Recent Advances in Supercritical CO2 Extraction of Pigments, Lipids and Bioactive Compounds from Microalgae.

Supercritical CO2 extraction is a green method that combines economic and environmental benefits. Microalgae, on the other hand, is a biomass in abundance, capable of providing a vast variety of valuable compounds, finding applications in the food industry, cosmetics, pharmaceuticals and biofuels. An extensive study on the existing literature concerning supercritical fluid extraction (SFE) of microalgae has been carried out focusing on carotenoids, chlorophylls, lipids and fatty acids recovery, as well as the bioactivity of the extracts. Moreover, kinetic models used to describe SFE process and experimental design are included. Finally, biomass pretreatment processes applied prior to SFE are mentioned, and other extraction methods used as benchmarks are also presented.

Extracts from Chlorella vulgaris Protect Mesenchymal Stromal Cells from Oxidative Stress Induced by Hydrogen Peroxide.

Microalgae as unicellular eukaryotic organisms demonstrate several advantages for biotechnological and biological applications. Natural derived microalgae products demand has increased in food, cosmetic and nutraceutical applications lately. The natural antioxidants have been used for attenuation of mitochondrial cell damage caused by oxidative stress. This study evaluates the in vitro protective effect of Chlorella vulgaris bioactive extracts against oxidative stress in human mesenchymal stromal/stem cells (MSCs). The classical solid-liquid and the supercritical extraction, using biomass of commercially available and laboratory cultivated C. vulgaris, are employed. Oxidative stress induced by 300 µM H2O2 reduces cell viability of MSCs. The addition of C. vulgaris extracts, with increased protein content compared to carbohydrates, to H2O2 treated MSCs counteracted the oxidative stress, reducing reactive oxygen species levels without affecting MSC proliferation. The supercritical extraction was the most efficient extraction method for carotenoids resulting in enhanced antioxidant activity. Pre-treatment of MSCs with C. vulgaris extracts mitigates the oxidative damage ensued by H2O2. Initial proteomic analysis of secretome from licensed (TNFα-activated) MSCs treated with algal extracts reveals a signature of differentially regulated proteins that fall into clinically relevant pathways such as inflammatory signaling. The enhanced antioxidative and possibly anti-inflammatory capacity could be explored in the context of future cell therapies.

Extraction of Bioactive Compounds from C. vulgaris Biomass Using Deep Eutectic Solvents.

C. vulgaris microalgae biomass was employed for the extraction of valuable bioactive compounds with deep eutectic-based solvents (DESs). Particularly, the Choline Chloride (ChCl) based DESs, ChCl:1,2 butanediol (1:4), ChCl:ethylene glycol (1:2), and ChCl:glycerol (1:2) mixed with water at 70/30 w/w ratio were used for that purpose. The extracts' total carotenoid (TCC) and phenolic contents (TPC), as well as their antioxidant activity (IC50), were determined within the process of identification of the most efficient solvent. This screening procedure revealed ChCl:1,2 butanediol (1:4)/H2O 70/30 w/w as the most compelling solvent; thus, it was employed thereafter for the extraction process optimization. Three extraction parameters, i.e., solvent-to-biomass ratio, temperature, and time were studied regarding their impact on the extract's TCC, TPC, and IC50. For the experimental design and process optimization, the statistical tool Response Surface Methodology was used. The resulting models' predictive capacity was confirmed experimentally by carrying out two additional extractions under conditions different from the experimental design.

Supercritical CO2 Extraction of High-Added Value Compounds from Chlorella vulgaris: Experimental Design, Modelling and Optimization.

Microalgae are well-known for their high-added value compounds and their recovery is currently of great interest. The aim of this work is the recovery of such components from Chlorella vulgaris through supercritical fluid extraction (SFE) with CO2. The effect of the extraction temperature (40-60 °C), pressure (110-250 bar), and solvent flow rate (20-40 g/min) was tested on yield, the extract's antioxidant activity, and the phenolic, chlorophyll and carotenoid content. Thus, data analysis indicated that the yield was mainly affected by temperature, carotenoids by pressure, while the extract's phenolics and antioxidant activity were affected by the synergy of temperature and pressure. Moreover, SFE's kinetic study was performed and experimental data were correlated using Sovová's mass transfer-based model. SFE optimization (60 °C, 250 bar, 40 g/min) led to 3.37% w/w yield, 44.35 mgextr/mgDPPH antioxidant activity (IC50), 18.29 mgGA/gextr total phenolic content, 35.55, 21.14 and 10.00 mg/gextr total chlorophyll, carotenoid and selected carotenoid content (astaxanthin, lutein and ß-carotene), respectively. A comparison of SFE with conventional aq. ethanol (90% v/v) extraction proved SFE's superiority regarding extraction duration, carotenoids, antioxidant activity and organoleptic characteristics of color and odor despite the lower yield. Finally, cosolvent addition (ethanol 10% w/w) at optimum SFE conditions improved the extract's antioxidant activity (19.46%) as well as yield (101.81%).

