A high performance liquid chromatographic (HPLC) method for simultaneous determination of sulfamonomethoxine (SMMX), sulfadimethoxine (SDMX), and their N4-acetylated forms in fish tissues was developed.
The sulfa drugs and their N4-acetylated forms extracted from samples were adsorbed on Sep-Pak Alumina B cartridge and eluted with 20ml of 70% acetonitrile. The eluate was evaporated to dryness. The residue was dissolved in 30% acetonitrile. The solution obtained was analyzed by HPLC. The chromatographic conditions were as follows: analytical cloumn, YMC-Pack C18; mobile phase, 0.05M phosphate buffer, pH 2.5-acetonitrile (65:35); wavelength, 265nm.
The average recoveries of SMMX, SDMX, and their N4-acetylated forms from serum, muscle, liver, kidney, and bile of rainbow trout were more than 80%. Their coefficients of variation were 1.1-6.0%. The detection limits for serum, muscle, liver, and kidney were 0.05ppm, and that for bile. was 0.1ppm.
