Alternative methods for RuBisCO extraction from sugar beet waste: A comparative approach of ultrasound and high voltage electrical discharge.

Ultrasound (US) and high voltage electric discharge (HVED) with water as a green solvent represent promising novel non-thermal techniques for protein extraction from sugar beet (Beta vulgaris subsp. vulgaris var. altissima) leaves. Compared to HVED, US proved to be a better alternative method for total soluble protein extraction with the aim of obtaining high yield of ribulose-1,5-bisphosphate carboxylase-oxygenase enzyme (RuBisCO). Regardless of the solvent temperature, the highest protein yields were observed at 100% amplitude and 9 min treatment time (84.60 ± 3.98 mg/gd.m. with cold and 96.75 ± 4.30 mg/gd.m. with room temperature deionized water). US treatments at 75% amplitude and 9 min treatment time showed the highest abundance of RuBisCO obtained by immunoblotting assay. The highest protein yields recorded among HVED-treated samples were observed at a voltage of 20 kV and a treatment time of 3 min, disregarding the used gas (33.33 ± 1.06 mg/gd.m. with argon and 34.89 ± 1.59 mg/gd.m. with nitrogen as injected gas), while the highest abundance of the RuBisCO among HVED-treated samples was noticed at 25 kV voltage and 3 min treatment time. By optimizing the US and HVED parameters, it is possible to affect the solubility and improve the isolation of RuBisCO, which could then be purified and implemented into new or already existing functional products.

Encapsulation of Rosemary Extracts using High Voltage Electrical Discharge in Calcium Alginate/Zein/Hydroxypropyl Methylcellulose Microparticles.

The increased demand for functional food with added health benefits is directing industrial procedures toward more sustainable production of naturally added bioactive compounds. The objective of this research was to investigate the potential of bioactive compounds from rosemary extract obtained using high-voltage electrical discharge as a green extraction method, for microencapsulation as a protective method for future application in functional food. Four types of microparticles were made via the ionic gelation method using alginate (Alg), zein (Z), and hydroxypropyl methylcellulose (HPMC) biopolymers and were analyzed considering the physicochemical properties. The diameter of dry microparticles ranged from 651.29 to 1087.37 µm. The shape and morphology analysis of microparticles showed that the obtained microparticles were quite spherical with a granular surface. The high encapsulation efficiency was obtained with a loading capacity of polyphenols up to 11.31 ± 1.47 mg GAE/g (Alg/Z microparticles). The microencapsulation method showed protective effects for rosemary polyphenols against pH changes during digestion. Specifically, the addition of both zein and HPMC to calcium-alginate resulted in microparticles with a prolonged release for better availability of polyphenols in the intestine. This research background indicates that the release of rosemary extract is highly dependent on the initial biopolymer composition with high potential for further functional food applications.

High Voltage Electrical Discharges as an Alternative Extraction Process of Phenolic and Volatile Compounds from Wild Thyme (Thymus serpyllum L.): In Silico and Experimental Approaches for Solubility Assessment.

The objective of this study was to evaluate the potential of green solvents for extractions of bioactive compounds (BACs) and essential oils from wild thyme (Thymus serpyllum L.) using theoretical and experimental procedures. Theoretical prediction was assessed by Hansen solubility parameters (HSPs) and conductor-like screening model for realistic solvents (COSMO-RS), to predict the most suitable solvents for extraction of BACs. An experimental procedure was performed by nonthermal technology high voltage electrical discharge (HVED) and it was compared with modified conventional extraction (CE). Obtained extracts were analyzed for chemical and physical changes during the treatment. Theoretical results for solution of BACs in ethanol and water, as green solvents, were confirmed by experimental results, while more accurate data was given by COSMO-RS assessment than HSPs. Results confirmed high potential of HVED for extraction of BACs and volatile compounds from wild thyme, in average, 2.03 times higher yield of extraction in terms of total phenolic content was found compared to CE. The main phenolic compound found in wild thyme extracts was rosmarinic acid, while the predominant volatile compound was carvacrol. Obtained extracts are considered safe and high-quality source reach in BACs that could be further used in functional food production.

The Potential of High Voltage Discharges for Green Solvent Extraction of Bioactive Compounds and Aromas from Rosemary (Rosmarinus officinalis L.)-Computational Simulation and Experimental Methods.

