Objective: To develop a simple, rapid and specific reversed phase High-Performance Liquid Chromatography (HPLC) method for comparative analysis of flavonoids, quercetin and kaempferol, in different plant parts (leaves, stem and roots) of Cissus quadrangularis Linn.Methods: The HPLC method which can be used effectively for separation of components from plants has been developed to perform a comparative analysis of flavonoids, quercetin and kaempferol, in different plant parts (leaves, stem and roots) of Cissus quadrangularis Linn. An endcapped C18 column at 370 nm, and water: acetonitrile (45:55) containing 0.1% o-phosphoric acid as mobile phase was used.Results: Quercetin and kaempferol were well resolved at about 5 min and 7 min respectively. Calibration curves for quercetin and kaempferol were linear in the range of 1-10 µg/ml (R2= 0.999) and 0.5-10 µg/ml (R2= 0.999) respectively. The sensitivity of the method was found to be higher, with a limit of detection (LOD) values of 0.42 μg/ml for kaempferol and 0.48 μg/ml for quercetin and limit of quantification (LOQ) values for kaempferol and quercetin 1.27 μg/ml and 1.47 μg/ml respectively. Percentage recovery values for kaempferol were found to be 99.18, 99.03 and 98.32 for the leaves, stem and root respectively, and for quercetin, they were 99.77, 100.12 and 100.54 for the leaves, stem and root respectively.Conclusion: The developed HPLC method for the analysis of flavonoids has enabled rapid, accurate and reproducible determination. The method can be applied successfully for analysis of quercetin and kaempferol in various plant parts of C. quadrangularis.Keywords: Cissus quadrangularis, HPLC, Kaempferol, Querceti