Isolation and Identification of Phenolic Antioxidants in Black Rice Bran

Black rice bran contains phenolic compounds of a high antioxidant activity. In this study, the 40% acetone extract of black rice bran was sequentially fractionated to obtain 5 fractions. Out of the 5 fractions, ethyl acetate fraction was subfractionated using the Sephadex LH‐20 chromatography. The antioxidant activity of phenolic compounds in the extracts was investigated by 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical assay, 2,2‐azino‐bis‐(3‐ethylenebenzothiozoline‐6‐sulfonic acid) (ABTS) radical cation assay, reducing power. The subfraction 2 from ethyl acetate fraction had the highest total phenolic contents (TPC) (816.0 μg/mg) and the lowest EC₅₀ values (47.8 μg/mL for DPPH radical assay, 112.8 μg/mL for ABTS radical cation assay, and 49.2 μg/mL for reducing power). These results were 3.1, 1.3, and 2.6 times lower than those of butylated hydroxytoluene (BHT), respectively. At a concentration of 100 μg/mL, the antioxidant activity and TPC of various extracts was closely correlated, with correlation coefficients (R²) higher than 0.86. The major phenolic acid in subfraction 2 was identified as ferulic acid (178.3 μg/mg) by HPLC and LC‐ESI/MS/MS analyses. Our finding identified ferulic acid as a major phenolic compound in black rice bran, and supports the potential use of black rice bran as a natural source of antioxidant.     

Phytochemicals and antioxidant capacities in rice brans of different color

Rice bran, a byproduct of the rice milling process, contains most of the phytochemicals. This study aimed at determining the concentrations of lipophilic, solvent-extractable (free), and cell wall-bound (bound) phytochemicals and their antioxidant capacities from brans of white, light brown, brown, purple, and red colors, and broccoli and blueberry for comparison. The concentrations of lipophilic antioxidants of vitamin E (tocopherol and tocotrienols) and γ-oryzanols were 319.67 to 443.73 and 3861.93 to 5911.12 μg/g bran dry weight (DW), respectively, and were not associated with bran color. The total phenolic, total flavonoid, and antioxidant capacities of ORAC (oxygen radical absorbance capacity), DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging, and iron-chelating in the free fraction were correlated with the intensity of bran color, while variations of these in the bound fraction were less than those in the free fraction among brans. Compounds in the bound fraction had higher antioxidant capacity of ORAC than DPPH, relative to those in the free fraction. The bound fraction of light-color brans contributed as much to its total ORAC as the free fraction. Total proanthocyanidin concentration was the highest in red rice bran, while total anthocyanin was highest in purple brans. The predominant anthocyanin was cyanidin-3-glucoside. Red and purple brans had several fold higher total phenolics and flavonoids as well as ORAC and DPPH, from both free and bound fractions, than freeze-dried blueberry and broccoli. These results indicate that rice brans are natural sources of hydrophilic and lipophilic phytochemicals for use in quality control of various food systems as well as for nutraceutical and functional food application.     

体外模拟胃、肠消化对6种黑色食品抗氧化成分及其活性的影响

通过体外模拟胃、肠消化法考察6 种黑色食品（黑米、黑苦荞、黑麦、黑豆、黑绿豆、黑小米）在胃、 肠道消化过程中抗氧化成分及其活性变化规律。结果发现,通过模拟胃消化2 h后,多酚和黄酮释放增长比例最高 的分别是黑小米和黑豆,分别增长28%和41%,进一步通过模拟肠消化2 h后,多酚和黄酮释放增长比例最高的分 别是黑麦和黑绿豆,分别增长113%和52%。1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自 由基清除能力、亚铁还原能力（ferric ion reducing antioxidant power,FRAP）和2’-联氨-双-3-乙基苯并噻唑啉-6-磺 酸（2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid,ABTS）自由基清除能力在模拟胃消化阶段的增长比例 最高的分别是黑豆（114%）、黑绿豆（31%）和黑米（28%）,在模拟肠消化阶段增长比例最高的分别是黑苦 荞（161%）、黑小米（124%）和黑米（199%）。通过模拟胃蛋白酶、胰酶和胆汁等消化作用后能显著提升黑色 食品的黄酮、多酚释放量和抗氧化活性（P＜0.05）,尤其是黑米,在消化前后均表现出最强的抗氧化活性,通过 胃、肠消化后,其DPPH自由基清除能力、FRAP和ABTS＋·清除能力分别为4 531.79、3 729.69 μmol Trolox/100 g md和 3 727.14 μmol Trolox/100 g md,分别较未消化时增长了200%、171%和383%,是较有应用前景的一种黑色食品。     

