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With the growing use of warm obturation techniques during endodontic treatment, more interest is directed toward sealers' compatibility with heat. This study aimed to evaluate the effect of heat application duration and temperature on epoxy resin- and calcium silicate-based sealers using chemical and thermogravimetric analyses.Freshly mixed samples (n = 5/group) of each sealer were heated at 200°C or 250°C for 30 or 60 seconds. Additional 2 sets of samples were examined directly after mixing or after setting without heat exposure. Raman spectroscopy was used to identify changes in the chemical structure, and a 2-way analysis of variance was performed to compare values of measurable peaks that exhibited changes. Additionally, Thermogravimetric Analysis (TGA) was used to evaluate the effect of heat on mass change where sealers were heated to 250°C at a rate of 20°C/min (11-minute duration) or maintained at 37°C for 8 hours.No differences were detected among all the spectra of calcium silicate samples of different groups, while TGA revealed 15% and 18% weight loss upon heating at 250°C and 37°C, respectively. For the resin sealer, significant differences were detected when samples were heated for 60 seconds, involving bonds of benzene rings and aromatic amines in the uncured resin. TGA revealed minimal changes in the sealer mass (1.2% and 1.8%) on heating at 250°C and 37°C, respectively.Heat application duration and temperature can affect the chemical structure of epoxy resin sealers. The consideration of endodontic sealer compatibility as well as the duration of heat application is essential when warm vertical obturation is used.No effect of heat on the chemical structure of calcium silicate sealer was detected.Calcium silicate sealer exhibited more weight loss due to water desorption.Several chemical bonds were affected by heat in the epoxy resin sealer.Heating duration had more impact on the resin sealer compared with temperature.