This study provided insight into the characterization of secondary effluent from a wastewater treatment plant located in northeastern China. The secondary effluent was separated into three fractions, the dissolved, the near-colloidal and the suspended, to study their individual characteristics. It revealed that most of the organics in the secondary effluent existed in the dissolved form, accounting for 78.1–86.5% of the total chemical oxygen demand and 82.6–86.6% of the total organic carbon. Results from the molecular weight distribution study further indicated that organics with MW < 1k Da constituted 56.3–62.7% of total organics. Moreover, the particle size distribution study suggested that particles between 2.0 and 6.8 μm in diameter made up 80.0% of the total suspended solids. Both biological oxygen demand/chemical oxygen demand and biological dissolved organic carbon/dissolved organic carbon were measured ranging from 0.2 to 0.3, suggesting the most secondary effluent organics were biologically refractory. This conclusion was further strengthened by the functional groups information obtained from the GC/MS (gas chromatography/mass spectrometry) analysis. The characteristics information revealed from this study will help the design and selection of water quality-specific tertiary treatment technologies for secondary effluent water purification and reuse.