Rapid HPLC analysis of the antiepileptic lamotrigine and its metabolites in human plasma

A liquid chromatographic method with diode array detection (DAD) has been developed for the analysis of the antiepileptic agent larnotrigine (LTG) and its metabolites, lamotrigine 2-N-glucuronide and 2-N-methylated in plasma samples. The analytes were separated on a C8 RP column, using a mobile phase composed of methanol and a 0.45 mM, pH 3.5 phosphate buffer containing 0.17% triethylamine (24:76 v/v). Melatonin was used as the internal standard (IS). The DAD detector was set at 220 nm for the detection of all the analytes. A simple protein precipitation with methanol guaranteed high extraction yield values (> 90%) and good purification from matrix interference. Good linearity was obtained in the 0.1 - 15.0 mu g/mL range for LTG and lamotrigine 2-N-glucuronide and in the 0.1-2.0 mu g/ml, range for lamotrigine 2-N-methylated. The analytical method was validated in terms of precision, extraction yield, and accuracy. These assays gave RSD% values for precision always lower than 4.3% and mean accuracy higher than 80%. The method seems to be suitable for the analysis of plasma samples from patients treated with Lamictal.