Ultrasound-assisted Aqueous Two-phase Extraction Research Articles

Ultrasound-assisted Aqueous Two-Phase Extraction of Flavonoids from Scutellariae Radix and Evaluation of their Bioactivities In Vitro

Background: Scutellariae Radix, one of the most widely used herbs in Traditional Chinese Medicine, exhibits various biological activities due to its chemical components, which stand out for a number of flavonoids. In this study, Ultrasound-assisted aqueous two-phase extraction (UAATPE) was employed for the first time to obtain a high extraction rate and high purity of flavonoids from Scutellariae Radix. Methods: The Box-Behnken response surface method (RSM) was utilized to optimize the extraction conditions with the application of the new aqueous two-phase system (ATPS) composed of ethanol and ammonium sulfate. The major influence factors, including ethanol concentration, ammonium sulfate concentration, liquid-to-solid ratio, sonication time, and extraction temperature, were investigated by the single-factor experiment. The compositional characterization of flavonoids was characterized with HPLC-UV. Scanning electron microscopy (SEM) was applied to research the surface morphology of raw material. Furthermore, the bioactivities of the extract obtained by UA-ATPE were studied in vitro. Results: The optimal extraction conditions were as follows: the ethanol content was 26.12% (w/w), the ammonium sulfate content was 20.02% (w/w), the liquid-to-solid ratio was 40 mL/g, the sonication time was 5 min with the ultrasonic power of 250 W, and the operating process was performed at room temperature. Compared with the traditional extraction methods, UA-ATPE exhibited higher extraction efficiency and better extraction selectivity. The DPPH and ABTS radical scavenging tests showed that enriched products possessed strong antioxidant activity. Conclusion: The study confirmed that the developed method of UA-ATPE could be used as an efficient, eco-friendly, and low-consumption method for the extraction and purification of flavonoids from Scutellariae Radix.

Comparison of an Ultrasound-Assisted Aqueous Two-Phase System Extraction of Anthocyanins from Pomegranate Pomaces by Utilizing the Artificial Neural Network-Genetic Algorithm and Response Surface Methodology Models.

As a by-product of pomegranate processing, the recycling and reuse of pomegranate pomaces (PPs) were crucial to environmentally sustainable development. Ultrasound-assisted aqueous two-phase extraction (UA-ATPE) was applied to extract the anthocyanins (ACNs) from PPs in this study, and the central composite design response surface methodology (CCD-RSM) and artificial neural network-genetic algorithm (ANN-GA) models were utilized to optimize the extraction parameters and achieve the best yield. The results indicated that the ANN-GA model built for the ACN yield had a greater degree of fit and accuracy than the RSM model. The ideal model process parameters were optimized to have a liquid-solid ratio of 49.0 mL/g, an ethanol concentration of 28 g/100 g, an ultrasonic time of 27 min, and an ultrasonic power of 330 W, with a maximum value of 86.98% for the anticipated ACN yield. The experimental maximum value was 87.82%, which was within the 95% confidence interval. A total of six ACNs from PPs were identified by utilizing UHPLC-ESI-HRMS/MS, with the maximum content of cyanidin-3-O-glucoside being 57.01 ± 1.36 mg/g DW. Therefore, this study has positive significance for exploring the potential value of more by-products and obtaining good ecological and economic benefits in the future.

Combined enzymatic and ultrasound-assisted aqueous two-phase extraction of antidiabetic flavonoid compounds from Strobilanthes crispus leaves

A novel green extraction method combining enzymatic and ultrasound-assisted aqueous two-phase extractions was employed to enhance and purify the flavonoid extract from Strobilanthes crispus leaves. Cellulase was used in the pretreatment, and ethanol-(NH4)2SO4 was selected as the solvent. A concentration of 7% (w/w) cellulase and a duration of 2 h were the optimal conditions for pretreatment. The optimal conditions for ultrasound-assisted aqueous two-phase extraction were 33% (w/w) ethanol and 14% (w/w) (NH4)2SO4 as they produced a yield (77.81%), partition coefficient (31.17), extraction efficiency (98.04%), and a high total flavonoid content (0.3666 mg QE/g dry leaf powder). Six compounds from the leaf extract were identified through liquid chromatography-mass spectrometry/mass spectrometry-quadrupole-time of flight (LCMS/MS-Q-TOF) analysis. The crude extract and three compounds in it (kaempferol, graveobioside A, and genistein) showed an antidiabetic activity with IC50 values of 390.35, 201.87, 292.73, and 431.82 mg/mL, respectively. These values are comparable to the standard drug acarbose.

