Bioactive extracts of orange (Citrus sinensis L. Osbeck) pomace obtained by SFE and low pressure techniques: Mathematical modeling and extract composition

The objective of this study was to obtain orange (Citrus sinensis L. Osbeck) pomace extract using supercritical fluid extraction (SFE) with CO₂ and with CO₂ and co-solvent. In order to evaluate the high pressure method in terms of process yield, extract composition and biological activity, low pressure methods were also applied to obtain orange extracts, such as ultrasound (UE) and soxhlet (SOX), with different organic solvents, and hydrodistillation (HD). The SFE conditions were temperatures of 313.15 K and 323.15 K and pressures from 100 to 300 bar. The SFE kinetics and mathematical modeling of the overall extraction curves (OEC) were also investigated. The antioxidant potential of the extracts was evaluated by the DPPH method, by the Folin-Ciocalteau method and by the β-carotene/linoleic acid bleaching method. The antimicrobial activity of the extracts was also studied. The main compounds identified were l-limonene, palmitic and oleic acids, n-butyl benzenesulfonamide and β-sitosterol.     

Extraction of antioxidant compounds from blackberry (Rubus sp.) bagasse using supercritical CO2 assisted by ultrasound

Supercritical carbon dioxide extraction (SFE) was performed to recover bioactive components from blackberry (Rubus sp.) industrial residues. Ultrasound was applied during the extractions in order to enhance rate and yield. Moreover, water and ethanol at different proportions were used as cosolvents to improve the extraction of polar compounds from the residues. The extraction global yields were measured at all performed conditions (pressure, temperature, ultrasound power and cosolvent). The extracts were evaluated in terms of their antioxidant activity, measured using two methods, phenolic content, monomeric anthocyanins, and anthocyanin profile. The application of ultrasound in SFE helped increasing the extraction rate at the beginning of the process, which could be observed on the extraction curves at 15 MPa, the lowest pressure applied. Scanning electron microscopy (FESEM) was used to analyze the blackberry bagasse undergoing SFE with and without ultrasounds and showed that ultrasound disturbs the cell walls, enhancing the release of the extractable compounds. The extracts have shown high antioxidant activity and phenolic contents when obtained at higher temperatures. Regarding anthocyanins, the use of water as cosolvent resulted in a significant increase. Four major anthocyanins were identified and quantified by ultra performance liquid chromatography (UPLC).     

Extraction of phenolic fraction from guava seeds (Psidium guajava L.) using supercritical carbon dioxide and co-solvents

In this work the supercritical fluid extraction (SFE) with carbon dioxide (CO₂) and with ethyl acetate (EtAc) and ethanol (EtOH) as co-solvents was applied to obtain the phenolic fraction from guava seeds (Psidium guajava L.). The extraction was explored at various operating conditions, using 10, 20 and 30 MPa and 40, 50 and 60 °C. The use of EtAc and EtOH as co-solvents in SFE was also studied. The supercritical process was compared with traditional techniques such as Soxhlet extraction using EtAc and EtOH as solvents. The quality of the different extracts, obtained using SFE and Soxhlet methods and different solvents, was evaluated through the antioxidant activity, obtained by the collection methods of scavenging DPPH and bleaching of β-carotene, and also through the total phenolic content (TPC) of the samples, by the Folin-Ciocalteu method. The antioxidant potential indicates the use of ethanol as co-solvent as the best modifier in SFE, used in concentration of 10% (w/w) at 50 °C and 30 MPa. The quality of the extracts obtained by SFE with EtOH varied with the operating conditions of temperature and pressure, with higher values obtained at 10 and 20 MPa for TPC results and also antioxidant methods. The process yield of the phenolic fraction was also evaluated for all the extraction procedures studied (SFE and Soxhlet), with results varying from 0.380 to 1.738% (w/w).     

