Optimization of extraction conditions of antioxidants from sunflower shells (Helianthus annuus L.) before and after enzymatic treatment

The effects of three independent variables: solvent polarity, temperature and extraction time on the antioxidant capacity, total phenolic content and phenolic acid composition in extracts obtained from sunflower shells before and after enzymatic treatment were studied. Response surface methodology based on three-level, three-variable Box-Behnken design was used for optimization of extraction parameters and evaluation of their effect on antioxidant capacity and total phenolic content in shell extracts.The average antioxidant capacities of extracts from sunflower shells without enzymatic treatment (368.1-1574.4 μmol TE/100 g) were higher than those for cellulolytic and pectolytic enzymes-treated shells (222.7-1419.0 and 270.7-1570.7 μmol TE/100 g, respectively). The content of total phenolic compounds ranged between 58.2-341.2 mg CGA/100 g, 26.7-277.3 mg CGA/100 g and 51.4-301.5 mg CGA/100 g for extracts obtained from shells without enzyme and treated with cellulolytic and pectolytic enzymes, respectively. Total phenolic content (TPC) in the studied shell extracts correlated significantly (p < 0.0001) positively with their antioxidant capacity determined by the ferric reducing antioxidant power (FRAP) method (r = 0.9275). Results of FRAP, TPC and phenolic acid composition in the studied shell extracts depend on the extraction conditions (solvent polarity, temperature, time), but they are independent on the addition of enzyme solutions. The antioxidant capacity and total phenolic content in the resulting extracts increased with a line in extraction temperature and solvent polarity.     

Response surface optimization of antioxidants extraction from chestnut (Castanea sativa) bur

The chestnut bur, a forest waste product from chestnut processing in the food industry, was studied as a potential source of natural antioxidants. Extractions were performed using aqueous solutions of methanol or ethanol. Experiments were planned according to an incomplete 3³ factorial design to study the influence of temperature (25-75 °C), time (30-120 min) and solvent concentration (50-90%) on extraction yield and on extract properties: total phenols content, antioxidant activity (using the FRAP, DPPH and ABTS methods) and average molecular weights. All dependent variables were influenced by temperature and solvent concentration whereas the influence of time was almost negligible. Using the response surface methodology the optimal extraction conditions were selected: the highest temperature assayed (75 °C), the lowest solvent concentration (50%) and an extraction time of 75 min for the methanolic extractions and of 30 min for the ethanolic ones. Under those conditions the values predicted for extraction yield and total phenols content were 18.95% and 36.32 g GAE/100 g extract for the methanolic extract and 17.95% and 26.11 g GAE/100 g extract for the ethanolic ones. Methanolic extracts showed superior total phenols content and antioxidant properties and slightly higher extraction yields than ethanolic extracts; however, ethanol is recommended for food applications due to its GRAS (Generally Recognized as Safe) qualification. Gallic acid esters of glucose, ellagic acid and small proportions of quercetin-3-β-d-glucoside, phenolic compounds with demonstrated antioxidant properties, were identified in chestnut bur extracts by RP-HPLC-ESI-TOF.     

Dyeing properties and colour fastness of wool dyed with indicaxanthin natural dye

This research work involves the dyeing of wool with indicaxanthin, a natural dye extracted from fruits of Opuntia ficus-indica. The optimal conditions for dye extraction were to mix 50 g of Juice from cactus pears with 100 mL of 80% aqueous ethanol as solvent for dye extraction. Liquid chromatography was applied for the separation. Two main dyes were obtained, which were identified as indicaxanthin (75 mg per 50 g) and betanin (5 mg per 50 g). The effect of dye bath pH, salt concentration, dyeing time and temperature were studied. The optimal conditions for wool dyeing with indicaxanthin dye were carried out at 70 °C for 90 min with the pre-treatment of various metal salts as mordant. The colour yields of the dye on the wool were found to be highly dependent of the pH, optimum results being obtained at pH 4. The K/S of wool increased in the order of the dyeing using KAl (SO₄)₂ > MnSO₄ > CoSO₄ > FeSO₄ > none > ZnSO₄ > CuSO₄. Un-mordanted samples have good properties of water and washing fastness. Mordants KAl (SO₄)₂ and CoSO₄ were found to give good light fastness (rating 5).     