黄色洋葱皮不同溶剂多酚提取物的总酚、总黄酮含量及抗氧化活性

以黄色洋葱皮为原料,分别利用70%甲醇、70%乙醇和70%丙酮作为提取溶剂进行多酚类物质提取。测定提取物中的总酚含量、总黄酮含量并研究其抗氧化活性(DPPH自由基清除能力、ABTS自由基清除能力以及铁还原能力)。研究结果显示:利用甲醇作为提取剂所得的多酚类物质提取率最高(12.35%),并且提取物中总酚和总黄酮含量最高,分别为83.17、34.31 mg/g。同时,甲醇提取物对DPPH自由基和ABTS自由基的半抑制浓度IC₅₀分别为0.059和0.031 mg/mL,低于抗坏血酸而高于乙醇提取物和丙酮提取物。     

黑小麦麸皮抗氧化活性成分提取及其抗氧化能力评价

黑小麦麸皮是黑小麦加工过程中的副产物,其富含多种生理活性物质,包括花青素、酚酸类物质、膳食纤维等。以黑小麦麸皮为原料,利用不同溶剂对其抗氧化活性成分进行提取并对其抗氧化能力评价研究。结果表明,75%乙醇提取物干物质得率最高,为10.72%;50%乙醇提取总酚含量最高,为2.9 mg/100 mL;50%乙醇提取物对1,1-二苯基-2-三硝基苯肼（DPPH）清除力最强,75%乙醇提取物对DPPH抗氧化物提取能力最强;75%丙酮提取物对2,2′-联氮-双-3-乙基苯并噻唑啉-6-磺酸（ABTS）清除力最强,75%甲醇提取物对ABTS抗氧化物提取能力最强。综合比较得出,50%乙醇更适于黑小麦麸皮抗氧化活性成分提取。     

Evaluation of antioxidant activities and phenolic subtype contents of ethanolic bran extracts of Thai pigmented rice varieties through chemical and cellular assays

This study revealed that the ethanolic bran extracts of 11 Thai pigmented (red and purple) and 2 nonpigmented rice varieties exerted scavenging activity against DPPH and ABTS radicals and ROS in HL‐60 cells in the following order: red > purple > nonpigmented rice. These rice extracts also showed the same order of phenolic and flavonoid contents, which were strongly correlated with their scavenging activity. Phenolic subtype analysis further indicated that proanthocyanidins as well as anthocyanins and protocatechuic acid contributed directly to antioxidant capacity in red and purple rice bran, respectively. In contrast, these pigmented rice bran extracts possessed moderate chelating activity partly attributed to their contents of phenolics and flavonoids, especially proanthocyanidins in red rice bran. Moreover, rice bran extracts significantly restored SOD and CAT activities in oxidative stress‐induced A549 cells. This study provides new insights on the intracellular potent antioxidant capacity of pigmented rice bran extracts in the cell‐based systems.     

