Validated HPLC fingerprint analysis for Simultaneous Determination of Quercetin and Kaempferol in Methanolic Extract of Tridax procumbens

Aim: A rapid, sensitive and accurate Reversed Phase-High Performance Liquid Chromatography method (RPHPLC)with Photo-diode array Detection (PDA) was developed and validated for the simultaneous determination of Quercetin and Kaempferol in the methanolic extract of Tridax procumbens plant. Method: Whole plant of Tridax procumbens was collected from Dhule District, Shirpur; dried and methanolic extract was prepared and analyzed for the presence of standards - Quercetin and Kaempferol using TLC and RPHPLC method and detected at λ=240nm.Results: The Rf values for Quercetin, and Kaempferol was 0.35 and 0.48 respectively. There was a linear relationship between chromatographic area and concentration of analytes over the concentration range 1-40 μg/ml for both, Quercetin and Kaempferol with the retention time of about 8.9 minutes and 10.6 minutesrespectively and correlation coefficient of linear regression analysis was found to be r2= 0.999 and detection limits were between 0.164-0.289μg/ml for Quercetin and 0.497-0.816 μg/ml for Kaempferol. The average percentage recovery was found to be 98.02% and 98.80% for Quercetin and Kaempferol respectively. Conclusion: The RP-HPLC method developed was found to be simple, sensitive, precise and accurate forroutine quality control of the powder of whole plant of Tridax procumbens.