Bioactive properties and chemical constituents of methanolic extract and its fractions from Jatropha curcas oil

The present work is designed to evaluate the bioactive properties of the crude methanolic extract of Jatropha curcas oil and its solvent fractions. The crude methanolic extract obtained was fractionated using a hydrophilic lipophilic balanced (HLB) cartridge and then eluted with different solvents in the order of hexane (F1), dichloromethane (F2), chloroform (F3), ethyl acetate (F4) and methanol (F5), respectively. Total phenolic content of the crude methanolic extract and its fractions was in the range of 0.19-4.5 mg/g as gallic acid equivalent. Antioxidant activity of the crude methanolic extract and its fractions were determined by two complementary test methods, namely, phenanthroline method and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging method. All samples demonstrated weak antioxidant activity (150-851 μmol Fe/100 g of the extract and IC₅₀ of 1.05-13.5 mg/mL). When compared to butylated hydroxytoluene (BHT), a reference synthetic antioxidant, both showed weaker antioxidative potential. The evaluation of antimicrobial activity of the extracts was performed using a disc diffusion method and a micro-well dilution method against six economic plant disease bacteria. The results showed that all extracts possessed strong to moderate antibacterial activity with varying degrees of growth inhibition against the test bacteria. The minimum inhibitory concentrations (MIC) were in the range of 14.92-428.6 μg/mL. In addition, the chemical constituents in each fraction of the extract were subjected to analyze by gas chromatography-mass spectrometry (GC-MS). The eleven constituents were identified. Among them, 2,4-di-tert-butylphenol, methyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionate and linoleic acid may be the main cause of its strong antibacterial activity. Therefore, this oil present in the methanolic extract had great potential as effective antibacterial sources.     

Isolation, identification and dyeing studies of betanin on modified acrylic fabrics

Mature red fruits of Opuntia ficus-indica contain two soluble pigment, betanin and indicaxanthin. The optimal conditions for dye extraction were to mix 50 g of juice from cactus pears with 100 mL of acidified water as solvent for dye extraction. Two main dyes were purified from the pigment extract by chromatography and identified by UV-vis, HPLC and LC-MS techniques as indicaxanthin (15 mg per 100 g) and betanin (280 mg per 100 g). The effect of dye bath pH, salt concentration, dyeing time and temperature was studied. The optimal conditions for dyeing modified acrylic fabrics with betanin dye were carried out at 50 °C for 45 min at pH 5. Un-mordanted samples have good properties of water and washing fastness. Mordant CoSO₄ was found to give good light fastness (rating 5).     

Supercritical carbon dioxide (SC-CO2) extraction of Nigella sativa L. oil using full factorial design

In this study, Nigella sativa L. oil was extracted using supercritical carbon dioxide with full factorial design to determine the best extraction condition (pressure, temperature and dynamic extraction time) for obtaining an extract with high yield, antioxidant activity and thymoquinone (TQ) quantity. The maximum thymoquinone content in the highest overall yield was achieved through SC-CO₂ extraction condition of 150 bar, 40 °C, 120 min with the value of 4.09 mg/ml. The highest SC-CO₂ extraction yield was 23.20% which obtained through extraction condition of 350 bar, 60 °C and 120 min. The extraction conducted at 350 bar, 50 °C, 60 min showed the lowest IC₅₀ value (highest antioxidant activity) of 2.59 mg/ml using DPPH radical scavenging activity method. Fatty acid composition of the extracted oil with highest radical scavenging activity was obtained by gas chromatographic analysis.     

Enzyme pretreatment and negative pressure cavitation extraction of genistein and apigenin from the roots of pigeon pea [Cajanus cajan (L.) Millsp.] and the evaluation of antioxidant activity

An enzyme pretreatment and negative-pressure cavitation extraction (E-NPCE) method was investigated for extraction genistein and apigenin from pigeon pea roots. The important parameters involved in E-NPCE process were optimized by single-factor experiments and then critical parameters were investigated by a 2³ full factorial central composite design (CCD) to optimize extract conditions. Under optimal conditions, the yields of genistein and apigenin were 0.628 mg/g and 0.359 mg/g, which represented an increase of 44.70% and 53.05%, respectively, compared to standard NPCE. Furthermore, from DPPH scavenging activity test the extract of E-NPCE showed better antioxidant activity than these of other methods. The results demonstrated that E-NPCE would have lower energy consumption, higher efficiency and could be an alternative method for natural compounds extraction.     

