Anaerobic digestion (AD) is a promising waste management strategy that reduces landfilling while generating biogas. Anaerobic co-digestion involves mixing two or more substrates to enhance the nutrient balance required for microorganism growth and thus improve the degradation. Monitoring AD is crucial for comprehending the biological process, optimizing process stability, and achieving efficient biogas production. In this work, we have used three dimensional excitation emission fluorescence spectroscopy and mass spectrometry metabolomics, two complementary techniques, to monitor the anaerobic co-digestion (AcoD) of cellulose, ash wood or oak wood with food waste. The two approaches were compared together and to the biogas production records. Results of this experiment demonstrated the complementarity of both analytical techniques with the measurement of the biogas production since 3D fluorescence spectroscopy and MS metabolomics revealed the earlier molecular changes occurring in the bioreactors, mainly associated with the hydrolysis step, whereas the biogas production data reflected the biological activity in the last step of the digestion. Moreover, in all cases, the three data sets effectively delineated the differences among the substrates. While the two wood substrates were poorly degradable as they were richer in aromatic compounds, cellulose was highly degradable and was characterized by the production of several glycolipids. Then, the three tested AcoDs resulted in a similar 3D EEM fluorescence and metabolomics profiles, close to the one observed for the AD of food waste alone, indicating that the incorporation of the food waste drove the molecular degradation events in the AcoDs. Substrate-specific differences were appreciated from the biogas production data. The overall results of this research are expected to provide insight into the design of guidelines for monitoring AcoD.