The invention provides a connection hole position determination device of a roller bed base. The device comprises a first connecting seat, a second connecting seat and a measuring tape; the first connecting seat is used to be detachably fixed at a start end of the roller bed base; the second connecting seat is used to be detachably disposed on the roller bed base and can move in the length direction of the roller bed base; the second connecting seat is provided with positioning holes, and the positioning holes are used to determine hole positions of connection holes in the roller bed base; the measuring tape comprises a ruler body and a measuring tape box; and the start end of the ruler body is connected with the first connecting seat, the zero graduation line of the ruler body coincides with an end face of the start end of the roller bed base, the tail end of the ruler body is accommodated in the measuring tape box, and the measuring tape box can move with the second connecting seat relative to the roller bed base so that a preset distance between the connection holes and the start end of the roller bed base is determined. The device can solve the problems of a large error and low efficiency which exist because hole positions of the connection holes are determined on the roller bed base through a steel plate ruler or a meter ruler currently.