The main focus of the present paper was to determine Quantitative Analysis, Antioxidant Assays and Thin layer Chromatography Analysis of the roots extracts of Withania somnifera, Bacopa monnieri, Terminalia arjuna, Rannunculus sceleratus and Acalypha indica. Extractions of the plant’s roots were carried out by Maceration method using three solvents Ethanol, Methanol and Chloroform. Phytochemical Quantitative analysis, consisting of Total Phenol Content, Total Flavonoid Content and Alkaloid Content were done using the standard phytochemical assays. The extracts were screened for Antioxidant Assays including DPPH (1,1-Diphenyl-2-picrylhydrazyl) Radical Scavenging Assay, ABTS(2,2'-azinobis-(3- ethylbenzothiazoline-6-sulfonicacid)) Radical Scavenging Assay, Hydrogen Peroxide Assay. Selected methanol and ethanol extracts were further used for particular compound separation process through Column Chromatography and Thin Layer Chromatography. The results of the Quantitative screening revealed the presence of the three important bioactive compounds i.e. Phenols, Flavonoids, and alkaloids in a high a very high concentration in Ethanol, methanol plant’s extracts but highly less concentration in Chloroform extracts. Similarly, the Antioxidant Scavenging Activity Of ethanol and methanol plant’s extracts were shown having a very high activity except in Bacopa monnieri and methanol extracts of Rannunculus sceleratus where moderate activity is shown. The Chloroform extracts had shown very less or no scavenging activity. The methanol and ethanol extracts were estimated having activity of DPPH:45%-65%, ABTS:45%-70%, H2O2:90%-97%. In this current investigation, only selected extracts i.e. ethanol and methanol extracts were subjected to column chromatographic separation, around 80 fractions of total extracts were separated. The TLC analysis of the ethanol and methanol extracts showed the presence of the three compounds as spots in specific fractions indicating the extracts as potent antioxidant agents, the existence of European Journal of Molecular & Clinical Medicine ISSN 2515-8260 Volume 7, Issue 11, 2020 4395 phytogenic chemical compounds which could exert several beneficial effects by virtue of their antioxidant activity.