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CENTENNIAL HONORS COLLEGE Western Illinois University Undergraduate Research Day 2017 Poster Presentation Quantification of Phenylbutazone in Equine Plasma for Doping Control in Horse Racing by Liquid Chromatography with Ultraviolet Detection Natasha Frig, Terrence Petry Faculty Mentor: Liguo Song Chemistry In this study, a liquid chromatography separation with ultraviolet detection (LC-UV) for the analysis of phenylbutazone (PBZ) in equine plasma has been developed. Baseline separation of PBZ from interfering compounds possibly found in equine plasma which include oxyphenbutazone (OPBZ), a hydroxylated metabolite of PBZ through the hepatic oxidative metabolism, and other commonly used non-steroidal anti-inflammatory drugs (NSAIDs) for horses, i.e. diclofenac, flunixin, ketoprofen, meclofenamic acid and naproxen, and possible internal standards to correct for any possible analyte loss during sample preparation, i.e. tolfenamic acid and ibuprofen, has been successfully achieved. Using a Kinetex C18 column from Phenomenex, three mobile phase systems containing 0.1% acetic acid, i.e. acetonitrile:water, methanol:water, and methanol:acetonitrile:water, have been systematically optimized for the separation under gradient and isocratic conditions. It has been discovered that while methanol provides the necessary selectivity for the separation, acetonitrile prevents specific interactions of the NSAIDs to the stationary phase. Consequently, the optimized mobile phase consists of 70% 95:5 methanol:acetonitrile and 30% water with 0.1% acetic acid under isocratic conditions. With the optimized mobile phase, the desired separation has been completed within 16 minutes. (191 words)
