Aqueous ionic liquid based ultrasonic assisted extraction of four acetophenones from the Chinese medicinal plant Cynanchum bungei Decne

In this study, an aqueous ionic liquid based ultrasonic assisted extraction (ILUAE) method for the extraction of the four acetophenones, namely 4-hydroxyacetophenone (1), 2,5-dihydroxyacetophenone (2), baishouwubenzophenone (3) and 2,4-dihydroxyacetophenone (4) from the Chinese medicinal plant Cynanchum bungei was developed. Three kinds of aqueous l-alkyl-3-methylimidazolium ionic liquids with different anion and alkyl chain were investigated. The results indicated that ionic liquids (ILs) showed remarkable effects on the extraction efficiency of acetophenones. In addition, the ILUAE, including several ultrasonic parameters, such as the ILs concentration, solvent to solid ratio, power, particle size, temperature, and extraction time have been optimized. Under these optimal conditions (e.g., with 0.6 M [C₄MIM]BF₄, solvent to solid ratio of 35:1, power of 175 W, particle size of 60–80 mesh, temperature of 25 °C and time of 50 min), this approach gained the highest extraction yields of four acetophenones 286.15, 21.65, 632.58 and 205.38 μg/g, respectively. The proposed approach has been evaluated by comparison with the conventional heat-reflux extraction (HRE) and regular UAE. The results indicated that ILUAE is an alternative method for extracting acetophenones from C. bungei.     

Ultrasound assisted enzyme catalyzed hydrolysis of waste cooking oil under solvent free condition

The present work demonstrates the hydrolysis of waste cooking oil (WCO) under solvent free condition using commercial available immobilized lipase (Novozyme 435) under the influence of ultrasound irradiation. The process parameters were optimized using a sequence of experimental protocol to evaluate the effects of temperature, molar ratios of substrates, enzyme loading, duty cycle and ultrasound intensity. It has been observed that ultrasound-assisted lipase-catalyzed hydrolysis of WCO would be a promising alternative for conventional methods. A maximum conversion of 75.19% was obtained at mild operating parameters: molar ratio of oil to water (buffer pH 7) 3:1, catalyst loading of 1.25% (w/w), lower ultrasound power 100 W (ultrasound intensity – 7356.68 W m⁻²), duty cycle 50% and temperature (50 °C) in a relatively short reaction time (2 h). The activation energy and thermodynamic study shows that the hydrolysis reaction is more feasible when ultrasound is combined with mechanical agitation as compared with the ultrasound alone and simple conventional stirring technique. Application of ultrasound considerably reduced the reaction time as compared to conventional reaction. The successive use of the catalyst for repetitive cycles under the optimum experimental conditions resulted in a loss of enzymatic activity and also minimized the product conversion.     

Ultrasound assisted enzymatic pre-treatment of high fat content dairy wastewater

This paper illustrates the application of ultrasound in a dairy waste water treatment for the removal of fat using enzyme as a catalyst. Lipase Z was used to perform the enzymatic pre-hydrolysis of a synthetic dairy wastewater containing around 2000 mg/L of fat content coupled with ultrasound irradiation. Different process parameters like effect of enzyme loading, temperature, ultrasound power, frequency, duty cycle and speed of agitation are optimized. The maximum hydrolysis of 78% is achieved at 0.2% enzyme loading (w/v), 30 °C temperature, 165 W of ultrasonication power at 25 kHz and 66% duty cycle. It was observed that the enzymatic pre-hydrolysis under the influence of ultrasound drastically reduces the reaction time from 24 h to 40 min as compared to conventional stirring with improved yield.     

Ultrasound assisted enzyme catalyzed synthesis of glycerol carbonate from glycerol and dimethyl carbonate

The present work illustrates the transesterification of glycerol to glycerol carbonate (GlyC) from dimethyl carbonate (DMC) using commercial immobilized lipase (Novozym 435) under ultrasonic irradiation. The experiments were performed in a batch reactor placed in an ultrasonic water bath using a sequence of experimental protocol to evaluate the effects of temperature, molar ratios of substrates, enzyme loading, duty cycle and ultrasound power on the conversion of glycerol to GlyC. It has been found that ultrasound-assisted lipase-catalyzed transesterification of glycerol would be a potential alternative to conventional alkali-catalyzed method, as high conversion (99.75%) was obtained at mild operating conditions: molar ratio of DMC to glycerol 3:1, catalyst amount of 13% (w/w), lower power input (100 W), duty cycle 50% and temperature (60 °C) in a relatively short reaction time (4 h) using Novozym 435 as catalyst. Ultrasound reduces the reaction time up to 4 h as compared to conventional stirring method (14 h) catalyzed by Novozym 435. The repeated use of the catalyst under the optimum experimental condition resulted in decay in both enzyme activity and product conversion.     

