Determination of pyrazinamide and its main metabolites in rat urine by high-performance liquid chromatography.
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.
Mehmedagic
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A
Department for Pharmaceutical Technology, Faculty of Pharmacy, Sarajevo, Bosnia.
Vérité
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P
Ménager
S
S
Tharasse
C
C
Chabenat
C
C
André
D
D
Lafont
O
O
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
D000995:Antitubercular Agents; C032191:5-hydroxypyrazinamide; D011718:Pyrazinamide; C005296:pyrazinoic acid; C054996:5-hydroxypyrazinoic acid