Development Of Analytical Method For Tyrosine Kinase Inibitors Based On HPLC-MS/MS Technnology And Research Of Therapeutic Drug Monitoring

[Objective]To develop the method for simultaneous determination of 18 tyrosine kinase inhibitors（TKIs）in human plasma by using LC-MS/MS analysis,and to provide reference for the adjustment of drug dose,individualized therapy and pharmacokinetic and/or pharmacodynamic parameters in vivo for tumor patients.[Methods]All the compounds were detected in positive ion mode on Agilent 6410 tandem mass spectrometry and two deuterated compounds 2H₃-regorafeinb and pazopanib-¹³C-D₃were used as internal standard of the analytes.The column was Waters XBridge?C18（2.1×100 mm,3.5μm,Waters Technology Corporation,USA）.The mobile phase A was acetonitrile and the mobile phase B was aqueous solution containing 0.1%formic acid.The gradient elution procedure was as follows:initial mobile phase ratio was 16:84（phase A:phase B）,and the organic phase increased from 16%to 86%in 0～0.5 min,then from 86%to 95%in 0.5～5 min,and finally remained at 95%in 5～6 min;the flow rate was 0.35 m L/min;Injection volume was 5μL;the analytical time was 6 min;and the column temperature was 35°C.According to the 2015 edition of the Chinese Pharmacopoeia biological sample quantitative analysis method verification guidelines,all the relevant verification were completed.the patients who were treated with TKIs in department of oncology of Shanghai Changzheng hospital were enrolled in this study,and 25 clinical samples conforming to drug therapy monitoring were collected,and this established and validated analytical method was used for blood drug concentration detection.And finally finding the range of therapeutic concentration through literature and combined with clinical efficacy,medication guidance and medication safety education for patients,to achieve the purpose of individualized drug.[Results]In this study,an LC-MS/MSmethod was developed and validated to detect the plasma concentration of small molecule TKIs,and the linear relationship of 18 TKIs was good,and the correlation coefficient r was all greater than 0.990.The matrix effects of all analysis compounds ranged from 67.04%to 117.57%,and the extraction recoveries ranged from 51.41%to 113.91%.The matrix factor and extraction recovery factor RSD%after internal standard correction were less than 15%.The method is simple,reproducible and sensitive and suitable for clinical monitoring of therapeutic drugs.Through measurement of 25 clinical samples,it was found that there were individualized differences in plasma drug concentration exposure under the same oral dose.Further studies are still needed on the relationship between drug exposure and clinical efficacy and side effects and then constructing the therapeutic window.Patients can obtain better treatment efficacy based on the vivo exposure of TKIs measured by LC-MS/MS.[Conclusion]In clinical practice,the new quantitative analysis method serves as a way of monitoring drug concentration in plasma of patients treated with 18 TKIs,which provides help for clinical pharmacy and is of great significance for further improving drug safety and efficacy.