DETERMINATION OF TROVAFLOXACIN, A NEW QUINOLONE ANTIBIOTIC, IN BIOLOGICAL SAMPLES BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

A simple, accurate and precise high-performance liquid chromatographic method was developed and validated for the determination of trovaflox acin, a new quinolone antibiotic, in serum and urine. Following solid- phase extraction, chromatographic separation was accomplished using a C-18 column with a mobile phase consisting of 0.04 M H3PO4-acetonitril e-tetrabutylammonium hydroxide-0.005 M dibutyl amine phosphate (D-4) r eagent (83:16.85:0.05:0.1, v/v), pH 3. Trovafloxacin and the internal standard (a methyl derivative of trovafloxacin) were detected by ultra violet absorbance at 275 nm. The lower limit of quantification for tro vafloxacin was 0.1 mu g/ml and the calibration curves were linear over a concentration range of 0.1 to 20.0 mu g/ml (r(2) = 0.9997). The ave rage recoveries were greater than 70% for both trovafloxacin and inter nal standard. The intra-day and inter-day coefficients of variation we re generally less than 5% in urine and serum over the concentration ra nge of 0.1 to 20.0 mu g/ml. Human serum samples could be stored for up to 12 months at -20 degrees C and urine samples could be stored up to 18 months at -80 degrees C.