A liquid chromatographic assay using a high-speed column for the determination of lamotrigine, a new antiepileptic drug, in human plasma

Abstract

A sensitive, specific and rapid liquid-chromatographic method for the determination of the new antiepileptic drug lamotrigine (LTG) in human plasma is described. The method involves the use of a commercially available 3-microns particle size normal-phase column and a microflow-cell-equipped ultraviolet detector. Extraction is carried out with ethyl acetate after alkalinization on a 100-microliters plasma sample containing LTG and 3,5-diamino-6-(2-methoxyphenyl)-1,2,4-triazine as internal standard. The residue is reconstituted with 50 microliters of ethanol, and 5 microliters of the final solution is injected into the column. Elution is carried out at 35 degrees C using n-hexane/absolute ethanol/35% ammonia (80/20/0.25 by volume) as mobile phase at a flow rate of 2.0 ml/min. Detection is at 313 nm. The chromatographic separation requires < 3 min and the sensitivity limit is < 0.1 mg/L. Recovery is 88-96.2%, whereas within-day and day-to-day coefficients of variation are between 4.1 and 7.7%.