Ultrasound assisted aqueous two-phase extraction of polysaccharides from corn stigma: Process optimization, structure characterization, and immunomodulatory activity

The two models of ultrasound assisted aqueous two-phase extraction (UATPE) of polysaccharides from corn stigma were established by response surface methodology (RSM) and artificial neural network (ANN), and compared the prediction ability of RSM and ANN models. Subsequently, the extraction process of corn stigma polysaccharides (CSPs) was optimized by the combination of genetic algorithm (GA) and ANN. The results show that two models could precisely predict the CSPs yield, whereas the prediction ability of ANN was better than RSM. The optimum combination of process parameters was as follows: the ultrasound power of 466 W, extraction temperature of 54 °C, liquid-to-solid ratio of 47 mL/g, and extraction time of 31 min. The highest CSPs yield was 9.89% ± 0.14% under the above parameters, and then the crude CSPs were further purified by DEAE-52 and Sephadex G − 100 chromatography to obtain a homogenous fraction (CSPs-2-SG with a molecular weight of 2.86 × 106 Da) that consisted of Man, GlcA, GalA, Glc, Gal, and Xyl with a molar ratio of 9.14:16.35:11.27:91.08:6.49:4.71. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) demonstrated that CSPs-2-SG showed irregular structures including uneven fragments, some holes, and spherical aggregates on the surface. Finally, the immunomodulatory activity of CSPs-2-SG was evaluated by RAW264.7 cells. CSPs-2-SG could improve viability, phagocytic rate, and NO content of RAW264.7 cells. Moreover, CSPs-2-SG could increase the levels of IL − 1β, IL-6, and TNF-α in RAW264.7 cells. The findings provide important references for utilizing corn stigma as a potential natural immune regulator.

Ultrasound assisted aqueous two-phase extraction of anthocyanins from blueberry and its anti-tumor activity

Blueberry contains abundant anthocyanins, polyphenols, polysaccharides, and other active compounds. Anthocyanins are one of the most principal active components in blueberry, which have high medical and medicinal values. Consequently, the aim of this paper is to optimize the ultrasound assisted aqueous two-phase extraction (UATPE) of anthocyanins from blueberry by response surface methodology (RSM). The optimal extraction process to achieve the highest yield of anthocyanins (5.96 ± 0.04) mg/g from blueberry via UATPE was obtained under the ultrasound power of 269 W, extraction temperature of 56 °C, ammonium sulfate mass fraction of 23 %, and ethanol concentration of 25%. Subsequently, we evaluated the anti-tumor activity of blueberry anthocyanins extract (BAE) obtained under the optimal process on breast cancer by MTT, confocal laser microscopy, and flow cytometer. The results show that BAE could memorably inhibit MCF-7 cells viability, increase the intensity of nuclear blue fluorescence, and accelerate the apoptosis of MCF-7 cells. The findings provide an effective and feasible method for the extraction of anthocyanins. Moreover, these results provide a material basis for the development of natural anti-tumor drugs.

Simultaneous extraction and preliminary purification of polyphenols from grape pomace using an aqueous two-phase system exposed to ultrasound irradiation: Process characterization and simulation.

In this study, an ultrasound-assisted aqueous two-phase (ATP) extraction method was used for the extraction and purification of phenolic compounds from grape pomace. The effect of acoustic energy densities (AED, 41.1, 63.5, 96.1, 111.2 W/L) and temperatures (20, 30, 40°C) on the yield of phenolics was investigated. An artificial neural network (ANN) was successfully used to correlate the extraction parameters with phenolic yield. Then, a diffusion model based on Fick's second law was used to model the mass transfer process during ultrasound-assisted ATP extraction and evaluate the effective diffusion coefficient of phenolics. The results revealed the increase in AED, and the temperature increased the effective diffusivity of phenolics. The HPLC analysis of anthocyanins and flavonols showed that ultrasound significantly increased the extraction yield of anthocyanins compared with the traditional method. High amounts of rutin and myricetin were recovered using the ATPS systems. Sugars were mainly distributed in the bottom phase, whereas phenolics were located in the top phase. Conclusively, ultrasound-assisted aqueous two-phase (ATP) extraction can be used as an effective method to achieve the simultaneous separation and preliminary purification of phenolics from grape pomace.

