Analytical pyrolysis of ovalbumin

In this study the thermal degradation of ovalbumin (OVA) under nitrogen atmosphere was investigated. For this scope, a multi instrumental approach based on thermogravimetry (TG), thermogravimetry coupled with infrared spectroscopy (TG/FTIR) and pyrolysis coupled with mass spectrometric detection, i.e. flash pyrolysis-coupled with gas chromatography-mass spectrometry (Py/GC/MS), evolved gas analysis coupled with mass spectrometry (EGA/MS) and double shot pyrolysis-coupled with gas chromatography-mass spectrometry (DSP/GC/MS), was used. The pyrolysis of a protein involves a combination of several complex mechanisms resulting in a very high number of products. The study highlighted that pyrolysis of OVA produces low-molecular weight gasses, such as CO2, H2O, HCNO, NH3 and CO, as main compounds. In addition, a series of organic compounds containing heteroatoms and unsaturations were also identified, whose formation occurred at different temperatures over the pyrolytic process. Among these, cyclic pyrolysis products were identified: dialkyl substituted 2,5-diketopiperazines (DKPs) and, for the first time, unsaturated-DKPs (un-DKPs), 3,5-alkyl-3,4-dihydro-2H-pyrrole-2,4-diones (ADPDs) and 3-alkenyl-5-alkyl-pyrrolidine-2,4-diones (AAPDs). These compounds are formed below 350°, and are produced by cyclisation reactions of two neighbouring amino acids. Pyroglutamic acid was also found among the main pyrolysis products of OVA, obtained as pyrolytic product of Glu, which is the most abundant amino acid in OVA. Aromatic compounds, such as pyridine, pyrrole, toluene, alkyl-benzenes and alkyl-pyrroles, phenol and alkyl-phenols, benzeneacetonitrile, benzenepropanenitrile, indole and alkyl-indoles, were detected, produced over a wide range of temperatures. This study highlighted for the first time that aromatic compounds produced below 320 °C are associated to the pyrolysis of specific amino acid side chains, while at higher temperatures, they are the pyrolysis products of the residual material remaining after condensation reactions, pyrolytic scissions and cyclization reactions.