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Room temperature ionic liquids (IL) are a class of ionic compounds consisting of an organic cation and either an organic or inorganic anion, whose melting temperature is below 100 °C, making them liquid at or near room temperature. Recently, the studies on the toxicity of IL toward living organisms has stimulated several chemical–physical investigation on the interaction between IL/water mixtures and basic biological systems, such as saccharides, membranes, peptides/proteins and nucleic acids. For instance, it has been found that certain IL exhibit several positive effects on proteins, including an increased thermal stability, the suppression of aggregation and an enhanced refolding ability. Other examples concern the observed unusual stability of deoxyribonucleic DNA stored at room temperature in IL/water solutions that has stimulated growing interest in using IL/water solutions as alternative solvents for DNA preservation and stabilization. Despite the increasing number of experimental and theoretical investigations on the chemical physics properties of systems made of IL and biomolecules, the deep knowledge on the basic principles able to organize and rationalize the vast variety of properties and phenomena displayed by these systems is still lacking. Synchrotron-based UV Resonance Raman spectroscopy (SR-based UVRR) is a powerful technique for revealing new insights on the nature of interactions between aqueous ionic liquids and bio-molecules on the most microscopic scales of their structure and dynamics. In this presentation, we will discuss selected case studies concerning the solvation properties of IL/water toward small peptides and DNA with particular emphasis on the role of water in altering the behavior of IL in case of biomolecules solvated in IL. Our results highlight the opportunity to use spectroscopic approaches for predicting properties of relatively extended families of IL/biomolecules combinations.