The evaluation of mental workload is measured by subjective ratings, physiological signals. It takes long time to analysis the measured signals and is very tedious and time-consumming work. Therefore, to evaluate the affect of workload effectively, real-time measurement system is required. In this paper, real-time mental workload measurement system using cardiac autonomic indiced which reflect well the mental workload was developed and evaluated. Analyzed indices were HR, IBI, Lorentz plot, CSI, CVI, and LF/HF ratio of heart rate variability. The system was applied to evaluate the affect of arithmetic task and showed good results. This system was consisted of ECG amplifier, A/D converter, and personal computer, and algorithm was implemented using LabVIEW.