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Solid-phase Extraction and TLC Quantification of Enrofoxacin, Oxytetracycline and Trimethoprim in Spiked Water Samples,

Antibiotics like enrofloxacin, oxytetracycline and trimethoprim are used routinely in veterinary medicine for prevention and control of disease, for therapeutic and prophylactic purposes in addition to their application as growth promoters. The excretion of these antibiotics together with improper waste disposal has led to considerable concentrations of these compounds. Even the processing of communal waste water in sewage treatment plants cannot avoid the entry of antibiotics into surface water because of their high stability. Finally, these compounds may even enter groundwater as well as drinking water produced from groundwater. Antibiotics analysis in surface water has mainly been carried out by gas (GC) or high-performance liquid chromatography (HPLC), usually in combination with mass spectrometric (MS) detection. In this work thin-layer chromatography (TLC) is presented as very simple and less expensive chromatographic method. TLC separation was performed on CN F254 s plates (Merck, Germany) with H2C2O4-methanol as mobile phase. Optimization of TLC separation was accomplished using genetic algorithm approach. The best separation was achieved using 0.5 M H2C2O4-methanol, 70+30 (v/v), as the optimum mobile phase. After air-drying, spots were detected and quantified at 254 nm by video-densitometry. Limit of detection (LOD) and limit of quantification (LOQ) for TLC determination were calculated for enrofloxacin, oxytetracycline, and trimethoprim being 400 ng/spot and 600 ng/spot for all compounds. The method was examined on spiked water samples. Antibiotics were extracted by solid-phase extraction (SPE) procedure using C18 Empore extraction disk (Varian, USA). The extraction efficiency was checked by recovery experiments.

enrofloxacin; oxytetracycline; trimethoprim; antibiotics; Solid-phase Extraction (SPE); Thin-layer chromatography (TLC)

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