Development of a sustainable method for the extraction of lycopene and β-carotene from food wastes

The main objective of the present work is the study of a profitable process not only in the extraction and selective separation of lycopene and β-carotene, two compounds present in tomato, but also in its potential application to food industry wastes. This is one of the industries that produce larger amounts of wastes, which are rich in high value biomolecules with great economic interest. However, the conventional methods used to extract this kind of compounds are expensive which limits their application at large scale. Lycopene and βcarotene are carotenoids with high commercial value, known for their antioxidant activity and benefits to human health. Their biggest source is tomato, one of the world’s most consumed fruits, reason for which large quantities of waste is produced. This work focuses on the study of diverse solvents with a high potential to extract carotenoids from tomato, as well as the search for more environmentally benign solvents than those currently used to extract lycopene and β-carotene from biomass. Additionally, special attention was paid to the creation of a continuous process that would allow the fractionation of the compounds for further purification. Thus, the present work started with the extraction of both carotenoids using a wide range of solvents, namely, organic solvents, conventional salts, ionic liquids, polymers and surfactants. In this stage, each solvent was evaluated in what regards their capacity of extraction as well as their penetration ability in biomass. The results collected showed that an adequate selection of the solvents may lead to the complete extraction of both carotenoids in one single step, particularly acetone and tetrahydrofuran were the most effective ones. However, the general low penetration capacity of salts, ionic liquids, polymers and surfactants makes these solvents ineffective in the solid-liquid extraction process. As the organic solvents showed the highest capacity to extract lycopene and βcarotene, in particular tetrahydrofuran and acetone, the latter solvent used in the development process of fractionation, using to this by strategic use of solvents. This step was only successfully developed through the manipulation of the solubility of each compound in ethanol and n-hexane. The results confirmed the possibility of fractionating the target compounds using the correct addition order of the solvents. Approximately, 39 % of the β-carotene was dissolved in ethanol and about 64 % of lycopene was dissolved in n-hexane, thus indicating their separation for two different solvents which shows the selective character of the developed process without any prior stage optimization. This study revealed that the use of organic solvents leads to selective extraction of lycopene and β-carotene, allowing diminishing the numerous stages involved in conventional methods. At the end, it was possible to idealize a sustainable and of high industrial relevance integrated process, nevertheless existing the need for additional optimization studies in the future.

Extraction and purification of immunoglobulin G with aqueous biphasic systems

The immune system is able to produce antibodies, which have the capacity to recognize and to bind to foreign molecules or pathogenic organisms. Currently, there are a diversity of diseases that can be treated with antibodies, like immunoglobulins G (IgG). Thereby, the development of cost-efficient processes for their extraction and purification is an area of main interest in biotechnology. Aqueous biphasic systems (ABS) have been investigated for this purpose, once they allow the reduction of costs and the number of steps involved in the process, when compared with conventional methods. Nevertheless, typical ABS have not showed to be selective, resulting in low purification factors and yields. In this context, the addition of ionic liquids (ILs) as adjuvants can be a viable and potential alternative to tailor the selectivity of these systems. In this work, ABS composed of polyethylene glycol (PEG) of different molecular weight, and a biodegradable salt (potassium citrate) using ILs as adjuvants (5 wt%), were studied for the extraction and purification of IgG from a rabbit source. Initially, it was tested the extraction time, the effect on the molecular weight of PEG in a buffer solution of K3C6H5O7/C6H8O7 at pH≈7, and the effect of pH (59) on the yield (YIgG) and extraction efficiency (EEIgG%) of IgG. The best results regarding EEIgG% were achieved with a centrifugation step at 1000 rpm, during 10 min, in order to promote the separation of phases followed by 120 min of equilibrium. This procedure was then applied to the remaining experiments. The results obtained in the study of PEGs with different molecular weights, revealed a high affinity of IgG for the PEG-rich phase, and particularly for PEGs of lower molecular weight (EEIgG% of 96 % with PEG 400). On the other hand, the variation of pH in the buffer solution did not show a significant effect on the EEIgG%. Finally, it was evaluated the influence of the addition of different ILs (5% wt) on the IgG extraction in ABS composed of PEG 400 at pH≈7. In these studies, it was possible to obtain EEIgG% of 100% with the ILs composed of the anions [TOS]-, [CH3CO2]-and Cl-, although the obtained YIgG% were lower than 40%. On the other hand, the ILs composed of the anions Br-, as well as of the cation [C10mim]+, although not leading to EEIgG% of 100%, provide an increase in the YIgG%. ABS composed of PEG, a biodegradable organic salt and ILs as adjuvants, revealed to be an alternative and promising method to purify IgG. However, additional studies are still required in order to reduce the loss of IgG.