Development and Validation of Second-Derivative Spectrophotometry Method for Simultaneous Estimation of Alprazolam and Fluoxetine Hydrochloride in Pure Powder and Tablet Formulation and Its Comparison with HPLC Method

Abstract

This paper describes validated Second Derivative Spectrophotometry (D2) method for simultaneous estimation
of Alprazolam (ALP) and Fluoxetine hydrochloride (FXT) in pure powder and formulation. D2 method,
applying the peak zero method, was developed for the determination of ALP and FXT in their combined tablet
formulations without prior separation. The solutions of standard and sample were prepared in 0.1M HCl.
Quantitative determination of the drugs was performed at 232.14 nm and at 225.25 nm (N = 4; ∆ λ = 2.8) for
ALP and FXT, respectively. Proposed D2 method was evaluated for the different validation parameters. The
specificity test showed that there was no interference from excipients commonly found in the commercial
pharmaceutical formulations at analytical wavelength of ALP and FXT. Quantification was achieved over the
concentration range 4-14 µg.mL-1 for both drugs with mean recovery of 99.36 ± 0.84 and 99.60 ± 0.93 % for
ALP and FXT, respectively. This method is simple, precise, and sensitive and applicable for the simultaneous
determination of ALP and FXT in pure powder and formulation. The method was compared to high-performance
liquid chromatography (HPLC) method, which was reported for the same drugs. No significant difference was
found between the methods for ALP and FXT quantitation.