PURPOSES : The objective of this study is to develop a methodology to estimate the minimum required separation distance (MRSD) between a vehicle and a bicycle when overtaking. METHODS : Three steps have been conducted to develop a methodology to estimate MSRD. First, a literature review has been conducted on the measurement of MSRD, and how it may be applied in Korea. Second, two assumptions have been made to develop a methodology to estimate the MSRD. The first assumption is that the maximum separation distance between a vehicle and a bicycle can be observed when they are at the same location longitudinally. In addition, it is assumed that the separation distance is invalid when the contra-flow exists. Finally, three cameras were installed at a height of 10 m to record the vehicle-bicycle interaction. Moreover, the vehicle trajectories as well as the separation distance were coded and analyzed. Through the hypothesis test and the interval estimation, the sample mean was tested and the confidence interval was estimated. RESULTS : 141 records of separation distance data were collected, and the hypothesis test demonstrated that the MSRD in Korea is significantly higher than other countries. In addition, the confidence interval of the population mean of MSRD is from 1.51 m to 1.65 m with 95% level of confidence. CONCLUSIONS : It is expected that the proposed methodology to estimate MSRD would be beneficial in studying road safety of vehicles and bicycles. Also, the proposed MSRD is expected to be designated in the act of road and transportation.