Optimization of ultrasound-assisted aqueous two-phase system extraction of polyphenolic compounds from Aronia melanocarpa pomace by response surface methodology

Aronia melanocarpa berries are abundant in polyphenolic compounds. After juice production, the pomace of pressed berries still contains a substantial amount of polyphenolic compounds. For efficient utilization of A. melanocarpa berries and the enhancement of polyphenolic compound yields in Aronia melanocarpa pomace (AMP), total phenolics (TP) and total flavonoids (TF) from AMP were extracted, using ultrasound-assisted aqueous two-phase system (UAE-ATPS) extraction method. First, the influences of ammonium sulfate concentration, ethanol–water ratio, ultrasonic time, and ultrasonic power on TP and TF yields were investigated. On this basis, process variables such as ammonium sulfate concentration (0.30–0.35?g?mL^?1), ethanol–water ratio (0.6–0.8), ultrasonic time (40–60?min), and ultrasonic power (175–225?W) were further optimized by implementing Box–Benhnken design with response surface methodology. The experimental results showed that optimal extraction conditions of TP from AMP were as follows: ammonium sulfate concentration of 0.324?g?mL^?1, ethanol–water ratio of 0.69, ultrasonic time of 52?min, and ultrasonic power of 200?W. Meanwhile, ammonium sulfate concentration of 0.320?g?mL^?1, ethanol–water ratio of 0.71, ultrasonic time of 50?min, and ultrasonic power of 200?W were determined as optimum extraction conditions of TF in AMP. Experimental validation was performed, where TP and TF yields reached 68.15?±?1.04 and 11.67?±?0.63?mg?g^?1, respectively. Close agreement was found between experimental and predicted values. Overall, the present results demonstrated that ultrasound-assisted aqueous two-phase system extraction method was successfully used to extract total phenolics and flavonoids in A. melanocarpa pomace.     

蒙古栎种壳多酚的超声辅助提取及抗氧化能力

采用超声辅助提取法对蒙古栎种壳多酚进行了提取。对超声提取过程中的各因素在单因素实验的基础上应用Box Behnken中心组合进行3因素3水平的实验设计法进行了得率与纯度双响应值的优化,得到蒙古栎种壳多酚最佳提取工艺条件为：乙醇体积分数为60.56%,液料比30.72∶1 mL·g^-1,提取时间42.39 min,在此条件下蒙古栎种壳多酚平均得率为0.38%,平均纯度为18.8%,该多酚清除DPPH自由基的EC₅₀值为176 μg·mL^-1,高于合成抗氧化剂BHA和BHT等。     

MODEL DEVELOPMENT AND PROCESS OPTIMIZATION FOR SOLVENT EXTRACTION OF POLYPHENOLS FROM RED GRAPES USING BOX–BEHNKEN DESIGN

The objective of the present study is to find out the optimum extraction conditions for extraction of polyphenols from red grapes using Box–Behnken design. Red grapes polyphenols were extracted using acid–ethanol solvent at various extraction temperature (40–60°C), extraction time (20–100 min) and different solid–liquid ratio (1:5–1:15 g:ml). The effect (main and interactive) of extraction conditions on total anthocyanin, phenolic and flavonoid content were studied using Box–Behnken design (three factors at three levels). The results showed that the contribution of the quadratic model was significant for all the responses. Second-order mathematical regression models were developed and were found to fit well with observed data. Derringer's desirability function methodology was performed to find out the optimal conditions based on both individual and combinations of all responses (extraction temperature: 57°C, time: 61 min, and solid–liquid ratio: 1:8.7 g:ml) were established. At this optimal condition, the anthocyanin yield, total phenolic and flavonoid content were 73.92 mg/100 g, 221.4 mg GAE/100 g, and 79.08 mg CE/100 g, respectively. A desirability value of 0.902 was achieved at this point.     

基于响应面法从连翘叶中提取连翘酯苷A和连翘苷的工艺优化

采用响应面分析法优化连翘叶中连翘酯苷A和连翘苷的最佳提取工艺条件,在单因素实验基础上,采用Box-Behnken中心组合设计原理研究液料比、乙醇浓度、浸提时间、浸提温度4个因素对连翘酯苷A得率和连翘苷得率的影响.利用Design Expert 8.0.5 b分析确定最佳提取工艺,根据实际条件,得到连翘酯苷A和连翘苷的最佳提取工艺为:乙醇浓度53％、提取温度74℃、浸提时间为53 min、液料比30 mL/g.回归模型预测最优条件下连翘酯苷A得率为7.53％,连翘苷得率为3.03％,验证值分别为7.56％,3.09％,与预测值的相对误差分别为0.4％,2.0％.     

