Research J. Pharm. and Tech. 15(1): January 2022 177 ISSN 0974-3618 (Print) www.rjptonline.org 0974-360X (Online) RESEARCH ARTICLE Identification of bioactive ingredients in Chenopodium murale L Chenopodiaceae by HPLC and GC/MS Rasha M.M. Mohasib 1 , A.Nagib 2 , A.Abdel Samad 2 , Zeinab A. Salama 1 *, Alaa A.Gaafar 1 , Hanan A.A. 1 , Taie, Sameh R. Hussein 3 1 Plant Biochemistry Department, National Research Centre, Dokki, Giza, Egypt. 2 Biochemistry Departement, Faculty of Agricultural, Cairo university, Dokki, Giza, Egypt. 3 Department of Phytochemistry and Plant Systematics, National Research Centre, 33 El Bohouth St., Dokki, Giza, Egypt. *Corresponding Author E-mail: dr.zeinabsalama70@gmail.com ABSTRACT: This study aims to evaluate the successive extraction of the active ingredients and their antioxidant activity, anti- arthritic as well as anticancer activity of aerial parts (stem, leaves, and flowers) of Chenopodium murale L. Therefore, C. murale plants were extracted using four solvents with a wide range of polarities: n-hexane, ethyl acetate, methanolic and aqueous extracts. Chemical analysis proved it to be a potential source of protein, fat, carbohydrates, the results showed the percentages were: moisture content (92.45%), ash content (18.19%), crude protein (30.42%), crude lipid (2.86%), and carbohydrate (48.3%) respectively of the C.murale. Methanol extracts showed the highest content of total phenolic (TP), total flavonoid (TF), and total tannin (TT). The active ingredients were assessed as well employing gas chromatography coupled to mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC). The HPLC analysis of phenolic compounds confirmed that the methanol extract of C.murale detected high amounts of coumarin, 3, 4, 5 methoxy-cinnamic, and pyrogallol respectively. The ethyl acetate extract of C.murale herb displayed a rise cytotoxic effects on MCF7 (89.30 %), aqueous HCT116 (81%), methanol (60.70%) as well as n-hexane (39.80) respectively at 37°C for 48 h of exposure and concentration 100 μg/ml. In anti-arthritic activity at a dose-dependent, the Ethyl acetate successive fraction proved to be the most significant where it produced a percentage of inhibition ranging from 51.73 to 95.77 % followed by methanol fraction 47.70 to 90.02 % at (P ≤ 0.05), compared to Diclofenac as standard 91.22 to 96.44 %. Thus, our findings highlight the potential of this plant for its possible clinical use to oppose malignancy development against especially breast and colon cell lines with anti-arthritic effects as a bioagent in pharmaceutical industries. KEYWORDS: Chenopodium murale; HPLC; GC/MS; antioxidant; anti-proliferative, anti-arthritic activities. INTRODUCTION: The Chenopodiaceae is a family compromised from 102 genera and 1400 species 1 . In general, the genus Chenopodium contains collections of weedy herbs nearly more than 200 species native to Europe, America, and Asia 2,3 . Chenopodium murale L. commonly called nettle leaf goosefoot is an essential annual herbaceous weed that grows in waste places 4,5 . It is green, alternate, broad lobed ad stalked long or short leaves with forked stem 6 . Received on 12.01.2021 Modified on 29.03.2021 Accepted on 15.06.2021 © RJPT All right reserved Research J. Pharm. and Tech 2022; 15(1):177-187. DOI: 10.52711/0974-360X.2022.00029 From the phytochemical point of view, the plants of Chenopodium L were reported to possess numerous medicinal activities used in popular medicine according to its variety of structural patterns of phytochemicals, theses chemical compounds are primary metabolites such as amino acids and proteins and secondary metabolites 7 , such as terpenoids 8,9 , sterols 10 , saponins 11 , alkaloids 12 , vitamins 13 , Flavonoids and cumarine 14,15 . A wide range of applications is well-known in folk medicine for the treatment of cough, abdominal pain, pulmonary obstruction, and have diuretic ad laxative activity 16 , as well as it is reputed to be good pharmacological activities of Chenopodium species such as anthelmintic, anti-inflammatory activity 17 ,