A novel enzyme-assisted ultrasonic approach for highly efficient extraction of resveratrol from Polygonum cuspidatum

Resveratrol is a promising multi-biofunctional phytochemical, which is abundant in Polygonum cuspidatum. Several methods for resveratrol extraction have been reported, while they often take a long extraction time accompanying with poor extraction yield. In this study, a novel enzyme-assisted ultrasonic approach for highly efficient extraction of resveratrol from P. cuspidatum was developed. According to results, the resveratrol yield significantly increased after glycosidases (Pectinex® or Viscozyme®) were applied in the process of extraction, and better extraction efficacy was found in the Pectinex®-assisted extraction compared to Viscozyme®-assisted extraction. Following, a 5-level-4-factor central composite rotatable design with response surface methodology (RSM) and artificial neural network (ANN) was selected to model and optimize the Pectinex®-assisted ultrasonic extraction. Based on the coefficient of determination (R²) calculated from the design data, ANN model displayed much more accurate in data fitting as compared to RSM model. The optimum conditions for the extraction determined by ANN model were substrate concentration of 5%, acoustic power of 150 W, pH of 5.4, temperature of 55 °C, the ratio of enzyme to substrate of 3950 polygalacturonase units (PGNU)/g of P. cuspidatum, and reaction time of 5 h, which can lead to a significantly high resveratrol yield of 11.88 mg/g.     

Ultrasound-assisted extraction and properties of polysaccharide from Ginkgo biloba leaves

Response surface methodology (RSM) was used to optimize the ultrasound-assisted extraction conditions of Ginkgo biloba leaves polysaccharide (GBLP). The optimum extraction conditions for the ultrasound-assisted extraction of GBLP were obtained as liquid to material ratio of 30 mL/g, ultrasonic power of 340 W, and extraction time of 50 min. Under these conditions, the yield of GBLP was 5.37 %. Two chemically modified polysaccharides, CM-GBLP and Ac-GBLP, were obtained by carboxymethylation and acetylation of GBLP. The physicochemical properties of these three polysaccharides were comparatively studied and their in vitro antioxidant activities were evaluated comprehensively. The results showed that the solubility of the chemically modified polysaccharides was significantly enhanced and the in vitro antioxidant activity was somewhat improved. This suggests that carboxymethylation and acetylation are effective methods to enhance polysaccharide properties, but the results exhibited some uncontrollability. At the same time, GBLP has also shown high potential for research and application.     

Ultrasound-assisted extraction,analysis and antioxidant activity of polysaccharide from the rinds of Garcinia mangostana L.

According to response surface methodology (RSM), the extraction conditions of ultrasound-assisted extraction of polysaccharide from the rinds of Garcinia mangostana L. (GMRP) were optimized and determined. The optimal conditions obtained through optimization were: the liquid to material ratio was 40 mL/g, ultrasonic power was 288 W and extraction time was 65 min. The average extraction rate of GMRP was 14.73%. Ac - GMRP was obtained by acetylation of GMRP, and the antioxidant activities of the two polysaccharides were compared in vitro. The results indicated that the antioxidant capacity of polysaccharide obtained after acetylation was significantly improved compared with that of GMRP. In conclusion, chemical modification of polysaccharide is an effective measure to improve its properties to a certain extent. Meanwhile, it implies that GMRP has great research value and potential.     

Ultrasound-assisted extraction,optimization, isolation,and antioxidant activity analysis of flavonoids from Astragalus membranaceus stems and leaves

Astragalus membranaceus is a medicinal and edible species in China, with a variety of biological activities. This study evaluated the reuse potential of A. membranaceus waste as a source of food antioxidants. Antioxidant and antifungal activities of flavonoids, polysaccharides, and saponins from A. membranaceus stems and leaves were evaluated. Results showed that inhibition rate of flavonoids on six tested fungi reaches 100 % at a concentration of 5 mg/mL, and the antioxidant test demonstrated satisfactory antioxidant activity. On this basis, an extremely economical ultrasonic-assisted extraction of flavonoids from A. membranaceus stems and leaves was developed and optimized via response surface methodology (RSM). Optimized conditions included an extraction time of 35 min, ethanol concentration of 75 %, liquid–solid ratio of 40 mL/g, and extraction temperature of 58 °C, in which the extraction yield of flavonoids was 22.0270 ± 2.5739 mg/g. The total flavonoids were separated and purified using activity-guided isolation technology, and frac. ccd with strong antioxidant activity were analyzed via HPLC-MS/MS. Results showed that main components are isoquercitrin and astragalin. This study can provide a potential innovative application for the development of natural food antioxidants from A. membranaceus waste.     

