Detection of the principal synthetic route indicative impurity in Lamotrigine

Abstract

An analytical method has been developed for the detection of trace amounts of the principal synthetic route indicative impurity in lamotrigine (3,5-diamino-6-(2,3-dichlorophenyl)-1,2,4-triazine). A sample extract was preconcentrated by normal-phase high-performance liquid chromatography (HPLC) and analysed by subsequent on-line reversed-phase HPLC–thermospray mass spectrometry (TSP-MS). During the sample extraction and concentration step, carried out by semipreparative normal-phase chromatography, the preliminary separation of the impurity from the lamotrigine takes place. The organic solvent (dichloroethane–methanol, 90:10, v/v) is evaporated from the collected fraction and the material is redissolved in a smaller volume of the reversed-phase mobile phase. The collected fraction is then subjected to reversed-phase HPLC–TSP-MS. The influence of an ultrasonic extraction step has been examined. When the method was applied to lamotrigine tablets, a shake flask partitioning step using 1 mg/ml EDTA in water–dichloroethane was used instead of the ultrasonic extraction. Detection limit and recovery measurements showed that the route indicative impurity formed during the synthesis could be detected in the 50–100 ppb (w/w) range.