Optimization of Microalga Chlorella vulgaris Magnetic Harvesting.

Harvesting of microalgae is a crucial step in microalgae-based mass production of different high value-added products. In the present work, magnetic harvesting of Chlorella vulgaris was investigated using microwave-synthesized naked magnetite (Fe3O4) particles with an average crystallite diameter of 20 nm. Optimization of the most important parameters of the magnetic harvesting process, namely pH, mass ratio (mr) of magnetite particles to biomass (g/g), and agitation speed (rpm) of the C. vulgaris biomass-Fe3O4 particles mixture, was performed using the response surface methodology (RSM) statistical tool. Harvesting efficiencies higher than 99% were obtained for pH 3.0 and mixing speed greater or equal to 350 rpm. Recovery of magnetic particles via detachment was shown to be feasible and the recovery particles could be reused at least five times with high harvesting efficiency. Consequently, the described harvesting approach of C. vulgaris cells leads to an efficient, simple, and quick process, that does not impair the quality of the harvested biomass.

Experimental Design and Optimization of Recovering Bioactive Compounds from Chlorella vulgaris through Conventional Extraction.

Microalgae contain an abundance of valuable bioactive compounds such as chlorophylls, carotenoids, and phenolics and, consequently, present great commercial interest. The aim of this work is the study and optimization of recovering the aforementioned components from the microalgae species Chlorella vulgaris through conventional extraction in a laboratory-scale apparatus using a "green" mixture of ethanol/water 90/10 v/v. The effect of three operational conditions-namely, temperature (30-60 °C), duration (6-24 h) and solvent-to-biomass ratio (20-90 mLsolv/gbiom), was examined regarding the extracts' yield (gravimetrically), antioxidant activity, phenolic, chlorophyll, and carotenoid contents (spectrophotometric assays), as well as concentration in key carotenoids, i.e., astaxanthin, lutein, and ß-carotene (reversed-phase-high-performance liquid chromatography (RP-HPLC)). For this purpose, a face-centered central composite design (FC-CCD) was employed. Data analysis resulted in the optimal extraction conditions of 30 °C, for 24 h with 37 mLsolv/gbiom and validation of the predicted models led to 15.39% w/w yield, 52.58 mgextr/mgDPPH (IC50) antioxidant activity, total phenolic, chlorophyll, and carotenoid content of 18.23, 53.47 and 9.92 mg/gextr, respectively, and the total sum of key carotenoids equal to 4.12 mg/gextr. The experimental data and predicted results were considered comparable, and consequently, the corresponding regression models were sufficiently reliable for prediction.

A Study on the Effect of Macro- and Micro- Nutrients on Nannochloropsis oceanica Growth, Fatty Acid Composition and Magnetic Harvesting Efficiency.

The effect of iron, manganese, phosphorus and nitrogen on growth and lipid synthesis of the microalgae Nannochloropsis oceanica CCMP1779, as well as their impact on the magnetic harvesting efficiency, are examined under their depriving cell culture conditions. Herein, it is demonstrated that nitrogen and manganese depletion primarily reduced cell growth while phosphorus and iron restriction led to higher dry biomass. Subsequently, the role of those nutrients on fatty acids profile was examined. Phosphorus and nitrogen restriction resulted in lower and higher lipid content, respectively. High amounts of polyunsaturated fatty acids like eicosapentaenoic acid are produced under iron and manganese depletion. Phosphorus deprivation favors monounsaturated fatty acids such as C18:1 and C16:1, while nitrogen restriction favors saturated fatty acid production like C14:0, C16:0 and C18:0. Since the presence/absence of macro- and micro-elements may affect the overall electrostatic charges on the outmost microalgae surface, it was also analyzed how these elements affect the magnetic harvesting efficiency. Results showed that phosphorus deprivation led to the best magnetic harvesting efficiency of N. oceanica cells (93%) as compared to other nutrient starvation as well as standard medium.