The behavior of the non-equilibrium grain boundary segregation of boron in low carbon steel was studied through a particle tracking autoradiography. The behavior of the non-equilibrium grain boundary segregation of boron during continuous cooling was compared with the isothermal kinetics of the non-equilibrium grain boundary segregation of boron at the holding temperature using an effective time method. On the basis of the experiments, the cooling rate dependence of the non-equilibrium segregation of boron was explained using the time dependence of the non-equilibrium segregation of boron in low carbon steel. The experimental observations for the cooling rate dependence of the grain boundary segregation of boron are in good agreement with the time dependence of the grain boundary segregation of boron. The mechanisms of the non-equilibrium segregation of boron during cooling in low carbon steel are also discussed.