Comprehensive two-step sequential extraction and pharmacological profiling of Emirati plant leaves: A multi-target natural product discovery approach

A green and efficient integrated extraction protocol was developed for the simultaneous enrichment and separation of bioactive compounds from the leaves of three Emirati plant species: mangrove ( Avicennia marina ), sidr ( Ziziphus spina-christi ), and ghaf ( Prosopis cineraria ). For the first time, a two-step extraction approach was implemented, using supercritical CO 2 extraction for the selective isolation of non-polar and low-polar compounds, followed by ultrasound-assisted extraction optimized with a Box–Behnken design. Optimization involved three variables: ethanol concentration (0–100 %), ultrasound amplitude (20–90 %), and extraction time (2–18 min). Both non-polar and polar fractions were enriched with bioactive constituents and exhibited significant antioxidant, antibacterial, antidiabetic, and anti-inflammatory activities. Among the studied species, sidr leaf extract demonstrated superior in vitro bioactivity, whereas mangrove and ghaf extracts showed comparatively limited pharmacological potential. Comprehensive LC-MS profiling of the sidr extract revealed quercetin, estradiol, and maslinic acid as the key metabolites. Based on its strong pharmacological effects, sidr was selected for detailed pathway and gene–metabolite network analysis. The results confirmed its multi-target actions across anti-inflammatory, antimicrobial, and antidiabetic pathways, supporting its potential as a natural therapeutic source. Overall, the developed protocol offers a sustainable, scalable, and highly efficient platform for the valorization of plant biomass. This approach facilitates the production of functional ingredients suitable for pharmaceutical, nutraceutical, and cosmetic applications. • Green two-step extraction efficiently isolated diverse plant leaf compounds. • Optimized UAE maximized yield and bioactivity while reducing extraction time. • Sidr showed strongest activity, rich in quercetin, estradiol and maslinic acid. • Network analysis confirmed sidr's multi-target therapeutic potential.