Effect of fermentation by Bacillus subtilis on antioxidant and cytotoxic activities of black rice bran

Black rice bran was fermented with Bacillus subtilis KU3 isolated from Korean traditional food, Kimchi. Antioxidant and cytotoxic activities of the fermented black rice bran were investigated. Total phenolic and anthocyanin contents decreased from 171.54 mg GAE g^?1 and 2.31 mg g^?1 to 139.13 mg GAE g^?1 and 2.12 mg g^?1, respectively, after fermentation. Antioxidant activities determined by 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical scavenging, β‐carotene bleaching and ferric thiocyanate assay were correlated with total phenolic and anthocyanin contents. Non‐fermented black rice bran extract (NFBE) showed greater antioxidant activities than fermented black rice bran extract (FBE). Cytotoxic activities measured by MTT assay showed that both NFBE and FBE had over 50% activities. The cytotoxic activities of FBE against MCF‐7 and HeLa cells were 71.65% and 68.07%, respectively, at 8.0 mg mL^?1, but those of NFBE were lower than 50%. These results suggested that the cytotoxic activity of black rice bran improved through fermentation, while antioxidant activity reduced.     

Antioxidant and tyrosinase inhibitory activities of different parts of guava (<Emphasis Type="Italic">Psidium guajava</Emphasis> L.)

The antioxidant and the tyrosinase inhibitory activities of 4 different solvents (acetone, ethanol, methanol, and water) for preparation of extracts from guava (branch, fruit, leaf, and seed) were evaluated by measuring total phenolic contents (TPC), DPPH radical scavenging activity, ABTS radical scavenging activity, reducing power (RP), and tyrosinase inhibitory activity. The extracts of branch and leaf showed relatively higher antioxidant properties than those of fruit and seed. The highest TPC (141.28 mg/g gallic acid equivalents), DPPH radical scavenging activity (IC₅₀=34.01 μg/mL), ABTS radical scavenging activity (IC₅₀=3.23 μg/mL), and RP (IC₅₀= 75.63 μg/mL) were found in acetone extract of leaf, while water extract of seed had the lowest antioxidant activity. The tyrosinase inhibitory activity of ethanol extract from guava leaf was 69.56%, which was the highest activity among the extracts. These results indicate that useful bioactive substances exist in the guava branch as well as leaf extracts.     

覆盆子不同极性溶剂提取物的抗氧化活性比较

为比较覆盆子不同极性溶剂提取物的抗氧化活性,分别用正丁醇、乙酸乙酯、丙酮、去离子水、50%乙醇和石油醚进行萃取,测定各提取物总多酚和总黄酮的含量,并通过清除DPPH自由基、ABTS自由基、羟基自由基和总还原能力4种体系对其体外抗氧化活性进行评价。结果显示,乙酸乙酯提取物中总多酚含量最高,50%乙醇提取物中总黄酮含量最高;50%乙醇提取物清除DPPH自由基能力和ABTS自由基能力最强,水提取物清除羟基自由基能力最强,丙酮提取物总还原能力最强。结果表明覆盆子不同极性溶剂的提取物均有抗氧化活性,可作为天然抗氧化剂开发。     

罗非鱼尾色素中抗氧化成分的提取及活性研究

为了研究鱼尾色素的抗氧化性,以提取率和DPPH自由基清除率为评价指标,采用溶剂浸提的方法对色素进行提取,利用DPPH法、ABTS法和铁氰化钾还原法对提取物进行抗氧化实验。结果表明,鱼尾色素抗氧化成分提取的较佳工艺为浸提溶剂无水乙醇、温度20℃、料液比(g/mL)1:5、时间24h、浸提1次;此时色素提取物的提取率达2.40%,DPPH自由基清除率为58.44%。抗氧化性实验表明,鱼尾色素提取物具有较强的清除DPPH自由基和ABTS+自由基能力,其IC50值为0.3891mg/mL和1.8654mg/mL。     

