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Enhancing Extraction and Detection of Veterinary Antibiotics in Solid and Liquid Fractions of Manure.

Analysis of veterinary antibiotics in separated liquid and solid fractions of animal manures is vital because of wide variations in the composition of agriculturally applied manure. Differentiation of antibiotic concentrations is important between liquid and solid manures, as their sorption onto the solid fraction depends on physicochemical properties of each antibiotic and manure composition (e.g., organic content, pH) and because each fraction may be treated and reused differently. Here, an efficient and sensitive method for the analysis of 22 veterinary antibiotics in the liquid and solid fractions of manure is reported. Tetracycline (TC), macrolide, and sulfonamide antibiotics were extracted from liquid manure by liquid-liquid extraction (LLE) with methanol following acidification with acetic acid. Extraction from solids was performed by sonication with acetonitrile, methanol, and 0.1 M EDTA-McIlvaine buffer. Cleanup of extracts was achieved by solid-phase extraction with hydrophilic-lipophilic balance (HLB) cartridges or tandem amino (NH2) and HLB cartridges. Quantification of antibiotics was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) under wrong-way-round (WWR) ionization for sulfonamides and TCs and right-way-round ionization for macrolides. Recoveries of 58 to 94.7% and 62 to 94.3% were obtained in liquid and solid manure, respectively. Method detection limits range from 1.2 to 12 ng L and 0.5 to 7.9 μg kg dry wt. in liquids and solids, respectively. This method allows for extraction and analysis of both mobile antibiotics in liquid phase and hydrophobic antibiotics adsorbed on the solids. Without separate analysis, antibiotic concentrations may be improperly estimated by analyzing whole manure, as reported in many studies to date.