蛋清蛋白酶解物抗氧化与血管紧张素转换酶抑制活性的研究

目的研究三种蛋白酶的蛋清蛋白酶解物(EWPH)的抗氧化及血管紧张素转换酶(ACE)抑制活性。方法分别采用Fenton体系、邻苯三酚自氧化体系和亚油酸自氧化体系测定胰蛋白酶、木瓜蛋白酶及alcalase碱性蛋白酶的EWPH清除羟自由基、超氧阴离子及抑制脂质过氧化的能力。利用高效液相色谱法测定ACE抑制活性。超滤分离上述三种蛋白酶的EWPH,检测各组分的抗氧化和ACE抑制活性。结果抗氧化活性最强的是木瓜蛋白酶的EWPH,在浓度10mg/ml时,对羟自由基和超氧阴离子清除率分别为53.14%和27.87%,在浓度0.8mg/ml时,对亚油酸自氧化的抑制率73.72%(D7)。ACE抑制效果最明显的是胰蛋白酶的EWPH,其IC50值为0.985mg/ml。低分子量(Mw3ku)组分酶解物的活性优于高分子量(Mw3ku)组分。结论三种蛋白酶的EWPHs均具有抗氧化和ACE抑制活性,但两种活性之间无明显的相关,且小分子组分的活性最强。     

大麦麦叶中黄酮类化合物清除自由基动态研究

目的 :　研究 4种大麦麦叶中提取的黄酮类化合物对超氧阴离子自由基 (O2· )、羟自由基 (· OH)的清除作用。方法 :　用氮蓝四唑 (NBT)光还原法测定 O2·的清除率 ,氧化还原法测定· OH的清除率。结果 :　随着黄酮在反应液中浓度的增加 ,清除率呈上升趋势 ,当黄酮浓度达到 1 2μg/ ml左右时 ,对 O2·和· OH的清除率可分别高达 95.56%和 94.1 2 %。结论 :体外实验证实 ,大麦麦叶提取物具有较强的抗氧化作用     

富硒螺旋藻清除氧自由基的初步研究

目的与方法:采用化学发光法研究富硒螺旋藻提取液、硒藻蓝蛋白、硒藻多糖、亚硒酸钠对超氧阴离子(_O2?)和羟自由基(·OH)的清除作用。结果:当体系中提取液浓度达400μg/ml时,富硒比未富硒螺旋藻提取液对_O2?的清除率提高30%,对·OH的清除率提高20%;硒藻蓝蛋白_比藻蓝蛋白对_O2?和·OH的清除作用有所提高,并呈现明显的剂量相关性,O2?50%清除率蛋白浓度为500μg/ml,·OH50%清除率蛋白浓度为125μg/ml;硒藻多糖对氧自由基具有极强的清除效率,_O2?和·OH50%清除率时的浓度分别为12μg/ml和60μg/ml;另外,在实验浓度范围,未检测到亚硒酸钠对超_O2?和·OH具有清除作用。结论:富硒螺旋藻可能因硒的代谢而产生活性有机硒态如硒藻蓝蛋白、硒多糖等,从而提高其清除自由基活性。     

山药多糖及其硒多糖抗氧化性的比较研究

目的:比较山药多糖和山药硒多糖的抗氧化活性。方法:采用邻苯三酚自氧化法测定两者对超氧阴离子自由基的清除作用,水杨酸法测定羟自由基清除作用,HPLC法测定DPPH浓度评价对DPPH自由基的清除作用,并与维生素C进行了比较。结果:山药多糖、山药硒多糖和Vc对超氧阴离子自由基清除的IC50值分别为1.852 mg/ml、1.515 mg/ml和0.6124 mg/ml。对羟基自由基清除IC50值分别为1.130 mg/ml、0.988 mg/ml和0.075 mg/ml;对DPPH基清除的IC50值分别为0.68 mg/ml、0.62 mg/ml和0.33 mg/ml。结论:山药硒多糖比山药多糖的抗氧化活性强。     

