Optimization of multi-stage countercurrent extraction of antioxidants from Ginkgo biloba L. leaves

Multi-stage countercurrent extraction (MCE) as a novel extraction technique was used to extract antioxidants from Ginkgo biloba leaves. Orthogonal array design (OAD) was employed to optimize the ratio of 60% ethanol to raw material (8–16 mL/g), extraction time (30–60 min) and extraction temperature (60–80 °C) to obtain a high yield of antioxidants from G. biloba leaves by MCE. The optimum conditions were a ratio of 60% ethanol to raw material of 16 mL/g and extraction time of 30 min at 80 °C. Under these conditions, the yields of flavonoids and total phenolics were 1.74% and 2.42%, respectively, and DPPH radicals scavenging activity of the extract was 89.97%. Compared with heat-reflux extraction, MCE had obvious advantages of less extraction time and lower solvent and energy consumption. It may be used as a promising technique for the extraction of bioactive compounds from plant materials.     

Optimization of supercritical fluid extraction of phytosterol from roselle seeds with a central composite design model

Recovery of phytosterol from roselle (Hibiscus sabdariffa L.) seeds via supercritical carbon dioxide extraction modified with ethanol was investigated at pressures of 200–400 bar, temperatures from 40 to 80 °C and at supercritical fluid flow rates from 10 to 20 ml/min. It was found that an entrainer such as ethanol could enhance the solubility and extraction yield of roselle seed oil from the seed matrix, compared to values obtained using supercritical CO₂. After a typical run (holding period of 30 min, continuous flow extraction of 3 h), the results indicate that the oil recovery was optimal with a recovery of 108.74% and a phytosterol composition of 7262.80 mg kg^?1 at relatively low temperature of 40 °C, a high pressure of 400 bar and at a high supercritical fluid flow rate of 20 ml/min in the presence of 2 ml/min EtOH as entrainer. The solubility of roselle seed oil increased with temperature at the operating pressures of 200, 300 and 400 bar. Supercritical fluid extraction involved a short extraction time and the minimal usage of small amounts of entrainer in the CO₂.     

Optimization of the supercritical fluid extraction of triterpenic acids from Eucalyptus globulus bark using experimental design

The supercritical CO₂ extraction of E. globulus deciduous bark was carried out at different temperatures (40–60 °C), pressures (100–200 bar), and ethanol contents (0.0–5.0 wt. %) to study triterpenic acids (TTAs) recovery. A factorial design of experiments and response surface methodology were implemented to analyze the influence of these variables upon extraction and perform its optimization. The best conditions were 200 bar, 40 °C and 5% ethanol, for which the statistically validated regression models provided: extraction yield of 1.2% (wt.), TTAs concentration of 50%, which corresponds to TTAs yield of 5.1 g/kg of bark and a recovery of 79.2% in comparison to the Soxhlet value. The trends of the free and acetylated TTAs were very different, due to their distinct CO₂-philic character caused by dissimilar polarities: the acetyl derivatives approached a plateau near 200 bar and 5% ethanol, while the free TTAs extraction always increased in the range of conditions studied.     

Supercritical fluid extraction of Prunus persica leaves and utilization possibilities as a source of phenolic compounds

Mediterranean countries contribute highly on world peach production and tonnes of waste leaves are released due to pruning. The aim of this study was to investigate the utilization possibilities of the leaves by supercritical fluid extraction. A statistical design was used to evaluate the effect of temperature (40–80 °C), pressure (150–300 bar) and concentration of ethanol as co-solvent (6–20%) at a flow rate of 15 g/min and for a duration of 60 min. The most effective variables were found as pressure and co-solvent ratio (p < 0.005). Optimum extraction conditions were elicited as 60 °C, 150 bar and 6% co-solvent yielding a total phenol content of 79.92 mg GAE/g extract, EC₅₀ value of 232.20 μg/ml and a radical scavenging activity of 53.25% which was higher than the value obtained by conventional solvent extraction method (32.23%). Consequently, Prunus persica L. leaves were found as a potential phenolic source for industrial applications.     

