Abstract Protein resistance against denaturant agent is a useful property in industrial applications. In the current research, the domains thermal unfolding of glycated human serum albumin (GHSA) and human serum albumin (HSA) were studied under incubation at physiological conditions for 35 days. The domains thermal unfolding of GHSA and HSA were evaluated using differential scanning calorimetry (DSC), circular dichroism (CD) and UV–vis spectroscopy. The results showed that the first energetic domain of GHSA remained after cooling back from 80 °C, while the first energetic domain of HSA disappeared at this temperature. Moreover, the second energetic domain of GHSA kept on after cooling back from 90 °C, but it disappeared in HSA at this temperate. Also, the secondary structure recovery after cooling back in GHSA was higher than HSA. Therefore, according to the obtained results, glucose can act as a stabilizer for HSA domains and prominently domain III because it has more lysine residue for glycation process.