玫瑰茄总黄酮超声波-微波协同提取及抗氧化活性

采用Box-Behnken响应面试验设计法对玫瑰茄总黄酮超声波-微波协同提取工艺进行优化,研究了乙醇浓度、液固比、提取时间、微波功率、提取次数5个单因素对总黄酮提取量的影响,建立了乙醇浓度、液固比和微波功率的三因素回归模型,并研究其抗氧化活性。结果表明,提取的最佳工艺参数为乙醇浓度40%、液固比50∶1（mL/g）、微波功率250 W、提取时间20 min、超声波功率50 W,在此条件下提取2次,总黄酮提取量为40.2 mg/g。同时以维生素C作对照,研究了玫瑰茄总黄酮粗提物和纯化物的体外抗氧化活性。结果表明,玫瑰茄总黄酮粗提物和纯化物都具有较强的抗氧化活性,能够有效清除O₂^-?自由基和DPPH自由基,清除能力与浓度呈较明显的量效关系,但抗氧化能力较维生素C弱。     

Evaluation of Ultrasound,Microwave, Ultrasound–Microwave,Hydrothermal and High Pressure Assisted Extraction Technologies for the Recovery of Phytochemicals and Antioxidants from Brown Macroalgae

This study aims to explore novel extraction technologies (ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE), ultrasound–microwave-assisted extraction (UMAE), hydrothermal-assisted extraction (HAE) and high-pressure-assisted extraction (HPAE)) and extraction time post-treatment (0 and 24 h) for the recovery of phytochemicals and associated antioxidant properties from Fucus vesiculosus and Pelvetia canaliculata. When using fixed extraction conditions (solvent: 50% ethanol; extraction time: 10 min; algae/solvent ratio: 1/10) for all the novel technologies, UAE generated extracts with the highest phytochemical contents from both macroalgae. The highest yields of compounds extracted from F. vesiculosus using UAE were: total phenolic content (445.0 ± 4.6 mg gallic acid equivalents/g), total phlorotannin content (362.9 ± 3.7 mg phloroglucinol equivalents/g), total flavonoid content (286.3 ± 7.8 mg quercetin equivalents/g) and total tannin content (189.1 ± 4.4 mg catechin equivalents/g). In the case of the antioxidant activities, the highest DPPH activities were achieved by UAE and UMAE from both macroalgae, while no clear pattern was recorded in the case of FRAP activities. The highest DPPH scavenging activities (112.5 ± 0.7 mg trolox equivalents/g) and FRAP activities (284.8 ± 2.2 mg trolox equivalents/g) were achieved from F. vesiculosus. Following the extraction treatment, an additional storage post-extraction (24 h) did not improve the yields of phytochemicals or antioxidant properties of the extracts.     

Optimisation of a solvent for the complete extraction of prolamins from heated foods

Wheat bread was lyophilised, ground, extracted and centrifuged. The supernatants were analysed for gluten content by RP-HPLC and a commercial sandwich ELISA. Prolamin extraction solvents contained tris(2-carboxyethyl)phosphine (TCEP; 1, 2, 5, 10, 20, 50 mmol/L), guanidine hydrochloride (GUA; 0 or 2 mol/L) and a buffered salt solution. A commercial cocktail solution (250 mmol/L mercaptoethanol (ME), 2 mol/L GUA, buffered salt solution) as well as 60% (v/v) ethanol were used as control solvents. Wheat flour was the control for the extractability of the native prolamins. 60% ethanol only extracted 37% of the prolamins from wheat bread (cocktail = 100%). When ME was replaced by TCEP the protein yield increased from 35% at the lowest TCEP-level to 95% when 20–50 mmol/L TCEP were used. The use of GUA was essential to extract prolamins quantitatively. Comparative protein analysis using RP-HPLC and ELISA showed that both methods provided comparable prolamin (gliadin) concentrations of the wheat flour (40.3–45.7 mg/g), when 60% ethanol was used as extraction solvent. The extraction yields from bread were considerably lower (16.7–24.7 mg/g). Cocktail and TCEP extracted almost the same amount of protein from flour and bread with TCEP showing slightly lower yields. Total extractable protein (gliadin + glutenin) as determined by RP-HPLC was 70.5–75.3 mg/g, and total gliadin as determined by ELISA was 42.7–44.2 mg/g. Thus, the study has shown that TCEP in combination with GUA extracts proteins from heated, gluten-containing foods as effective as the commercial cocktail solution.     

