Fluorometric determination of isoniazid and its metabolites in urine by high-performance liquid chromatography using in-line derivatization.
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.
Kohno
H
H
Pharmaceutical Division, National Tokyo Chest Hospital, Japan.
Kubo
H
H
Furukawa
K
K
Yoshino
N
N
Nishikawa
T
T
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
D007539:Isonicotinic Acids; D007538:Isoniazid