Ultrasound assisted extraction of β-carotene from Spirulina platensis

This paper illustrates the Ultrasound Assisted Extraction (UAE) of β-carotene from Spirulina platensis. Various parameters such as extraction time, solvent type, biomass to solvent ratio, temperature, electrical acoustic intensity, length of the probe tip dipped into the solvent, duty cycle and pre treatment effect were explored for the extraction of β-carotene. From economic point of view, the optimal conditions for the extraction of β-carotene from Spirulina were 1.5 g Spirulina (2 min pre soaked in methanol) in 50 ml n-heptane at 30 °C temperature, 167 W/cm² electrical acoustic intensity and 61.5% duty cycle for 8 min with probe tip length of 0.5 cm dipped into the extracting solvent from the surface. The maximum extraction achieved under the above mentioned optimum parameters was 47.10%. The pre-treatment time showed a promising effect on the yield as pre-treating the biomass with methanol for 2 min before ultrasonication showed 12 times increase in extraction yield of β-carotene.     

Alkaline and ultrasound assisted alkaline pretreatment for intensification of delignification process from sustainable raw-material

Alkaline and ultrasound-assisted alkaline pretreatment under mild operating conditions have been investigated for intensification of delignification. The effect of NaOH concentration, biomass loading, temperature, ultrasonic power and duty cycle on the delignification has been studied. Most favorable conditions for only alkaline pretreatment were alkali concentration of 1.75 N, solid loading of 0.8% (w/v), temperature of 353 K and pretreatment time of 6 h and under these conditions, 40.2% delignification was obtained. In case of ultrasound-assisted alkaline approach, most favorable conditions obtained were alkali concentration of 1 N, paper loading of 0.5% (w/v), sonication power of 100 W, duty cycle of 80% and pretreatment time of 70 min and the delignification obtained in ultrasound-assisted alkaline approach under these conditions was 80%. The material samples were characterized by FTIR, SEM, XRD and TGA technique. The lignin was recovered from solution by precipitation method and was characterized by FTIR, GPC and TGA technique.     

Aqueous ionic liquid based ultrasonic assisted extraction of eight ginsenosides from ginseng root

We developed an aqueous ionic liquid based ultrasonic assisted extraction (ILUAE) method for the extraction of the eight ginsenosides (ginsenoside-Rg1, -Re, -Rf, -Rb1, -Rc, -Rb2, -Rb3 and -Rd) from ginseng root. A series of l-alkyl-3-methylimidazolium ionic liquids differing in composition of anions and cations were evaluated for extraction efficiency. The results indicated that the ILUAE method has a remarkable ability to improve the extraction efficiency of ginsenosides. In addition, the ILUAE procedure was also optimized on some ultrasonic parameters, such as the IL concentration, solvent to solid ratio and extraction time. Under these optimal conditions (e.g., with 0.3 M [C₃MIM]Br, solvent to solid ratio of 10:1 and extraction time of 20 min), this approach gained the highest extraction yields of total ginsenosides 17.81 ± 0.47 mg/g. Compared with the regular UAE, the proposed approach exhibited 3.16 times higher efficiency and 33% shorter extraction time, which indicated that ILUAE has a broad prospect for sample preparation of medicinal plants.     