Antioxidant activity of phenolic compounds obtained by autohydrolysis of corn residues

This study aimed to evaluate the effect of autohydrolysis temperature of corn residues in the antioxidant activity of the phenolic compounds extracted from the liquid phase. The treatments were carried out at 160, 180, 190 and 200 °C for 30 min in a pressurized batch reactor. Two different methods for phenolic compounds extraction from the autohydrolysis liquors were investigated. For that purpose, solvent extraction with ethyl acetate and acidic precipitation were performed for phenolic compounds recovery. These methods have been compared in terms of extraction yield, physicochemical properties of obtained polyphenols (characterization by Fourier Transform Infrared Spectroscopy, Thermogravimetric Analysis and Gel Permeation Chromatography), total phenolic content, total antioxidant capacity and Trolox equivalent antioxidant capacity (TEAC) values, measured in DPPH (2,2-diphenyl-1-picrylhydrazyl) test system. The maximum phenolic contents ranged from 6.04 mg GAE/100 mg extract in acidic precipitated samples to 16.45 mg GAE/100 mg extract in ethyl acetate soluble fractions. The results indicated that the ethyl acetate fractions possessed the highest antioxidant activity, reaching after 30-60 min the same capacity reported for the reference synthetic antioxidants (Trolox).     

Fractionation of flax shives with pressurized aqueous ethanol

Fractionation of flax shives into major biopolymer constituents, such as cellulose, hemicelluloses, and lignin, was carried out with pressurized aqueous ethanol in a pressurized low-polarity water extractor. The effect of processing parameters such as temperature, ethanol concentration, flow rate, sample size and solvent/feed ratio on the simultaneous extraction of hemicelluloses and lignin was determined. More than 80% of total hemicelluloses and ∼78% of total lignin were removed simultaneously in a single step under the following conditions: 180 °C, 30% (v/v) ethanol concentration, 3 mL/min flow rate, and 45 mL/g solvent/feed ratio. Under these extraction conditions, cellulose degradation was negligible. Further, the separation of lignin from hemicelluloses was carried out using two simple alternative methodologies based on precipitation. Since no acidic or alkali catalysts were used, the degradation of biopolymers was negligible and the oligomer/monomer ratio of sugars was 825:1. Characterization of fractionated biopolymers was carried out with scanning electron microscopy and a Fourier-transform infrared spectrometer (FT-IR). FT-IR spectra of isolated lignin and hemicelluloses showed that both polymers were comparable to commercially available products.     

超声辅助提取澳洲坚果青皮总黄酮工艺优化及抗氧化性能研究

以澳洲坚果青皮为原料,研究了超声辅助提取总黄酮工艺条件以及抗氧化活性。通过L₉(3 ⁴)正交试验得到超声功率为300 W条件下的最佳提取工艺:提取时间为45 min、乙醇浓度为50%、提取温度为50 ℃、料液比为1:60 （g/mL）,在此条件下总黄酮提取量为(1638.59±44.26) mg/100 g。通过抗氧化实验发现,澳洲坚果青皮总黄酮ABTS自由清除能力约为Trolox的2.48倍,总抗氧化还原能力约为Trolox的1.90倍,由此表明澳洲坚果青皮总黄酮具有较强的抗氧化活性。     

