The main aim of this thesis is to investigate the glycation products, especially glycation intermediate products, generated in the glycation reaction model in vitro extracellular matrix (ECM) and ribose-5-phosphate using solid-state nuclear magnetic resonance (SSNMR) spectroscopy. The background and motivations of this project are outlined in chapter 1.

Chapter 2 reviews the current understanding of the composition and organisation of the ECM, the structure of collagen and the critical roles collagen plays in the ECM. Chapter 3 introduces the glycation reaction and summarises the glycation products identified to date, with a discussion of the most widely-used characterisation techniques and in vitro model systems for studying the glycation reaction and glycation products. Chapter 4 covers the basic theory of SSNMR spectroscopy, which is the major analytical technique used in this thesis, and the principles behind the SSNMR experiments used in this project. Above three chapters provide the essential background knowledge for studying the glycation reaction chemistry and the glycation products in in vitro collagenous ECM model using SSNMR spectroscopy.

Protocols for preparing, glycating and characterising in vitro ECM samples as well as all relevant experimental details are included in chapter 5, which is followed by a discussion in chapter 6 on the development of a method to extract collagen from in vitro ECM. In chapters 7 and 8, glycation products identified in different in vitro glycation models using SSNMR are outlined, where chapter 7 focuses on glycation products derived from the glycation reaction between non-isotope-enriched collagenous materials and 13C-enriched glycating agents, while chapter 8 concentrates on the effects of glycation on lysine and arginine residues in collagenous samples using 13C, 15N-enriched collagenous materials and 13C-enriched glycating agents. Chapter 9 discusses the development of a protocol to extract cellular metabolites, specifically carbonyl metabolites, and estimate their glycation potential, in order to understand the contribution that cell necrosis may have on glycation in patients. Conclusion and future work are outlined in chapter 10.