A simple, sensitive, precise, and accurate RP-HPLC assay method was developed for simultaneous estimation of Inulin and Esculetin in a formulation containing Chicory. Inulin is a polysaccharide and due to a lack of chromophore, it is to be derivatized before analyzing. Hence was opted to derivatized Inulin using Seliwinoff’s reagent. The chromatographic separation was achieved using C18 Phenomenex Hyperclone BDS column (250×4.6mm, 5μ). The mobile phase containing a mixture of 10 mM of phosphate buffer (pH 3.0) and acetonitrile (35: 65 v/v) at a flow rate of 1ml/min was used and the response was measured at 364nm (isosbestic point) using a PDA detector. The retention time of Esculetin and Inulin is found to be 3.7 and 7.1 minutes respectively with a resolution of 8.5. The method was linear over the concentration range of 0.05-0.8 μg/ml for Esculetin and 0.4-3 μg/ml for derivatized Inulin with a correlation constant of ≥ 0.999. The method was found to be precise, and robust within the acceptable limits. The LOD and LOQ were to be 0.006308 μg/ml and 0.01912 μg/ml for Esculetin and 0.06131 μg/ml and 0.1857 μg/ml for derivatized Inulin which