Abstract: A simple high performance liquid chromatography (HPLC) method was developed for the simultaneous determination of rutin,quercetin,kaempferol in Amaranthus spinosus L.using methanol as extractant with ultrasonic assisted extraction.Chromatographic separation was carried out using a Phenomenex C18 column with a gradient elution,the mobile phase was methanol-0.2% acetic acid solution with the UV detection wavelength set at 360 nm and the flow rate of 1.0 mL/min.The linearity range for rutin,quercetin,kaempferol were found to be 00.002562.5000 μg/mL,and the correlation coefficients were 0.99999, 0.99997, 0.99999, respectively.The limits of detection (LOD,S/N=3) were 0.1411 ng/mL,0.0795 ng/mL,0.0188 ng/mL,respectively.The limit of quantification (LOQ,S/N=10) were 0.4703 ng/mL,0.2652 ng/mL,0.0627 ng/mL,respectively.Accuracy of the method was checked by conducting recovery studies at three different concentration levels for all the three compounds and the average recoveries were ranged from 95.79% to 101.41%.The developed HPLC method was found to be simple,precise,specific,sensitive and accuracy.It provided a new and scientific mean for routine quality control of Amaranthus spinosus L.

Key words: rutin, quercetin, kaempferol, ultrasonic-assisted extraction, high performance liquid chromatography, Amaranthus spinosus L.