Optimization of ultrasound-assisted aqueous two-phase extraction of polysaccharides from seabuckthorn fruits using response methodology, physicochemical characterization and bioactivities.

Seabuckthorn fruits contains many active subtances, among them, the seabuckthorn polysaccharide is one of the main active ingredients, and exhibits diverse bioactivities. The extraction of polysaccharides from seabuckthorn fruits is the most important step for their wide applications. Ultrasound-assisted aqueous two-phase extraction (UA-ATPE) is a promising green method for extracting polysaccharides. Additionally, physicochemical characterization and antioxidant activities can evaluate the potential functions and applications in the food and medicine industries. Based on the single-factor experiments, 20.70% (w/w) ammonium sulfate ((NH4 )2 SO4 ) and 27.56% (w/w) ethanol were determined as the suitable composition for aqueous two-phase. The optimum conditions of UA-ATPE obtained by response surface methodology were as follows: ultrasonic power (390 W), extraction time (41 min), liquid-to-material ratio (72: 1mL/g), and the total yield of the polysaccharides reached 34.14 ± 0.10%, The molecular weights of the purified upper-phase seabuckthorn polysaccharide (PUSP) and the purified lower-phase seabuckthorn polysaccharide (PLSP) were 65 525 and 26 776 Da, respectively. PUSP and PLSP contained the same six monosaccharides (galacturonic acid, rhamnose, xylose, mannose, glucose and galactose), but with different molar ratios. Furthermore, PUSP and PLSP displayed certain viscoelastic property, had no triple helical structure, possessed different thermal stability, surface morphology and conformation in aqueous solution. PUSP and PLSP displayed strong antioxidant properties by the assays of scavenging ability of ABTS+ ·, the protection of DNA damage and erythrocyte hemolysis. UA-ATPE significantly increased the yield of seabuckthorn polysaccharides. PUSP and PLSP were different in many aspects, such as molar ratio, surface shape and antioxidant activities. Seabuckthornpolysaccharides possess great potential in medicine and functional foods. © 2022 Society of Chemical Industry.

Enhanced extraction of bioactive natural products using ultrasound-assisted aqueous two-phase system: Application to flavonoids extraction from jujube peels

The ultrasound-assisted aqueous two-phase extraction (UA-ATPE) was employed to develop an effective technique for the extraction of flavonoids from jujube peels (JPs). The extraction conditions were further optimized as K2HPO4 35% (w/w), ethanol 20% (w/w), solid–liquid ratio 1:30 g/mL (w/v), ultrasonic power 200 W, and extraction time 50 min. Moreover, rutin, quercetin 3-β-d-glucoside, and kaempferol-3-O-rutinosid were identified as the main flavonoids by UPLC-MS/MS. Finally, the extraction mechanism of UA-ATPE was explored, which is salting out effect, hydrogen bonding, van der Waals force, and ultrasound promoted the mass transfer of solvent to cells of JPs, then extraction behavior occurred. The mechanical destruction of JPs cells by ultrasound also further accelerated the release of flavonoids. Flavonoids were captured by W/W emulsion in the bottom phase and distributed to the top phase. Overall, this study proposes a green and clean method, UA-ATPE, to extract flavonoids from JPs, while revealing the mechanism of UA-ATPE.

Modeling, optimization, purification, and characterization of polysaccharides from Lilium lancifolium Thunb

Ultrasound assisted aqueous two-phase extraction of polysaccharides from Lilium lancifolium Thunb was modeled by response surface methodology (RSM) and artificial neural network (ANN), and optimized using genetic algorithm coupled with ANN (GA-ANN). Statistical analysis showed that the models obtained by RSM and ANN could accurately predict the Lilium lancifolium Thunb polysaccharides (LLPs) yield. However, ANN prediction was more accurate than RSM. The optimum extraction parameters to achieve the highest LLPs yield (15.17 ± 0.21)% was obtained at the ultrasound power of 250 W, extraction temperature of 63 °C, liquid-to-solid ratio of 21 mL/g, and extraction time of 32 min. Subsequently, the crude LLPs were further purified via DEAE-52 and Sephadex G-100 chromatography to obtain a homogenous fraction (LLPs-2-SG, 421.41 kDa) that contained mannose, glucose, and galactose in a molar ratio of 10.52:23.06:7.19. The structure of LLPs-2-SG was characterized with UV–vis, FT–IR, AFM, and SEM. The findings provide a critical method for the extraction, separation and purification of polysaccharides from natural resources.

