Determination of busulfan and methotrexate in children with small volume of plasma by HPLC-MS/MS
Objective To develop a high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)to determine busulfan and methotrexate in the plasma and monitor the therapeutic drugs in children with hematological disease.Methods The plasma samples were pretreated by protein precipition by methanol.The analytes were eluted on Phenomenex Kinetex EVO C1 8 column with the mobile phase consisting of water(2 mmol·L-1 ammonium acetate+0.07％formic acid)-methanol at the flow rate of 0.5 mL·min-1.Ions monitored in the multiple reaction monitoring(MRM)mode were m/z 264.1 → 151.0 for busulfan,m/z 272.0 → 159.0 for busulfan-D8(internal standard),m/z 455.1 → 308.2 for methotrexate,and m/z 458.1 → 311.1 for methotrexate-D3(internal standard).Results Calibration plots were established at the concentration range of 62.50～4000 ng·mL-1 for busulfan(r=0.9984)and 15.63～1000 ng·mL-1 for methotrexate(r=0.9990).Accuracy,precision,matrix effect and stability all met the requirements.Conclusion The method is simple,rapid and suitable for therapeutic drug monitoring of busulfan and methotrexate in the plasma of pediatric patients with neoplastic hematological disorder.
busulfanmethotrexateHPLC-MS/MStherapeutic drug monitoring
万方数据
维普