黑色小扁豆中不同结合态酚类含量、花青素组成及其抗氧化活性研究

黑色小扁豆因含有丰富的营养素、微量营养素和植物化学物成分而受到研究者的关注。为深入了解小扁豆的营养价值,对小扁豆中不同结合形态的酚类化合物含量及其抗氧化活性(亚铁离子还原能力、DPPH自由基清除能力、ABTS自由基清除能力)进行测定。结果表明,黑色小扁豆中不同存在形式的酚类提取物含量差异显著。酯键合态酚类提取物中含有较高的酚酸含量(1.61 mg/g),碱水解结合态酚类提取物具有较高的黄酮含量(1.07 mg/g)及抗氧化活性【亚铁离子还原能力(43.80 mmol/g)、DPPH自由基清除能力(2.13 mg/g)、ABTS清除能力(3.92 mg/g)】。对黑色小扁豆中存在的花青素进行提取和分析鉴定,其含有的花青素主要为飞燕草-3-O-(2-O-β-D-吡喃葡萄糖基-α-L-吡喃阿拉伯糖苷)和矢车菊素衍生物。     

黑米与欧洲越橘花色苷抗氧化活性比较研究

为探究黑米与欧洲越橘花色苷抗氧化活性的差异,以抗坏血酸为阳性对照,通过比较其总还原力及清除超氧阴离子、羟自由基、过氧化氢、DPPH·、ABTS+·的能力,对其体外抗氧化作用进行了比较研究。结果表明,在测试浓度范围内,黑米花色苷与欧洲越橘花色苷总还原能力、清除过氧化氢、DPPH·、ABTS+·能力均高于阳性对照,且黑米花色苷具有更好地清除DPPH·、ABTS+·能力(IC₅₀分别为0.88、0.75 μg/mL);两种花色苷清除羟自由基、超氧阴离子自由基能力均低于阳性对照,但黑米花色苷清除超氧阴离子能力是欧洲越橘的3.63倍,清除羟自由基能力与欧洲越橘花色苷相当。研究表明,黑米花色苷具有更好的抗氧化性,是良好的天然抗氧化剂。     

Antioxidant and antimicrobial activities of Smilax china L. leaf extracts

Antioxidant and antimicrobial activities of Smilax china L. leaf extracts obtained with methanol, ethanol, acetone, and water were investigated. Antioxidant activity was evaluated by determining the DPPH radical scavenging activity, ABTS radical scavenging activities, total phenol content (TPC), and reducing power (RP). The highest DPPH, ABTS radical scavenging activity, and RP were found in the ethanol extract, which also showed the highest TPC (105.81±0.48 μg gallic acid equivalents/mL). The antimicrobial activity of all the extracts against foodborne microorganisms was determined by paper disc method. All the extracts inhibited the growth of Listeria monocytogenes, Staphylococcus aureus, and Salmonella Typhimurium, however, no antimicrobial activity was observed against Escherichia coli O157:H7. The results indicated that Smilax china L. possessed antioxidant and antimicrobial substances, and suggested that the ethanol extract can be applied into food and cosmetic industry.     

雪莲果体外抗氧化和自由基清除能力

抗氧化能力是衡量果蔬营养及保健价值的重要指标之一。雪莲果是一种药食两用植物,对多种慢性疾病有缓解作用。分别采用5 种方法(DPPH、ABTS、FRAP、SASR 和MCC)对雪莲果块根甲醇提取液的自由基清除能力及抗氧化活性进行体外评价。结果表明：DPPH 法测定的Trolox 当量抗氧化能力(TEAC)为410.263mg Trolox/g md,抗坏血酸当量抗氧化能力(AEAC)为230.485mg VC/g md,IC50 值为1.464mg/mL;ABTS 法测定的TEAC 为267.584mg Trolox/g md,AEAC为41.597mg VC/g md,IC50 值为1.269mg/mL;SRSA 法测定的TEAC 为652.816mg Trolox/g md,AEAC 为101.451mg VC/g md,IC50 值为7.720mg/mL。说明雪莲果块根提取物对DPPH 自由基、ABTS+·和O2·三种不同的自由基均有一定的清除活性。此外,雪莲果块根提取物还具有较高的总抗氧化(FRAP 值为131.723 mgFeSO4/g md)和较强的金属螯合能力(73.193%)。     

Effect of storage period on physiochemical,total phenolic content and antioxidant properties of bran enriched snacks