Arbutus unedo L. leaves as source of phytochemicals with bioactive properties

In recent years the strawberry tree (Arbutus unedo L.) is being gradually replaced by other species with higher economic value. With the ultimate goal of selecting superior genotypes, the present work was initiated to study the antioxidant and antimicrobial activities, and total phenolic content in 19 different genotypes of A. unedo leaves from the Trás-os-Montes region of Portugal.The genotype Bragança 1 contains higher total phenolic content (215.0 mg GAE/g_extract) whereas the Vila Boa 4 genotype shows lower total phenolic content (148.0 mg GAE/g_extract). In both methods tested to evaluate the antioxidant activity, Vila Verde and Donai displayed the highest antioxidant capacity (EC₅₀ values of 0.088 and 0.090 mg/mL, respectively, for DPPH; EC₅₀ values of 0.233 and 0.245 mg/mL, respectively, for reducing power assay) while Vila Boa 2 reported the lowest antioxidant potential (EC₅₀ values of 0.142 and 0.378 mg/mL, respectively, in DPPH and reducing power methods). Linear negative correlations were established between the total phenol contents and the EC₅₀ values for both of the antioxidant activity methods tested. Preliminary assays for antimicrobial potential showed that extracts from A. unedo leaves display antibacterial activity, with MIC values of 1 and 5 mg/mL for some Gram-positive and Gram-negative bacteria, respectively. Taken together, the results suggest that A. unedo leaves are a potential source of natural compounds with valuable bioactive properties that could be explored by the pharmaceutical, chemical and food industries.     

Ultrasound-Assisted Water Extraction of Mastocarpus stellatus Carrageenan with Adequate Mechanical and Antiproliferative Properties

Ultrasound-assisted water extraction was optimized to recover gelling biopolymers and antioxidant compounds from Mastocarpus stellatus. A set of experiments following a Box–Behnken design was proposed to study the influence of extraction time, solid liquid ratio, and ultrasound amplitude on the yield, sulfate content, and thermo-rheological properties (viscoelasticity and gelling temperature) of the carrageenan fraction, as well as the composition (protein and phenolic content) and antiradical capacity of the soluble extracts. Operating at 80 °C and 80 kHz, the models predicted a compromise optimum extraction conditions at ~35 min, solid liquid ratio of ~2 g/100 g, and ultrasound amplitude of ~79%. Under these conditions, 40.3% carrageenan yield was attained and this product presented 46% sulfate and good mechanical properties, a viscoelastic modulus of 741.4 Pa, with the lowest gelling temperatures of 39.4 °C. The carrageenans also exhibited promising antiproliferative properties on selected human cancer cellular lines, A-549, A-2780, HeLa 229, and HT-29 with EC₅₀ under 51.9 μg/mL. The dried soluble extract contained 20.4 mg protein/g, 11.3 mg gallic acid eq/g, and the antiradical potency was equivalent to 59 mg Trolox/g.     

Effect of particle size of rice bran on gamma-oryzanol content and compounds

The reduction of particle size can facilitate the extraction of phytochemical compounds. In this study was evaluated the effect of rice bran particles size in the yield and characteristics of γ-oryzanol compound. The γ-oryzanol extraction was realized with hexane and isopropanol solvents and quantification by spectrophotometric method. The γ-oryzanol extracts were characterized in relation of theirs majority components in HPLC-UV and the antioxidant capacity verified by the free radical DPPH^● consumption. The γ-oryzanol yield varied of 0.10–1.54 mg/g of bran, and the highest yield was obtained in particles smaller than 0.39 mm. The γ-oryzanol majority components presence (cycloartenyl ferulate, 2,4-methylenecycloartanyl ferulate, campesteryl ferulate e β-sitosteryl ferulate) in the extracts was confirmed and verified differences in the profile of this components in function of different particles sizes. The γ-oryzanol extract obtained from particle sizes between 0.73 and 1.67 mm demonstrated most specific inhibition of DPPH radical (6.7%) and IC₅₀ 6.63 μg/mL. When the particle size is reduced, the access surface to the extraction solvent is increased resulting in more γ-oryzanol extraction, however the extract from larger particles was more efficient as antioxidant.     

