This work explores the heterogeneous reaction between HUmic-LIke Substances (so-called HULIS) and ozone. Genuine atmospheric HULIS were extracted from aerosol samples collected in Chamonix (France) in winter and used in coated flow tube experiments to evaluate heterogeneous uptake of O₃ on such mixtures. The uptake coefficient (γ) was investigated as a function of pH (from 2.5 to 10), O₃ concentration (from 8 to 33 × 10¹¹ molecules cm⁻³), relative humidity (20 to 65%) and photon flux (from 0 to 1.66 × 10¹⁵ photons cm⁻² s⁻¹). Reactive uptake was found to increase in the irradiated experiment with pH, humidity and photon flux. The extract was characterized before and after exposure to O₃ and/or UV light in the attempt to elucidate the effect of the photochemical aging. Carbon content measurements, UV-vis spectroscopy and functional groups analysis revealed a decrease of the UV absorbance as well as of the carbon mass content, while the functionalization rate (COOH and C═O) and therefore the polarity increased during the simulated photochemical exposure.