ABSTRACT
There has been growing interest in microalgal biomolecules for health and cosmetics, as well as in the use of microalgae as aquaculture feed due to the need to replace fishmeal and fish oil with sustainable yet equally nutritious alternatives. Aim of this study is to evaluate the potential of five marine microalgal species, namely Chlorella minutissima, Dunaliella salina, Isochrysis galbana, Nannochloropsis oculata and Tisochrysis lutea, for the co-production of antioxidants and aquaculture feed. Batch cultivation was performed under saturating light intensity and continuous aeration. Freeze-dried biomass was extracted sequentially with water and methanol and evaluated for phenolic content and antioxidant activity, as well as proximate composition and fatty acid profile. Methanolic extracts of C. minutissima presented the highest phenolic content, measured with the Folin-Ciocalteu assay, and antioxidant activity. However, HPLC and LC-MS showed the presence of non-pigment compounds only in T. lutea. Total phenolic content and antioxidant activity were correlated to chlorophyll content. N. oculata and T. lutea were rich in eicosapentaenoic acid and docosahexaenoic acid, respectively, as well as in protein. In conclusion, N. oculata and T. lutea are suitable candidates for further optimization, while the data presented suggest that pigment effects on the Folin-Ciocalteu method require reconsideration.
ABSTRACT
The aim of this study was the development of an efficient "green" extraction method of Nannochloropsis oculata to produce antioxidant extracts and nutritious residual biomass. Twenty-one extraction methods were evaluated by measuring the reactivity with the Folin-Ciocalteu reagent: ultrasonication or maceration at different temperatures with different organic solvents, extraction at different pH values, enzyme-assisted extraction, encapsulation with ß-cyclodextrin, and the use of natural deep eutectic solvents. Ultrasound-assisted extraction with ethanol or betaine: 1,2-propanediol in a molar ratio of 2:5 (BP) had optimal extractive capacity. Both extracts were evaluated with antioxidant assays and the ethanol extract exhibited significantly higher (at least twofold) values. The determination of carotenoids by LC-MS and HPLC-DAD revealed the dominance of violaxanthin and antheraxanthin and their fourfold higher concentrations in the ethanol extract. The 1H-NMR characterization of the ethanol extract confirmed the results of the colorimetric and chromatographic assays. The microalgal biomass was characterized before and after the extraction in terms of humidity, ash, carbohydrates, proteins, chlorophyll-a, carotenoids, and lipids; the identity and content of the latter were determined with gas chromatography. BP caused a smaller depletion of the lipids from the biomass compared to ethanol, but proteins, carbohydrates, and ash were at a higher content in the biomass obtained after ethanol extraction, whereas the biomass was dry and easy to handle. Although further optimization may take place for the scale-up of those procedures, our study paves the way for a green strategy for the valorization of microalgae in cosmetics without generating waste, since the remaining biomass can be used for aquafeed.
ABSTRACT
Salvia fruticosa and S. pomifera subsp. calycina are native to Eastern Mediterranean and S. pomifera subsp. pomifera is endemic to Greece. The primary aim of this study was to develop an analytical methodology for metabolomic profiling and to study their efficacy in combating glycation, the major biochemical complication of diabetes. After sequential ultrasound-assisted extraction of 2 g of leaves with petroleum ether and 70% methanol, the volatile metabolites in the petroleum ether extracts were studied with GC-MS (Gas Chromatography-Mass Spectrometry), whereas the polar metabolites in the hydroalcoholic extracts were determined and quantified by UHPLC-DAD-ESI-MS (Ultra-High Performance Liquid Chromatography-Diode Array Detector-Mass Spectrometry). This methodology was applied to five populations belonging to the three native taxa. 1,8-Cineole was the predominant volatile (34.8-39.0%) in S. fruticosa, while S. pomifera had a greater content of α-thujone (19.7-41.0%) and ß-thujone (6.0-39.1%). Principal Component Analysis (PCA) analysis of the volatiles could discriminate the different taxa. UHPLC-DAD-ESI-MS demonstrated the presence of 50 compounds, twenty of which were quantified. PCA revealed that not only the taxa but also the populations of S. pomifera subsp. pomifera could be differentiated. All Salvia samples inhibited advanced glycation end-product formation in a bovine serum albumin/2-deoxyribose assay; rosmarinic and carnosic acid shared this activity. This study demonstrates the antiglycation activity of S. fruticosa and S. pomifera extracts for the first time and presents a miniaturized methodology for their metabolomic profiling, which could aid chemotaxonomic studies and serve as a tool for their authentication and quality control.
