RESUMO Submitted on February 29th, 2016 and accepted on August 22nd, 2017The objective of this study was to develop and validate the methodology of carbon dioxide concentration in the evaluation of vigor of Triticum aestivum L. seeds based on infrared spectroscopy. The proposed method quantifies CO 2 content using a drag system and exhaustion to the gas released by the seeds. Samples of 50 seeds of six lots of cultivar CD 123 were incubated at temperatures of 15, 25 and 40 °C. The CO 2 content released after 1, 3, 6, 9, 12 and 24 h of incubation were quantified. Additionally, the percentage of normal seedlings emerged in the field were evaluated. The simple correlation coefficients were evaluated among tests. After determining the reading conditions, the precision and accuracy of the proposed method were evaluated, using 15 lots of seeds. For the evaluation of the respiratory activity in Triticum aestivum seeds, a sample of 25 seeds, incubated at 25 °C for a minimum of 12 h is recommended since it allows to classify lots with different levels of vigor and predict the establishment of seedlings in the field, being the appropriate method for measuring CO 2 as it externalizes precision between successive measurements and agreement with the reference method.