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TITLE:  Determination of non-protein bound phenylbutazone in bovine plasma using ultrafiltration and liquid chromatography with ultraviolet detection
 
AUTHORS:  De Veau EJ;
 
YEAR:  1999
 
JOURNAL ABBREV:  J Chromatogr B Biomed Sci Appl
 
MONTH:  Jan
 
TYPE:  JOUR
 
REFMAN INDEX:  184
 
JOURNAL FULL:  Journal of chromatography B, Biomedical sciences and applications
 
VOLUME:  721
 
ISSUE:  1
 
START PAGE:  141
 
END PAGE:  145
 
KEYWORDS:  Animals;Anti-Inflammatory Agents;Anti-Inflammatory Agents,Non-Steroidal;blood;Cattle;Chromatography;Chromatography,Liquid;Food;methods;Phenylbutazone;Sodium;Spectrophotometry,Ultraviolet;Ultrafiltration;veterinary;Veterinary Medicine;
 
ABSTRACT:  A liquid chromatographic procedure using UV detection was coupled with ultrafiltration for the quantitation of free phenylbutazone in bovine plasma, in the range of 20 ng/ml to 2.0 microg/ml. Whole plasma samples (0.5 to 1 ml) were placed in a 2-ml centrifugal concentrator with a molecular-mass cut-off membrane of 10000 and centrifuged at 4500 g for 2 h at 4 degrees C using a fixed angle rotor. The ultrafiltrate was transferred to an LC vial with a 200-microl insert and 100 microl was injected into an LC system. The chromatographic system used a C18 reversed-phase column connected to a UV detector set at 264 nm. The mobile phase was 0.2 M sodium phosphate buffer (pH 7)-methanol (1:1). Recoveries of phenylbutazone from protein-free plasma water fortified at levels of 20 ng/ml to 2 microg/ml ranged from 91 to 93%, with relative standard deviations (R.S.D.s) ranging from 1 to 4%. The concentration of incurred non-protein bound phenylbutazone obtained from a cow intravenously dosed twice with 2 g phenylbutazone, 8 h apart, was 111, 26 and 11 ng/ml for 2, 72 and 104 h post first phenylbutazone dose, respectively
 
AFFILIATIONS:  U.S. Food and Drug Administration, Center for Veterinary Medicine, Laurel, MD 20708, USA. edeveau@bangate.fda.gov
 
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