Development Of LC-MS/MS Methods For Hematologic Tumor Drugs In TDM Based On Dry Blood Spot And Multi-component Analysis

BackgroundCancers are the leading causes of death in all countries around the world,and hematological malignancies are the fourth most common malignancy in both men and women.Currently,the use of targeted drugs,such as proteasome inhibitors bortezomib and tyrosine kinase inhibitors(TKI),play important roles in clinical therapy.Since patients of hematological malignancies are mostly children and the elderly,their physiological,biochemical and pathological mechanisms are quite different from those of ordinary patients,with different pharmacokinetic and pharmacodynamic characteristics.Therefore,clinical rational drug use should be guided in the treatment process.The most powerful tool is therapeutic drug monitoring(TDM).The treatment window of bortezomib and TKIs is narrow,drug effectiveness or toxicity is correlated with drug exposure,drug metabolism has a large individual difference,and treatment failure will bring serious consequences,which are in line with the principle of clinical selection and development of TDM drugs.However,due to the poor adaptability of bortezomib and the low efficiency of TKI drugs,there are still some obstacles in the development of clinical TDM of these two kinds of drugs,due to the fact that traditional TDM is usually carried out by plasma samples and analysis methods for single drug.With the continuous development of analytical and testing technology,the methods applied to TDM on the basis of liquid chromatography-tandem mass spectrometry(LC-MS/MS)are also constantly innovating,creating a better choice for the development of TDM of bortezomib and TKIs.As a simple sampling technique,The dried blood spot(DBS)is suitable for improving the adaptability of TDM of bortezomib due to its advantages of no venipuncture,small sample size,excellent stability,convenient storage and transportation,etc.And multicomponent analysis greatly expanded the number of TKIs that could be measured simultaneously without increasing the complexity of the entire sample processing process and prolonging the overall analysis time,making it feasible for a large number of hematological tumor patients to carry out TDM simultaneously after the application of TKIs.PurposeThis study aims to establish one LC-MS/MS method for bortezomib determination using dried blood spots with low volume and high sensitivity,for the determination of bortezomib concentration in DBS samples,and to determine the correlation between concentration in plasma samples and DBS samples,and another LC-MS/MS method for TKIs determination,for the simultaneous determination of 40 TKIs in plasma samples,which is promising to provide technical support for the further development of TDM and related research of these two kinds of drugs.MethodsWhatman 903 paper was used to prepare bortezomib DBS samples,and the determination method of bortezomib DBS samples was established and verified with LC-MS/MS.At the same time,the stability of bortezomib plasma samples and DBS samples was compared,and the influence of hematocrit on the determination of bortezomib DBS concentration was investigated.Clinical plasma and DBS samples of bortezomib were collected and the consistency test of the determination results was conducted.Establish a multi-component analysis method for the determination of 40 TKIs in plasma samples with LC-MS/MS,and the method validation was conducted based on the recommendations published by the U.S.Food and Drug Administration,by accessing the following parameters:selectivity,linearity and the lower limit of qualification,accuracy and precision,stability,matrix effect and recovery.ResultsThe concentration of bortezomib had a good linear relationship in the range of 0.2-20 ng/mL in DBS matrix,and the accuracy of all quality control samples ranged from 94.7%to 112.5%,and the precision was lower than 10.7%.The matrix effect of DBS was lower than 13.2%and the recovery was between 87.3%-100.2%.Bortezomib DBS samples showed significantly better stability when stored at low and room temperature compared to plasma samples.Different hematocrit values had no significant effect on the accuracy of concentration determination.There was no significant difference in the measured concentrations of bortezomib in clinical plasma samples and DBS samples.The concentrations of 40 kinds of TKIs showed good linearity within the range of their respective standard curves in plasma matrix,and the accuracy of all quality control samples ranged from 85.9%-114.1%,and the precision was lower than 11.0%.The extraction recovery ranged from 92.6%-114.7%,and the matrix effect of plasma was lower than 11.3%.This method can be used for the determination of drug concentration in multiple patients with different kinds of TKI,which was rapid,convenient,and suitable for clinical TDM.ConclusionBased on LC-MS/MS technology,this study established and verified the determination method of bortezomib in DBS and the multi-component analysis method for determination of the concentration of forty TKIs simultaneously in plasma for the first time,which made up for the poor adaptability of bortezomib and the lack of high efficiency of TKI drugs in the traditional TDM process.It can offer technical means for the formulation of the treatment window of these two kinds of hematological malignancy drugs,provide guidance for the adjustment of drug administration plan and dose,and reduce toxicity and side effects on the premise of ensuring efficacy,so as to achieve precision medicine.