Long-chain quinones are essential components of both bacterial and eukaryotic respiratory chains, and some of the main unsolved questions on energy transduction in membranes are complicated by the lack of consistent information on the physical state of the quinones in membrane bilayers. We have recorded, at various temperatures and under different conditions, the infrared spectra of ubiquinone-10 (the main species in mitochondria) and several analogues. The C = 0 stretching Vibration band located at 1663 -1670 cm-I has been identified as the most sensitive one to phase and environmental changes. Three distinct phases have been characterized in which pure ubiquinone-10 may exist: crystalline (LC,), isotropic liquid (IL) and liquid crystalline (L& The only allowed thermotropic transitions are LCl --t IL, IL + L, and L, -+ LC1. Our investigations with pure quinones provide a simpler and more detailed description of their phase changes than any of the previous studies and shed light on their behaviour in membranes. When incorporated into phospholipid bilayers, ubiquinone-10 appears to be removed from the aqueous environment and is found to exist, in the 4-70°C range, in an isotropic liquid phase, in the form of small aggregates.