丹酚酸抗氧化活性及其对DNA损伤保护作用

目的探讨丹酚酸抗氧化活性及其对DNA氧化损伤的保护作用机制。方法运用自由基反应模型观察丹酚酸对二苯代苦味酰肼自由基（DPPH）和羟自由基的淬灭效应,使用邻菲罗啉（Phen）-CuSO4-VC-H2O2-DNA化学发光体系测定丹酚酸对DNA损伤的抑制作用。高效液相色谱（HPLC）测定丹酚酸的组成。结果丹酚酸水提物（AE）、乙酸乙酯萃取物（EE）、大孔吸附树脂纯化物（RE）和对照维生素（VC）、二丁基羟基甲苯（BHT）对DPPH自由基的半数清除浓度（IC50）分别为141.2,15.3,14.6,15.8和71.5μg/ml。化学发光体系中,丹酚酸的加入能明显抑制并延迟DNA损伤。AE、EE、RE抑制发光50%浓度（IC50）值分别为1 293.6,62.0,49.2μg/ml。丹酚酸清除DPPH自由基能力和对羟自由基攻击的DNA损伤保护作用受丹酚酸组成和丹酚酸B含量的影响。结论丹酚酸属于预防型和断链型抗氧化剂,并可有效清除DPPH和羟自由基,对DNA损伤有明显保护作用。丹酚酸的抗氧化活性及对DNA损伤的保护作用与丹酚酸B含量有关。     

石榴籽提取物体外抗氧化活性研究

目的研究石榴籽提取物的体外抗氧化作用。方法采用普鲁士蓝法、邻二氮菲-Fe2+氧化法和邻苯三酚自氧化法分别测定石榴籽提取物的总抗氧化能力、石榴籽提取物对羟自由基(.OH)和超氧阴离子(.O2-)的清除作用,并与维生素C抗氧化活性相比较。结果石榴籽提取物对.OH和.O2-具有很强的清除作用,其IC50分别为0.157和0.027mg/ml,与维生素C抗氧化作用相似。结论石榴籽提取物具有较强的清除自由基能力,是一种天然有效的抗氧化剂。     

虾青素抗氧化活性机制研究进展

虾青素是一种氧化型类胡萝卜素,因其特殊的分子结构而具有较强的抗氧化活性,在食品添加剂、保健食品和医药等工业领域有着广阔的应用前景。本文综述了虾青素淬灭单线态氧和清除自由基、降低膜流动性、提高内源性抗氧化酶系统的防御作用、减少DNA氧化损伤和抑制脂质过氧化等近年来抗氧化活性方面的研究进展。     

甘露寡糖对小鼠肝脏中自由基的清除及抗氧化酶活性的影响

目的:研究微波合成的甘露寡糖(MOS)在体外清除羟自由基(·OH)及小鼠肝脏清除自由基和抗氧化酶活性。方法:以甘露糖为原料合成甘露寡糖。体外:测定其对于羟自由基(·OH)的清除率。体内:昆明种雄性小鼠(体重:20±2g)36只,按体重随机分为6组,每组6只,每日灌胃一次,分别为正常对照组(2g/kg生理盐水)、葡萄糖组(2g/kg葡萄糖)、甘露糖组(2g/kg甘露糖),MOS加葡萄糖组(MOS1g+葡萄糖)1g/kg,低浓度MOS组(MOS2g/kg),高浓度MOS组(MOS5g/kg),灌胃14d后,无菌取肝脏组织待测。结果:补充1～2g/kg的MOS可提高小鼠肝脏组织SOD、CAT、GSH-Px活性,提高GSH水平,提高肝脏组织总抗氧化能力及对·OH的清除率,降低MDA含量及ROS水平。结论:MOS体外可有效清除羟自由基(·OH);小鼠适量添加MOS有益于提高肝脏组织的抗氧化能力,保护肝脏免受各种氧化应激和损伤。     

