Simple accurate and precise validated UV spectrophotometric methods have been described here for the simultaneous determination of Tretinoin (TN) Eusolex (EX) Hydroquinone (HQ) & Hydrocortisone acetate (HC) in their dosage form. Simultaneous determination of these four drugs was a major challenge till now. Each spectrum was filtered alone to its zero-order absorption spectrum (D0) form getting spectra typical to each pure component separately. Four methods were applied depending upon constants obtained from extended regions of partially overlapped spectra in the zero or first derivative forms. The methods applied are constant multiplication coupled with spectrum subtraction method (CM-SS) derivative transformation coupled with spectrum subtraction method (DT-SS) constant value method (CV) and concentration value (conc.value) method. The partially overlapped spectra of TN and EX in the mixture were obtained by CM-SS in their zero-order form allowing direct measurement at their λmax while the resolved binary mixture of HQ and HC obtained by SS; was determined by derivatization and transformed to their zero order by DT-SS. Also Ex and HQ concentrations were determined by the graphical representation of data only without regression equation by concentration value method and the results were compared to the conventional constant value method using a regression equation. The methods applied to the quaternary mixture under study were successfully applied for the simultaneous determination of the four drugs in synthetic mixtures and in their combined dosage form Tritospot® cream. Comparing the acquired results statistically together and to official methods demonstrated no significant difference.
Keywords: Concentration value; Constant multiplication; Constant value; Derivative transformation; Spectrum subtraction.
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