Objective. To investigate biological effectiveness of 252Cf brachytherapy source using Monte Carlo-calculated microdosimetric distributions. Approach. 252Cf source capsule was placed at the center of the spherical water phantom and phase-space data were scored as a function of radial distance in water (R = 1–5 cm) using TOPAS Monte Carlo code. The phase-space data were used to calculate microdosimetric distributions at 1 μm site size. Using these distributions, Relative Biological Effectiveness (RBE), mean quality factor () and Oxygen Enhancement Ratio (OER) were calculated as a function of R. Main results. The overall shapes of the microdosimetric distributions are comparable at all the radial distances in water. However, slight variation in the bin-wise yield is observed with R. RBE, and OER are insensitive to R over the range 1–5 cm. Microdosimetric kinetic model based RBE values are about 2.3 and 2.8 for HSG tumour cells and V79 cells, respectively, whereas biological weighting function-based RBE is about 2.8. ICRP 60 and ICRU 40 recommendation-based values are about 14.5 and 16, respectively. Theory of dual radiation action based RBE is 11.4. The calculated value of OER is 1.6. Significance. This study demonstrates the relative insensitivity of RBE, and OER radially away from the 252Cf source along the distances of 1–5 cm in water.