title: Determination of pyrazinamide and its main metabolites in rat urine by high-performance liquid chromatography.
abstract: A new high-performance liquid chromatograhic procedure for simultaneous determination of pyrazinamide (PZA) and its three metabolites 5-hydroxypyrazinamide (5-OH-PZA), pyrazinoic acid (PA), and 5-hydroxypyrazinoic acid (5-OH-PA), in rat urine was developed. 5-OH-PZA and 5-OH-PA standards were obtained by enzymatic synthesis (xanthine oxidase) and checked by HPLC and GC-MS. Chromatographic separation was achieved in 0.01 M KH2PO4 (pH 5.2), circulating at 0.9 ml/min, on a C18 silica column, at 22 degrees C. The limits of detection were 300 microg/l for PZA, 125 microg/l for PA, 90 microg/l for 5-OH-PZA and 70 microg/l for 5-OH-PA. Good linearity (r2>0.99) was observed within the calibration ranges studied: 0.375-7.50 mg/l for PZA, 0.416-3.33 mg/l for PA, 0.830-6.64 mg/l for 5-OH-PZA and 2.83-22.6 mg/l for 5-OHPA. Accuracy was always lower than +/- 10.8%. Precision was in the range 0.33-5.7%. The method will constitute a useful tool for studies on the influence of drug interactions in tuberculosis treatment.
authors.affiliation: Department for Pharmaceutical Technology, Faculty of Pharmacy, Sarajevo, Bosnia.
authors.forename: A
authors.initials: A
authors.lastname: Mehmedagic
authors.forename: P
authors.initials: P
authors.lastname: Vérité
authors.forename: S
authors.initials: S
authors.lastname: Ménager
authors.forename: C
authors.initials: C
authors.lastname: Tharasse
authors.forename: C
authors.initials: C
authors.lastname: Chabenat
authors.forename: D
authors.initials: D
authors.lastname: André
authors.forename: O
authors.initials: O
authors.lastname: Lafont
mesh_terms: D000818:Animals; D000995:Antitubercular Agents / Q000652:urine*; D002851:Chromatography, High Pressure Liquid; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry; D011718:Pyrazinamide / Q000031:analogs & derivatives / Q000652:urine*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results
chemical_list: D000995:Antitubercular Agents; C032191:5-hydroxypyrazinamide; D011718:Pyrazinamide; C005296:pyrazinoic acid; C054996:5-hydroxypyrazinoic acid