Several medicinal plants have been used from the traditional period of times to cure different diseases but there is little scientific evidence. The phytochemicals of plants can reduce cardiovascular and other diseases. The present study analyzed the five different medicinal plants of the Gulmi and Rupandehi districts of Nepal using in vitro studies. They were Crateva Unilocularis, Aegle marmelos, Nyctanthes arbor-tristis, Urtica dioica and Justicia adhatoda. The antioxidant potential of the methanolic extract of plants was evaluated by DPPH radical scavenging assay, total phenolic content was determined by using the Folin-Ciocalteu method and total flavonoid content was determined by using the Aluminium chloride colorimetric method. Results revealed that the methanolic extract of plants contained phytochemicals such as alkaloids, flavonoids, polyphenols, saponins, quinones, terpenoids, etc. The extract of Nyctanthes arbor-tristi showed the highest % of radical scavenging activity up to 64.931±0.032% with an IC50 value of 70.506±1.55μg/ml followed by Aegle marmelos and the lowest in Urtica dioica. Nyctanthes arbor-tristis revealed the highest TPC (97.647±7.01mgGAE/g) and lowest in Urtica dioica. Crateva unilocularis had the highest TFC 31.99±2.345mgQE/g and followed by Nyctanthes arbortristis and lowest in Justicia adhatoda. These parameters were analyzed from the period 5 September 2021 to 10 October 2021.