A bioactivity based comparison of Echinacea purpurea extracts obtained by various processes

Echinacea species is provided as dietary supplements for various infectious and immune related disorders and has a potential role in cancer prevention. The aim of this study was to optimize the extraction of total flavonoids using different extraction methods and investigate the cytotoxic effects on various cancer cell lines (CaCo-2, MCF-7, A549, U87MG, and HeLa) and VERO (African green monkey) as a non-cancerous cell line. Box-Behnken statistical design was used to evaluate the effect of pressure (100–200 bar), temperature (40–80 °C) and ethanol as co-solvent (6–20 wt%) at a flow rate of 15 g/min for 60 min in supercritical CO₂ extraction and the effect of temperature (60–100 °C), time (5–15 min) and power (300–900 W) in microwave-assisted extraction. Optimum extraction conditions were elicited as 300 bar, 80 °C and 13% co-solvent yielding 0.472 mg rutin equivalent total flavonoids/g extract in SC-CO₂ extraction, whereas 60 °C, 10 min and 300 W yielded the highest (0.202 mg rutin equivalent) total flavonoids in microwave-assisted extraction. Additional trials with subcritical water (0.022 mg/g) and Soxhlet extraction with methanol (0.238 mg/g) yielded lower flavonoid contents. The exposures upto 50 μg/ml of extracts revealed no significant inhibition on the proliferation of both tested cancer cells and healthy VERO cells.     

Optimization of subcritical water extraction of antioxidants from Coriandrum sativum seeds by response surface methodology

Subcritical water extraction (SWE) of antioxidants from Coriandrum sativum seeds (CSS) was optimized by simultaneous maximization of the total phenolics (TP) and total flavonoids (TF) yield and antioxidant activity, using IC₅₀ value. Box–Behnken experimental design (BBD) on three levels and three variables was used for optimization together with response surface methodology (RSM). Influence of temperature (100–200 °C), pressure (30–90 bar) and extraction time (10–30 min) on each response was investigated. Experimentally obtained values were fitted to a second-order polynomial model and multiple regression. Analysis of variance (ANOVA) was used to evaluate model fitness and determine optimal conditions. Moreover, three-dimensional surface plots were generated from employed mathematical model. The optimal SWE conditions obtained in simultaneous optimization were temperature of 200 °C, pressure of 30 bar and extraction time of 28.3 min, while obtained values of TP and TF yields and IC₅₀ value at this experimental point would be 2.5452 g GAE/100 g CSS, 0.6311 g CE/100 g CSS and 0.01372 mg/ml, respectively. Moreover, good and moderate linear correlation was observed between antioxidant activity (IC₅₀ value) and total phenolics content (R² = 0.965), and total flavonoids content (R² = 0.709) which indicated that these groups of compounds are responsible for antioxidant activity of C. sativum extracts.     

Optimization of the supercritical fluid coextraction of oil and diterpenes from spent coffee grounds using experimental design and response surface methodology

The reported work aimed at the optimization of operating conditions of the supercritical fluid extraction (SFE) of spent coffee grounds (SCG) using pure or modified CO₂, with particular emphasis on oil enrichment with diterpenes like kahweol, cafestol and 16-O-methylcafestol. The analysis comprised the application of Box–Behnken design of experiments and response surface methodology, and involved three operating variables: pressure (140–190 bar), temperature (40–70 °C) and cosolvent (ethanol) addition (0–5 wt.%). The best conditions to maximize total extraction yield are 190 bar/55 °C/5 wt.% EtOH, leading to 11.97% (g_oil/100 g_SCG). In terms of the concentration of diterpenic compounds in the supercritical extracts, the best operating conditions are 140 bar/40 °C/0 wt.% EtOH, providing 102.90 mg g⁻¹_oil. The measurement of extraction curves near optimized conditions (140 bar/55 °C/0 wt.% EtOH and 190 bar/55 °C/0 wt.% EtOH) confirmed the trends of the statistical analysis and revealed that SFE enhances diterpenes concentration by 212–410% at the expenses of reducing the extraction yield between 39% and 79% in comparison to n-hexane extraction.     

