Achyranthes aspera Linn. has been broadly used as traditionally medicine in several countries. All parts of this plant have been used in treatment and prevention of several complications. Taxonomically Achyranthes aspera Linn. belong to family Amaranthaceae. In the present study, stem of Achyranthes aspera Linn. were subjected to Pharmacognostic evaluation. Various methods including macroscopic, microscopic, physiochemical and phytochemical methods were applied to determine the features for the identification and standardization of intact and powdered drug of stem of Achyranthes aspera Linn. This analysis is proved useful to differentiate it from other drug material. The powdered plant material was removed by Soxhlet extractor in Ethyl acetate and ethanol after defatting with petroleum ether.Phytochemical testing of extracts discovered presence of glycosides, flavonoids, carbohydrates, etc. The total phenolic and flavonoid content of plant extracts was expressed as Gallic acid equivalents and as quercetin equivalents separately. Antioxidant activity was carried out by 2,2-diphenyl-1-picrylhyrazyl (DPPH) assay. DPPH scavenging assay were performed to evaluate the antioxidant activity which was found maximum at 100 microgram concentration for both extracts. The results of acute oral toxicity studies of plant extracts as per standard references revealed median lethal dose (LD50) could be greater than 2000mg/kg body weight in rats. Accordinglysafe experimental dose was calculated as 100 and 200mg/kg. The evaluation of in-vivo anxiolytic activity in rats was done using elevated plus maze model. The results showed increases locomotor activity in both extracts at dose of 200mg/kg compared to vehicle group.