Artificial neural network model- and response surface methodology-based optimization of Atractylodis Macrocephalae Rhizoma polysaccharide extraction,kinetic modelling and structural characterization

Atractylodis Macrocephalae Rhizoma (AMR) is the dried rhizome of Atractylodes macrocephala Koidz, which is widely used in the development of health products. AMR contains a large number of polysaccharides, but at present there are fewer applications for these polysaccharides. In this study, the effects of different extraction methods on the Atractylodis Macrocephalae Rhizoma polysaccharide (AMRP) yield were investigated, and the conditions for ultrasound-assisted extraction were optimized by response surface methodology (RSM) and three neural network models (BP neural network, GA-BP neural network and ACO-GA-BP neural network). The best conditions were a liquid-to-solid ratio of 17 mL/g, ultrasonic power of 400 W, extraction temperature of 72 °C, and extraction time of 40 min, which yielded 31.31% AMRP. The kinetic equation of AMRP was determined and compared with the results predicted by three neural network models. It was finally determined that the extraction conditions, kinetic processes and kinetic equation predicted by the GA-ACO-BP neural network were optimal. In addition, AMRP was characterized using SEM, FTIR, HPLC, UV, XRD, and NMR, and the structural study revealed that AMRP has a rough exterior and a porous interior; moreover, it contains high levels of glucose (5.07%), arabinose (0.80%), and galactose (0.74%). AMRP has three crystal structures, consisting of two β-type monosaccharides and one α-type monosaccharide. Additionally, the effectiveness of AMRP as an antioxidant was demonstrated in an in vitro experiment.     

Ultrasound-assisted polysaccharide extraction from Cercis chinensis and properites,antioxidant activity of polysaccharide

The purpose of the study was to improve the extraction of polysaccharide from the leaves of Cercis chinensis Bunge using ultrasound, and compare the difference between boiling and ultrasound extraction in terms of polysaccharide content, monosaccharide compounds, and evaluate how the factors affected the bioactivity. The best conditions, according to the single factor experiments and the Box-Bohnken design (BBD), were an intensity of ultrasound of 180 W, duration of extraction of 40 min, proportion of water to material of 15:1 (g/g), and a higher polysaccharide yield of 20.02 ± 0.55 (mg/g) than in boiling extraction (16.09 ± 0.82 mg/g). The antioxidative experiment suggested the polysaccharide by ultrasound exhibited higher DPPH, hydroxyl radical scavenging capacities, and reducing power at 1.2–1.4 mg/mL, which was superior to the boiling polysaccharide. Further analysis showed that the ultrasonic purified polysaccharides like Gla, N-Glu, and GluA contained more total sugar and uronic acids than the boiling method did. This may indicate that the ultrasonic isolation of the polysaccharides increase the antioxidant activity of the polysaccharides.     

Modeling of process intensification of biodiesel production from Aegle Marmelos Correa seed oil using microreactor assisted with ultrasonic mixing

