A New Method Study Of High-throughput Determination Of Blood Drug Concentration By HPLC-MS/MS

PartⅠ Determination of nine mental drugs in human plasma using solid-phase supported liquid-liquid extraction and HPLC-MS/MSObjective:A rapid and reliable method using solid-phase supported liquid-liquid extraction（SLE）coupled with high-performance liquid chromatography tandem mass spectrometry was established for the analysis of nine mental drugs（aripiprazole,olanzapine,sertralin,moclobemide,paroxetine,maprotiline,iproniazid,risperidone and fluoxetine）in human plasma.Method:The plasma samples were pre-treated with 96-well alkaline diatomite plate and quantified by isotope internal standard method.In this experiment,the conditions affecting the pretreatment of solid support liquid-liquid extraction were optimized,including the type and dosage of diluent,the type and dosage of eluent,and the addition of eluent.The collected eluate was dissolved by nitrogen blowing and determined by high performance liquid chromatography-mass spectrometer.Results:A good linearities was obtained in the range of 1～100 ng·m L^-1for aripiprazole,olanzapine,sertralin,moclobemide,paroxetine,maprotiline and iproniazid;0.1～10 ng·m L^-1 for risperidone and fluoxetine with a correlation coefficient above 0.995.The method matrix effect（ME）was80～120%,negligible;intraday and interday precision（RSD）was less than20%;accuracy was 77.24%～127.71%.Conclusion:This method can quickly and sensitively determine the concentration of nine mental drugs in human plasma,provide support for the clinical monitoring of depression patients,and promote the treatment of depression.PartⅡ Determination of six immunosuppressant drug concentrations in human whole blood using Qu ECh ERS and HPLC-MS/MSObjective:A high performance liquid chromatography tandem mass spectrometry method was established to simultaneously determine the concentrations of six immunosuppressant drugs（mycophenolic acid,mycophenolate mofetil,tacrolimus,rapamycin,everolimus and pimecrolimus）in human whole blood by optimizing the Qu ECh ERS（rapid,simple,inexpensive,effective,stable and safe）pretreatment method.Method:Human whole blood samples were extracted by ethyl acetate,anhydrous magnesium sulfate salting out and purified by PSA adsorbent.After centrifugation,the supernatant nitrogen was blown and methanol was redissolved.The conditions optimized in this experiment include the type and dosage of extractant,the amount of salting out agent,the type and dosage of purifying agent.Results:Under the optimal conditions,a good linearities was obtained in the range of 1～100 ng·m L^-1 for tacrolimus,rapamycin,everolimus and pimecrolimus;0.1～10 ng·m L^-1 for mycophenolate mofetil;0.1～10 ng·m L^-1 for mycophenolic acid with a correlation coefficient above 0.99.The intraday and interday precision（RSD）was less than 15%,and the recovery rate was between 85.07%～114.95%.Conclusion:This method is rapid,sensitive,specific and reproducible.It is suitable for simultaneous determination of mycophenolic acid,mycophenolate mofetil,tacrolimus,rapamycin,everolimus and pimecrolimus in human whole blood.It provides support for clinical monitoring of blood drug concentration and promotes the treatment of diseases.PartⅢ Determination of twelve weight-reducing medicines concentrations in human plasma using 96-well protein precipitation plates and HPLC-MS/MSObjective:A specific and sensitive method was developed by HPLC-MS/MS to simultaneous determination of twelve weight-reducing medicines in plasma,including methylephedrine,amphetamine,bupropion and fenfluramine.Method:Human plasma samples were pretreated with 96-well protein precipitation plate,and the collected filtrate was dissolved by methanol after nitrogen blowing.In this experiment,the types and dosage of precipitator were optimized and compared with the common protein precipitation method.Results:The detection sensitivity of various drugs was high.Under the optimal conditions,a good linearities was obtained in the range of 0.1～20ng·m L^-1 for bupropion,fluoxetine,bisacodyl,fenfluramine,sibutramine and rimonabant;0.5～20 ng·m L^-1 for methylephedrine,amphetamine,bumetanide,lovastatin,simvastatin and fenofibrate with a correlation coefficient above0.995.Matrix effect was negligible,the intraday and interday precision（RSD）was less than 15%,and the recovery rate was between 75.63%～108.21%.Conclusion:This method is fast,simple,specific and sensitive.It is suitable for the simultaneous determination of 12 weight-reducing drugs such as methylephedrine,amphetamine,bupropion and fenfluramine in human blood,and provides a reference for the detection of other drugs.