Determination of sabarubicin and its metabolite M3 in human urine using UHPLC-MS/MS

Abstract：Objective:To establish an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for determining the concentrations of sabarubicin and its metabolite M3 in human urine for a phase Ⅰ clinical pharmacokinetic study in Chinese small cell lung cancer (SCLC) patients.Methods:Liquid-liquid extraction by chloroform was used to isolate sabarubicin and M3,using doxorubicin hydrochloride as the internal standard (IS).The optimal chromatographic behavior was achieved on an ACQUITY UPLC BEH Shield RP18 (100 mm× 2.1 mm,1.7 μm) column using a mixture of 0.1％ formic acid water and acetonitrile (90∶ 10,v/v) as the mobile phase.The flow rate was 0.3 mL· min-1 ESI + detection was performed in MRM mode using target ions at m/z 644.2→130.2 for sabarubicin,m/z 646.2→130.1 for M3 and m/z 544.3 →397.1 for doxorubicin.Results:The calibration curve exhibited excellent linearity over the range of 2 ～ 200 ng·mL-1 for both sabarubicin and M3,with reproducible coefficients of correlation greater than 0.99.No endogenous interference was observed at the retention time of the analytes and the carry-over effect was found to be negligible from previous concentrated samples.The method was validated with respect to selectivity,extraction recovery,matrix effect,intra-and inter-day precision,accuracy,and stabilities,all of which met the requirements of FDA and CFDA guidelines,and this method has already been used to detect the concentrations of sabarubicin and M3 in Chinese patients.Conclusion:We developed a sensitive,accurate and rapid method to quantify sabarubicin and M3 in human urine,which can fulfill the requirements of the clinical pharmacokinetic study of sabarubicin.