Effects of Drying and Extraction Methods on the Quality and Antioxidant Activity of Sea Buckthorn (Hippophae rhamnoides) Berries and Leaves

Sea buckthorn is a promising source of bioactive compounds. However, there is limited information on the effect of post harvest drying and extraction of sea buckthorn on its antioxidant capacity. The effect of freeze, air, and solar dryingon the extraction yield of sea buckthorn berries and leaves was evaluated with respect to their bioactive content. Sequential extraction with solvents of different polarity and solvent-free microwave-assisted extraction were applied. Freeze-drying has better performance in the berries’ extracts, while air-drying has better performance in the leaves’ extracts. All data were analyzed by Multi Factor Analysis of Variance (ANOVA) and Duncan's post-hoc tests at a level of α = .05. Although the extraction yield in the sequential extraction of sea buckthorn berries is significantly better, the leaves’ extracts exhibit superior radical scavenging ability. Accelerated solar drying appears to preserve sea buckthorn leaves without degrading their antioxidant content. The components responsible for the high antioxidant activity of leaves’ extracts were found to be several flavonoids and polyphenols. The extract obtained by the microwave extraction of fresh berries exhibits moderate antioxidant activity compared to the polar extracts of freeze-dried berries.     

桉树叶片提取物的杀虫活性及应用初步研究

采用3种溶剂对桉树叶片进行加热回流提取,分别采用喷雾法和浸渍法测定提取物对鳞翅目害虫黏虫、半翅目害虫蚜虫及蜱螨目害虫朱砂叶螨生物活性。在初筛的基础上,对甲醇、丙酮和石油醚提取物进行了烟蚜复筛及温室盆栽试验。试验结果表明:各提取物对黏虫及朱砂叶螨在10000mg/L的浓度均未表现出杀虫活性,甲醇、丙酮和石油醚提取物对蚜虫均表现出一定的毒杀活性,对烟蚜的LC_(50)值分别为4 644.61、4 276.94、507.47 mg/L,其中石油醚提取物表现出了较高的生物活性。进行了进一步的温室盆栽验证试验,发现药后7 d,10000 mg/L的石油醚提取物对烟蚜的防效达到80.31%,有进一步研究的价值。     

An evaluation of supercritical fluid extraction as an analytical tool to determine fat in canola,flax, solin,and mustard

Supercritical fluid extraction (SFE) with carbon dioxide was used to extract oil from soft oilseeds (flax, solin, canola, and mustard). Oil content determinations from the SFE method AOCS Am 3–96, with and without ethanol as a modifier, were compared to results obtained with an exhaustive extraction using petroleum ether (FOSFA as in AOCS Am 2–93). Without the modifier, oil recoveries using SFE were 10 to 15% lower than oil contents by the FOSFA method for the flax and canola samples. For mustard, the oil recoveries by SFE were about 20 to 30% lower than oil contents by the FOSFA method. In the presence of the modifier, oil recoveries for flax and canola were about 3% lower than the FOSFA recoveries. Varying the time, temperature, and amount of modifier (ethanol) showed that recoveries increased with time, pressure, temperature, and amount of modifier independently of the oilseeds tested. Kinetics of the SFE extraction showed that the oil recoveries increased with the extraction time and reached a plateau after 60 min. Multiple extractions (2×30 min), however, gave better recoveries than a single extraction for the same amount of time (60 min). The best results were obtained using multiple extractions without modifier or a combination of multiple extractions first without and then with 15% modifier. Under these last two conditions, oil recoveries were close to 100% for flax, solin, and canola, but mustard oil recoveries were still 10% lower than recoveries using the FOSFA method. Mustard samples gave the lowest oil recovery from SFE when compared to FOSFA method recoveries whatever conditions were tested, suggesting a matrix effect on the oil recovery. The acyl lipid content of the various extracts was studied using the sum of all FA expressed as TAG as a measure of acyl lipid extraction. The acyl lipid contents of the extracts were close to 100% when no modifier was used during the SFE. In the presence of modifier, the acyl lipid contents of the extracts were 10 to 15% lower than the results obtained without modifier. The amount of acyl lipid in the extract decreased as the quantity of modifier increased. This suggests that increasing the ethanol modifier increased the amount of polar compounds extracted without significantly increasing the total amount of lipids. The FA profiles were constant throughout the various extraction procedures.     

