Coupling of liquid-liquid extraction and mathematical filtration techniques for the separation and quantification of five components in semisolid dosage form with severely overlapped spectra.

Quadriderm cream was a combination of four components; Clioquinol (CLIO), Betamethasone (BETA), Tolnaftate (TOL), Gentamicin (GEN) in addition to the preservative Chlorocresol (CC). Four components CLIO, TOL, BETA, and CC were extracted in methanol and determined by mathematic filtration spectrophotometric techniques. The partially overlapped spectrum of CLIO was determined by constant value, constant multiplication, and concentration value methods then eliminated via spectrum subtraction (SS) to get the resolved ternary mixture of TOL, BETA, and CC with severely overlapping spectra. TOL was determined by derivative ratio at zero crossing point of BETA using CC as a divisor. While, BETA could be determined using TOL as a divisor at zero crossing of CC. BETA and CC were obtained using novel (DDFS) followed by SS. By applying these novel procedures, the DD spectrum of each component alone was recovered where P was directly proportional to its concentration. Liquid-liquid extraction technique was used for the semisolid dosage form where GEN was extracted with a mixture of chloroform: water (50:50, v/v); and the induced fluorescence obtained by derivatization with o-phthalaldehyde was measured at 419 nm after excitation at 359 nm. Accuracy and precision testing of the developed methods showed good results. Specificity of the methods was ensured and was successfully applied for the analysis of pharmaceutical formulation of the five components in combination. ICH guidelines were used for validation of the proposed methods. Statistical data were calculated, and the results were satisfactory revealing no significant difference regarding accuracy and precision.