title: High-performance liquid chromatographic method for the routine determination of sulphadimidine, its hydroxy metabolites and N4-acetylsulphadimidine in body fluids and cell culture media.
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.
authors.affiliation: Department of Veterinary Basic Sciences, University of Utrecht, Netherlands.
authors.forename: G A
authors.initials: GA
authors.lastname: Van 't Klooster
authors.forename: P B
authors.initials: PB
authors.lastname: van Seeventer
authors.forename: H J
authors.initials: HJ
authors.lastname: Kolker
authors.forename: L A
authors.initials: LA
authors.lastname: Smit
authors.forename: R F
authors.initials: RF
authors.lastname: Witkamp
mesh_terms: D000818:Animals; D002250:Carbon Radioisotopes; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003470:Culture Media / Q000737:chemistry*; D004285:Dogs; D006041:Goats; D008099:Liver / Q000737:chemistry / Q000166:cytology; D008297:Male; D008862:Microsomes, Liver / Q000737:chemistry; D011874:Radiometry; D013418:Sulfamethazine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000097:blood / Q000652:urine; D014466:Ultraviolet Rays
chemical_list: D002250:Carbon Radioisotopes; D003470:Culture Media; C032633:N-acetylsulfamethazine; D013418:Sulfamethazine