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Efficacy of the medicinal plants depends on the nature of primary and secondary metabolites present and assay of metabolites in raw botanical parts used for the drugs. Quantity and nature of metabolites that are presented are correlated with the growth of the plants, herbs and the season in which they are collected. Hence, it is important for a search of the best time and season for collection, based on the part used of medicinal plants, the assay of the secondary metabolites present and other morphological features for assess the maturity etc., In this study, it is aimed to evaluate the seasonal variation in terms of assay of wedelolactone through RP-HPLC and validate the best procurement time for the whole plant of Eclipta prostrata L. besides botanical verification and pharmacognostic authentication. The developed method was validated, results of analytical findings were summarized by all three triplicate tests for the three different solvent extracts of whole plant material of Bhringraj for all collected six seasons and found more abundance of wedelolactone in the range of 0.146-0.209 % w/w in Sharad ritu (Sep-Oct)/ Autumn season.