Conventionally, the batch type reactors were used for the production of biodiesel. However, in recent years, the usage of microreactors has started emerging as a significant substitute for biodiesel production due to its higher conversion rate at a short duration. These microreactors have a significantly high surface to volume ratio and high heat-mass transfer rate. The disadvantage of this type of reactors is its low mixing rate of the reagents. This can be overcome with the assistance of ultrasonic mixing. The main objective of this paper is to study the interlaced effect of a continuous flow microreactor and ultrasonic mixing on trans-esterification of Aegle Marmelos Correa seed oil using sodium methoxide catalyst. Results of microreactors with 0.3 mm and 0.8 mm diameter were compared. The effects of process parameters namely, flow rate (2–10 mL/min), reaction temperature (45–65 °C), catalyst amount (0.5–2.5 wt%), oil to methanol molar ratio (1:6–1:18) and ultrasonic mixing time (30–150 s) were studied using response surface methodology (RSM). The biodiesel yield of 98% and 91.8% were obtained for 0.3 mm and 0.8 mm microreactors, respectively. The maximum biodiesel yield observed in 0.3 mm reactor under following optimum conditions: 6.8 mL/min flow rate, 48 °C reaction temperature, 1.3 wt% catalyst, 1:9 oil to methanol molar ratio and 83 s ultrasonic mixing time. The predictive and generalization abilities of RSM and artificial neural network (ANN) models were evaluated and compared. The study showed that ANN and RSM models could predict the yield with an R² value of 0.9955 and 0.9900 respectively. However, the ANN model predicted the yield with the least mean square error value of 0.00001294, which is much lower than RSM.     

Multimode ultrasonic extraction of polysaccharides from maca (Lepidium meyenii): Optimization,purification, and in vitro immunoregulatory activity

This study evaluates the effect of multimodal ultrasound on the extraction efficiency and immunoregulatory activity of polysaccharides from Lepidium meyenii Walp. (LMP). The separation and purification of maca polysaccharides were investigated by the DEAE-52 cellulose column, and the monosaccharide compositions were identified by HPGPC. Their immune activity was analyzed by the secretion of cytokines (TNF-α and IL-6) from RAW 264.7 macrophage. The results showed that the optimal extraction conditions were energy aggregation alternation dual-frequency ultrasound (EADU) with frequency combinations of 20/35, extraction time of 15 min, material/water ratio of 1:10 g/mL, ultrasonic power intensity of 150 W/L, intermittent time ratio of 4 s/3 s, and extraction temperature of 50 ℃. The extraction rates of purified polysaccharides (US3) increased by 44.90%. The LMP extracted by EADU contained arabinose, galactose, and glucose in the molar ratios of 2.9:2.72:5.05. In addition, US3 promoted the release of TNF-α and IL-6 from RAW 264.7 better than RS3 (purified polysaccharides extracted by hot water), which indicated that US3 exerted remarkable immune activity. It could be an excellent functional additive in food or medicine.     

Ultrasound-assisted extraction of phenolic acids,flavonols, and flavan-3-ols from muscadine grape skins and seeds using natural deep eutectic solvents and predictive modelling by artificial neural networking

The objective of this study was to investigate the extraction efficiency of 9 natural deep eutectic solvents (NDES) with the assistance of ultrasound for phenolic acids, flavonols, and flavan-3-ols in muscadine grape (Carlos) skins and seeds in comparison to 75% ethanol. Artificial neural networking (ANN) was applied to optimize NDES water content, ultrasonication time, solid-to-solvent ratio, and extraction temperature to achieve the highest extraction yields for ellagic acid, catechin and epicatechin. A newly formulated NDES (#1) consists of choline chloride: levulinic acid: ethylene glycol 1:1:2 and 20% water extracted the highest amount of ellagic acid in the skin at 22.1 mg/g. This yield was 1.73-fold of that by 75% ethanol. A modified NDES (#3) consisting of choline chloride: proline: malic acid 1:1:1 and 30% water extracted the highest amount of catechin (0.61 mg/g) and epicatechin (0.89 mg/g) in the skin, and 2.77 mg/g and 0.37 mg/g in the seed, respectively. The optimal yield of ellagic acid in the skin using NDES #1 was 25.3 mg/g (observed) and 25.3 mg/g (predicted). The optimal yield of (catechin + epicatechin) in seed using NDES #3 was 9.8 mg/g (observed) and 9.6 mg/g (predicted). This study showed the high extraction efficiency of selected NDES for polyphenols under optimized conditions.     

Kinetic modeling and optimization of maceration and ultrasound-extraction of resinoid from the aerial parts of white lady’s bedstraw (Galium mollugo L.)

