Simultaneous determination of four vasoactive phytochemicals in different pharmaceutical preparations by a simple HPLC-DAD method

A new, simple isocratic HPLC method was developed and validated for the simultaneous estimation of four vasoactive phytochemicals: ascorbic acid (ASC), rutin (RUT), hesperidin (HSP), and diosmin (DSM) in different pharmaceutical preparations. Separation of the four compounds was achieved in about 8 min on a phenomenex-C₁₈ column (250 mm x 4.6 mm, 5 μm particle size) using a mobile phase consists of 50 mM phosphate buffer (pH 4.0):acetonitrile (75:25, v/v) eluted at 1.0 mL min^-1 with diode array detection at 275 nm. Linear correlation was obeyed over the concentration ranges of 2.0-60.0, 1.0-20.0, 2.0-50.0, and 5.0-100.0 μg mL^-1 for ASC, RUT, HSP, and DSM, respectively. The developed method is efficient, reproducible and appropriate as a versatile method for quality control of various combinations of these drugs in different pharmaceutical preparations with high values of %recoveries which agreed well with the labeled claim. The method is also a stability-indicating one for DSM in its pure state and single-component tablets. A comparison of the results obtained by the developed method with those of the comparison methods confirmed no significant differences between them with respect to accuracy and precision.