In order to meet increasingly stringent total nitrogen limits, supplemental carbon must be added to improve the performance of the biological nutrient removal process. An industrial by-product that contained ethylene glycol and propylene glycol was used as a substitute carbon source for methanol in this study. The objectives of this study were to investigate the efficiency of using the glycol mixture as carbon source, including the calculation of denitrification rate and yield at two different initial concentrations of glycols. Possible inhibition effect on nitrification was also investigated. Three SBR reactors were operated by adding methanol, a low dosage of glycol, and a high dosage of glycol into the reactors. The low dosage glycol reactor exhibited the best performance, with the highest denitrification rate of 11.55 mg NOx-N/g MLVSS"h and the lowest yield of 0.21 mg VSS/mg COD. Small nitrite accumulation was observed in the low dosage glycol reactor (COD=185"•15 mg/L), but effluent quality was not influenced. Excess glycol in the reactor caused deteriorated performance. The high dosage glycol reactor (COD=345"•20 mg/L) performed with the lowest denitrfication rate of 8.56 mg NOx-N/g MLVSS"h and the highest yield of 0.55 mg VSS/ mg COD. The reactor with the high dosage of glycol also inhibited the lowest nitrification rate of 1.15 mg NH3-N oxidized/g MLVSS"h, which indicated that excess glycol may cause nitrification inhibition.