拐枣中总黄酮提取工艺优化及其抗黄嘌呤氧化酶活性研究

为了研究拐枣总黄酮(TF)最佳提取工艺及其体外抗黄嘌呤氧化酶(xanthine oxidase,XO)活性。在单因素试验基础上,实验选择料液比、提取时间和乙醇浓度为自变量,应用Box-Benhnken中心组合法进行3因素3水平试验设计,以拐枣总黄酮得率为响应值,进行响应面分析,并采用D101大孔树脂对总黄酮进行进一步富集纯化,在此基础上研究拐枣总黄酮体外抗XO活性。结果表明,拐枣总黄酮的最佳提取条件为料液比15.0 mL/g、提取时间1.85 h、乙醇浓度53.0%,在此条件下,拐枣总黄酮的平均提取率可达到2.56%。体外活性结果表明,我们首次发现拐枣总黄酮具有显著的抗XO活性,其IC_(50 )为18.63±1.67μg/mL。本研究可为从拐枣中寻找和发现天然抗XO活性分子提供参考,同时可为拐枣的开发利用提供思路。     

无花果果皮中花青素的提取工艺的建立

本研究使用单因素方法考察了无花果(Ficus carica L.)果皮中花青素的最佳提取条件,并考察了7种参数对花青素提取率的影响。参数设置如下:溶剂性质(水,甲醇,乙醇和丙酮)、提取次数(1~3次)、固液比(1/50,1/100,1/150和1/200)、提取时间(60 min,120 min,180 min和240 min)、甲醇浓度(0,20%,40%,60%,80%和100%)、酸类型(盐酸,乙酸,柠檬酸和酒石酸)和酸浓度(0,1%,2%,5%和10%)。使用pH-示差法测量无花果果皮中单体花色素的含量。研究显示,无花果果皮中花青素的最佳提取条件为:溶剂为甲醇溶剂,提取次数为2次,固液比为1/100,提取时间为180 min,甲醇浓度为80%,酸类型为柠檬酸,柠檬酸浓度为5%。该最佳提取条件下的花青素的提取率达到最高(345.62 mg/100g DS)。     

效应面法优化石榴皮抑菌物的提取工艺

采用单因素和效应面法对石榴皮抑菌物提取条件进行优化,并对结果进行验证。结果发现:最佳提取温度为59℃、时间为73 min、液料比为40∶1(mL/g)。实验证明石榴皮提取物对枯草杆菌具有很强的抑制作用,且所用工艺简单易行,说明效应面优化法建立的数学模型预测结果可靠。     

响应面设计法优化仙鹤草总多酚的超声提取工艺

在单因素实验的基础上,采用响应面分析法对影响超声辅助提取仙鹤草总多酚得率的主要因素(超声温度、料液比和乙醇浓度)进行优化,建立了影响因素与总多酚之间的函数关系。获得最佳工艺条件为:71%乙醇,料液比1∶24,超声提取温度60℃,提取时间20 min。在此最佳条件下,总多酚得率为3.56%,试验结果与模型预测值相符。     

Optimization of the extraction of polyphenols and antioxidant activity from Malva parviflora L. leaves using Box–Behnken design

Abstract

Box–Behnken Design (BBD) was used to optimize the extraction conditions of polyphenols from Malva (Malva parviflora L.) leaves. The effect of ethanol concentration (20–80%), solvent/leaf powder ratio (10:1 to 30:1, v/w) and extraction time (5–45?min) on the polyphenols yield and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of the obtained extracts were investigated. Quadratic models fit well. The optimal conditions (53.40% ethanol, solvent/leaf powder ratio 20:1 (v/w), and 15?min) resulted in an extract with a maximum yield of polyphenols (1098.4?mg GAE/100?g leaf powder) and high inhibition percentage of DPPH radical (33.31%) with desirability 0.742. High Performance Liquid Chromatography (HPLC) analysis indicated that the major identified polyphenol compounds extracted at the optimal conditions were naringenin, ρ-coumaric acid, apigenin-7-glucoside, luteolin, and cinnamic acid. These findings indicate that M. parviflora leaf extracts possess DPPH radical scavenging activity and could be used as a natural source for bioactive products.     

