# Ozone-Assisted Green Upgrading of Lactuca sativa Oil: Characterization and Bioactivity for Clean-Label Functional Applications

## TL;DR

Ozonation improves the chemical and biological properties of lettuce oil, making it a promising functional food ingredient with biomedical potential.

## Contribution

This study demonstrates that ozonation enhances the bioactivity of lettuce oil, offering a green method for functional food development.

## Key findings

- Ozonation increases the number and diversity of chemical constituents in lettuce oil.
- Ozonated lettuce oil shows enhanced anti-Helicobacter pylori and antioxidant activities.
- Molecular docking supports the interaction of ozonation-derived compounds with human and bacterial enzymes.

## Abstract

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.