Abstract
We developed and validated a stability-indicating assay method for the simultaneous determination of enrofloxacin and piroxicam in combination and in the presence of degradation products. Reversephase high-performance liquid chromatography analyses were carried out on a Vertisep C18 column and acetonitrile-water (48:52 v/v, pH 3.0) mobile phase with a 1.00 mL min−1 flow rate. The efficient chromatographic separation of these drugs and their forced degradation products was achieved in less than 5 min with a peak purity match factor higher than 950. The method used showed linearity in the concentration ranges of 0.25 to 16.0 μg mL−1 for enrofloxacin (r = 0.9997) and 0.125 to 8.0 μg mL−1 for piroxicam (r = 0.9999) as well as precision (relative standard deviation lower than 2%), accuracy (mean recovery 100 ± 2%), and robustness, according to ICH (International Conference on Harmonization) and AOAC (Association of Official Analytical Chemists) guidelines. This method can simultaneously determine the combination of these drugs in a veterinary formulation and separate the drug peaks from their forced degradation products. Additionally, its optimized chromatographic conditions can contribute to the quality control of this formulation in pharmaceutical manufacturing plants and minimize waste from the organic solvent.
Highlights
Fluoroquinolones are a group of synthetic antibacterial agents that are widely used for the treatment of multiple bacterial infections in humans and in veterinary medicine because of their safety, broad antibacterial spectrum, and good tolerance (Chamseddin & Jira, 2011)
Enrofloxacin (ENRO; Figure 1A) was the first of the fluoroquinolones developed exclusively for use in veterinary medicine and was approved in the late 1980s (Bimazutube et al, 2009). This drug is chemically known as 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-4oxo-1,4-dihydroquinoline-3-carboxylic acid, with the molecular formula C19H22FN3O3 and a molecular weight of 359.36 g mol−1 (British Pharmacopoeia, 2012)
high-performance liquid chromatography (HPLC)-grade acetonitrile (Vetec, Brazil), analytical-grade phosphoric acid (Synth, Brazil), and fresh ultrapure water from a Milli-Q® Plus system were used in all analyses
Summary
Fluoroquinolones are a group of synthetic antibacterial agents that are widely used for the treatment of multiple bacterial infections in humans and in veterinary medicine because of their safety, broad antibacterial spectrum, and good tolerance (Chamseddin & Jira, 2011). These agents act by inhibiting DNA gyrase. Enrofloxacin (ENRO; Figure 1A) was the first of the fluoroquinolones developed exclusively for use in veterinary medicine and was approved in the late 1980s (Bimazutube et al, 2009).
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