Ultrasound-assisted extraction of bioactive pigments from Spirulina platensis in natural deep eutectic solvents

Abstract

Spirulina platensis biopigments have been documented as a potential source of nutritional, physiological, and pharmacological purposes due to the presence of bioactive pigments, total phenolic content (TPC) and the consequent antioxidant activity that these compounds present. Bioextracts market has increased in the last decades and is a key option for replacing fossil-derived products and promote the transition for a bio-based economy. To take advantage of these compounds more effectively, optimized extraction processes must be researched and used in biomass sources. The present study focused on optimizing the ultrasound-assisted extraction (UAE) using response surface methodology. Three factor and three level Box–Behnken design was used to optimize the extraction of bioactive pigments, and to investigate the effects of three independent variables, $x$₁: extraction temperature (40–60 °C), $x$₂: extraction cycle time (20–40 min), and $x$₃: solvent-to-biomass ratio (50–70 mL/mg) on total pigment yield, antioxidant assay, and TPC (dependent variables). A second-order polynomial model was used for predicting the responses. Statistically, the model was validated using an analysis of variance. Results revealed that ultrasound-assisted temperature, time, and solvent-to-biomass ratio had a significant (p < 0.05) influence on the total pigment yield, while temperature and solvent-to-biomass ratio had a significant influence in the antioxidant activity, and temperature significantly influenced the total pigment yield. For total pigment yield, antioxidant activity, and total phenol content, the ${R}^{2}$ values of the models generated were 0.8627, 0.8460, and 0.9003, respectively, indicating that the models developed based on second-order polynomials were satisfactorily accurate for analyzing interactions between parameters. Desirability functions showed that the optimal extraction parameters were temperature: 60 °C, extraction cycle time: 20 min; and a solvent-to-biomass ratio of 70 mL/mg. Under optimal conditions, experimental values for total pigment yield, total phenol content expressed as gallic acid equivalent (GAE), and antioxidant activity expressed as Trolox equivalent (TRE) were: 165.19 ± 1.01 mg/g Dry Matter (DM), 36.50 ± 0.98 mg GAE/g DM, and 37.98 ± 0.58 mg TRE/g DM, respectively. The experimental values showed a good agreement with the predicted values with residual standard low 1% under optimum conditions. This optimized ultrasound-assisted method in natural eutectic solvents is effective and scalable to a green extraction of the bioactive pigments from Spirulina platensis with potential application to food, pharmaceutical, functional materials, and packaging.

Keywords

Green chemistry / Green extraction / Microalgae / Optimal / Bioeconomy

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Funding

FCT(UIDB/00195/2020)

Programa Operacional Regional do Centro(CENTRO-01-0247-FEDER-072630)