海滩植物厚藤（Ipomoea Pes-caprae）抗氧化活性研究

采用加速溶剂萃取技术(ASE)快速制备厚藤活性提取物,并以DPPH法对厚藤茎叶不同溶剂提取物抗氧化活性进行评价。抗坏血酸抗氧化能力乙酸乙酯提取物抗氧化能力乙醇提取物抗氧化能力甲醇提取物抗氧化能力石油醚提取物抗氧化能力,其IC50值分别为0.203 4、0.836 2、0.868 8、1.867 5、6.628 3 g/L。结果显示,厚藤茎叶提取物具有一定的抗氧化活性,且以乙酸乙酯为提取溶剂时,效果最佳。     

Supercritical Fluid Extraction Of Lemon Verbena (Aloysia triphylla): Process Kinetics and Scale-Up,Extract Chemical Composition and Antioxidant Activity,and Economic Evaluation

The extract of lemon verbena ( Aloysia triphylla ) was obtained by supercritical fluid extraction (SFE) using laboratory and pilot scale equipments. The scale-up criterion selected (maintaining solvent to feed ratio constant) was successfully used for a 14-fold scale-up. The extract obtained in pilot scale was separated in three fractions, which were characterized as for their phytochemical profile, total flavonoids content, and antioxidant activity. The extracts obtained by SFE were compared to classical Soxhlet extraction method. Maximum yield obtained for SFE was 1.8%, and for Soxhlet, 7.1%. The chemical composition revealed different phytochemical profiles for SFE and Soxhlet extracts; the last ones presented more flavonoids, while SFE extracts were more concentrated in volatile compounds. The major compounds identified in the volatile fraction of the extracts were spathulenol, phytol and octadecatrienal. Some extracts presented pro-oxidant activity and others presented antioxidant activity. The SFE process was shown to be economically feasible for obtaining lemon verbena extracts; the minimum manufacturing cost (COM) obtained was US$ 1070.00/kg, with a payback time of 2 years.     

Antioxidant and radical-scavenging activities of buckwheat seed components

The search for endogenous components in food ingredients exhibiting antioxidant activity has been intensified in order to eliminate synthetic antioxidants. Tocopherols are widely used as natural antioxidants, although their protective ability is not always sufficient. Buckwheat seed components were evaluated for antioxidant and free radical-scavenging activities using solvents of different polarities to isolate components from hulls and groats. Components extracted from buckwheat hulls were pro-oxidant in canola oil. Antioxidant activity of extracts from buckwheat groats increased when more polar solvents were used for extraction. The highest activity was observed for the methanolic extract. Radical-scavenging activity of buckwheat extracts was analyzed with DPPH (2,2-diphenyl-2-picryl-hydrazyl). This activity increased when the more polar solvents were used for extraction, with the highest activity observed for the methanolic extract. It was also observed that the radical scavenging effectiveness of extracts was concentration dependent. Analysis revealed the presence of tocopherols in the hexane extract, while methanolic extracts were rich in phenolic acids and flavonoids.     

Radical-scavenging activity of extracts from Cordia verbenacea DC obtained by different methods

The possibility of using the leaves of Cordia verbenacea as a new source of natural antioxidant compounds was investigated. In the present work, extracts from C. verbenacea were obtained using different extraction methods: supercritical fluid extraction (SFE), Soxhlet (SE), hydrodistillation and maceration, with the objective to evaluate the methods in terms of yield and antioxidant potential. The high-pressure technique was applied using pure CO₂ and CO₂ with co-solvent at different temperatures and pressures (30, 40 and 50 °C and 100, 200, and 300 bar). Organic solvents with different polarities were used to obtain extracts by low-pressure extraction processes. The extracts were evaluated according to their antioxidant activity using total phenolic content, scavenging abilities on DPPH radical, total antioxidant activities (ABTS^•+), superoxide anion radical-scavenging (O₂^•) and protection against lipid peroxidation in vitro (LPO). Ethyl acetate fraction obtained by maceration and extract isolated by SE using 25% aqueous mixture of ethanol possessed the highest scavenger activity against DPPH radical (IC₅₀ = 9.2 ± 0.4 μg/ml, IC₅₀ = 27.4 ± 0.1 μg/ml, respectively). The SFE with 8% ethanol as a co-solvent produced extracts with distinguished increase in the antioxidant activity. The Soxhlet extract with ethyl acetate exhibited a strong reduction of lipid peroxidation (IC₅₀ = 209 ± 3 μg/ml) value comparable to the standard rutin (IC₅₀ = 203 ± 2 μg/ml). The results indicate that extracts of C. verbenacea have important potential as a source of bioactive compounds with antioxidant activity.     