The objective of this investigation was to compare total phenolic content and antioxidant activities of various types of cereal brans enriched snack as affected by storage conditions. Extruded snacks were prepared by the addition of cereal brans (wheat, oat and rice) singly and in combination (w:r:o::2:1.5:1.5) to rice flour at 10, 20 and 30 % bran supplementation. The content of total phenolics in the extracts (conventional and microwave assisted extraction) was determined spectrometrically according to the Folin–Ciocalteu procedure and calculated as gallic acid equivalents. Radical scavenging capacity measured by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) methods was studied. The effect of the storage on antioxidant content and capacity showed that total phenolic content and antioxidant activities of bran enriched snacks decreased during storage at ambient temperature. However, a little increase in moisture content, water activity and free fatty acid was observed in six of month’s storage period.     

不同品种苹果非浓缩还原汁的多酚组成及与抗氧化能力的关系

目的:通过分析15种非浓缩还原(NFC)苹果汁的多酚组成、抗氧化能力及二者相关性,研究多酚组成对抗氧化能力的贡献,为果汁生产企业在原料果选择方面提供借鉴。方法:制备15个品种的NFC苹果汁,利用超高效液相色谱(UPLC)和体外抗氧化试验分析多酚组成和抗氧化能力(DPPH和ABTS自由基清除能力),通过单因素方差和相关性分析处理试验结果。结果:NFC苹果汁中,单体酚以绿原酸含量最高,表儿茶素没食子酸酯含量最低;大类多酚有原花青素、酚酸和黄酮类3种,其中酚酸含量最高(102~331μg/mL),黄酮含量最低(5~30μg/mL)。不同品种苹果中,“乔纳金”苹果的DPPH自由基清除能力最高为89.1%,“秋香”苹果的ABTS自由基清除能力最强为92.6%。结论:原花青素大类物质是NFC苹果汁体外抗氧化能力的主要贡献者,原花青素B2、表儿茶素和表儿茶素没食子酸酯3种单体酚有较强的DPPH自由基清除能力,而ABTS自由基清除能力更多地依赖于总酚含量。     

溶剂提取对青稞中不同形态多酚组成及抗氧化活性的影响

为研究不同极性溶剂对青稞全谷物中不同形态酚类化合物含量、组成及抗氧化性的影响,建立适宜于青稞全谷物中多酚提取的方法。以4?个不同品种青稞为原料,比较4?种不同溶剂及酸法、碱法分别对青稞中游离酚和结合酚含量、组成与抗氧化活性的影响。结果表明,所有提取试剂中80%丙酮溶液提取的游离态总酚含量（139.79～235.96?mg/100?g）及总黄酮含量（9.88～15.52?mg/100?g）最高,酸法提取的青稞结合态总酚含量是碱法的1.9～3.1?倍,结合态黄酮含量是碱法的1.3～2.9?倍;80%丙酮溶液提取物中检测到8～18?种青稞游离酚类化合物,且酚类化合物含量显著高于其他溶剂,绿原酸、苯甲酸、儿茶素、槲皮素、芦丁是其主要的游离酚类化合物。与碱法相比,酸法能释放出更多的结合酚类化合物类型及含量,没食子酸、p-香豆酸、丁香酸、苯甲酸、藜芦酸、橙皮苷是其主要的结合酚类化合物;参试青稞80%丙酮溶液提取物显示出最高的1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基清除能力（852.56～1?484.18?μmol/100?g）,2,2’-联氮-双-（3-乙基苯并噻唑啉-6-磺酸）二铵盐（2,2’-azinobis-(3-ethylbenzthiazoline-6-sulphonate),ABTS）自由基清除能力（358.93～518.09?μmol/100?g）及铁离子还原能力（ferric ion reducing antioxidant power,FRAP）（1?250.55～2?041.16?μmol/100?g）。酸法水解参试青稞结合酚DPPH自由基清除能力、FRAP、ABTS＋?清除能力分别是碱法水解的7.6～10.3、1.2～1.8?倍和1.1～1.3?倍。因此,80%丙酮溶剂和酸法分别是青稞中游离酚与结合酚的适宜提取溶剂,且本研究表明青稞全谷物中富含丰富的酚类物质,是一种潜在的天然抗氧化剂来源。     

