Abstract

DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF LAMOTRIGINE AND CLOZAPINE

Namratha A.*, Channabasavaraj K. P. and Madhusudana D. N.

ABSTRACT

In the present research work a simple, sensitive, precise, rapid and accurate reverse phase high performance liquid chromatography (RP- HPLC) method was developed and validated for simultaneous estimation of Clozapine and Lamotrigine. The Chromatographic separation was performed using LiChrospher RP-18e (250 mm×4.6 mm, 5 μm) column as stationary phase and Methanol: Acetonitrile: 15 mM phosphate buffer (pH 2.5) (40:30:30 % v/v/v) as mobile phase with a flow rate of 1 mL/min. The analysis was performed at ambient temperature and the eluent was monitored at 223nm using UV detector. The retention time of Lamotrigine and Clozapine were found to be 3.4 min and 4.4 min, respectively. The calibration curves were linear (r2 = 0.9991 and 0.9994) over a concentration range of 5-40 μg/mL for Lamotrigine and Clozapine, respectively. The Limit of detection (LOD) for Lamotrigine and Clozapine were observed to be 1 μg/mL and 1.5 μg/mL, respectively. The limits of quantitation (LOQ) were found to be 2 μg/mL and 2.5 μg/mL, respectively. The developed method was validated as per ICH guidelines using parameters like linearity, specificity, system suitability, precision, ruggedness, robustness, accuracy, sensitivity. All the validation parameters were found to be well within the acceptance criteria. Hence the proposed method can be used for the simultaneous analysis of Clozapine and Lamotrigine in bulk and tablet dosage forms.

Keywords: RP-HPLC, Clozapine (CLOZ), Lamotrigine (LAMO), Validation, tablet dosage form.