Supercritical fluid extraction of lipids from broccoli leaves

The supercritical fluid extraction (SFE) and fractionation of lipids from broccoli leaves is presented in this work. For this purpose the effect of the different variables on the extraction was studied, obtaining the best results at 60°C, 300 bar and 3 mL/min. Two different fractions were obtained: First, the samples were extracted with pure CO₂, and afterward the residual material was extracted using CO₂ modified with 15% of methanol. The total fatty acid content of the extracts was determined by GC‐MS and compared with those results obtained by Soxhlet extraction with hexane and a chloroform/methanol (2:1) mixture. The SFE extracts presented a higher percentage of unsaturated fatty acids, especially the polyunsaturated 18:3 n ? 3. The methodology was successfully applied to the analysis of the fatty acid composition of the leaves from five different cultivars of broccoli. In all the samples the main fatty acids were α‐linolenic (18:3 n ? 3), linoleic (18:2 n ? 6), and palmitic (16:0). Among the different cultivars analyzed, Naxos variety presented the highest levels in fatty acids, while Parthenon and Viola the lowest. Practical applications: The proposed method allows the fractionation of lipids from broccoli leaves using a small volume of organic solvent and mild conditions. This is advantageous compared to conventional methods where large volumes or organic solvents are used, and the cost and time for the removal of these solvents, along with the possibility of degradation and toxicity, are the major disadvantages. The results obtained contribute to a better compositional characterization and a possible revaluation of this by‐product as a source of biologically active compounds.     

Optimization of experimental parameters for the quantification of polymer additives using SFE/HPLC

Analysis of low concentration additives; for example, antioxidants, in polymeric materials remains a difficult task. In the usual analytical methods, additives are extracted using large quantities of solvents first followed by concentrating the resulting solution for making possible the analysis. The supercritical fluid extraction (SFE) technique eliminates the use of large quantities of solvents and simplifies the analytical procedure. This work has been done with the goal of extracting the antioxidants Irganox 1010 and Irgafos 168 from a polypropylene matrix by using the SFE technique and by subsequent analysis using high‐performance liquid chromatography (HPLC). The experimental parameters; that is, temperatures, pressure, and modifiers have been varied to find the best extraction conditions. The optimum temperature and pressure for extraction of above‐mentioned polymer additives were found to be 120°C and 384 bar, with methanol as the modifier. The quantitative extractions are significantly faster than those reported earlier in the literature. The results point out that the technique used in these experiments—SFE combined with HPLC—is a reliable and environmentally friendly alternative to the commonly used liquid extraction and analytical methods. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 76: 938–946, 2000     

Extraction of anthocyanins and luteolin from Arrabidaea chica by sequential extraction in fixed bed using supercritical CO2, ethanol and water as solvents

Phenolic compounds from Arrabidaea chica Verlot leaves, besides conferring staining properties to their extracts, also have various biological activities including anti-inflammatory and wound-healing properties. To evaluate new possibilities for obtaining extracts with differentiated yield and composition, sequential extractions in fixed bed were performed at 40 and 50 °C, and 300 and 400 bar, using as extracting solvents pure supercritical carbon dioxide (scCO₂) in a first step, acidified ethanol in a second step and acidified water in a third extraction step. Four flavonoids of interest were investigated in the extracts, one of them being flavone (luteolin), and three anthocyanin compounds of type 3-desoxyanthocyanidins which were quantified by high performance liquid chromatography (HPLC). The extraction curves, the global yields and the concentration and yield of the compounds under study were evaluated. The results indicated that the cumulative total yields in the three steps ranged from 22% to 27% in all conditions of temperature and pressure, with the highest global yield at 50 °C and 300 bar. Although the lowest extraction yield was obtained using pure scCO₂, this step was highly selective, since only carajurin in its aglycone form was extracted among the compounds of interest and this was confirmed by analysis of MS/MS.     

