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.
[Full Text Article]
[Download Certificate]
Impact Factor : 8.181 
