title: Fluorometric determination of isoniazid and its metabolites in urine by high-performance liquid chromatography using in-line derivatization.
abstract: A rapid, simple, and accurate method has been developed for the determination of isoniazid and its metabolites (isonicotinic acid, isonicotinylglycine, and acetylisoniazid) in human urine by high-performance liquid chromatography. Isoniazid and its metabolites are separated by reversed-phase ion-exchange chromatography with a mobile phase containing hydrogen peroxide as a fluorogenic reagent and butanesulfonate as a hydrophobic ion exchanger, and are detected by fluorometry (excitation at 317 nm and emission at 415 nm) using in-line derivatization at high temperature (160 degrees C). The detection limits are isonicotinic acid, 0.5 mumol/L; isonicotinylglycine, 1 mumol/L; acetylisoniazid, 1 mumol/L; and isoniazid, 1.5 mumol/L. This method can be applied for acetylator phenotyping.
authors.affiliation: Pharmaceutical Division, National Tokyo Chest Hospital, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Kohno
authors.forename: H
authors.initials: H
authors.lastname: Kubo
authors.forename: K
authors.initials: K
authors.lastname: Furukawa
authors.forename: N
authors.initials: N
authors.lastname: Yoshino
authors.forename: T
authors.initials: T
authors.lastname: Nishikawa
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange; D005470:Fluorometry / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007538:Isoniazid / Q000031:analogs & derivatives / Q000652:urine*; D007539:Isonicotinic Acids / Q000652:urine; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013696:Temperature
chemical_list: D007539:Isonicotinic Acids; D007538:Isoniazid