This study aimed to identify chemical compounds in raw commercial laundry wastewater and after biodegradation in an anaerobic fluidized bed reactor (FBR) on an increased scale. Solid-phase extraction (SPE) and solid-phase microextraction (SPME) techniques were used to concentrate and extract the compounds from the complex matrices that were identified in gas chromatography coupled to mass spectrometry. By using SPE, it was possible to identify the hydrophobic and hydrophilic compounds diversity in the commercial laundry wastewater, FBR influent and effluent, and to concentrate more quantity of analytes when compared with the SPME technique. The major compounds identified were oleamide, hexadecanol, dodecanol, nonadecanol, eicosanol, ethylene glycol monolauryl ester, chlorododecane, tridecanol, tert-octyl phenol, and nonylphenol. In the FBR effluent was observed a lower compound diversity, probably due to the degradation process and change in chemical structure of these compounds. The linear alkylbenzene sulfonate (LAS) concentration in commercial laundry wastewater ranged from 290.3 to 1.2 mg/L. By SPE technique was observed greater extraction of toxic and recalcitrant compounds from commercial laundry wastewater, FBR influent and efluente when compared with SPME. Even though the SPE technique is more expensive, this methodology is more interesting for the mapping of toxic compounds to a complex matrix.