Correlation between iodide dosimetry and terephthalic acid dosimetry to evaluate the reactive radical production due to the acoustic cavitation activity

Acoustic cavitation plays an important role in sonochemical processes and the rate of sonochemical reaction is influenced by sonication parameters. There are several methods to evaluate cavitation activity such as chemical dosimetry. In this study, to comparison between iodide dosimetry and terephthalic acid dosimetry, efficacy of sonication parameters in reactive radical production has been considered by iodide and terephthalic acid dosimetries. For this purpose, efficacy of different exposure parameters on cavitations production by 1 MHz ultrasound has been studied. The absorbance of KI dosimeter was measured by spectrophotometer and the fluorescence of terephthalic acid dosimeter was measured using spectrofluorometer after sonication. The result of experiments related to sonication time and intensity showed that with increasing time of sonication or intensity, the absorbance is increased. It has been shown that the absorbance for continuous mode is remarkably higher than for pulsing mode (p-value < 0.05). Also results show that with increasing the duty cycles of pulsed field, the inertial cavitation activity is increased. With compensation of sonication time or intensity in different duty cycles, no significant absorbance difference were observed unless 20% duty cycle. A significant correlation between the absorbance and fluorescence intensities (count) at different intensity (R = 0.971), different sonication time (R = 0.999) and different duty cycle (R = 0.967) were observed (p-value < 0.05). It is concluded that the sonication parameters having important influences on reactive radical production. These results suggest that there is a correlation between iodide dosimetry and terephthalic acid dosimetry to examine the acoustic cavitation activity in ultrasound field.     

Impact of ultrasound on solid–liquid extraction of phenolic compounds from maritime pine sawdust waste. Kinetics,optimization and large scale experiments

Maritime pine sawdust, a by-product from industry of wood transformation, has been investigated as a potential source of polyphenols which were extracted by ultrasound-assisted maceration (UAM). UAM was optimized for enhancing extraction efficiency of polyphenols and reducing time-consuming. In a first time, a preliminary study was carried out to optimize the solid/liquid ratio (6 g of dry material per mL) and the particle size (0.26 cm²) by conventional maceration (CVM). Under these conditions, the optimum conditions for polyphenols extraction by UAM, obtained by response surface methodology, were 0.67 W/cm² for the ultrasonic intensity (UI), 40 °C for the processing temperature (T) and 43 min for the sonication time (t). UAM was compared with CVM, the results showed that the quantity of polyphenols was improved by 40% (342.4 and 233.5 mg of catechin equivalent per 100 g of dry basis, respectively for UAM and CVM). A multistage cross-current extraction procedure allowed evaluating the real impact of UAM on the solid–liquid extraction enhancement. The potential industrialization of this procedure was implemented through a transition from a lab sonicated reactor (3 L) to a large scale one with 30 L volume.     

Effects of various factors of ultrasonic treatment on the extraction recovery of drugs from fish tissues

In the present research, a combined extraction method of ultrasound-assisted extraction (UAE) in conjunction with solid phase extraction (SPE) was applied to isolation and enrichment of selected drugs (metoprolol, ticlopidine, propranolol, carbamazepine, naproxen, acenocumarol, diclofenac, ibuprofen) from fish tissues. The extracted analytes were separated and determined by ultra-high performance liquid chromatography with UV detection (UHPLC–UV) technique. The selectivity of the developed UHPLC–UV method was confirmed by comparison with ultra-high performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) analysis.The important parameters, such as composition of type and pH of extraction solvent, solid/liquid rate volume of extraction solvent and number of extraction cycles were studied. The ultrasonic parameters, such as time, power and temperature of the process were optimized by using a half-fraction factorial central composite design (CCD). The mixture of 10 mL of methanol and 7 mL of water (pH 2.2) (three times) was chosen for the extraction of selected drug from fish tissues. The results showed that the highest recoveries of analytes were obtained with an extraction temperature of 40 °C, ultrasonic power of 300 W, extraction time of 30 min.Under the optimal conditions, the linearity of method was 0.12–5.00 μg/g. The determination coefficients (R²) were from 0.979 to 0.998. The limits of detection (LODs) and limits of quantification (LOQs) for the extracted compounds were 0.04–0.17 μg/g and 0.12–0.50 μg/g, respectively. The recoveries were between 85.5% and 115.8%.     

