ETD

• filter-aided sample preparation
• heart failure
• liquid chromatography tandem mass spectrometry
• microextraction by packed sorbent
• natriuretic peptides
• urine proteomics

This PhD project developed and validated an ultra-high-performance liquid chromatography-tandem mass spectrometry method to determine natriuretic peptides (NPs) in human plasma. These hormones are known as golden biomarkers for the diagnosis and the prognosis of heart failure as plasmatic concentrations increase with the severity of the disease (tenths-hundreds of pg/mL). The method solves issues related to cross-reactions with species sharing homologous molecular structures that affect quantifications by commercially available immunoassays used in clinical settings.
Since NPs are prone to degradation and adsorption on material surfaces, their stability was studied both in aqueous solutions and plasma samples. Sample collection procedures, sample pre-treatment, and storage protocols were optimized to guarantee sample representativeness and avoid bias. Finally, the analytical platform was employed to monitor heart failure patients.
During the period abroad, my research activity aimed at the application of filter-aided sample preparation coupled with untargeted mass spectrometry to monitor urinary proteome in heart failure patients. Bioinformatic and statistical tools were employed for the identification of differentially expressed proteins related to the disease. The finding of a specific pattern could help physicians in the diagnosis of heart failure and the assignment of a more specific therapy to the patient.