Abstract

A simple and rapid high performance liquid chromatographic method was developed, validated and applied for the simultaneous determination of marbofloxacin (MBX) and trovafloxacin (TVX) in sheep plasma. Samples were extracted with 20% perchloric acid and MBX and TVX were separated on a C18 column using a gradient mobile phase system consisting of 17.5mM of NaH2PO4 and 1.5mM of tetrabutylammonium hydroxide aqueous solution, pH 3 (A) and 50% acetonitrile and 50% methanol (B), with UV detection at 293 and 270nm. The retention times of MBX and TVX were 4.9 and 6.6min respectively. The detection and quantification limits for MBX and TVX were 2ng/mL and 10ng/mL respectively for both compounds. The calibration curves were linear over a concentration range of 10–50,000ng/mL for both antibiotics. The linearity, precision, accuracy, recovery and stability of the assay were evaluated from spiked sheep plasma. The method was successfully applied to sheep plasma samples obtained from MBX and TVX pharmacokinetic studies.

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