Brazilian Ginseng extraction via LPSE and SFE: Global yields, extraction kinetics, chemical composition and antioxidant activity

Brazilian Ginseng extracts of two species, Pfaffia paniculata and Pfaffia glomerata, were obtained by supercritical fluid extraction (SFE) with CO₂ and by low-pressure solvent extraction (LPSE) with methanol, hexane and ethanol. The SFE assays were conducted at pressures of 100, 200 and 300 bar, and temperatures of 30 and 50 °C. The qualitative chemical compositions of the extracts were determined by thin layer chromatography (TLC). One of the active principles of interest from P. glomerata extract, β-ecdysone, was identified and quantified by HPLC. The antioxidant activities of Brazilian Ginseng extracts were determined by the coupled reaction of linolenic acid and β-carotene. For P. paniculata, the highest SFE yield was obtained at 200 bar/50 °C (0.22%, dry basis—d.b.), while the best extraction condition for P. glomerata was obtained at 200 bar/30 °C (0.18%, d.b.). The higher extract yields obtained by LPSE were 2.0% and 5.8% (w/w, d.b.) for P. paniculata and P. glomerata, respectively, both obtained with methanol as extraction solvent. From the overall extraction curve of P. glomerata, it was possible to obtain the kinetic parameters of extraction; the duration of the CER (constant extraction rate) period was determined as 134 min. The TLC plates showed the possible presence of flavonoids in the ethanolic extract for both Pfaffia species. The antioxidant activity analysis detected that LPSE extracts had higher activity than SFE extracts.     

Extraction of phenolic compounds and anthocyanins from blueberry (Vaccinium myrtillus L.) residues using supercritical CO2 and pressurized liquids

This work explored the potential of subcritical liquids and supercritical carbon dioxide (CO₂) in the recovery of extracts containing phenolic compounds, antioxidants and anthocyanins from residues of blueberry (Vaccinium myrtillus L.) processing. Supercritical CO₂ and pressurized liquids are alternatives to the use of toxic organic solvents or extraction methods that apply high temperatures. Blueberry is the fruit with the highest antioxidant and polyphenol content, which is present in both peel and pulp. In the extraction with pressurized liquids (PLE), water, ethanol and acetone were used at different proportions, with temperature, pressure and solvent flow rate kept constant at 40 °C, 20 MPa and 10 ml/min, respectively. The extracts were analyzed and the highest antioxidant activities and phenolic contents were found in the extracts obtained with pure ethanol and ethanol + water. The highest concentrations of anthocyanins were recovered with acidified water as solvent. In supercritical fluid extraction (SFE) with CO₂, water, acidified water, and ethanol were used as modifiers, and the best condition for all functional components evaluated was SFE with 90% CO₂, 5% water, and 5% ethanol. Sixteen anthocyanins were identified and quantified by ultra performance liquid chromatography (UPLC).     

Supercritical CO2 and low-pressure solvent extraction of mango (Mangifera indica) leaves: Global yield,extraction kinetics,chemical composition and cost of manufacturing

Low-pressure solvent extraction (LPSE) and supercritical fluid extraction (SFE) were used to obtain extracts from mango (Mangifera indica) leaves. Kinetics curves were determined for both methodologies. The extracts chemical compositions and manufacturing costs were determined for both processes. Global yield isotherms for SFE process were determined at 10–40 MPa and 313–323 K. The highest yield was 2.24% at 30 MPa and 323 K; the LPSE yield (9.3%) was almost three times higher than that of SFE (3.6%). Thin layer chromatography showed that mango leaves extracts have several classes of compounds as alkaloids, flavonoids and terpenoids, recovered by both methods. The cost of manufacturing (COM) mango leaves extracts were US$ 32/kg and US$ 92/kg for LPSE and SFE, respectively.     

Supercritical fluid extracts from tamarillo (Solanum betaceum Sendtn) epicarp and its application as protectors against lipid oxidation of cooked beef meat