Ultrasound-assisted aqueous extraction of total flavonoids and hydroxytyrosol from olive leaves optimized by response surface methodology

Abstract

Olive leaves were often extracted with methanol or ethanol at different proportions. In this study, ultrasound-assisted aqueous extraction was adopted for olive leaf extraction. The yields of total flavonoids (TF) and hydroxytyrosol (HT) were optimized by central composite experimental design. Two second-order polynomial equations were established to quantify the relationship between the responses and the processing parameters. Under the optimal condition of extracting at 60?°C for 60?min with the solvent-to-material ratio of 40, TF and HT amounted to 57.31?±?0.35 and 1.80?±?0.02?mg/g dry leaves (DL), respectively. The scavenging rate of all extracts against α, α-diphenyl-β-picrylhydrazyl (DPPH) and hydroxyl free radicals was screened. The integrated scores, representing both active ingredients and antioxidant capacity of the extracts, were calculated by principle component analysis (PCA). The optimal extract gained the highest score in PCA. In addition, compared to the extracts from 80% methanol to 44% ethanol, the ultrasound-assisted aqueous extract was richer in TF, HT, and polyphenols, while it also presented stronger ferric reducing antioxidant power (FRAP), but poorer strength to quench hydroxyl radicals. The study indicated that the aqueous extract of olive leaves may present broad potential opportunities in health-care sector.     

鹿蹄草中2″-O-没食子酰基金丝桃苷的提取工艺研究

以东北野生鹿蹄草为原料,利用超声辅助提取技术进行提取,在单因素实验的基础上对提取条件进行了考察,根据CCD(Central Composite Design)实验设计原理,采用3因素3水平的响应面分析法,以鹿蹄草中主要黄酮2″-O-没食子酰基金丝桃苷(2″-O-galloylhyperin)为指标,对提取过程进行优化,得到最佳工艺参数为：提取时间45 min,超声功率40 kHz,提取次数3次,提取温度60℃,液固比33∶1,乙醇浓度50%。在最佳提取条件下,2″-O-没食子酰基金丝桃苷的提取率可达5.024 mg·g^-1。     

均匀设计法优化高丽参多糖的提取

利用均匀设计法优化了高丽参多糖的提取过程.主要考察料液质量比、浸泡时间、提取温度和提取时间四个因素对提取率的影响.通过数学模型得到最优提取状况和3D曲面图.结果表明,提取温度对高丽参多糖得率影响最大,在料液质量比1:35、浸泡时间135 min、水提取温度81℃条件下提取156 min,高丽参多糖最大理论提取率为49.51%.最优化模型下实际多糖得率49.29%与理论得率相符.     

超声辅助低共熔溶剂提取沙棘籽粕多酚的工艺优化

以一系列低共熔溶剂为提取剂,采用超声波辅助法从沙棘籽粕中提取多酚。在单因素试验结果基础上,利用Box-Behnken实验设计,运用响应面分析法对影响沙棘籽粕多酚得率的主要因素（超声功率、超声时间、超声温度）进行优化。结果表明,沙棘籽粕多酚最佳提取工艺条件为：以含水量为30%的氯化胆碱-草酸低共熔溶剂为最佳提取剂,液料比为14∶1,超声功率420 W,超声时间56 min,超声温度44℃。在此条件下,多酚得率为3.31±0.008%。对比试验发现：氯化胆碱-草酸低共熔溶剂对沙棘籽粕多酚的得率明显优于传统溶剂;与热回流提取相比,超声提取法具有明显的优势。     

有柄石韦总多酚含量测定及其超声提取工艺优化

为优选有柄石韦Pyrrosia petiolos中总多酚超声提取工艺,采用Folin-Ciocalteus比色法对总多酚进行含量测定,通过单因素和正交试验,对提取工艺参数进行优化。结果表明,没食子酸对照品在浓度为1～10 mg·L-1范围内,与吸光度呈良好线性关系（y=2.406x-0.012,R=0.9994）,有柄石韦总多酚的最佳提取工艺条件为：55%乙醇,料液比1∶10（W/V）,超声提取60 min,超声提取温度75℃,总多酚提取率为21.47 mg·g-1。     

Effect of polyphenol extract on performance,serum biochemistry,skin pigmentation and carcass characteristics in broiler chickens fed with different cereal sources under hot-humid conditions

This study investigated the interaction between polyphenols with different cereal sources and their effects on performance, serum biochemistry, corticosterone levels and carcass characteristics in broiler chickens reared under hot-humid environment. Newly hatched coloured broiler chicks (n = 240) were randomly divided into six groups with five replicates of two different cereal sources, namely corn and broken rice-sorghum combined, and three levels of pomegranate peel polyphenol extracts (PPP) 0, 50 and 100 mg/kg in each cereal groups. Birds were maintained under standard management conditions for six weeks during hot-humid environment (Temperature: 29–36 °C; Relative Humidity: 69–80%). Fortnight body weight and feed consumption were recorded and serum biochemical constituents were estimated at 28 and 42 days of age. The body weight gain was significantly (p < 0.05) higher in broken rice-sorghum diet than in corn diet. The supplementation of polyphenols increased the skin and shank pigmentation. Serum protein, lipids and minerals showed significant difference due to cereals, polyphenols and their interactions. Corn-fed birds had significantly higher visceral organs weight than the alternate grain-fed broilers. The serum corticosterone levels were significantly reduced in the PPP supplemented groups than the control, it could be concluded that, supplementation of 50 mg/kg polyphenol extract from pomegranate peels improved production performance in broilers fed broken rice-sorghum as cereal source under hot-humid conditions.     

