High-performance liquid chromatographic method for the routine determination of sulphadimidine, its hydroxy metabolites and N4-acetylsulphadimidine in body fluids and cell culture media. |
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Authors: | G A Van 't Klooster P B van Seeventer H J Kolker L A Smit R F Witkamp |
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Institution: | Department of Veterinary Basic Sciences, University of Utrecht, Netherlands. |
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Abstract: | A simple high-performance liquid chromatographic method is presented for the determination of trace amounts of sulphadimidine (SDD), its hydroxylated metabolites and N4-acetyl-SDD in blood plasma, urine, hepatocyte culture media and microsomal incubations. The synthesis of 5-hydroxy-SDD and an improved method for the isolation of 4-methylhydroxy-SDD from urine are described and their respective specific absorption coefficients at 265 nm are calculated by on-line radiochemical and ultraviolet detection. The limit of detection of the analytical method is 0.05 micrograms/ml for SDD and its hydroxy metabolites and 0.2 micrograms/ml for N4-acetyl-SDD. Linear calibration graphs for SDD and its metabolites were constructed from 0.2 to 50 micrograms/ml. The method has been applied to biotransformation studies in vivo and in vitro. |
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