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The objective was to determine suitable Near-Infrared (NIR) wavelength regions to identify ethanol, total soluble sugars, glycerol, and organic acids in the fermentation broth for a corn dry grind process. Effects of temperature on spectral absorption were identified and related to loadings of principal component scores. The wavenumber region between 12000 to 6400 cm-1 had low correlation coefficients for the 0.5-mm short pathlength transflectance accessory. Wavenumbers having high correlation coefficients for ethanol were found in the region between 4712 to 4200 cm-1, while that for total sugars were from 5000 to 4286 cm-1. High RPD and low RMSEP values for both ethanol and total soluble sugars validated the suitability of the wavelength regions found. Lactic acid and acetic acid were present at very low levels and did not appear to separate from the other fermentation constituents for the given pathlength and concentrations observed.