The power battery thermal management problem in EV (electric vehicle) and HEV (hybrid electric vehicle) has been widely discussed, and EIS (electrochemical impedance spectroscopy) is an effective experimental method to test and estimate the status of the battery. Firstly, an electrochemical-based impedance matrix analysis for lithium-ion battery is developed to describe the impedance response of electrochemical impedance spectroscopy. Then a method, based on electrochemical impedance spectroscopy measurement, has been proposed to estimate the internal temperature of power lithium-ion battery by analyzing the phase shift and magnitude of impedance at different ambient temperatures. Respectively, the SoC (state of charge) and temperature have different effects on the impedance characteristics of battery at various frequency ranges in the electrochemical impedance spectroscopy experimental study. Also the impedance spectrum affected by SoH (state of health) is discussed in the paper preliminary. Therefore, the excitation frequency selected to estimate the inner temperature is in the frequency range which is significantly influenced by temperature without the SoC and SoH. The intrinsic relationship between the phase shift and temperature is established under the chosen excitation frequency. And the magnitude of impedance related to temperature is studied in the paper. In practical applications, through obtaining the phase shift and magnitude of impedance, the inner temperature estimation could be achieved. Then the verification experiments are conduced to validate the estimate method. Finally, an estimate strategy and an on-line estimation system implementation scheme utilizing battery management system are presented to describe the engineering value.