Supercritical Carbon Dioxide and Microwave-Assisted Extraction of Functional Lipophilic Compounds from Arthrospira platensis.

Arthrospira platensis biomass was used in order to obtain functional lipophilic compounds through green extraction technologies such as supercritical carbon dioxide fluid extraction (SFE) and microwave-assisted extraction (MAE). The temperature (T) factor was evaluated for MAE, while for SFE, pressure (P), temperature (T), and co-solvent (ethanol) (CS) were evaluated. The maximum extraction yield of the obtained oleoresin was (4.07% ± 0.14%) and (4.27% ± 0.10%) for SFE and MAE, respectively. Extracts were characterized by gas chromatography mass spectrometry (GC-MS) and gas chromatography flame ionization detector (GC-FID). The maximum contents of functional lipophilic compounds in the SFE and MAE extracts were: for carotenoids 283 ± 0.10 µg/g and 629 ± 0.13 µg/g, respectively; for tocopherols 5.01 ± 0.05 µg/g and 2.46 ± 0.09 µg/g, respectively; and for fatty acids 34.76 ± 0.08 mg/g and 15.88 ± 0.06 mg/g, respectively. In conclusion, the SFE process at P 450 bar, T 60 °C and CS 53.33% of CO2 produced the highest yield of tocopherols, carotenoids and fatty acids. The MAE process at 400 W and 50 °C gives the best extracts in terms of tocopherols and carotenoids. For yield and fatty acids, the MAE process at 400 W and 70 °C produced the highest values. Both SFE and MAE showed to be suitable green extraction technologies for obtaining functional lipophilic compounds from Arthrospira platensis.

Rapid Lipid Content Screening in Neochloris oleoabundans Utilizing Carbon-Based Dielectrophoresis.

In this study, we carried out a heterogeneous cytoplasmic lipid content screening of Neochloris oleoabundans microalgae by dielectrophoresis (DEP), using castellated glassy carbon microelectrodes in a PDMS microchannel. For this purpose, microalgae were cultured in nitrogen-replete (N+) and nitrogen-deplete (N-) suspensions to promote low and high cytoplasmic lipid production in cells, respectively. Experiments were carried out over a wide frequency window (100 kHz-30 MHz) at a fixed amplitude of 7 VPP. The results showed a statistically significant difference between the dielectrophoretic behavior of N+ and N- cells at low frequencies (100-800 kHz), whereas a weak response was observed for mid- and high frequencies (1-30 MHz). Additionally, a finite element analysis using a 3D model was conducted to determine the dielectrophoretic trapping zones across the electrode gaps. These results suggest that low-cost glassy carbon is a reliable material for microalgae classification-between low and high cytoplasmic lipid content-through DEP, providing a fast and straightforward mechanism.

Evaluation and Predictive Modeling of Removal Condition for Bioadsorption of Indigo Blue Dye by Spirulina platensis.

Among the different chemical and physical treatments used to remove the color of the textile effluents, bioremediation offers many benefits to the environment. In this study, we determined the potential of Spirulina platensis (S. platensis) for decolorizing indigo blue dye under different incubation conditions. The microalgae were incubated at different pH (from 4 to 10) to calibrate for the optimal discoloration condition; a pH of 4 was found to be optimal. The biomass concentration in all experiments was 1 g/L, which was able to decolorize the indigo blue dye by day 3. These results showed that S. platensis is capable of removing indigo blue dye at low biomass. However, this was dependent on the treatment conditions, where temperature played the most crucial role. Two theoretical adsorption models, namely (1) a first-order model equation and (2) a second-order rate equation, were compared with observed adsorption vs. time curves for different initial concentrations (from 25 to 100 mg/L). The comparison between models showed similar accuracy and agreement with the experimental values. The observed adsorption isotherms for three temperatures (30, 40, and 50 °C) were plotted, showing fairly linear behavior in the measured range. The adsorption equilibrium isotherms were estimated, providing an initial description of the dye removal capacity of S. platensis.

How to use Nile Red, a selective fluorescent stain for microalgal neutral lipids.

The use of Nile Red for rapid monitoring of the neutral lipid content in microalgae has gained interest over the last decade, since neutral lipids are feedstock for renewable transportation fuel. In this review, we discuss the main considerations needed to make an NR protocol reliable for staining neutral lipids in microalgae. Cell wall permeability must be enhanced by using stain carriers: DMSO (5% v/v to 25% v/v), glycerol (0.1 to 0.125mg/mL), or EDTA (3.0 to 3.8mg/mL). Temperatures between 30 and 40°C facilitate the diffusion of NR through the cell wall without incurring excess quenching. Good NR-lipid interaction requires using a low NR/cell ratio; the NR concentration must be between 0.25µg/mL and 2.0µg/mL, and the cell concentration >5×10(4)cells/mL. In order to have the maximum and stable NR fluorescence, it is necessary to scan the excitation/emission wavelengths for up to a 40-min of incubation time. We outline a five-step method to customize the Nile Red protocol to a specific strain: 1) Evaluate the strain's suitability by checking for the presence of neutral lipid, 2) Select of the best excitation/emission wavelength, 3) Optimization of incubation time, stain carrier, dye concentration, and temperature, 4) Prepare single-strain algal cultures with different lipid contents to calibrate NR fluorescence with neutral-lipid content, and 5) Correlate NR fluorescence intensity to neutral lipid content for the same strain. Once the protocol is customized, the NR method allows for rapid and reliable monitoring of neutral lipid content of a microalgae strain.