Potential of Choline Chloride – Based Natural Deep Eutectic Solvents (NaDES) in the Extraction of Microalgal Metabolites

In a typical chemical process, the solvents are widely used for the dissolution of the reagents, to favor the kinetics and the thermodynamics of a chemical reaction, for the extraction of products, for the separation of mixtures. However most of the currently used organic solvents are characterized by different properties harmful to human health and the environment. Among the principles of Green Chemistry are that solvents should be innocuous to Man and to the Environment (safer solvents) and that the substances used in a chemical process should be chosen to minimize the potential for chemical accidents (intrinsically safe processes). Biorefining, the biomass Era counterpart of oil refining is most likely going to be extraction-based, and thus heavily solvent-dependent, much as the Oil Era was based on distillation and hence heat-dependent. Ionic Liquids (ILS) and eutectic mixtures exploited as solvents (DES) are two major classes of solvents that are making their way in Green Chemistry and, in particular, in biomass processing research. NaDES ('Natural Deep Eutectic Solvents'), i.e. mixtures formed by natural primary metabolites present in all organisms, such as sugars, polyols, amino acids, organic acids, derivatives of choline, form intermolecular hydrogen bonds and, when mixed in a certain ratio, change their state from solid to liquid forming a eutectic system. The most interesting NaDESs are those in which water is one of a ternary system since the degree of dilution with water modifies such physical properties of the NaDES as the density, the viscosity, and the polarity. By modulating the water content the solvation power can be adjusted to specific needs. In this work, the PCH (1,2-propanediol, choline chloride, water 1:1:1) NaDES was used to treat microalgal biomass and carry out the extraction of cellular components, such as lipids, proteins, carbohydrates and photosynthetic pigments (chlorophylls and carotenoids) from the biomass itself. Three sets of experiments were carried out based on different contact time between biomass and PCH: 24 and 72 hours, with and without pre-treatment with ultrasound. Biomass was shaken together with the PCH solvent in the presence of glass beads to promote the extraction efficiency. The analysis of the extract composition was carried out spectrophotometrically for pigments (chlorophylls and carotenoids), with biochemical assays for proteins and carbohydrates and gravimetrically for the determination of lipids. The results showed the ability of PCH, coupled with the mechanical destruction of cell walls, to solubilize a wide range of polar biomolecules at room temperature.