Herbicide safener, a diverse group of chemicals, is an important tool used to protect plants from herbicidal injury. With the aim of decreasing drift injury of haloxyfop-P-methyl to sensitive plants, the protective effect of four safeners (R-28725, 3-dichloroacetyl oxazolidine and its two optical isomers) was evaluated. Physiological and biochemical tests were conducted under laboratory conditions in Northeast Agricultural University, China, by using seed treatment with safener and soil treatment with haloxyfop-P-methyl, respectively. The maize seeds treated with these safeners were safe from haloxyfop-P-methyl treatment. A positive correlation between growth level and endogenous glutathione (GSH) content, glutathione S-transferases (GST) activity was observed in this research. Enhancement of GSH content, GST activity and affinity of GST to 1-chloro-2,4-dinitrobenzene (CDNB) in maize treated by R-28725 was maximum. However, the detoxification of herbicide was not accompanied by the increase of acetyl-CoA-carboxylase (ACC) activity in maize.