新疆树莓果实营养成分及其提取物抗氧化性研究

目的分析新疆野生树莓果实营养成分,并对其提取物进行抗氧化活性测定。方法进行营养保健成分含量分析,并对果实95%乙醇提取物经石油醚脱脂后,依次用乙酸乙酯、正丁醇萃取,观察不同萃取部位对DPPH·和·OH的清除能力。结果树莓鲜果中必需氨基酸含量高达320mg%,非必需氨基酸中谷氨酸含量较高(180mg%);富含维生素C(28.04mg%)和钾(147.32mg%);有机酸含量高达1.72%,以柠檬酸为主;其它如总酚、总黄酮、鞣化酸、单宁含量分别为498、125、2.2和290mg%,SOD活性高达606.93U/ml。树莓不同提取物均显示了较好的抗氧化活性,以乙酸乙酯部分最高,与清除DPPH自由基相比,清除羟自由基能力强。结论新疆野生红树莓果富含多种营养保健成分和抗氧化活性,有很好的开发价值。     

三七多糖抗氧化活性研究

目的:探讨三七多糖的体外抗氧化活性。方法:三七粗多糖经Sevage法纯化后用苯酚—硫酸法测定其含量,并采用还原/抗氧化能力、清除DPPH自由基、OH自由基和ABTS+自由基能力考察三七多糖的抗氧化能力。结果:三七多糖含量为80.49 mg/g;与对照品BHA和茶多酚相比较,还原/抗氧化能力和清除DPPH自由基能力(FRAP=11586.3μmol/g、IC50=0.055 mg/ml)都比茶多酚(FRAP=6507.5μmol/g、IC50=0.143mg/ml)强,但都比BHA(FRAP=1536.5μmol/g、IC50=0.030 mg/ml)弱;清除羟基自由基能力比BHA(IC50=0.028 mg/ml)及茶多酚(IC50=0.002 mg/m L)都强;清除ABTS+自由基能力(IC50=0.018 mg/ml)比BHA(IC50=0.175 mg/ml)强,但比茶多酚(IC50=0.002 mg/ml)弱。结论:三七多糖具有较强的抗氧化能力,为其生物活性的深入研究提供了参考依据。     

The antioxidant and radical scavenging activities of black pepper (Piper nigrum) seeds

Water and ethanol crude extracts from black pepper (Piper nigrum) were investigated for their antioxidant and radical scavenging activities in six different assay, namely, total antioxidant activity, reducing power, 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, and metal chelating activities. Both water extract (WEBP) and ethanol extract (EEBP) of black pepper exhibited strong total antioxidant activity. The 75?µg/ml concentration of WEBP and EEBP showed 95.5% and 93.3% inhibition on peroxidation of linoleic acid emulsion, respectively. On the other hand, at the same concentration, standard antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and α-tocopherol exhibited 92.1%, 95.0%, and 70.4% inhibition on peroxidation of linoleic acid emulsion, respectively. Also, total phenolic content in both WEBP and EEBP were determined as gallic acid equivalents. The total phenolics content of water and ethanol extracts were determined by the Folin-Ciocalteu procedure and 54.3 and 42.8?µg gallic acid equivalent of phenols was detected in 1?mg WEBP and EEBP.     

