Spectrophotometric and chromatographic methods for the simultaneous determination of rutin and ascorbic acid in their pharmaceutical formulation

Kareem M Younes; Mai A Basha; Maissa Y Salem

Abstract

Spectrophotometric and chromatographic methods for the simultaneous determination of rutin and ascorbic acid in their pharmaceutical formulation

Author(s): Kareem M Younes*, Mai A Basha and Maissa Y Salem

Four different methods namely, first-derivative of ratio spectra, bivariate, thin layer chromatography and high performance liquid chromatography were used to determine rutin and ascorbic acid simultaneously in their pharmaceutical dosage form. The derivative ratio spectra method was based on measuring the peak amplitudes for ascorbic acid at 275 and 286 nm using 20.0μg.ml-1 rutin as a divisor over a concentration range of 1.0 – 10.0 μg.ml- 1 for ascorbic acid, while rutin can be measured directly at 363 nm without interference from ascorbic acid over a concentration range of 4.0-20.0 μg.ml-1 for rutin. Bivariate method is used for simultaneous determination of both drugs over a concentration range of 2.0 – 12.0 and 2.0 – 16.0 μg.ml-1 for ascorbic acid and rutin, respectively. The method was based on measuring the absorbance at the selected wavelengths. A TLC separation with densitometric detection of both drugs was achieved using ethyl acetate: methyl isobutyl ketone: formic acid: glacial acetic acid: water [20:12:2.8:1.2:20, v/v/v/v/v] as developing solvent. The method allowed determination of rutin and ascorbic acid in concentration ranges of 2.0-6.0 and 4.0-8.0 μg.spot-1, respectively. Furthermore, a high performance liquid chromatographic procedure with ultraviolet detection at 270 nm was developed for the separation and determination of the studied drugs using a C8 column over a concentration range of 5.0-40.0 and 0.5 – 10.0 μg.ml-1 for rutin and ascorbic acid, respectively. The mobile phase is composed of ammonium acetate buffer: acetonitrile [80: 20, v/v], pH was adjusted to 6.3 by orthophosphoric acid. The proposed methods were successfully applied for the determination of the studied drugs in their mixtures and in pharmaceutical formulations containing them.

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