Characterization and quantification of livestock odorants is one of the most challenging analytical tasks because odor‐causing gases are very reactive, polar and often present at very low concentrations in a complex matrix of less important or irrelevant gases. The objective of this research was to develop a novel analytical method for characterization of the livestock odorants including their odor character, odor intensity, and hedonic tone and to apply this method for quantitative analysis of the key odorants responsible for livestock odor. Sorbent tubes packed with Tenax TA were used for field sampling. The automated one‐step thermal desorption module coupled with multidimensional gas chromatography‐mass spectrometry∕olfactometry system was used for simultaneous chemical and odor analysis. Fifteen odorous VOCs and semi‐VOCs identified from different livestock species operations were quantified. Method detection limits ranges from 40 pg for skatole to 3590 pg for acetic acid. In addition, odor character, odor intensity and hedonic tone associated with each of the target odorants are also analyzed simultaneously. We found that the mass of each VOCs in the sample correlates well with the log stimulus intensity. All of the correlation coefficients (R2) are greater than 0.74, and the top 10 correlation coefficients were greater than 0.90.