Simultaneous HPLC-DAD analysis of five flavonoids in diabetic rat plasma and its application in the study of pharmacokinetics

A HPLC-DAD method for simultaneous analysis of five flavonoids (rutin, quercitrin, quercetin, kaempferol, and isorhamnetin) in diabetic rat plasma has been developed and validated. Separation of the five flavonoids was accomplished on a C 18 column (250 mm × 4.6 mm i.d., 5-µm particle) and detection was performed at 350 nm. The best resolution was achieved with a methanol-0.1% formic acid gradient at a flow rate of 1.0 mL min −1 . The correlation coefficients for all the calibration plots ( r > 0.999) showed linearity was good over the range tested. The relative standard deviation of the method was less than 7% and 10% for intra- and inter-day assays, and average recovery was between 77.2 and 99.2%. Sensitivity was high and detection limits were between 0.006 and 0.02 µg mL −1 . The method has been successfully used to determine drug concentrations in diabetic rat plasma samples and the pharmacokinetics of the drugs.