Extraction of caffeine from natural matter using a bio-renewable agrochemical solvent

This paper reports experimental data on the pressurized liquid extraction of caffeine from green coffee beans and green tea leaves using ethyl lactate (ethyl 2-hydroxy-propanoate). This solvent is a new bio-renewable agrochemical solvent, naturally produced by fermentation from corn derived feedstock, which has been recently considered as a very suitable and environmental benign solvent for food industrial applications.Static extraction assays (one step during 10 min) were carried out in an Accelerated solvent extraction (ASE) system at three different extraction temperatures, namely 100, 150 and 200 °C. Extraction yield and caffeine recovery were determined and compared with those obtained when using other liquid solvents, such as ethyl acetate or ethanol. High recovery of caffeine (≈60%) was found in the extracts produced using ethyl lactate, which demonstrates the potential use of this green solvent for the extraction of caffeine from different vegetable sources.     

Sequential extraction of bioactive compounds from Melia azedarach L. in fixed bed extractor using CO2, ethanol and water

Melia azedarach L. is a plant with wide use in folk medicine since it contains many bioactive compounds of interest. The present study aimed to extract bioactive compounds from M. azedarach fruits by a sequential process in fixed bed using various solvent mixtures. Extractions were performed at 50 °C and 300 bar in four sequential steps using supercritical CO₂ (scCO₂), scCO₂/ethanol, pure ethanol, and ethanol/water mixture as solvents, respectively. The efficacy of the extraction process was evaluated by extraction yield and kinetics, and analysis of extracts by: (1) thin layer chromatography (TLC), (2) phenolics content, (3) reduction of surface tension of water, (4) gas chromatography (GC–MS), (5) electrospray ionization mass spectrometry (ESI–MS) and (6) antiviral activity. The overall extraction yield reached 45% and TLC analysis showed extracts with different composition. extract obtained from CO₂/ethanol mixture (SCEE) exhibited the greatest ability to reduce surface tension of water from 72.4 mN m⁻¹ [1] of pure water to 26.9 mN m⁻¹ of an aqueous solution of 40 g L⁻¹. The highest phenolics contents were observed in both the hydroalcoholic extract and scCO₂/ethanolic extract. Volatile oils were not detected in the supercritical extracts by GC–MS. MS analyses identified the fatty acids: linoleic, palmitic and myristic acid in the supercritical extract (SCE), and the phenolics: caffeic acid and malic acid in the other extracts. In addition, SCE and SCEE extracts showed significant inhibition percentage against Herpes Simplex Virus Type 1. The extraction process proposed in the present study produced extracts with significant potential for application in food and pharmaceutical industries.     

Supercritical carbon dioxide extraction of bioactive flavonoid from Strobilanthes crispus (Pecah Kaca)

The bioactive flavonoid compounds of Strobilanthes crispus (Pecah Kaca) leaves obtained by using supercritical carbon dioxide (SC-CO₂) extraction were investigated and the obtained crude extract yields were compared in order to select the best operation parameters. Since carbon dioxide is a non-polar solvent, ethanol was used as co-solvent to increase the polarity of the fluid. The studied parameters were pressure (100, 150 and 200 bar), temperature (40, 50 and 60 °C) and dynamic extraction time (40, 60 and 80 min). The optimum extraction condition occurred at 200 bar, 50 °C and 60 min. Based on the mean value, pressure had dominant effect on the extraction yield. Apart from the optimum SFE conditions two other conditions namely at minimum (100 bar, 40 °C, 40 min) and maximum (200 bar, 60 °C, 80 min) levels of each studied parameters as control runs were analyzed by HPLC to determine the major bioactive flavonoid compounds from S. crispus. Under the optimum conditions eight flavonoid compounds were identified; they were (+)-catechin, (?)-epicatechin, rutin, myricetin, luteolin, apigenin, naringenin and kaempferol.     