Intensification of extraction of curcumin from Curcuma amada using ultrasound assisted approach: Effect of different operating parameters

Curcumin, a dietary phytochemical, has been extracted from rhizomes of Curcuma amada using ultrasound assisted extraction (UAE) and the results compared with the conventional extraction approach to establish the process intensification benefits. The effect of operating parameters such as type of solvent, extraction time, extraction temperature, solid to solvent ratio, particle size and ultrasonic power on the extraction yield have been investigated in details for the approach UAE. The maximum extraction yield as 72% was obtained in 1 h under optimized conditions of 35 °C temperature, solid to solvent ratio of 1:25, particle size of 0.09 mm, ultrasonic power of 250 W and ultrasound frequency of 22 kHz with ethanol as the solvent. The obtained yield was significantly higher as compared to the batch extraction where only about 62% yield was achieved in 8 h of treatment. Peleg’s model was used to describe the kinetics of UAE and the model showed a good agreement with the experimental results. Overall, ultrasound has been established to be a green process for extraction of curcumin with benefits of reduction in time as compared to batch extraction and the operating temperature as compared to Soxhlet extraction.     

Purification of serratiopeptidase from Serratia marcescens NRRL B 23112 using ultrasound assisted three phase partitioning

The ultrasound assisted three phase partitioning (UATPP) is a novel bioseparation method for separation and purification of biomolecules. In the present work, UATPP was investigated for the first time for purification of serratiopeptidase from Serratia marcescens NRRL B 23112. Effect of various process parameters such as ammonium sulphate saturation, t-butanol to crude extract ratio, pH, ultrasonic frequency, ultrasonic intensity, duty cycle and irradiation time were evaluated and optimized. The optimized conditions were found to be as follows: ammonium sulphate saturation 30% (w/v), pH 7.0, t-butanol to crude ratio 1:1 (v/v), ultrasound frequency 25 kHz, ultrasound intensity 0.05 W/cm², duty cycle 20% and irradiation time 5 min. The maximum purity and recovery obtained from UATPP was 9.4-fold and 96% respectively as compared to the three phase partitioning (TPP) (4.2-fold and 83%). Also the process time for UATPP was significantly reduced to 5 min from 1 h as compared to TPP. The results indicate that, UATPP is an efficient technique for the purification of serratiopeptidase with maximum purity, recovery and reduced processing time.     

Effects of various factors of ultrasonic treatment on the extraction yield of all-trans-lycopene from red grapefruit (Citrus paradise Macf.)

The effects of various factors, including the extraction time, temperature, solvent/material ratio, the ultrasonic intensity and duty cycle of ultrasonic irradiation on the extraction yield of all-trans-lycopene from red grapefruit by ultrasound-assisted extraction (UAE) were investigated. In comparison with conventional solvent extraction (CSE), UAE showed a pronounced greater extraction yield and reduced extraction time effectively with a peak value at 30 min. The extraction yield was significantly influenced by temperature and the optimum condition was 30 °C. The extraction yield increased with increasing of solvent/material ratio until equilibrium was arrived at the optimal ratio of 3:1 (mL/g). The extraction yield increased first and then decreased with an increase in ultrasonic intensity. The extraction yield of UAE increased with the increase of duty cycle, whereas pulsed ultrasound with proper intervals was more efficient than continuous ultrasonication. The degradation via isomerisation of all-trans-lycopene under ultrasonic treatment was also observed with the formation of 9,13′-di-cis-, 9,13-di-cis-, 15-cis-, 13-cis- and 9-cis-lycopene isomers which were tentatively identified by HPLC-PAD.     

Hydrolytic enzymes in activated sludge: Extraction of protease and lipase by stirring and ultrasonication

Hydrolytic enzymes released by the microorganisms in activated sludge are responsible for the organic matter degradation; however, the optimal extraction procedure of this valuable resource has not been well established until now. The present study evaluates the recovery of protease and lipase from the activated sludge by using stirring and ultrasonication, varying different parameters such as extraction time, concentration of additives (Triton X100, Cation Exchange Resin and Tris buffer), stirring velocity, ultrasonic power and sludge source. Sludge was collected from two urban wastewater treatment plants located in Prague (Czech Republic) and Reus (Spain). It was found that stirring using 2% v/v Triton X100 for 1 h was enough to extract 57.4 protease units/g VSS, and that the same method using a combination of 10 mM Tris pH 7.5 + 0.48 g/mL CER + 0.5% TX100 as an additive allowed to extract 15.5 lipase units/g VSS from sludge collected from Reus Wastewater Treatment Plant. Ultrasonication allowed reducing the extraction time to 10 min for protease (using 2% v/v Triton X100 yielding 52.9 units/g VSS) and to 20 min for lipase (without any additive yielding nearly 21.4 units/g VSS), which makes this method appropriate for the extraction of enzymes from the activated sludge, and suitable to be scaled up for its application in the industry.     