核桃仁皮不同极性提取物的体外抗氧化活性分析

采用乙酸乙酯、正丁醇、氯仿、石油醚对核桃仁皮多酚进行提取,通过清除DPPH自由基、OH自由基、ABTS自由基法与铁离子还原能力初步评价其体外抗氧化活性。结果表明,不同极性溶剂对核桃仁皮多酚提取能力为乙酸乙酯>正丁醇>氯仿>石油醚;乙酸乙酯提取物具有最强的DPPH自由基清除能力和ABTS自由基清除能力,其IC₅₀值分别为5.004和29.654 μg/mL,其次为正丁醇(9.596、23.681 μg/mL)、氯仿提取物(62.719、144.475 μg/mL)以及石油醚提取物(48.316、146.424 μg/mL);正丁醇提取物具有最强的OH自由基清除能力,其IC₅₀值为393.578 μg/mL,其次为乙酸乙酯(510.186 μg/mL)、氯仿提取物(627.003 μg/mL)与石油醚提取物(840.315 μg/mL);在铁离子还原能力实验中,当样品浓度均为100 μg/mL时,各提取物的吸光度为乙酸乙酯(0.821)>正丁醇(0.509)>氯仿(0.406)>石油醚(0.142)。总之,不同极性提取物其抗氧化能力有差异,依次为乙酸乙酯>正丁醇>氯仿>石油醚,且与质量浓度呈正相关。     

青海藜麦资源酚类物质及其抗氧化活性分析

为了明确青海藜麦资源酚类物质含量及其抗氧化活性的差异,筛选出酚类物质和抗氧化活性高的藜麦资源,采用化学法分析了90份藜麦资源中游离酚类、结合酚类含量,通过DPPH·、ABTS+·清除力及FRAP铁还原力指标评价各组分的体外抗氧化活性。结果表明:不同藜麦酚类物质含量均存在较大差异,且结合酚类物质含量差异较大,游离酚类物质含量相对较小,游离酚类和结合酚类物质在清除DPPH·、ABTS+·、FRAP方面存在明显差异,其中结合酚类具有较强的清除DPPH·、ABTS+·能力,分别达4732.56、3371.97 μmol Trolox Eq/100 g DW;游离酚类的FRAP还原能力相对较强,达1408.64 μmol Trolox Eq/100 g DW。白色、黄色藜麦的结合酚类物质清除DPPH·、ABTS+·能力显著高于红色藜麦,分别达4657.41、5065.66 μmol Trolox Eq/100 g DW,3532.37、3350.72 μmol Trolox Eq/100 g DW;红色藜麦的酚类物质FRAP铁还原力最强(3264.42 μmol Trolox Eq/100 g DW)。聚类分析表明,参试的大部分青海藜麦资源结合酚类物质含量较低,DPPH·、ABTS+·清除力较弱,但FRAP铁还原力较强,其中有6个藜麦资源的结合酚类物质含量最高,5个藜麦资源的抗氧化活性最强。此结果为功能性藜麦新品种的选育提供了优质资源和指导意义。     

溪黄草多酚的抗氧化活性

对溪黄草中的多酚提取物的抗氧化性进行了研究。通过DPPH自由基清除实验、过氧化氢清除实验、还原力实验、抑制β-胡萝卜素褪色实验以及抑制亚油酸过氧化实验,分别研究了溪黄草多酚与抗坏血的抗氧化能力。结果表明:溪黄草多酚提取物具有良好的抗氧化能力,其清除DPPH自由基、清除过氧化氢、抑制亚油酸过氧化的IC50值分别为5.00、42.51与66.58μg/mL,高于抗坏血酸相应的IC50值,即5.99、56.74与94.83μg/mL。表明在试验范围内,溪黄草多酚的抗氧化活性强于抗坏血酸。