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United States Department of Agriculture

Agricultural Research Service

Title: Determination of Fluoroquinolones in Serum Using An on-Line Cleanup Column Coupled to High Performance Immunoaffinity/reversed-Phase Liquid Chromatography

item Holtzapple, Carol
item Buckley, Sandra
item Stanker, Larry

Submitted to: The Journal of Chromatography B: Biomedical Applications
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: November 2, 2000
Publication Date: April 1, 2001

Interpretive Summary: Fluoroquinolones are antibiotics that are used in the livestock industry to prevent bacterial diseases. Although these drugs are useful for keeping animals healthy, drug residues must not be present in animal products, such as meats, that are brought to market. In order to detect the presence of fluoroquinolones in cattle before slaughter, a simple, automated test was developed to detect residues of this drug in serum. The test is as sensitive as a previously developed test and has the added advantage that it does not require the use of environmentally unfriendly chemicals. Rapid tests such as the one described here should help producers, as well as government agencies, to screen cattle for the presence of fluoroquinolone residues.

Technical Abstract: A simple, rapid, and reliable method for the simultaneous analysis of ciprofloxacin, enrofloxacin, sarafloxacin, and difloxacin in serum has been developed. A high performance immunoaffinity chromatography (HPIAC) column containing covalently bound anti-sarafloxacin antibodies was used to capture the fluoroquinolones while allowing matrix components of the serum to elute to waste. Due to the highly selective nature of the antibodies, the only sample preparation required was passage of the samples through a 0.2 micrometer filter prior to injection to remove particulate matter. After binding to the HPIAC column, the fluoroquinolones were eluted directly onto a reversed phase (RP) column for final separation of the compounds prior to fluorescence detection at excitation and emission wavelengths of 280 and 450 nm, respectively. No significant interference from the sample matrix were observed, indicating good selectively with the HPIAC column. The method yielded high recoveries of all four fluoroquinolones from fortified bovine serum that were greater than 95 percent with good reproducibility (CV < 7.0 percent). The on-line, automated method described here provides simple, sensitive, and specific method for multi-residue detection of fluoroquinolones in serum.

Last Modified: 4/22/2015
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