Ultrasonically enhanced extraction of luteolin and apigenin from the leaves of Perilla frutescens (L.) Britt. using liquid carbon dioxide and ethanol

The present study reports on the ultrasonic enhancement of the liquid carbon dioxide (CO₂) extraction of luteolin and apigenin from the leaves of Perilla frutescens (L.) Britt., to which ethanol is added as a cosolvent. The purpose of this research is also to investigate the effects of the particle size, temperature, pressure, irradiation power, irradiation time, and ethanol content in the liquid CO₂ solution on the extraction yield using single-factor experiments. We qualitatively and quantitatively analyzed the yields in the extract using HPLC (high-performance liquid chromatography). The liquid CO₂ mixed with ethanol was used at temperatures of 5, 20 and 25 °C with extraction pressures from 8 to 14 MPa. The yields of luteolin and apigenin in the extraction were clearly enhanced by the ultrasound irradiation, but the selectivity of the extract was not changed. The yields of luteolin and apigenin in the extract were also significantly improved by adjusting the operating temperature, the irradiation time, and the ethanol content in the liquid CO₂ solution, but no change in the selectivity of the extract was observed.     

Ultrasound-assisted extraction of phenolic compounds from Laurus nobilis L. and their antioxidant activity

Bay leaves (BL) (Laurus nobilis L., Family: Laureceae) are traditionally used to treat some symptoms of gastrointestinal problems, such as epigastric bloating, impaired digestion, eructing and flatulence. These biological properties are mainly attributed to its phenolic compounds. In this paper, ultrasound-assisted extraction of phenolic compounds from Laurus nobilis L. (Laureceae) was studied.Effects of several experimental factors, such as sonication time, solid/liquid ratio and concentration of solvent on extraction of phenolic compounds were evaluated through a randomized complete block design with factorial treatment arrangement (3³). The best extraction conditions were: 1 g plant sample with 12 mL of 35% ethanol, for 40 min, obtaining a yield of phenolic compounds of 17.32 ± 1.52 mg g^?1 of plant. In addition, free radical-scavenging potential of DPPH and lipid oxidation inhibition, by linoleic acid peroxidation of the selected extract was measured in order to evidence their antioxidant properties. Results indicated that high amounts of phenolic compounds can be extracted from L. nobilis by ultrasound-assisted extraction technology.     

Ultrasound stimulated production of a fibrinolytic enzyme

The present study is aimed at enhanced production of a fibrinolytic enzyme from Bacillus sphaericus MTCC 3672 under ultrasonic stimulation. Various process parameters viz; irradiation at different growth phases, ultrasonication power, irradiation duration, duty cycle and multiple irradiation were studied for enhancement of fibrinolytic enzyme productivity. The optimum conditions were found as follows, irradiation of ultrasonic waves to fermentation broth at 12 h of growth phase with 25 kHz frequency, 160 W ultrasound power, 20% duty cycle for 5 min. The productivity of fibrinolytic enzyme was increased 1.82-fold from 110 to 201 U/mL compared with the non sonicated control fermentation. Drop in glucose concentration from 0.41% to 0.12% w/v in ultrasonicated batch implies that, ultrasonication increases the cell permeability, improves substrate intake and progresses metabolism of microbial cell. Microscopic images before and after ultrasonic stimulation clearly signifies the impact of duty cycle on decreasing biomass concentration. However, environmental scanning electron micrograph does not show any cell lysis at optimum ultrasonic irradiation. Offshoots of our results will contribute to fulfill the demand of enhancement of microbial therapeutic enzyme productivity, through ultrasonication stimulation.     

