Development and validation of a reversed-phase HPLC method for analysis of tetrahydrozoline hydrochloride in eye drop formulations

Abstract

A simple,precise,accurate,andstability-indicatingmethodisdevelopedandvalidated for analysisoftetrahydrozolinehydrochlorideineyedropformulations.Separationwasachieved on areversed-phaseC8 column (125mm 4.6 mmi.d.,5 mm) using a mobile phase consisting of acetonitrile/phosphate bufferofpH3.0(20:80,v/v)ataflowrateof1.0mL/minandUVdetection at 240nm.ThismethodisvalidatedaccordingtoUnitedStatesPharmacopeiarequirementsfor new methods,whichincludeaccuracy,precision,selectivity,robustness,andlinearityandrange. This methodshowsenoughselectivity,accuracy,precision,andlinearityandrangetosatisfy Federal DrugAdministration/InternationalConferenceonHarmonizationregulatoryrequire- ments. Thecurrentmethoddemonstratesgoodlinearityovertherangeof0.025–0.075mg/mLof tetrahydrozoline with r2 0.999. Theaveragerecoveryofthemethodis100.8%witharelative standard deviationof0.47%.Thedegreeofreproducibilityoftheresultsobtainedasaresultof small deliberatevariationsinthemethodparametersandbychanginganalyticaloperatorshas proven thatthemethodisrobustandrugged.