An experimental and computational protocol was established for the simultaneous determination of several key gasoline properties from a single Fourier transform infrared (FT-IR) spectrum. The study has shown that midband FT-IR spectroscopy combined with multivariate calibration analysis is a versatile, efficient, and accurate technique for the simultaneous estimation of key gasoline properties within about 1 min with less than 2 mL of sample. The FT-IR-derived values of gasoline properties include research and motor octane numbers, aromatic, olefinic, and saturated hydrocarbon content, benzene content, and concentrations of ethanol, methyl tert-butyl ether, and total oxygen. Concentrations of other oxygenated compounds are expected to be equally predictable. However, since these oxygen-containing species have not been adequately represented among the currently commercially available gasoline samples, their calibration may only be achieved using laboratory fuel blends. Midrange boiling point data may also be estimated. Fuel properties determined by minor concentrations of fuel components, e.g., flash point, sulfur content, etc., may not be modeled because the corresponding FT-IR signals are below detection limits of presented experimental protocol. The precision of this procedure was shown to be comparable to reproducibility of the standard laboratory analyses used for direct measurement of specific fuel properties, with squared correlation coefficient (R(2)) ranging from 0.94 to 0.99 between the two sets of measurements. This new methodology could increase the corresponding output of the petroleum laboratories by a factor of over 200 to 1 while maintaining data integrity and minimizing sample requirements, environmental hazards, and cost.