Extraction of Epigallocatechin-3-gallate from green tea via supercritical fluid technology: Neural network modeling and response surface optimization

In this study the extraction of (−)-Epigallocatechin-3-gallate (EGCG) from Iranian green tea was investigated by supercritical CO₂ with ethanol as co-solvent. Design of experiments and modeling were carried out with response surface methodology by Minitab software. The HPLC analysis of the extracted samples was used in conjunction with response surface design to optimize four operating variables of supercritical CO₂ extraction (pressure, temperature, CO₂ flow rate and extraction dynamic time). Optimum recovery of EGCG (0.462 g/g) was obtained at 19.3 MPa, 43.7 °C, 106 min (dynamic) and 1.5 ml/min (CO₂ flow rate). Moreover, a three-layer artificial neural network was developed for modeling EGCG extraction from green tea. In this regard, different networks (by changing the number of neurons in the hidden layer and algorithm of network training) were compared with evaluation of networks accuracy in extraction recovery prediction. Finally, the Levenberg–Marquardt algorithm with the six neurons in the hidden layer has been found to be the most suitable network.     

Extraction of low-molar-mass phenolics and lipophilic compounds from Pinus pinaster wood with compressed CO2

Near-supercritical and supercritical CO₂ was used to extract low-molar-mass phenolics and lipophilic compounds from Pinus pinaster wood. Extraction of samples containing sapwood and knotwood was carried out at 10⿿25 MPa and 30⿿50 °C to assess the influence of the operational conditions on the yields of total extracts and phenolics, as well as on the radical scavenging capacity of extracts. The use of ethanol as a co-solvent increased both the extraction yields and the concentration of phenolics in extracts. Operating under selected conditions (25 MPa, 50 °C, 10% ethanol), the extraction yield accounted for 4.1 wt% of the oven-dry wood. The extracts contained up to 7.6 g of phenolic compounds (measured as gallic acid equivalents) per 100 g extract, and showed one third of the radical scavenging capacity of Trolox. Native resin acids accounted for about 24 g per 100 g extracts, whereas flavonoids, lignans, stilbenes and juvabiones were found at lower proportions.     

Black sesame (Sesamum indicum L.) seeds extracts by CO2 supercritical fluid extraction: Isotherms of global yield,kinetics data,total fatty acids,phytosterols and neuroprotective effects

In Brazilian folk medicine, black sesame (Sesamum indicum L.) seeds are one of the most important ingredients present in the tea used to treat stroke victims. Nevertheless, extracts by supercritical extraction of black sesame seeds have not been applied in neuroprotection studies. The objective of this work was to investigate some process variables of supercritical fluid extraction in black sesame to generate extracts applicable in stroke research. Two isothermal (40 and 60 °C) were explored, combined with pressures that ranged from 200 to 400 bar, at a constant mass flow rate of 5.9 × 10⁻⁵ kg/s. The global yields ranged from 37 to 53% in dry basis. The highest mass yield was obtained at 60 °C and 400 bar. The fatty acid composition comprised a high unsaturated/saturated ratio. Chromatographic analysis of phytosterol content in the high global yield extract revealed higher amounts of β-sitosterol + sitostanol, cholesterol, campesterol + campestanol + 24-methylene cholesterol, Δ-5 avenasterol and stigmasterol, while lower levels of Δ-5,24 stigmastadienol, brassicasterol, clerosterol + Δ-5-23 stigmastadienol, Δ-7 avenasterol, eritrodiol and Δ-7 stigmasterol were present in the black sesame extract. Overall extraction curves from the supercritical fluid extraction (SFE) in lower and higher global yields (200 and 400 bar at 60 °C) fitted the Tan and Liou, Goto et al. and Sovová models. These models presented the best residual sum of squares values. Pilot experiments suggest that black sesame extract is neuroprotective following endothelin-1-induced focal ischemia into the motor cortex of adult male rat, observing a decrease in leukocyte infiltration in the group treated with SFE of black sesame seeds when compared with control group.     

Supercritical extraction of carob kibbles (Ceratonia siliqua L.)

