Date of Award

2016

Degree Type

Dissertation

Degree Name

Doctor of Philosophy in Clinical-Bioanalytical Chemistry

Department

Chemistry

First Advisor

Guo, Baochuan

Subject Headings

Analytical Chemistry

Abstract

In the past decade, bioanalytical method development has become an integral part of the clinical diagnosis, biomarker discovery, and drug discovery and development. The new and emerged bioanalytical techniques allow the quantitative and qualitative analysis of bio molecules with remarkably high sensitivity and specificity. Specifically, these bioanalytical methods based on LC-MS and methylation-specific PCR are well suited for detecting low-abundance metabolites in various biological fluids and DNA in plasma and tissues for biomarker investigation. They offer great clinical promise for early disease diagnosis and therapeutic intervention. This dissertation broadly organized into two parts, part one talks about the application of LC-MS/MS for drug studies and part two converses about cancer diagnostics using PCR. Part I, The LC-MS/MS quantitative method is essential for the study of pharmacokinetic and toxicological properties in drug screening. Dependent on the type of molecule analyzed, different methods were established to achieve accurate and reliable detection. LC-MS/MS methods were developed and validated for quantitative analysis of an anti-cancer agent CSUOH0901, and an anti-parasitic agent BMCL26. Part II, a Panel of methylated DNA biomarkers for early detection of hepatocellular carcinoma (HCC) was developed by using methylation-specific PCR method. We also developed combination biomarkers for HCC screening. This marker combination enabled sensitive and specific detection of DNA hypermethylation on several tumor-associated genes. The studies with FFPE tissue samples successfully differentiated between HCC and normal tissue samples.

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