Ultrasound assisted aqueous two-phase extraction of polysaccharides from Cornus officinalis fruit: Modeling, optimization, purification, and characterization

Ultrasound assisted aqueous two-phase extraction of polysaccharides from Cornus officinalis fruit was modeled by response surface methodology (RSM) and artificial neural network (ANN), and optimized using genetic algorithm coupled with ANN (GA-ANN). Statistical analysis showed that the models obtained by RSM and ANN could accurately predict the Cornus officinalis polysaccharides (COPs) yield. However, ANN prediction was more accurate than RSM. The optimum extraction parameters to achieve the highest COPs yield (7.85 ± 0.09)% was obtained at the ultrasound power of 350 W, extraction temperature of 51 ℃, liquid-to-solid ratio of 17 mL/g, and extraction time of 38 min. Subsequently, the crude COPs were further purified via DEAE-52 and Sephadex G-100 chromatography to obtain a homogenous fraction (COPs-4-SG, 33.64 kDa) that contained galacturonic acid, arabinose, mannose, glucose, and galactose in a molar ratio of 34.82:14.19:6.75:13.48:12.26. The structure of COPs-4-SG was also characterized with UV–vis, fourier-transform infrared spectroscopy (FT–IR), atomic force microscopy (AFM), scanning electron microscopy (SEM), Congo-red test, and circular dichroism (CD). The findings provide a feasible way for the extraction, purification, and optimization of polysaccharides from plant resources

Ultrasound-assisted aqueous two-phase extraction of total flavonoids from Tremella fuciformis and antioxidant activity of extracted flavonoids

Here, the extraction of total flavonoids from Tremella fuciformis (TF) by aqueous two-phase extraction combined with ultrasound is presented. The extraction was optimized via a single-variable approach using the variables of mass fraction of ethanol and NaH2PO4, ultrasonication time, and TF sample amount. Response surface methodology (RSM) was then applied to determine the best conditions for extraction using the following three key parameters: ultrasonication time, ethanol mass fraction, and NaH2PO4 mass fraction. The optimum conditions were as follows: an aqueous two-phase system of ethanol/NaH2PO4, an ethanol mass fraction of 23%, a NaH2PO4 mass fraction of 27.18%, and an extraction time of 8.25 min. Under these conditions, the maximum extraction rate of total flavonoids was 0.158 mg/g. The scavenging rates for superoxide anion and hydroxyl radical, as well as the antioxidant activity of total flavonoids in TF, were studied. Total flavonoids from TF showed some scavenging ability, but the scavenging effects were lower than those of vitamin C (Vc) at the same concentrations. Nevertheless, ultrasound-assisted two-phase extraction was considered an efficient and rapid method that can be used to extract total flavonoids from TF.

Optimization of ultrasound-assisted aqueous two phase extraction of polyphenols from olive leaves

In this study, polyphenols from olive leaves was extracted by ultrasound-assisted aqueous two phase extraction (UAATPE). Based on single factor experiment and response surface methodology (RSM), the optimum extraction conditions of polyphenols contained 29% (w/w) (NH4)2SO4, 35% (w/w) ethanol, pH 6.7, and 45 °C. The maximum extraction yield of polyphenols and oleuropein content were 34.06 mg/g and 44.13 mg/L, respectively. Compared with ultrasound-assisted extraction (UAE) and aqueous two phase extraction (ATPE), the extraction yield of polyphenols by UAATPE was 9.48 and 61.19% higher, respectively. In addition, the extract of UAATPE had higher purity. The results of antioxidant activity showed that polyphenols extracted by UAATPE had stronger DPPH and hydroxyl radicals scavenging ability and reducing power. Therefore, UAATPE is an efficient method for extracting polyphenols from olive leaves.

