RP-HPLC Method Development and Its Validation for Simultaneous Estimation of Alprazolam and Fluoxetine Hydrochloride in Pharmaceutical Dosage Form

Abstract

A simple, specific, accurate and precise reverse phase high pressure liquid chromatographic method
has been developed for the simultaneous determination of Alprazolam(Alp) and Fluoxetine
Hydrochloride(Flx) in tablet dosage form by reverse phase C18 column, Phenomenex (250mm x
4.60mm), Particle Size 5 . The samples were analyzed by using Acetonitrile: Water in the ratio of
75:25 (pH adjusted to 2.75 with 0.1% orthophosphoric acid) as a mobile phase at the flow rate of 1.1
ml/min in isocratic mode and detection wavelength 224 nm. Both the drugs were eluted within 5
minutes and give sharp peak with high theoretical plate count and low tailing factor. The retention
time for Fluoxetine Hydrochloride and Alprazolam was found to be 2.06 and 3.14 min respectively.
The validation was carried according to ICH guidelines. In linearity curve correlation coefficients for
Alprazolam and Fluoxetine Hydrochloride were found to be 0.9989 and 0.9994 respectively. The
percent recovery was 99.56 for Alprazolam and 99.96 for Fluoxetine Hydrochloride indicating
accuracy and reliability of method. So the method can be used for estimation of combination of these
drugs in tablet dosage form.