This study was conducted two kinds of aims: 1) to modify the analytical methods(conditions) by high performance liquid chromatography-fluorescence detector for the detection of residual ivermectin in raw milk, 2) to provide basic information for the evaluation of standard of the residual ivermectin in raw milk. It could be considerable that negative ion spectra can be better method in the LC/MS analysis for the detection of residues. Characteristic daughter ions were observed in negative ion spectra, however, linear line was not formed in positive ion one. Three Holstein cows (500±10kg) were applied to commercial ointment of ivermectin just one time at the first day of test, and residues in raw milk were examined for 20day after administration. The limit of detection(LOD) was 0.65ng(n=5) by HPLC/FLD, and recovery rates were 87.85%~99.47%. The peak was observed at the 4th day, and residues lasted to the end. Thus ivermectin was prohibited when lactating.