Steam assisted gravity drainage (SAGD) boiler blow-down (BBD) water contains high concentrations of dissolved organic matter (DOM) and total dissolved solids (TDS). A detailed understanding of the BBD chemistry, particularly the DOM composition, is important for better management and recycle of this water. In this study, we fractionated the dissolved organic matter in the BBD using DAX-8, Dowex, and Duo lite resins into hydrophobic and hydrophilic fractions of acid, base, and neutral compounds. Additionally, the DOM was fractionated on the basis of size by filtering the BBD through a series of membranes with progressively tighter molecular weight cutoffs of 10, 3, and 0.5 kDa. Fluorescence excitation-emission matrix spectroscopy (EEMs), specific UV absorbance (SUVA), and FTIR were used to characterize the water samples and the different fractions. The ion exchange fractionation revealed that the DOM contained a high percentage of hydrophobic acids (39%) and hydrophilic neutrals (28.5%). The different ion exchange fractions had distinct fluorescence excitation-emission signatures. The permeate samples from the membrane fractionation, on the other hand, did not reveal any significant difference in the fluorescence EEM spectra, indicating that the hydrophilic and hydrophobic constituents of the DOM could not be separated on the basis of pore size by these membranes. The SAGD boiler blow-down water was found to be significantly concentrated in DOM compared to oil sands mining process affected water.