Carob pulp kibbles, a by-product of carob been gum production, was studied as a source of bioactive agents. Firstly, the carob kibbles were submitted to an aqueous extraction to extract sugars, and supercritical fluid extraction (SFE) was applied to the solid residue of that aqueous extraction, by using compressed carbon dioxide (SC-CO₂) as the solvent and a mixture of ethanol and water (80:20, v/v) as a co-solvent. Pressure and temperature were studied in the ranges 15–22 MPa, and 40–70 °C. Particle diameter, and co-solvent percentage in ranges of 0.27–1.07 mm, and 0–12.4%, respectively, were also studied, as well as the flow rate of SC-CO₂ between 0.28 and 0.85 kg h⁻¹, corresponding, respectively, to 0.0062 and 0.0210 cm s⁻¹ of superficial velocity. The extracts were characterised in terms of antioxidant capacity by DPPH method, and total phenolics content by the Folin–Ciocalteu method. The central composite non-factorial design was used to optimise the extraction conditions, using the Statistica, version 6 software (Statsoft). The best results, in terms of yield and antioxidant capacity, were found at 22 MPa, 40 °C, 0.27 mm particle size, about 12.4% of co-solvent and a flow rate of 0.29 kg h⁻¹ of SC-CO₂. The phenolics profile of the extracts obtained at these conditions was qualitatively evaluated by HPLC-DAD. The solid residue of the supercritical extraction was also studied showing to be a dietary fiber, which can be compared to Caromax™, a carob fiber commercialised by Nutrinova Inc.     

Subcritical water extraction of steviol glycosides from Stevia rebaudiana leaves and characterization of the raffinate phase

The objectives of this work were to obtain steviol glycosides of S. rebaudiana leaves, possessing natural and noncaloric sweetener properties, using subcritical water extraction; to assess optimum extraction conditions; to determine biological activities of Stevia extracts and to characterize the raffinate phase. A Box–“Bhenken” statistical design was used to evaluate the effects of various values of temperature (100–150 °C), time (30–60 min) and flow rate (2–6 ml/min) at a pressure of 230 bar applying a solid/liquid ratio of 1:10 (m:v). The most effective parameter was temperature (p < 0.005). Optimum extraction conditions were elicited as 125 °C, 45 min, 4 ml/min flow rate which yielded 38.67 mg/g stevioside and 35.68 mg/g rebaudioside A. The total phenolic, flavonoid contents and DPPH free radical scavenging activity were found as 48.63 mg gallic acid/g extract, 29.81 mg quercetin/g extract and 92.50%, respectively. After extraction, total chlorophyll, carotenoid contents and dietary fibers were quantified as 31.91 mg/100 g, 5.71 mg/100 g and 4.98% in the raffinate phase. Hence, both extract and raffinate phases of S. rebaudiana leaves can be utilized as sources of natural sweeteners, fibers and coloring agents in the industry.     

Removal of contaminants from polluted drilling mud using supercritical carbon dioxide extraction

Liquid drilling fluid is often called drilling mud is heavy, viscous fluid mixtures use to carry rock cuttings to the surface and lubricate and cool the drill bit. During carrying cuttings they contaminated which not only reduce their functionality but also make them a hazardous and dangerous wastes which cannot be discharged anywhere without treatment. Due to this fact, in the present study, supercritical extraction process was used to remove contaminants from the drilling mud. Regarding this, effect of different parameters including extraction temperature (313–338 K) and pressure (100–200 bar), flow rate of CO₂ (0.05–0.36 cm³/s) and static time (20–130 min) on the removal of contaminations from drilling mud was examined using the design of experiment of changing one factor at a time. The obtained results revealed that the optimum operational conditions that lead to the highest removal degree of contaminations are temperature and pressure of 333 K and 180 bar, respectively, flow rate of lower than 0.1 cm³/s and the static time of 110 min. In addition, to examine the effect of the supercritical extraction on the crystalline structure modification and removal contaminations X-ray diffraction (XRD) and scanning electron microscopy (SEM) analyses were performed which confirmed the successful removal of contaminations from the drilling mud without significant crystalline modification.     

