Different types of municipal solid waste incinerator (MSWI) fly and bottom ash were extracted by TCLP and PBET procedures. The biotoxicity of the leachate of fly ash and bottom ash was evaluated by Vibrio fischeri light inhibition test. The results indicate the following: (1) The optimal solid/liquid ratio was 1:100 for PBET extraction because it had the highest Pb and Cu extractable mass from MSWI fly ash. (2) The extractable metal mass from both fly ash and bottom ash by PBET procedure was significantly higher than that by TCLP procedure. (3) The metal concentrations of fly ash leachate from a fluidized bed incinerator was lower than that from mass-burning and mass-burning combined with rotary kiln incinerator. (4) The TCLP and PBET leachate from all MSWI fly ash samples showed biotoxicity. Even though bottom ash is regarded as a non-hazardous material, its TCLP and PBET leachate also showed biotoxicity. The pH significantly influenced the biotoxicity of leachate.