Simultaneous determination of primidone and its three major metabolites in rat urine by high-performance liquid chromatography using solid-phase extraction.
A new high-performance liquid chromatographic method for simultaneous determination of primidone (PRM) and of its three major metabolites, phenobarbital (PB), p-hydroxyphenobarbital (p-HO-PB) and phenylethylmalonamide (PEMA), in rat urine, was developed. After acid hydrolysis, these compounds were extracted from urine by means of a Bond Elut Certify LRC column with good clean-up. The extracts were chromatographed on a C18 reversed-phase column using isocratic elution at 40 degrees C, with UV detection at 227 nm. The limit of detection was 0.5 mg/ml for the four compounds. Good linearity (r2>0.99) was observed within the calibration ranges studied: 37.4-299.3 microg/ml for PRM, 26.4-211.2 microg/ml for PB, 12.5-100.2 microg/ml for p-HO-PB and 12.1-97.0 microg/ml for PEMA. Repeatability was in the range 3.1-6.8%. This method constitutes a useful tool for studies on the influence of various parameters on primidone metabolism.
Ferranti
V
V
Laboratoire de Pharmacochimie, Faculté de Médecine et de Pharmacie de Rouen, Saint-Etienne-du-Rouvray, France.
Chabenat
C
C
Ménager
S
S
Lafont
O
O
D000818:Animals; D000927:Anticonvulsants / Q000378:metabolism / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005069:Evaluation Studies as Topic; D010634:Phenobarbital / Q000031:analogs & derivatives / Q000652:urine; D010657:Phenylethylmalonamide / Q000652:urine; D011324:Primidone / Q000378:metabolism / Q000652:urine*; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results
D000927:Anticonvulsants; D011324:Primidone; C017414:4-hydroxyphenobarbital; D010657:Phenylethylmalonamide; D010634:Phenobarbital