Box-Behnken响应面法优化超声波辅助提取辣木叶多酚工艺研究

采用超声波辅助提取辣木Moringa oleifera叶多酚,通过单因素试验考察乙醇浓度、料液比、提取时间和提取次数对辣木叶多酚得率的影响,进一步利用四因素三水平Box-Behnken组合设计响应面试验对提取工艺参数进行优化。结果表明,超声波辅助提取辣木叶多酚的最佳工艺条件为乙醇浓度62%、料液比1∶30(W/V)、提取时间30 min、提取3次,在此条件下辣木叶多酚得率为27.32 mg·g-1,所得回归模型拟合情况良好。     

微波辅助提取木豆根中染料木素工艺

以木豆根为原料,利用微波辅助提取技术进行提取,在单因素实验的基础上对提取条件进行了考察,根据BBD(Box-Behnken design)实验设计原理,采用3因素3水平的响应面分析法,以木豆根中主要异黄酮染料木素(genistein)为指标,对提取过程进行优化,得到最佳工艺参数为：提取温度为68℃,固液比为32∶1 mL·g^-1,乙醇浓度为78%,提取功率700 W,提取时间15 min。在最佳提取条件下染料木素的提取率可达到0.465±0.032 mg·g^-1。本研究对于微波提取技术的应用及木豆根的开发和利用都具有显著的意义。     

超声法提取山里红叶总黄酮及其抗氧化活性研究

采用超声辅助提取法从山里红叶中提取总黄酮。通过Box Behnken实验设计(BBD)结合响应面法(RSM)来优化超声提取的条件。影响山里红叶总黄酮提取效率的4个主要变量为液固比,温度,乙醇浓度和时间,得到的最佳值分别为15,40℃,40%,32 min。在此条件下,总黄酮的产率为15.50 mg·g^-1。体外抗氧化实验表明,山里红叶提取物的DPPH自由基清除能力为0.69 mg·mL^-1(以IC₅₀值表示) ,与传统的浸渍提取和热回流提取方法相比,超声提取的方法具有更好的抗氧化活性。实验结果表明超声提取法适用于提取山里红叶中的总黄酮,并且其提取物具有较好的抗氧化活性。     

深山含笑叶片总酚超声波提取工艺的优化

采用单因素实验方法研究了超声波辅助提取过程中乙醇浓度、料液比、提取温度和提取时间对深山含笑（Michelia maudiae Dunn）叶片总酚提取率的影响,并采用正交实验方法确定了最佳提取工艺条件。结果表明,深山含笑叶片总酚超声波辅助提取的最佳提取工艺为：按1：30（质量-体积比）的料液比加入体积分数70％的乙醇,于65℃条件下用超声波辅助提取30min。采用最佳的超声波提取工艺,深山含笑叶片的总酚提取率可达到11．41％。定性分析结果显示,深山含笑叶片的总酚提取物具有典型的酚类化合物特性,并显示出鞣质类成分、黄酮类成分和香豆素类成分的定性反应特征。     

银杏叶多酚的机械力辅助提取及其体外抗氧化活性研究

为优化银杏叶多酚提取工艺,通过单因素试验考察填充率、球磨转速、球磨时间、乙醇浓度、料液比、提取温度、提取时间七个因素对机械力辅助提取银杏叶多酚得率的影响,以银杏叶多酚得率为响应值,采用Box-Benhnken三因素三水平响应面设计优化工艺,同时比较了4种提取方法对银杏叶多酚提取得率和抗氧化活性的差异。结果表明,机械力辅助提取银杏叶多酚的最佳工艺条件为:填充率26%、球磨转速为400rpm、球磨时间为15min。在此条件下,银杏叶多酚的得率为7.33%。机械力辅助乙醇提取银杏叶多酚得率低于碱水提取法,但是抗氧化活性高于碱水法提取的银杏叶多酚;抗氧化活性与乙醇回流法提取的银杏叶多酚相当,但是提取得率高于乙醇回流法。此提取工艺高效可行,具有一定的参考价值。