In this paper, extraction of resinoid from the aerial parts of white lady’s bedstraw (Galium mollugo L.) using an aqueous ethanol solution (50% by volume) was studied at different temperatures in the absence and the presence of ultrasound. This study indicated that ultrasound-assisted extraction was effective for extracting the resinoid and gave better resinoid yields at lower extraction temperature and in much shorter time than the maceration. A phenomenological model was developed for modeling the kinetics of the extraction process. The model successfully describes the two-step extraction consisting of washing followed by diffusion of extractable substances and shows that ultrasound influences only the first step. The extraction process was optimized using response surface methodology (RMS) and artificial neural network (ANN) models. For the former modeling, the second-order polynomial equation was applied, while the second one was performed by an ANN-GA combination. The high coefficient of determination and the low MRPD between the ANN prediction and the corresponding experimental data proved that modeling the extraction process in the absence and the presence of ultrasound using ANN was more accurate than RSM modeling. The optimum extraction temperature was determined to be 80 and 40 °C, respectively for the maceration and the ultrasound-assisted extraction, ensuring the highest resinoid yield of 22.0 g/100 g in 4 h and 25.1 g/100 g in 30 min, which agreed with the yields obtained experimentally for the same time (21.7 and 25.3 g/100 g, respectively).     

Sequential extraction and enrichment of flavonoids from Euonymus alatus by ultrasonic-assisted polyethylene glycol-based extraction coupled to temperature-induced cloud point extraction

A green method for simultaneous extraction and enrichment of flavonoids from Euonymus alatus was developed by ultrasonic-assisted extraction (UAE) and temperature-induced cloud point extraction (TICPE) using PEG-base aqueous solution as the extractant. Based on screening different molecular weights of PEGs, PEG-400/water was used as the extractant, and the effects of key factors on extraction yields of flavonoids were investigated by single-factor experiments and response surface methodology (RSM). The optimum conditions of UAE were as follows: PEG-400 concentration of 16% (w/w), particle size of 80 mesh, solvent-to-material ratio of 60:1, extraction temperature of 90 °C and extraction time of 15 min. The results obtained by validation experiments were consistent with the values predicted by RSM. Temperature-induced formation of the aqueous two-phase system (ATPS) and TICPE process were further investigated by controlling temperature and adding (NH₄)₂SO₄. In the presence of (NH₄)₂SO₄, the ATPS formed at 75 ℃ and pH 3.5 could effectively improve separation and recovery of flavonoids with enrichment factor of above five times. Gallic acid, catechin, dihydromyricetin and ellagic acid in the extract were identified and confirmed by UPLC-Q-TOF-MS and the corresponding standards. The UAE-TICPE coupled to HPLC was successfully applied for extraction and determination of flavonoids in two batches of Euonymus alatus. The extraction yields of catechin, dihydromyricetin and total flavonoids were 0.377–0.684 mg/g, 1.091–1.353 mg/g and 2.612–3.146 mg/g, respectively. Compared to conventional extraction methods, PEG-based UAE integrated with TICPE in one-step procedure exhibited higher extraction efficiency and better extraction selectivity.     

Optimization of ultrasonic-assisted extraction of pomegranate (Punica granatum L.) seed oil

The effectiveness of ultrasonic-assisted extraction (UAE) of pomegranate seed oil (PSO) was evaluated using a variety of solvents. Petroleum ether was the most effective for oil extraction, followed by n-hexane, ethyl acetate, diethyl ether, acetone, and isopropanol. Several variables, such as ultrasonic power, extraction temperature, extraction time, and the ratio of solvent volume and seed weight (S/S ratio) were studied for optimization using response surface methodology (RSM). The highest oil yield, 25.11% (w/w), was obtained using petroleum ether under optimal conditions for ultrasonic power, extraction temperature, extraction time, and S/S ratio at 140 W, 40 °C, 36 min, and 10 ml/g, respectively. The PSO yield extracted by UAE was significantly higher than by using Soxhlet extraction (SE; 20.50%) and supercritical fluid extraction (SFE; 15.72%). The fatty acid compositions were significantly different among the PSO extracted by Soxhlet extraction, SFE, and UAE, with punicic acid (>65%) being the most dominant using UAE.     