Effect of extraction methods on characteristic and composition of Indonesian cashew nut shell liquid

Long chain phenols contained in cashew nut shell liquid (CNSL) are found to have important pharmaceutical applications, such as antitumor, antimicrobial, urease inhibitory and lipoxygenase activities, and also are well known in coating and resin industry. The impact of different extraction methods on CNSL yield, selectivity towards preferable compounds, composition and characteristic of extracts was investigated. Four different methods employed in extracting CNSL from CNS were: supercritical carbon dioxide (SC-CO₂) extraction (40 °C, 300 bar, 4 h), subcritical water (SCW) extraction (140 °C, 22 bar, 1 h), soxhlet extraction (solvent boiling point, atmospheric pressure, 30 h), and two-step extraction, which comprises a solvent extraction followed by a SCW extraction. Characteristic of the extracts differed significantly. Methanol and two-step extraction resulted in darker and more turbid extracts, while n-hexane and SC-CO₂ extracts were clearer and lighter in color. GC-FID/MS chromatograms showed differences in compositions of the extracts obtained by different methods. Two-step extraction yielded extracts that contain 81.17-82.98% total long chain phenols (around 50% based on dry CNS) with monounsaturated cardanol as the major compound, producing higher amount of total phenols than other methods. SCW extraction showed high selectivity towards monounsaturated cardanol and stigmasterol, while high concentration of monounsaturated anacardic acid and cardol appeared in SC-CO₂ extract.     

Polyol production by chemical modification of date seeds

Polyol production from chemical modification of date seeds has been investigated through oxypropylation and liquefaction techniques (using organic solvents in the presence of a catalyst). The obtained products were characterized using infrared spectroscopy analysis, ¹H NMR, thermogravimetric analysis and other parameters such as hydroxyl number (I_OH) and viscosity. Results showed that 93% of the solid substance was converted into polyol in the oxypropylation reaction at the date seeds/propylene oxide ratio of 0.25 and 10% potassium hydroxide at 160 °C. The oxypropylated product has I_OH of 779 mg KOH/g and viscosity of 6.9 Pa s. Regarding the liquefaction technique, results show that a yield of 95% was obtained at the date seeds/liquefying solvent ratio of 0.25 in 60 min of reaction at 160 °C. The liquefied product shows I_OH of 336 mg KOH/g and viscosity of 0.9 Pa s.     

工业大麻中大麻二酚提取工艺优化和生物活性分析

为优化工业大麻中CBD提取工艺,建立CBD高效液相分析方法,评估CBD体外酶抑制活性和抗氧化活性,拟通过设计单因素实验,以溶剂种类、超声时间、料液比、提取次数为影响因素,CBD含量为评价指标优化提取工艺,并通过检测CBD对黄嘌呤氧化酶、多酚氧化酶的抑制率及ABTS自由基的清除率监测其体外酶抑制活性。结果表明:甲醇为提取溶剂、提取2次、超声10 min、料液比1:20 g/mL为最佳提取条件。生物活性实验表明CBD能明显抑制酶活,其对XOD、PPO、ABTS的C50分别为1.18μg/mL(40s)、1.03μg/mL (60s)、4.99μg/mL和1.68μg/mL。该工艺操作简单、稳定可靠、成本较低,为工业大麻花叶中CBD的提取及定性定量提供一种新的技术方法,且该方法提取的CBD酶抑制活性和抗氧化活性好,具有重要的药用价值。     

响应面法优化枇杷花总黄酮超声波辅助提取工艺的研究

采用超声波辅助提取枇杷花中的总黄酮,研究超声波功率、乙醇浓度、提取温度、提取时间、液料比及提取次数对总黄酮提取得率的影响。根据单因素试验,选择在超声波功率80 W、一次浸提条件下优化提取工艺;采用Box-Behnken中心组合试验设计原理,设计4因素3水平试验,以响应面分析法优化乙醇浓度、提取温度、提取时间及液料比4个因素对总黄酮提取得率的影响。结果表明：当超声波功率为80 W时,枇杷花中总黄酮超声波辅助提取的优化工艺参数为：乙醇浓度47.33％、提取温度58.42 ℃、提取时间12.01 min、液料比（mL ∶ g）45.56 ∶ 1,在此条件下枇杷花总黄酮一次浸提的提取得率可达10.48％。