Novel characterization of the thermal properties of batteries have been introduced by defining their frequency-dependent thermal impedance functions. The thermal impedance function can be approximated as a thermal impedance spectrum by analyzing the experimental temperature transient which is related to the thermal impedance function through Laplace transformation. In order to obtain the temperature transient, a process has been devised to generate an external heat pulse with heating wire and to measure the response of the battery. This process is used to study several commercial Li-ion batteries of cylindrical type. Thermal impedance measurements have been performed using a potentiostat/galvanostat controlled by a digital signal processor, which is more commonly available than a flow-meter usually applied for thermal property measurements. Thermal impedance spectra obtained for batteries produced by different manufacturers are found to differ considerably. Comparison of spectra at different states-of-charge indicates an independence of the thermal impedance on the charge state of the battery. It is shown that the thermal impedance spectrum can be used to obtain simultaneously the thermal capacity and the thermal conductivity of the battery by non-linear complex least-squares fit of the spectrum to the thermal impedance model.