Fires in terrestrial ecosystems produce large amounts of charcoal that persist in the environment and represent a substantial pool of sequestered carbon in soil. The objective of this research was to investigate the effect of charring on mid-infrared spectra of materials likely to be present in forest fires in order to determine the feasibility of determining charred organic matter in soils. Four materials (cellulose, lignin, pine bark, and pine wood) and char from these materials, created by charring for various durations (1 to 168 h) and at various temperatures (200 to 450 °C), were studied. Mid-infrared spectra and measures of acidity (total acids, carboxylic acids, lactones, and phenols as determined by titration) were determined for 56 different samples (not all samples were charred at all temperatures/durations). Results showed spectral changes that varied with the material, temperature, and duration of charring. Despite the wide range of spectral changes seen with the differing materials and length/temperature of charring, partial least squares calibrations for total acids, carboxylic acids, lactones, and phenols were successfully created (coefficient of determination and root mean squared deviation of 0.970 and 0.380; 0.933 and 0.227; 0.976 and 0.120; and 0.982 and 0.101 meq/g, respectively), indicating that there is a sufficient commonality in the changes to develop calibrations without the need for unique calibrations for each specific material or condition of char formation.