Systematic optimization of an SPE with HPLC-FLD method for fluoroquinolone detection in wastewater

•WCX is better suited than HLB for solid-phase extraction of fluoroquinolones.•Real wastewater matrix had minimal impact on recovery and fluorescence response.•HPLC-FLD method fully resolves 11 fluoroquinolones.•SPE with HPLC-FLD method demonstrates sensitivity equivalent to LC–MS/MS.•FQs detected in raw wastewater, wastewater effluent, and surface water. This paper describes a selective and ultra-sensitive analytical method for simultaneous determination of 11 fluoroquinolone (FQ) antibiotics in environmental and wastewater samples. The method employs offline solid-phase extraction (SPE) and reversed-phase high performance liquid chromatography with fluorescence detection (HPLC-FLD). A weak cation exchange SPE protocol was developed with a novel loading volume optimization algorithm and a methanol cleanup step to remove background organic matter. Various parameters were optimized to recover FQs from water/wastewater and analyte recovery was generally greater than 80%. Chromatographic separation of the 11 FQs was achieved on a 150mm pentafluorophenyl column using a gradient elution scheme with methanol, acetonitrile, and 20mM phosphate buffer (pH=2.4). Excitation and emission wavelengths were individually optimized for each FQ using fluorescence spectroscopy; the excitation and emission wavelengths were 276–296nm and 444–506nm, respectively. Instrumental quantitation limits were 20–100pg of mass injected. Of the 11 FQs investigated, seven (i.e., ciprofloxacin, difloxacin, enrofloxacin, fleroxacin, norfloxacin, moxifloxacin, and ofloxacin) were detected during a four-month sampling campaign of wastewater and wastewater-impacted surface water. Concentrations of FQs in raw wastewater, wastewater effluent, and wastewater-impacted surface water were 5–1292, 2–504, and 4–187ng/L, respectively.