Ultrasound-assisted aqueous two-phase extraction of synephrine, naringin, and neohesperidin from Citrus aurantium L. fruitlets

The ultrasound-assisted aqueous two-phase extraction (UA-ATPE) was first employed to develop an effective technique for simultaneous extraction and preliminary purification of synephrine, naringin, and neohesperidin from Citrus aurantium L. fruitlets. Five types of ethanol/salts of aqueous two-phase system (ATPS) were investigated and then the extraction conditions were further optimized using single-factor experiments and response surface methodology (RSM) via Box–Behnken Design (BBD). The optimum process parameters were concluded as follows: 20.60% (w/w) K2CO3, 27% (w/w) ethanol, solvent-to-material ratio of 45.17:1 (g:g), 120-mesh particle size of fruitlets powder, extraction temperature of 50 °C, extraction time of 30 min, and ultrasonic power of 80 W. Under these conditions, the extraction yields of synephrine, naringin, and neohesperidin were up to 11.17 mg/g, 7.39 mg/g, and 89.27 mg/g, respectively. The yield of neohesperidin extracted by the optimal UA-ATPE was over eight times higher than that extracted by the ultrasound-assisted extraction (UAE) using conventional solvents, and the total yield of target compounds was over twice higher while the impurity content in the extract was much lower. Therefore, UA-ATPE appeared to be a highly effective and promising approach for the extraction of synephrine, naringin, and neohesperidin from C. aurantium fruitlets.

Ultrasound-assisted aqueous two-phase extraction of resveratrol from the enzymatic hydrolysates of Polygonum cuspidatum

An efficient ultrasound-assisted aqueous two-phase extraction (UAATPE) and enrichment process for resveratrol from the enzymatic hydrolysates of Polygonum cuspidatum using an ethanol/ammonium sulfate system was developed. After enzymatic hydrolysis using polydatin-β-glucosidase (PBG) which was a wild extracellular enzyme, 99.7% of polydatin was converted to resveratrol. The maximum yield of resveratrol was 10.7 mg/g and the corresponding recovery rate was 99.1% at 21% (w/w) ethanol/22% (w/w) (NH4)2SO4. The extraction yield of resveratrol in UAATPE was increased by 43.6 and 79.4% compared with ultrasound-assisted extraction and solvent extraction with stirring, respectively. Meanwhile, the total sugars in UAATPE were only 46.8% of ultrasound-assisted extraction and 56.0% of solvent extraction with stirring. The process of UAATPE is a promising technique for extraction and enrichment of bioactive components.

Extraction and physicochemical properties of polysaccharides from Ziziphus Jujuba cv. Muzao by ultrasound-assisted aqueous two-phase extraction

In this paper, ultrasound-assisted aqueous two-phase extraction (UAATPE) performed via a one-step procedure was applied to extract polysaccharides from Ziziphus Jujuba cv. Muzao. Using an ethanol/ammonium sulfate system as a multiphase solvent, effects of UAATPE on extraction of ZMP, such as solvent-to-material ratio, composition of aqueous two-phase system, and extraction time and temperature, were investigated using response surface methodology on basis of results of single-factor experiment. The final optimal conditions were ATPS composition: 29% ethanol and 15% (NH4)2SO4, extraction time: 38min, extraction temperature: 48°C, solvent-to-material ratio: 30, and microwave power: 70W. Under these conditions, the experimental extraction yield was 8.18%. Chemical analysis revealed that ZMP were composed of rhamnose, arabinose, xylose, mannose, glucose, galactose, and galacturonic acid, at a ratio of 1.46:2.47:2.27:1.12:1.00:1.57:5.40, respectively. The structures were also characterized with UV, FTIR, and SEM. The DPPH and ABTS radical scavenging activities showed ZMP had moderately antioxidant activity.

Aqueous two-phase assisted by ultrasound for the extraction of anthocyanins from Lycium ruthenicum Murr.

