A simple method for the reduction of aflatoxins B1 (AFB1), B2 (AFB2), G1 (AFG1), G2 (AFG2) and ochratoxin A (OTA) in white pepper was studied. Response surface methodology (RSM) was applied to determine the effect of four variables, which included time (20–60 min), temperature (30–70°C), calcium hydroxide (Ca(OH)2) (0–1%) and hydrogen peroxide (H2O2) (1–3%) during the washing step of white pepper. The efficacy of the method was evaluated by the determination of mycotoxins by HPLC with fluorescence detection (FD). Statistical analysis showed that the experimental data could be adequately fitted into a second-order polynomial model, with a multiple regression coefficient (R 2) in the range of 0.805–0.907 for AFG2 and AFG1, respectively. The optimal condition was 57.8 min, 62.0°C, of 0.6% (w/v) and 2.8% (v/v) for time, temperature, Ca(OH)2 and H2O2 respectively. By applying the optimum condition, the mycotoxins reduction was found to be in the range of 68.5–100% for AFB2 and AFG1 respectively.