Ultrasound-assisted extraction of polysaccharides from Rhododendron aganniphum: Antioxidant activity and rheological properties

In this study, we aimed to optimize the extraction of polysaccharides from the leaves of Rhododendron aganniphum and investigate its rheological properties and antioxidant activity. After optimizing the operating parameters using a Box-Behnken design (BBD), the results showed that the optimal ultrasound-assisted extraction conditions were as follows: extraction temperature, 55 °C; liquid-solid ratio, 25:1; extraction time, 2.2 h; and ultrasound treatment power, 200 W. The optimized experimental yield of polysaccharides by ultrasound-assisted extraction (PUAE) was 9.428%, higher than that obtained by hot water extraction (PHWE) for 12 h at the same liquid-solid ratio and extraction temperature. In the in vitro antioxidant activity tests, PUAE had higher positive radical scavenging activity for hydroxyl, superoxide and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals than PHWE. However, PUAE and PHWE solutions had similar intermolecular interactions in the steady-shear flow and dynamic viscoelasticity tests, resulting in similar macroscopic behaviour. With respect to the apparent viscosity, storage modulus (G′) and loss modulus (G″) of PUAE were lower at the same shear rate or angular frequency. All PUAE solutions exhibited non-Newtonian shear-thinning pseudoplastic behaviour that was accurately described by the Carreau model but was better fit by the power-law model at high shear rates (≥1/s), which demonstrated that the variation in the apparent viscosity dependence was greater at higher concentrations and shear rates. The G′ and G″ of the solutions increased as the experimental frequency increased from 0.05 to 500 rad/s under all experimental concentrations, and the modulus crossover point decreased gradually with increasing PUAE concentration. The above results demonstrated that the ultrasound-assisted extraction methods gave a higher yield of polysaccharides from the leaves of R. aganniphum with a shorter extraction time than the hot water extraction method, which could affect the apparent viscosity and dynamic viscoelasticity. PUAE presented good radical scavenging activity for DPPH, superoxide and hydroxyl radicals in vitro and could be used as a natural antioxidant in the food and medical industries.     

Ultrasonic-assisted extraction,characteristics and activity of Ipomoea batatas polysaccharide

Ipomoea batatas polysaccharides (IBPs) have many important physiological functions. The optimal extraction conditions were extraction time of 40 min, solid–liquid ratio of 1:8 and ultrasonic power of 240 W. 1D/2D nuclear magnetic resonance (1D/2D NMR) analysis showed that the main chain of IBP-1A was mainly composed of →4)-α-D-Glcp-(1→ and →3, 6)-β-D-Glcp-(1→ residues. In vivo experiments showed that polysaccharide significantly increased the levels of antioxidation-related enzymes and metabolites in older mice. It could significantly relieve oxidative stress injury and delay aging. Therefore, this study provided a new theoretical basis for the development of IBPs as antioxidant food.     

Optimisation of ultrasound assisted extraction of phenolic compounds from Sparganii rhizoma with response surface methodology

The present study reports on the extraction of phenolic compounds from sparganii rhizome. Box–Behnken Design (BBD), a widely used form of response surface methodology (RSM), was used to investigate the effect of process variables on the ultrasound-assisted extraction (UAE). Three independent variables including ethanol concentration (%), extraction time (min) and solvent-to-material ratio (mL/g) were studied. The results showed that the optimal UAE condition was obtained with an ethanol concentration of 75.3%, an extraction time of 40 min and a solvent-to-material ratio of 19.21 mL/g for total phenols, and an ethanol concentration of 80%, an extraction time of 33.54 min and solvent-to-material ratio of 22.72 mL/g for combination of ρ-hydroxybenzaldehyde, ρ-coumaric acid, vanillic acid, ferulic acid, rutin and kaempferol. The experimental values under optimal conditions were in good consistent with the predicted values, which suggested UAE is more efficient process as compared to solvent extraction.     

Optimization of ultrasonic cell grinder extraction of anthocyanins from blueberry using response surface methodology

Ultrasonic cell grinder extraction (UCGE), using water as the solvent, was firstly applied to extract anthocyanins from blueberry. Extraction yield was related with four variables, including ratio of solution to solid, extraction power, buffer time, and extraction time. On the basis of response surface methodology (RSM), the optimal conditions were determined to be the ratio of solution to solid as 25:1 (mL/g), the extraction power as 1500 W, the buffer time as 3.0 s, and the extraction time as 40 min. The experimental yield of anthocyanins using UCGE was 2.89 mg/g higher than that of conventional ultrasound-assisted extraction (CUAE). This study indicated that UCGE was an innovative, efficient, and environment friendly method in ultrasonic extraction fields, and had a potential to effectively extract other bioactive constituents.     

