In vitro and in silico biological activities and comprehensive metabolomic profiling using UHPLC-ESI-QTOF-MS/MS of Himalrandia tetrasperma: A novel source of bioactive metabolites from Pakistan

This study is an inclusive assessment of the metabolite profile and biological potential of Himalrandia tetrasperma leaf extract. The crude extract was prepared using 80% methanol and fractionated with hexane, ethyl acetate, acetone, and distilled water. The assessed biological activities include antioxidant (DPPH), phytochemical, antimicrobial, and in vitro and in silico AChE inhibitory and cytotoxic effects against the leukemia U937 cell line. Metabolomic analyses were performed with ATR-FTIR and UHPLC-ESI-QTOF-MS/MS. The crude extract showed significant antioxidant activity (IC 50 : 395 ± 3 μg/mL). The MIC values ranged from 500 to 1375 μg/mL against bacteria and from 625 to 1375 μg/mL against fungi. The ethyl acetate fraction exhibited the uppermost AChE inhibitory activity (IC 50 : 620 ± 2 μg/mL), and the hexane fraction displayed the most significant cytotoxic effect (GI 50 : 531 ± 1 μg/mL). FTIR study recognized various functional groups indicative of phytochemicals such as phenolics, carboxylic acids, aldehydes, ketones, ethers, esters, and amines. UHPLC-ESI-QTOF-MS/MS detected 182 metabolites, including terpenoids (49.28%), lipids (17.77%), steroids (11.13%), flavonoids (4.94%), carbohydrates (3.67%), and phenols and phenolic acids (3.65%). The most abundant individual metabolite was Oleanolic acid (10.11%), followed by Ursolic acid (4.67%), Coriolic acid (3.52%), and Gulonic acid (2.82%). Docking studies confirmed that Quercetin 3-glucoside-7-rhamnoside (BE -12.11 kcal/mol) and Alcesefoliside (-11.6 kcal/mol) showed significant inhibitory potential against AChE and DNA Topoisomerase II (for cytotoxicity), respectively. Overall, this research represents a sustainable approach, identifying numerous potent bioactive metabolites with promising antioxidant, antimicrobial, AChE-inhibitory, and anticancer activities, which may potentially lead to new drug development through further advanced studies. • H. tetrasperma is a novel plant of medicinal and nutritional value from Pakistan. • Metabolomic profiling by UHPLC-ESI-QTOF-MS/MS identified 182 metabolites. • Extract was rich in terpenoids, lipids, steroids, polyphenols, and carbohydrates. • Many of the identified compounds support the evaluated biological activities. • In-silico study validates the potential against AChE and leukemia U937 cell line.