The possibility of using the tamarillo (Solanum betaceum (Cav.) Sendtn (syn. Cyphomandra betacea)) epicarp as source of compounds with antioxidant activity in cooked beef meat (CBM) was explored. Extracts from tamarillo by supercritical fluid extraction (SFE) and Soxhlet extraction (SE) were obtained. The SFE was performed using pure CO₂ at different temperatures and pressures (40 and 50 °C; 10, 20 and 30 MPa) and CO₂ added with ethanol (CO₂/EtOH) as co-solvent (2, 5 and 8%, w/w). The SFE kinetics and mathematical modeling of the overall extraction curves (OEC) were also investigated. EtOH and hexane were used in the SE. The antioxidant activity (AA) of extracts was evaluated in CBM as well as the protection against lipid oxidation was determined by measuring lipid hydroperoxides (LHP) and thiobarbituric acid reactive species (TBARS). The extract obtained by SFE with CO₂/EtOH (50 °C/30 MPa and 2% of EtOH) showed the highest AA. In SFE, the co-solvent addition improved considerably the AA and the extraction yield. The extracts obtained by SFE with CO₂/EtOH showed a better AA compared with the synthetic antioxidant TBHQ. The highest yield values were achieved by SE with ethanol (7.7 ± 0.4%) and by SFE with 5% EtOH (1.9 ± 0.1%). The results indicate that extracts of tamarillo epicarp are a potential source of antioxidant compounds.     

不同萃取技术获得的Artemisia sp的甲醇提取物其抗氧化和抗菌活性的比较

The polyphenol content,antioxidant and antimicrobial activities of the extracts obtained by classical, ultrasonic and Soxhlet extractions from dry aerial parts of two Artemisia species(Artemisia vulgaris and Artemisia campestris)were compared.Ultrasound positively affected the yield of extractive substance and the kinetics of extraction,but the extract obtained by the classical extraction showed the highest antioxidant activities and contained higher total contents of phenolic compounds and flavonoids than the extracts obtained by two other extraction techniques.Both flavonoid aglycones(apigenin,quercetin,quercetin 3,3′-dimethyl ether)and flavonoid glycosides(rutin,hyperoside and kaempferol 3-rhamnoside)were identified by thin layer chromatograph(TLC)analysis in the extracts from both species.A.campestris extracts were richer in quercetin than A.vulgaris and its antimicrobial activity was also better than A.vulgaris.Extracts obtained from both species were found to be more effective on the tested yeasts than bacteria.The kinetics of the total extractive substances,such as phenolic,flavonoids and quercetin extraction,was successfully described by the model of unsteady-state diffusion.     

Simultaneous extraction of rosemary and spinach leaves and its effect on the antioxidant activity of products

Mixed vegetal extracts are interesting target of new products as nutraceuticals, superior ingredients for the design of functional food, singular ingredients for cosmetics, etc. In this work the extraction of a mixture of spinach and rosemary leaves (50 wt.% of each plant) was investigated in terms of its antioxidant activity, and compared with the extraction of the separate species. Phenolic diterpenes of rosemary and carotenoids of spinach were target compounds due their recognized biological activities. Two different extraction techniques were applied, namely pressurized liquid extraction using hexane at two different temperatures (100 and 150 °C) and supercritical fluid extraction with pure carbon dioxide at 40 °C and two different pressures (20 and 30 MPa). For each extraction technique and conditions three different raw materials were employed: spinach leaves, rosemary leaves and the mixture 50:50 of spinach and rosemary leaves.The antioxidant activity of the samples produced was evaluated with the ABTS assay and showed to be enhanced when the species are simultaneously extracted, with antioxidant values around 20% higher than the values corresponding to mixing the extracts obtained by separate. A possible synergic effect between carotenoids and phenolic diterpenes was studied, although no specific synergic activity could be observed. However, the enhanced antioxidant activity could be attributed to a definite increase of the concentration of carnosic acid, which was observed in the samples produced by the simultaneous extraction.     

Screening the Antioxidant and Antimicrobial Properties of the Extracts from Plantain (Plantago Major L.) Leaves

Abstract

Antioxidant and antimicrobial activities of the extracts obtained by classical (maceration) and ultrasonic (40 kHz) extraction from dry Plantago major leaves were compared. The antioxidant activities of extracts obtained by ultrasonic and classical extraction were 0.87±0.02 and 0.85±0.02 µg/µg DPPH, respectively. Ultrasound positively affected the extractive substance yield and the kinetics of extraction, but the extract obtained by classical extraction contained higher total contents of phenolic compounds and flavonoids than that obtained by ultrasonic extraction. Extracts of P. major showed better antimicrobial activity against the yeasts than against the bacteria.