Novel modified ultrasonication technique for the extraction of lycopene from tomatoes

Lycopene extraction was carried out via the ultrasonic assisted extraction (UAE) with response surface methodology (RSM). Sonication enhanced the efficiency of relative lycopene yield (enhancement of 26% extraction yield of lycopene in 6 replications at 40.0 min, 40.0 °C and 70.0% v/w in the presence of ultrasound), lowered the extraction temperature and shortened the total extraction time. The extraction was applied with the addition of oxygen-free nitrogen flow and change of water route during water bath sonication. The highest relative yield of lycopene obtained was 100% at 45.0 °C with total extraction time of 50.0 min (30:10:10) and ratio of solvent to freeze-dried tomato sample (v/w) of 80.0:1. Optimisation of the lycopene extraction had been performed, giving the average relative lycopene yield of 99% at 45.6 min, 47.6 °C and ratio of solvent to freeze-dried tomato sample (v/w) of 74.4:1. From the optimised model, the average yield of all-trans lycopene obtained was 5.11 ± 0.27 mg/g dry weight. The all-trans lycopene obtained from the high-performance liquid chromatography (HPLC) chromatograms was 96.81 ± 0.81% with 3.19 ± 0.81% of cis-lycopenes. The purity of total-lycopene obtained was 98.27 ± 0.52% with β-carotene constituted 1.73 ± 0.52% of the extract. The current improved, UAE of lycopene from tomatoes with the aid of RSM also enhanced the extraction yield of trans-lycopene by 75.93% compared to optimised conventional method of extraction. Hence, the current, improved UAE of lycopene promotes the extraction yield of lycopene and at the same time, minimises the degradation and isomerisation of lycopene.     

Ultrasound-assisted extraction of saffron bioactive compounds; separation of crocins,picrocrocin, and safranal optimized by artificial bee colony

In this work, a four-factor five-level full factorial central composite design (CCD) was used to optimize the ultrasound-assisted extraction (UAE) of saffron major components, namely picrocrocin, safranal and crocin. The process parameters included ethanol concentration (0–100%), extraction time (2–10 min), duty cycle (0.2–1.0) and ultrasonic amplitude (20–100%). The extracted compounds were measured both by spectrophotometry and chromatography techniques. The results revealed that the middle concentrations of ethanol and relatively long process durations along with high duty cycles and ultrasonic amplitudes had the most profound impact on the yields of the extracted bioactives. UAE was optimized using response surface methodology (RSM) and artificial bee colony (ABC); a comparison between these methods indicated their optimization power was approximately the same. According to the RSM analysis, an ethanol concentration of 58.58%, extraction time of 6.85 min, duty cycle of 0.82 and amplitude of 91.11% were the optimum extraction conditions, while the optimal conditions resulting from ABC were 53.07%, 7.32 min, 0.93 and 100% for the UAE variables respectively. Finally, HPLC analysis was carried out on the UAE optimum extract resulting from RSM. Four crocetin esters were detected in the optimal extract.     

Ultrasound assisted extraction of pectin from waste Artocarpus heterophyllus fruit peel

Four factors three level face centered central composite response surface design was employed in this study to investigate and optimize the effect of process variables (liquid-solid (LS) ratio (10:1–20:1 ml/g), pH (1−2), sonication time (15–30 min) and extraction temperature (50–70 °C)) on the maximum extraction yield of pectin from waste Artocarpus heterophyllus (Jackfruit) peel by ultrasound assisted extraction method. Numerical optimization method was adapted in this study and the following optimal condition was obtained as follows: Liquid-solid ratio of 15:1 ml/g, pH of 1.6, sonication time of 24 min and temperature of 60 °C. The optimal condition was validated through experiments and the observed value was interrelated with predicted value.