Pyrolysis—gas chromatography/mass spectrometry in the characterization of glycated albumin

Probing Protein Glycation by Chromatography and Mass Spectrometry: Analysis of Glycation Adducts

“…In the simplest way, it can be achieved by pyrolysis GC-MS (Py-GC-MS). Thus, based on the experiments with polylysine [ 94 ], Lapolla and co-workers proposed this technique as a potential tool for DM diagnostics, and demonstrated clear differences of pyrolysis profiles obtained for glycated albumin from those, acquired with untreated protein [ 95 ] ( Table 1 ). However, this approach has at least two essential disadvantages: from one hand, high temperatures, applied for degradation of the polypeptide chain, result in degradation of glycated amino acid side chains (that might lead to the loss of structural information), from another, the pattern of resulted pyrolysis cleavage products is difficult to interpret.…”

The role of mass spectrometry in the study of non-enzymatic protein glycation in diabetes

“…This approach, employed to characterize glycated polylysine, led to interesting results: a spectral library search identi®ed six compounds that originated from thermal degradation of the structures present only in the glycated polylysine (Lapolla et al, 1992a). A similar investigation on glycated albumin identi®ed a series of oxygen-containing heterocycles, identi®ed as originating from glycation products (Lapolla et al, 1992b).…”

Investigations on Protein Nonenzymatic Glycation by a New and Effective Mass Spectrometric Method

“…Such investigations, even if allowed to determine the structure of some glyco-oxidated products, showed severe limitations, due the presence in the reaction mixture of artifact products originated by the chemical treatment of the protein sample (10). In parallel, pyrolysis GC/MS experiments were performed to obtain a fingerprint somehow related to the glycation level (11). Also in this case, the aim was partially reached: clear differences were present between pyrolysis products of glycated and not glycated substrates, but either the quite low reproducibility of pyrolysis or the impossibility to relate the structure of pyrolysis products with the glycated species present in specific proteins loci made this approach of scarce analytical relevance.…”