ISO pneumatic cylinder reliability test requires air leakage measurement. Air cylinder has many parts and the leakage shall be measured before, during and after endurance test, and, the leakage should smaller than the specified value. The existing measurement method needs complex operation and the calibration of leak detector, and, has to separate the testing cylinder from endurance test device, which causes the change of contact condition of seal in the cylinder. Therefore, it is hard to evaluate the air leakage during endurance test, and guarantee the reliability of the conventional measurement method. In this paper, a new method for air leakage measurement using isothermal chamber, which does not requires calibration or temperature compensation, and, can measure air leakage accurately with quite simple operations, is proposed. As a result, reliability of air leakage measurement can be improved because the proposed method does not have to separate the testing cylinder from the endurance test device for air leakage measurement. The effectiveness of the proposed method is proved by error analysis of leak measurement from experimental result.