL. The method has satisfactory sensitivity (lower limit of quantification, 20ng/mL) and precision (relative standard deviation less than 15%) for all the concentrations tested with a good trueness (100 ± 5%). This method was applied to measure Su from pediatric patients with samples collected 4h after dose 1 (n = 46), before dose 7 (n = 56), and after dose 9 (n = 54) infusions of Bu. Su (mean ± SD) was detectable in plasma of patients 4h after dose 1, and higher levels were observed after dose 9 (249.9 ± 123.4ng/mL). This method may be used in clinical studies investigating the role of Su on adverse events and drug interactions associated with Bu therapy. Figure Overall sample preparation procedure for quantification of sulfolane and busulfan in plasma from patients receiving higher doses of busulfa