Aqueous two-phase system coupled with ultrasound for the extraction of lignans from seeds of Schisandra chinensis (turcz.) Baill

In this study the potential use of an aqueous two phase system (ATPS) coupled with ultrasound for the extraction of lignans from Schisandra chinensis seeds was evaluated and optimized using response surface methodology (RSM). The main bioactive components, schizandrin (SA), schisantherin A (SAA) and deoxyschizandrin (DSA) were selected as markers. The partitioning behavior of lignans in different salt-types of ATPS was compared. The optimization ATPS of 25% (w/w) (NH₄)₂SO₄ and 19% (w/w) ethanol were selected based on their higher upper phase partitioning coefficient (>74) and the recovery (>93%) for three markers. Using the optimized ATPS solvent, the RMS results showed 20:1 of solvent:solid, 800 W and 61.1 min were the optimal ultrasound assisted extraction conditions, under which 13.10 mg/g SA, 1.87 mg/g SAA and 1.84 mg/g DSA were recovered in the upper phase, whereas the wasted stigmas accumulated in the lower phase. Compared with 80% ethanol (v/v) ultrasonic extraction, similar yields were obtained, but the present method exhibited higher extraction purity for the selective extraction of lignans from S. chinensis seeds.     

Esterification of sodium 4-hydroxybenzoate by ultrasound-assisted solid–liquid phase-transfer catalysis using dual-site phase-transfer catalyst

The catalytic esterification of sodium 4-hydroxybenzoate with benzyl bromide by ultrasound-assisted solid–liquid phase-transfer catalysis (U-SLPTC) was investigated using the novel dual-site phase-transfer catalyst 4,4′-bis(tributylammoniomethyl)-1,1′-biphenyl dichloride (BTBAMBC), which was synthesized from the reaction of 4,4′-bis(chloromethyl)-1,1′-biphenyl and tributylamine. Without catalyst and in the absence of water, the product yield at 60 °C was only 0.36% in 30 min of reaction even under ultrasound irradiation (28 kHz/300 W) and 250 rpm of stirring speed. When 1 cm³ of water and 0.5 mmol of BTBAMBC were added, the yield increased to 84.3%. The catalytic intermediate 4,4′-bis(tributylammoniomethyl)-1,1′-biphenyl di-4-hydroxybenzoate was also synthesized to verify the intrinsic reaction which was mainly conducted in the quasi-aqueous phase locating between solid and organic phases. Pseudo-first-order kinetic equation was used to correlate the overall reaction, and the apparent rate coefficient with ultrasound (28 kHz/300 W) was 0.1057 min⁻¹, with 88% higher than that (0.0563 min⁻¹) without ultrasound. The esterification under ultrasonic irradiation using BTBAMBC by solid–liquid phase-transfer catalysis was developed.     

Ultrasound assisted acid catalyzed lactose hydrolysis: Understanding into effect of operating parameters and scale up studies

The current work deals with the value addition of lactose by transforming into hydrolyzed lactose syrup containing glucose and galactose in major proportion using the novel approach of ultrasound assisted acid catalyzed lactose hydrolysis. The hydrolysis of lactose was performed in ultrasonic bath (33 kHz) at 50% duty cycle at different temperatures as 65 °C and 70 °C and two different hydrochloric acid (HCl) concentrations as 2.5 N and 3 N. It was observed that acid concentration, temperature and ultrasonic treatment were the major factors in deciding the time required to achieve ∼90% hydrolysis. The ultrasonic assisted approach resulted in reduction in the reaction time and the extent of intensification was established to be dependent on the temperature, acid concentration and time of ultrasonic exposure. It was observed that the maximum process intensification obtained by introduction of ultrasound in the lactose hydrolysis process performed at 70 °C and 3 N HCl was reduction in the required time for ∼90% hydrolysis from 4 h (without the presence of ultrasound) to 3 h. The scale-up study was also performed using an ultrasonic bath with longitudinal horn (36 kHz as operating frequency) at 50% duty cycle, optimized temperature of 70 °C and acid concentration of 3 N. It was observed that the reaction was faster in the presence of ultrasound and stirring by axial impeller at rpm of 225 ± 25. The time required to complete ∼90% of hydrolysis remained almost the same as observed for small scale study on ultrasonic bath (33 kHz) at 50% duty cycle. The use of recovered lactose from whey samples instead of pure lactose did not result in any significant changes in the progress of hydrolysis, confirming the efficacy of the selected approach. Overall, the work has presented a novel ultrasound assisted approach for intensified lactose hydrolysis.