Each pollutant concentration was followed by high performance

Each pollutant concentration was followed by high-performance liquid chromatography (HPLC, Waters instrument equipped with 600E Controller, photo diode array 996 detector (λdet = 255 nm), Waters 717 Autosampler; Waters, USA). A reversed-phase Nova-Pack C 18 (150 × 3.9 mm, 4 μm) column at 25 °C was used. Isocratic elution was performed with methanol–phosphoric FK-228 water solution 0.1% (50:50, v/v) with a flow rate 0.5 mL min−1 for the individual pHBA (tR 3.257 min), MP (tR 4.452 min), EP (tR 5.559 min) and 0.9 mL min−1 for PP (tR 4.590 min), BuP (tR 7.799 min), BeP (tR 8.223 min). The compounds’ mixture was analyzed with a flow rate of 0.5 mL min−1 (tRPP 8.502 min, tRBuP 14.051 min, tRBeP 14.690 min). The injection volume of the standards and the samples was 50 μL. The limits of detection (LOD) and quantification (LOQ) under the present chromatographic conditions were established at a tube-within-a-tube system signal-to-noise ratio using Empower Pro software (build 1154; Waters, USA). The stock solution containing each EDCs (50 μg mL−1) was prepared and diluted to eight appropriate concentrations in the range of 0.05–50 μg mL−1 and each solution was injected in duplicate. The LOD and LOQ values were experimentally verified by injections of the mixture of six compounds at the LOD and LOQ concentrations. The LOD and the LOQ were calculated to be 0.05 and 0.103 μg mL−1, respectively.