Rosemary (Rosmarinus officinalis L.) is a Mediterranean medicinal and aromatic plant widely used due to valuable bioactive compounds (BACs) and aromas. The aim of the study was to evaluate the extraction of intracellular compounds from rosemary combining experimental procedure by means of high voltage electrical discharge (HVED), with a theoretical approach using two computational simulation methods: conductor-like screening model for real solvents and Hansen solubility parameters. The optimal HVED parameters were as follows: frequency 100 Hz, pulse width 400 ns, gap between electrodes 15 mm, liquid to solid ratio 50 mL/g, voltage 15 and 20 kV for argon, and 20 and 25 kV for nitrogen gas. Green solvents were used, water and ethanol (25% and 50%). The comparison was done with modified conventional extraction (CE) extracted by magnetic stirring and physicochemical analyses of obtained extracts were done. Results showed that HVED extracts in average 2.13-times higher total phenol content compared to CE. Furthermore, nitrogen, longer treatment time and higher voltage enhanced higher yields in HVED extraction. HVED was confirmed to have a high potential for extraction of BACs from rosemary. The computational stimulation methods were confirmed by experimental study, ethanol had higher potential of solubility of BACs and aromas from rosemary compared to water.

Phenolic and Antioxidant Analysis of Olive Leaves Extracts (Olea europaea L.) Obtained by High Voltage Electrical Discharges (HVED).

BACKGROUND: The aim of this study was to evaluate high voltage electrical discharges (HVED) as a green technology, in order to establish the effectiveness of phenolic extraction from olive leaves against conventional extraction (CE). HVED parameters included different green solvents (water, ethanol), treatment times (3 and 9 min), gases (nitrogen, argon), and voltages (15, 20, 25 kV). METHODS: Phenolic compounds were characterized by ultra-performance liquid chromatography-tandem mass spectrometer (UPLC-MS/MS), while antioxidant potency (total phenolic content and antioxidant capacity) were monitored spectrophotometrically. Data for Near infrared spectroscopy (NIR) spectroscopy, colorimetry, zeta potential, particle size, and conductivity were also reported. RESULTS: The highest yield of phenolic compounds was obtained for the sample treated with argon/9 min/20 kV/50% (3.2 times higher as compared to CE). Obtained results suggested the usage of HVED technology in simultaneous extraction and nanoformulation, and production of stable emulsion systems. Antioxidant capacity (AOC) of obtained extracts showed no significant difference upon the HVED treatment. CONCLUSIONS: Ethanol with HVED destroys the linkage between phenolic compounds and components of the plant material to which they are bound. All extracts were compliant with legal requirements regarding content of contaminants, pesticide residues and toxic metals. In conclusion, HVED presents an excellent potential for phenolic compounds extraction for further use in functional food manufacturing.

Nutritional Status and Nutrition Quality in Patients with Non-Alcoholic Fatty Liver Disease.

Non-alcoholic fatty liver disease (NAFLD) is becoming a major health burden with increasing prevalence worldwide due to its close association with the epidemic of obesity. Currently there is no standardized pharmacological treatment, and the only proven effective therapeutic strategy is lifestyle modification, therefore it is important to determine the potential dietary targets for the prevention and treatment of NAFLD. We assessed nutritional status in 30 patients diagnosed with NAFLD using anthropometric parameters, hand grip strength, and lifestyle and dietetic parameters (physical activity, NRS2002 form and three-day food diary). The mean body mass index was 29.62±4.61 kg/m2, yielding 86.67% of obese or overweight patients. Physical activity results indicat-ed poorly active subjects. Excessive energy intake was recorded in 27.78% of patients. The mean in-take of macronutrients was as follows: 15.5% of proteins, 42.3% of carbohydrates and 42.2% of fat, with -deficient micronutrient intake of calcium, magnesium, iron, zinc, and vitamins A, B1 and B2. The -results showed that the quality of nutrition in study subjects was not accordant to current rec-ommendations and that they consumed a high proportion of fat, especially saturated fatty acids, along with low micronutrient intake. The results obtained might point to the importance of unbalanced diet as a contributing factor in NAFLD development.