The Study On TDM Of Tri-azole Antifungal Agents And Tyrosine Kinase Inhibitors And Identification Of Endogenous Keto-steroids In Human Plasma Using LC-MS/MS

LC-MS/MS was widely used in drug analysis,metabonomics and therapeutic drug monitoring due to its excellent selectivity and sensitivity.We used low-resolution mass spectrometry and high-resolution mass spectrometry to perform therapeutic drug monitoring for tri-azole antifungal agents and tyrosine kinase inhibitors and identify endogenous keto-steroids,respectively.1.The study for therapeutic drug monitoring of tri-azole antifungal agents and tyrosine kinase inhibitors using LC-MS/MS.Tyrosine kinase inhibitors are first line therapy in patients with chronic myelogenous leukemia or acute lymphoblastic leukemia with Philadelphia chromosome.Tri-azole antifungals were recommended as first-line therapy for the treatment of invasive aspergillosis.Tri-azole antifungals may result in significantly increased plasma levels and thereby increased risks of toxicities from tyrosine kinase inhibitors.We developed and validated an LC-MS/MS method for simultaneous determination of imatinib,dasatinib,nilotinib,voriconazole and itraconazole in human plasma.The method was linear over the concentration range of 20.0 to 4000 ng/m L for imatinib and nilotinib,50.0 to 10000 ng/m L for voriconazole and itraconazole,and 2.00 to 400 ng/m L for dasatinib.The intra-and inter-day precision and accuracy,selectivity,recovery,matrix effect and stability were validated for each analyte.Dose adjustments of tyrosine kinase inhibitors and antifungal agents based on therapeutic drug monitoring and clinical efficacy was applied to decrease the risk of adverse drug reactions.Besides,therapeutic drug monitoring study using this method revealed the degree of drug interactions between tyrosine kinase inhibitors,antifungal agents and other commonly co-administered medications,which demonstrated the clinical influence of drug interactions on the treatment outcome in patients.2.Identification of endogenous keto-steroids in human plasma using LC-MS/MS.Steroids are a family of tetracyclic aliphatic compounds with cyclopentane polyphenanthrene,which are important signaling molecules in organisms.They are associated with the occurrence of various diseases.The low content of steroids in body fluids,poor ionization efficiency,and large interferences with other components in biological matrices made the detection of endogenous steroids challenging.In our study,QAO derivatization was used to enhance the detection of keto-steroids by LC-MS/MS.We summarized characteristic product ions of QAO derivatives of keto-steroids and established an analysis strategy based on Orbitrap LC-MS.The analytical strategy was used to analyze the human plasma samples derivatized with QAO reagents.We identified a total of 90 keto-steroids in human plasma,including 30 sulfate conjugates and 8 glucuronide conjugates.The analytical strategy established in this project provides some reference for the study of other compounds containing carbonyl group.