ABSTRACTIn this study, an efficient ultrasound-assisted aqueous two-phase extraction method was used for the extraction of anthocyanins from Lycium ruthenicum Murr. An ethanol/ammonium sulfate system was chosen for the aqueous two-phase system due to its fine partitioning and recycling behaviors. Single-factor experiments were conducted to determine the optimized composition of the system, and the response surface methodology was used for the further optimization of the ultrasound-assisted aqueous two-phase extraction. The optimal conditions were as follows: a salt concentration of 20%, an ethanol concentration of 25%, an extraction time of 33.7 min, an extraction temperature of 25°C, a liquid/solid ratio of 50:1 w/w, pH value of 3.98, and an ultrasound power of 600 W. Under the above conditions, the yields of anthocyanins reached 4.71 mg/g dry sample. For the further purification, D-101 resin was used, and the purity of anthocyanins reached 25.3%. In conclusion, ultrasound-assisted aqueous two-phase extraction was an efficient, ecofriendly, and economical method, and it may be a promising technique for extracting bioactive components from plants.

Ultrasound assisted extraction in aqueous two-phase system for the integrated extraction and separation of antioxidants from wheat chaff

Abstract A procedure for simultaneous extraction and separation of xylooligosaccharides and phenol compounds from wheat chaff by ultrasound assisted ethanol/ammonium sulfate aqueous two-phase extraction was studied. In aqueous two-phase system 23.8% (w/w) salt, 24.3% (w/w) ethanol and 1.2% (w/w) solids load using ultrasound waves (30 Hz, 500 W, 10 min), extraction yields of sugars and phenols were 16 mg/g and 2.67 mg/g dry material, respectively. Under these conditions xylooligosaccharides and phenol compounds partitioned toward the bottom and the top phase, respectively, and selectivity of their separation was almost 50. The bottom phase with recovered 96% extracted sugars and the top phase containing 64% extracted phenols expressed 0.45 μmol TE/g and 1.1 μmol TE/g ABTS radical-scavenging activities, respectively. Result indicated that ultrasound assisted aqueous two-phase extraction in ethanol/ammonium sulfate system could be a promising technique for integrated extraction and separation of bioactive components from wheat chaff.

Ultrasound-assisted aqueous two-phase extraction of phenylethanoid glycosides from Cistanche deserticola Y. C. Ma stems.

An efficient ultrasound-assisted aqueous two-phase extraction and enrichment process for phenylethanoid glycosides from Cistanche deserticola Y. C. Ma stems was developed in this work. An ethanol/ammonium sulfate system was chosen for the aqueous two-phase system due to its fine partitioning and recycling behaviors. Single-factorexperiments and response surface methodology were used to optimize the process parameters of the ultrasound-assisted aqueous two-phase extraction. The optimal conditions were as follows: a salt concentration of 23.5%, an ethanol concentration of 20%, an extraction time of 37 min, an extraction temperature of 30°C, a liquid/solid ratio of 30:1 w/w, and an ultrasound power of 300 W. Under the above conditions, the extraction yields of echinacoside and acteoside (the main components of phenylethanoid glycosides) reached 5.35 and 6.22 mg/g dry material weight, respectively. The contents of echinacoside and acteoside in the extracts reached 27.56 and 30.23 mg/g, respectively, which were 2.46- and 2.58-fold higher than the amounts obtained in ultrasound-assisted extraction. In conclusion, ultrasound-assisted aqueous two-phase extraction was an efficient, ecofriendly, and economical method, and it may be a promising technique for extracting and enriching bioactive components from plants.

Ionic liquid-based ultrasound-assisted aqueous two-phase extraction of the pyrethroid insecticides in tea drinks

Ionic liquid-based ultrasound-assisted aqueous two-phase extraction was established and applied to the extraction of four pyrethroid insecticides from tea drinks prior to their determination by high-performance liquid chromatography. The main parameters affecting the extraction efficiency, including the type and amount of ionic liquid, type of salt, ultrasonic time, pH value of the sample solution, type of reverse extraction solvent, concentration of (NH4)2SO4 and volume of cyclohexane, were investigated. Under the optimal experimental conditions, good linearity was obtained (r > 0.998). The recoveries of pyrethroids obtained by analyzing the four spiked tea samples were from 70.22 to 122.1%. The intra-day and inter-day precisions were in the range of 2.65–11% and 1.95–10.83% (n = 5), respectively. Limits of detection for determining the analytes were in the range of 4.86–9.53 ng mL−1. The results showed that the present method was a simple, efficient and feasible method for the determination of pyrethroids in the tea drinks.