Simultaneous determination of rutin and ascorbic acid mixture in their pure forms and combined dosage form

• A simple, sensitive and selective HPLC-UV method has been developed for simultaneous analysis of ascorbic acid and rutin
• The proposed HPLC-UV method has been validated according to ICH guidelines
• The method has been applied for analysis of studied drugs in laboratory prepared mixture and pharmaceutical formulations

A simple, rapid, sensitive and selective high performance liquid chromatographic (HPLC) method with ultraviolet detection has been developed for simultaneous determination of ascorbic acid and rutin in pure forms and pharmaceutical dosage form. HPLC separation was performed on Phenomenex C18 analytical column with 0.1% v/v acetic acid in water and acetonitrile (75:25, v/v), as mobile phase. The separation was done at ambient temperature with flow rate of 1 mL·min− 1 in isocratic mode. HPLC measurements were carried out using ultraviolet detection wavelength at 257 nm. The average retention times were 2.72 and 7.00 min for ascorbic acid and rutin, respectively. The calibration plots were constructed over the concentration range of 5.0–30.0 for ascorbic acid and 10.0–60.0 μg·mL− 1 for rutin. The limits of detection were 1.06 and 1.89 μg·mL− 1 and limits of quantification were 3.54 and 6.31 μg·mL− 1 for ascorbic acid and rutin, respectively. The proposed HPLC-UV method was successfully applied for determination of ascorbic acid in its tablets and for simultaneous determination of the studied drugs in their laboratory prepared mixtures and in pharmaceutical formulation. Statistical comparisons of the results with the reference method show an excellent agreement and indicate no significant difference in respect to accuracy and precision.

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