Determination of tropicamide in pharmaceutical formulations using high-performance liquid chromatography.

An isocratic, reversed-phase liquid chromatographic method was developed for determination of tropicamide using atropine as an internal standard in a pharmaceutical dosage form. Tropicamide and atropine sulfate were separated using a microBondapak ODS (C18) column by isocratic elution of mobile phase with flow rate of 2.0 ml/min. The mobile phase composition was methanol-50 mM phosphate buffer (pH 4; 30:70, v/v). The eluate was monitored at 257 nm with detector range setting fixed at 0.01 AUFS. Under these conditions, the retention times were 4.81 min for atropine and 11.89 min for tropicamide. The standard calibration curve was linear over a sample concentration range from 2 to 300 microg/ml, with limit of detection of 0.15 microg/ml. The assay linearity was good (typically r2 = 0.9992) and the standard curves were linear in the detection range. The precision of the method (expressed by relative standard deviation) and the accuracy (mean error in percent) were <5% for both intra- and inter-day assays. Recovery at 80-120% of labeled claim ranged from 98.4 to 100.7% for tropicamide. The proposed method was satisfactorily applied to the determination of tropicamide in pharmaceutical preparation and stability indicating studies.