The Establishment Of Analytical Methods For The Therapeutic Drug Monitoring Of Aminoglycosides And Vancomycin And Its Clinical Application

Amikacin,isepamicin and tobramycin are important aminoglycosides for treatment of multidrug-resistant gram-negative nosocomial infections.Vancomycin is glycopeptide,which is preferred antibiotic for treatment of infections caused by Methecillin-resistant Staphlococcus aureus(MRSA).Data from the CHINET surveillance of bacterial resistance in China showed that both of these antibiotics had have high antimicrobial activities for drug-resistant gram-negative bacteria or gram-positive bacteria,the monotherapy or multiple chemotherapy are suggested in clinicals.Neurotoxicity of these antibiotics is obvious due to renal excretion,clinical efficacy and neurotoxicity are related to blood concentration.To improve their safety and efficacy,therapeutic drug monitoring(TDM)is suggested to be conducted in clinics.The dosage regimen can be adjusted according to drug concentrations in patients' blood,treatment guidelines and their deseases.Accurate and sensitive methods to quantify the drugs concentrations in human serum are neccessary.The major methods for quantification of aminoglycosides and vancomycin are immunoassay.The analytical immunoassay kit for amikacin,isepamicin and tobramycin are supported in foreign countries,while the kits are unvailable in China due to the insufficiency of registration certificate and TDM for aminoglycosides can't be carried out.There are immunoassay,high performance liquid chromatography(HPLC),LC-MS/MS to analyzer vancomycin..It's reported that lower limit of quantitation of different instruments from different manufacturers would be different,and deviation exists among different instruments.The accuracy of detection would be challenged.Detective bias from large volume data of analysing clinical samples and its influence on judgments of adjustingdosage regimen havn't been researched.This study are divied into two parts:Part one: The establishment of three LC-MS/MS methods to analyze the concentrations of aminoglycosides in human serum.We developed LC-MS/MS methods to quantify three aminoglycosides concentrations in human serum,and validated the methods in specificity,precision and accuracy,matrix effect and recovery,standard curve,stabilities of samples stored at different temperature,reproducibility,residual effect and interaction.Samples from patients infected gram-negative bacteria and dosed with amikacin were collectedandanalyzed.The trough and peak concentrations in Chinese patients' serum were collected and detected for the first time,which can provide the basis for the clinical study on amikacin therapeutic drug monitoring for Chinese patients infected gram-negative bacteria.Part two:Based on former experiment,this study assayed detective bias from large volume data of analysing clinical samples and its influence on judgments of adjustingdosage regimen,which would reference for the choice of analytical method.