Simultaneous measurement of ethosuximide, primidone, phenobarbital, phenytoin, carbamazepine, and their bioactive metabolites by liquid chromatography.

A simple, isocratic liquid-chromatographic method was developed for simultaneously measuring ethosuximide, primidone, phenobarbital, phenytoin, carbamazepine, and their bioactive metabolites within 10 min. The chromatographic system involves a Waters' Radial-NOVA PAK C18 reversed-phase column and acetone/methanol/acetonitrile/10 mmol/L phosphate buffer (10/21/8/61 by vol, pH adjusted to 7.95 with NaOH) as mobile phase. The antiepileptic drugs are extracted from 50 microL of serum by mixing with 50 microL of acetonitrile containing 10 mg of tolybarb per liter as internal standard. After centrifugation, 20 microL of the supernate is injected onto the column and eluted with mobile phase at the rate of 2.8 mL/min at ambient temperature. The column effluent is monitored at 200 nm. The method can detect the five antiepileptic drugs in concentrations as low as 0.5 mg/L. Analytical recovery ranges from 98 to 102%. Within-run CV ranged from 2.9 to 5.8% and between-run CV from 4.7 to 7.1%. The method can also be used to measure N-desmethyl-methsuximide, chloramphenicol, and pentobarbital.