Abstract
DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF HYDROXYAMPHETAMINE HYDROBROMIDE AND TROPICAMIDE IN OPHTHALMIC FORMULATION

Yajnesh P., Mubeen G., Ritu Vivek K.*, Lalitha N. and Krishnasis C.

ABSTRACT

A simple and rapid RP-HPLC method was developed and validated for the simultaneous estimation of Hydroxyamphetamine hydrobromide and Tropicamide in ophthalmic formulation using Phenomenex BDS C-18 (250 × 4.6 mm, 5 μ) column and Acetonitrile: Phosphate buffer (50 mM, pH 5.6) in the ratio of 40: 60 v/v as mobile phase with 1 mL/min flow rate and UV detection at 257 nm. Retention time for Hydroxyamphetamine hydrobromide and Tropicamide was observed to be 2.40 and 4.10 minutes respectively. The content in the formulation was found to be about 104.83 and 105.91% for Hydroxyamphetamine hydrobromide and Tropicamide respectively. The mean percentage recovery at three different levels was found to be 100.75 – 102.09% for Hydroxyamphetamine hydrobromide and 99.32 – 101.89% for Tropicamide. Intraday and Interday precision was found to be below 2% with no significant difference in reproducibility between two different analysts. Linearity (R2) was found to be above 0.99 for Hydroxyamphetamine hydrobromide and Tropicamide in the concentration range of 40-400 μg/mL and 20-250 μg/mL respectively. All the ICH parameters Q2 (R1) were found to be well within the acceptance criteria and hence can be applied for routine analysis for Hydroxyamphetamine hydrobromide and Tropicamide in ophthalmic formulation.

Keywords: Hydroxyamphetamine hydrobromide, Tropicamide, RP-HPLC, Ophthalmic formulation.


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