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Abstract
In this paper high-performance liquid chromatography with diode array UV detection (HPLC/DAD) after SPE sample pretreatment for simultaneous analysis of pharmaceuticals from three different classes: enrofloxacin, oxytetracycline and trimethoprim, has been developed, optimised and validated. The chromatographic separation was developed on spiked wellspring water samples and checked on model wastewater samples of veterinary pharmaceuticals before and after RO/NF membrane treatment (feed and permeate streams) and on process wastewaters of industrial origin. Chromatographic separation was performed on Varian ProStar HPLC/DAD with C-18 column (Microsorb-MV 100 C18, 150 × 4.6 mm; 5 µm, Varian, USA). Detection and quantification was performed at 254 nm. The best separation was achieved with mobile phase 0.5% formic acid and 1% trifluoroacetic acid in 0.05 M ammonium acetate-methanol, 70 + 30, (v/v) after extraction procedure on polystyrenedivinylbenzene Varian Empore extraction disks. The extraction efficiency was checked by recovery experiments.
Acknowledgements
This work has been supported by the Unity Through Knowledge Fund (UKF), which was established by the Croatian Ministry of Science, Education and Sports through the World Bank Loan No. 7320-HR: Reduction of environmental risks posed by pharmaceuticals and their degradation products in process wastewaters, through RO/NF membrane treatment (REPHAD), and Croatian Ministry of Science, Education and Sports Projects: 125-1253008-1350, 125-1253008-3009, 125-2120898-3148.