Herein, the methanolic extract of Rumex hastatus was prepared and subjected to silica gel column chromatographic separation for purification. The chromatographic analysis yielded four bioactive compounds namely, 1,8-dihydroxy-3-methyl-anthraquinone (C1, chrysophanol), 3-methoxy-7-methyl-1,5-dihydroxy-anthraquinone (C2), 6-methyl-1,3,7-trihydroxy-anthraquinone (C3), and 4-hydroxycinnamic acid (C4). The structures of all the isolated bio-entities were elucidated by spectroscopic analysis (1D, 2D-NMR, and MS). The biological potentialities of bioactive compounds were evaluated by determining their antioxidant and anti-urease activities. The results revealed that compound 1 showed the highest nitrite radical scavenging activity (IC₅₀ = 0.39 mM) followed by C2 and C4 (IC₅₀ = 0.45) mM and C3 (IC₅₀ = 0.47 mM). Similarly, the determined IC₅₀ values for anti-urease activity were C2 (IC₅₀ = 0.39 mM), C1 (IC₅₀ = 0.40), C4 (IC₅₀ = 0.41), and C3 (IC₅₀ = 44). Molecular docking study revealed maximum interactions among the hydroxyl group of all compounds. In conclusion, Rumex hastatus extract could be a promising alternative to chemical additives in pharmaceutical industries due to its noteworthy biological activities. PRACTICAL APPLICATIONS: As we have shown in this study that Rumex hastatus is a prolific source of biologically active compounds with potent antioxidant and anti-urease activities, therefore, R. hastatus extract could be used as a promising alternative to chemical additives in pharmaceutical industries due to its unique compounds and noteworthy biological activities.

Keywords: Rumex hastatus; anti-oxidant activities; anti-urease; bioactive compounds; chemical composition; methanolic extract; molecular docking; structure elucidation.