Ozonation is an emergent green technology that modifies the chemical composition and bioactivity of natural oils, creating new opportunities for functional and biomedical use. In this study, the chemical changes and in vitro activities of lettuce (Lactuca sativa) oil before and after ozonation were evaluated. Gas chromatography–mass spectrometry (GC–MS) revealed an increase in both the number and diversity of constituents in ozonated oil, with (Z)-13-docosenamide and trans-13-octadecenoic acid as predominant components. Fourier-transform infrared (FTIR) spectra showed overall similarity between native and ozonated oils, but with three additional characteristic bands in the ozonated sample. Bioassays demonstrated that ozonation enhanced anti-Helicobacter pylori activity (inhibition zone 21.3 ± 0.3 mm), supported bactericidal effects, and improved antibiofilm and antihemolytic properties. The antioxidant capacity of ozonated oil was modestly increased (IC₅₀ = 3.95 ± 0.4 µg/mL), while butyrylcholinesterase inhibition was more markedly enhanced (IC₅₀ = 2.58 ± 0.6 µg/mL), compared to that of the non-ozonated oil (IC₅₀ = 6.14 ± 0.3 µg/mL and IC₅₀ = 4.38 ± 0.4 µg/mL, respectively). Molecular docking suggested strong interactions of major ozonation-derived compounds with human BuChE and H. pylori urease, providing mechanistic support for the observed activities. Overall, these results indicate that ozonation modestly but consistently enhances the biological potential of lettuce oil through compositional shifts, highlighting its promise for development as a safe functional food ingredient with possible biomedical applications.

Natural products may address infection, oxidative stress and metabolic disorders. This study used gas chromatography-mass spectrometry (GC-MS), bioassays and molecular docking to characterize Delonix regia seed oil and evaluate its antimicrobial, antibiofilm, radical-scavenging and pancreatic lipase-inhibitory activities. The most abundant tentatively identified peaks in the GC-MS total ion chromatogram were β-sitosterol (26.35%), stigmasterol (14.57%), linoleic acid (10.39%), and oleic acid (5.82%). Minimum inhibitory concentrations (MICs) were 15.62 µg/mL for Listeria monocytogenes, Salmonella typhi, and Candida albicans, and 62.5 µg/mL for methicillin-resistant Staphylococcus aureus. The corresponding inhibition zones were 28 ± 0.7, 21 ± 0.3, 31 ± 0.1, and 15 ± 0.5 mm, respectively. At 75% of the minimum bactericidal/fungicidal concentration, the oil inhibited biofilm formation by 94.95% for both L. monocytogenes and C. albicans. DPPH and ABTS radical-scavenging IC₅₀ values were 17.04 ± 0.25 and 13.17 ± 0.40 µg/mL, respectively, while the pancreatic lipase IC₅₀ was 26.29 ± 0.08 µg/mL. Time-kill assays demonstrated ≥3 log₁₀ CFU/mL reductions, with no recoverable colonies for any of the five tested pathogens within 3 h. Docking predicted stronger stigmasterol binding to L. monocytogenes Sortase A (−6.60 kcal/mol) than to C. albicans lanosterol 14α-demethylase (CYP51; −3.41 kcal/mol), involving hydrogen bonds with Met63 and Arg381, respectively, and hydrophobic contacts. These findings identify a phytosterol-enriched GC-MS-detectable fraction of Delonix regia seed oil with in vitro activities warranting further mechanistic and in vivo investigation.

This study explored the impact of ethanol as a co-solvent in supercritical fluid extraction on the recovery of bioactive compounds from carrot seeds and assessed the resulting extracts for antimicrobial, α-amylase and α-glucosidase, and butyrylcholinesterase inhibitory potential. Ethanol supplementation significantly improved extraction performance, with the yield increasing from 110 mg in the absence of ethanol to 134 mg at 5% ethanol, followed by a slight decrease to 132 mg at 10%. High-performance liquid chromatography (HPLC) revealed pronounced phytochemical enrichment at 5% ethanol, particularly for chlorogenic acid (1541.24 µg/g), gallic acid (1279.27 µg/g), and hesperetin (1513.68 µg/g), indicating enhanced recovery of phenolic and flavonoid constituents. The 5% ethanol extract demonstrated superior antimicrobial activity, producing inhibition zones of 19 mm against Enterococcus faecalis, 26 mm against Klebsiella pneumoniae, 25 mm against Staphylococcus aureus, and 29 mm against Candida albicans. Values of both minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were markedly reduced, while antibiofilm activity reached 93.11% for E. faecalis and 91.00% for K. pneumoniae. The extract also exhibited potent inhibitory effects with IC₅₀ values of 7.74 and 13.37 µg/mL, against α-amylase and α-glucosidase, correspondingly, as well as strong butyrylcholinesterase inhibition (IC₅₀ = 2.51 µg/mL), highlighting promising α-amylase/α-glucosidase and butyrylcholinesterase inhibitory potential. Molecular docking further supported these findings, showing that chlorogenic acid bound more strongly than vanillin to OmpK36, lysosomal acid-α-glucosidase, and butyrylcholinesterase, with docking scores ranging from −6.1 to −6.9 kcal/mol. These findings identify ethanol-modified supercritical fluid extraction as a sustainable and effective green strategy for improving the recovery of carrot seed bioactives and enhancing their multifunctional in vitro biological properties. Notably, this study provides the first comprehensive evidence that 5% ethanol modification selectively enriches key phenolic constituents, including chlorogenic acid, gallic acid, and hesperetin, in carrot seed extracts, with corresponding enhancement of α-amylase, α-glucosidase, and butyrylcholinesterase inhibitory activities.

Background/Objectives: Delonix regia seeds contain phytochemicals with therapeutic potential, but their activity against Helicobacter pylori and biomedical properties remain incompletely characterized. This study aimed to characterize the phenolic and amino acid composition of D. regia seed extract (DRSE) and evaluate its anticancer, wound-healing, anti-inflammatory, anticoagulant, antibacterial, antibiofilm and urease-targeted docking activities. Methods: DRSE was analyzed by high-performance liquid chromatography (HPLC) and amino acid analysis. Biological effects were assessed using MTT cytotoxicity, scratch wound-healing, bovine serum albumin (BSA) denaturation, prothrombin time (PT) and partial thromboplastin time (PTT), antibacterial and crystal violet antibiofilm assays. Gallic acid and vanillin were docked against H. pylori urease (PDB ID: 1E9Y). Results: Gallic acid was the predominant phenolic compound (1417.90 µg/g), while aspartic and glutamic acids dominated the amino acid fraction. DRSE showed preferential cytotoxicity toward A431 carcinoma cells (IC₅₀ = 108.97 ± 0.68 µg/mL) compared with HFB4 fibroblasts (IC₅₀ = 318.56 ± 2.06 µg/mL), yielding a selectivity index of 2.92. Scratch closure was comparable to the control (81.71% versus 80.85%), although the migration rate increased to 16.12 µm. DRSE inhibited protein denaturation by 91.20% (IC₅₀ = 6.39 ± 0.19 µg/mL) and prolonged PT and PTT to 27.37 and 90.50 s, respectively. It inhibited H. pylori with minimum inhibitory and bactericidal concentrations of 31.25 µg/mL and suppressed biofilm formation by 95.39%. Gallic acid and vanillin showed comparable urease docking scores of −4.72 and −4.71 kcal/mol. Conclusions: DRSE showed selective anticancer, anti-H. pylori, antibiofilm, anti-inflammatory, anticoagulant, and moderate pro-migratory activities. Mechanistic, safety and in vivo studies are warranted.