Extraction of caffeine from tea stalk and fiber wastes using supercritical carbon dioxide

The purpose of this study was to investigate obtaining of caffeine from tea plant wastes by supercritical carbon dioxide extraction. Experiments were carried out with tea stalk and fiber wastes of Turkish tea plants that has no economical value. Stalk and fiber wastes were supplied from tea factories. These wastes were ground, sieved and dried at 105 °C temperature in an oven. Parameters affecting caffeine leaching from tea wastes were determined to be, extraction time, extraction temperature, carbon dioxide flow rate, process pressure and particle size. The maximum yield of caffeine from tea stalk wastes and fiber wastes were 14.9 mg/g tea stalk and 19.2 mg/g tea fiber, respectively. The yield increase had been recorded as 61.9% and 65.5%, respectively, in comparison with the chloroform extraction of tea stalk and fiber wastes.     

Decaffeination of guaraná seeds in a microextraction column using water-saturated CO2

Experimental data on the reduction of the caffeine content of guaraná seeds with water-saturated supercritical carbon dioxide were obtained in order to provide information on the ability of supercritical fluids for the decaffeination of widely consumed caffeine-rich natural products. The extraction was performed using a semi-continuous flow high-pressure micro-extraction apparatus at 40 and 70 °C and pressures of 100, 200 and 400 bar. Carbon dioxide flow rates of 5.7 and 9.4 g min⁻¹ were used. The extraction curves obtained showed the existence of thermodynamic solubility dependent, intermediate and diffusion controlled regions. Extraction at 400 bar and 70 °C using water-saturated supercritical carbon dioxide at a flow rate of 5.7 g min⁻¹ allowed the removal of almost 98% of the initial caffeine content in wet ground guaraná seeds, in a period of 240 min. When extractions were performed at lower pressures or temperatures, additional time and larger amounts of carbon dioxide were necessary to achieve the same yield. Increasing carbon dioxide flow rate did not present any economic advantage unless the extraction was limited to the thermodynamic solubility region. For total extraction of caffeine, the use of low flow rates resulted in a similar final product yield but at a much lower solvent consumption. A retrograde behavior for the extraction of caffeine from guaraná seeds was also observed at 100 bar for the 40 and 70 °C isotherms.     

Pressurized hot water extraction of β-glucans from waxy barley

Pressurized hot water (PHW) is a new and promising solvent for the extraction of β-glucans from cereals. The effect of temperature, extraction time and pressure has been studied for the extraction of β-glucans from waxy barley (6.0% in β-glucans). Extraction yield, molecular weight and degradation products have been evaluated. Temperature (110–180 °C) and extraction time (15–75 min) showed a strong influence on extraction process. Extraction yield and MW are optimized for 155 °C, 18 min and 50 bar, leading to 53.7% extraction yield and 200 kDa. The presence of degradation products, such as HMF, is negligible under these conditions. The comparison between these optimal results and that from a conventional process (55 °C, 3 h and ambient pressure) reveals a big difference in MW (200 kDa vs. 55 kDa, respectively). PHW causes a significantly reduction in extraction time and an increase of β-glucan MW of almost 4 times as major benefits.     

Relevance of ions in pressurized fluid extraction of carbohydrates and phenolics from barley hull

The removal of total carbohydrates and phenolics from the hull of a new barley variety BT 584 using solid-liquid batch and aqueous pressurized fluid (ethanol or ionic liquid) extractions were evaluated. Using pressurized fluids, temperature was the most significant variable for the extraction of biocompounds from barley hull. The highest phenolics extraction (189.1 ± 3.1 mg/g hull) was obtained using pressurized aqueous ionic liquid while the highest carbohydrates extraction (450.3 ± 7.8 mg/g hull) was obtained using pressurized aqueous ethanol. The predicted solubility of ferulic acid and glucose in water, ionic liquid or carbonic acid, and the acid dissociation constant of the compounds studied allowed proposing an extraction mechanism based on biomass interaction with ionic species formed during pressurized fluid extraction. The anionic species facilitated removal of phenolics while the interaction of cationic/anionic species facilitated removal of carbohydrates from barley hull biomass.