正相高效液相色谱-荧光检测法同时测定植物油中的四种维生素E异构体

目的:建立正相高效液相色谱-荧光检测法测定植物油中的四种维生素E异构体。方法:采用Waters Sunfire硅胶柱(4.6 mm×250 mm,5μm),流动相为正己烷∶叔丁基甲醚∶2-甲基环丙烷=92∶8∶0.8,流速1.2 ml/min,柱温25℃,荧光检测器检测。结果:该方法测定四种维生素E的线性范围均为0.5μg/ml～100.0μg/ml,四种维生素E的检出限均为0.1μg/ml,加标回收率为92%～98%。结论:该方法操作简单、灵敏度高、回收率和重现性良好,结果准确可靠。     

甲醇提取柑橘皮总黄酮及其体外抗氧化活性研究

目的:以柑橘皮为原料,利用甲醇为溶剂,采用浸提法从柑橘皮提取生物活性物质总黄酮,通过体外实验评价柑橘皮总黄酮的抗氧化活性,以提高柑橘资源的开发利用效率。方法:通过单因素实验和正交实验确定柑橘皮总黄酮的最佳提取工艺条件。以芦丁为对照,体外测定柑橘皮总黄酮提取物对超氧阴离子自由基(O-2.)、羟自由基(.OH)及二苯代苦味酰基苯肼(DPPH.)自由基的清除率。结果:以甲醇为溶剂提取柑橘皮总黄酮的最佳提取工艺条件为:料液比1:30、溶剂浓度70%、温度60℃、浸提时间4 h的提取条件下,总黄酮产量为4.96%。体外抗氧化活性研究结果表明,柑橘皮总黄酮具有较高的自由基清除能力,其清除效果随提取物浓度的增加而增强。柑橘皮总黄酮对O-2.和DPPH.自由基的清除率高于芦丁,而对于.OH自由基的清除率比芦丁稍低。结论:柑橘皮总黄酮具有明显的抗氧化能力,为综合开发利用柑橘资源提供了科学依据。     

Free radical scavenger and antioxidant capacity correlation of alpha-tocopherol and Trolox measured by three in vitro methodologies

The objective of this study was to correlate the free radical scavenging and antioxidant activity of two known substances (Trolox and alpha-tocopherol), using three in vitro methods (linoleic acid emulsion, brain homogenate and 2,2-diphenyl-1-picryl-hydrazyl [DPPH]). At steady state, alpha-tocopherol showed a greater inhibition of spontaneous oxidation of brain homogenate (59.42%+/-1.91) than Trolox (38.50%+/-2.38), while the latter showed a better antioxidant activity performance regarding inhibition of linoleic acid peroxidation (100% versus 84.02%+/-1.98) and free radical scavenging activity (93.56%+/-5.71 versus 66.72%+/-6.28). When the IC50 value was used as a parameter, alpha-tocopherol presented greater antioxidant activity than Trolox evaluated in brain homogenate and DPPH, without a significant difference when using linoleic acid emulsion. Both compounds showed the same antioxidant efficiency measured by DPPH kinetics (0.37 mM). Antioxidant activity significantly changed according to the substrate, the parameter adopted to compare the substances in the same method and the form used to express antioxidant concentration.     

In vitro antioxidant and hypoglycemic activities of Ethiopian spice blend Berbere

The metal chelating activity, antioxidant properties, and the effect on carbohydrate-hydrolyzing enzymes of Ethiopian spice blend Berbere have been investigated. Berbere contains a total amount of phenols corresponding to 71.3 mg chlorogenic acid equivalent per gram of extract and a total flavonoid content of 32.5 mg quercetin equivalent per gram of extract. An increase of the resistance towards forced oxidation was obtained when Berbere was added to sunflower oil. In order to evaluate the bioactivity of the non-polar constituents, an n-hexane extract was obtained from Berbere. The gas chromatography-mass spectrometry analysis revealed the presence of 19 fatty acids constituents (98.1% of the total oil content). Among them, linoleic acid was the major component (72.0% of the total lipids). The ethanolic extract had the highest ferric-reducing ability power (35.4 μM Fe(II)/g) and DPPH scavenging activity with a concentration giving 50% inhibition (IC(50)) value of 34.8 μg/ml. Moreover, this extract exhibited good hypoglycemic activity against α-amylase (IC(50) = 78.3 μg/ml). In conclusion, Ethiopian spice blend Berbere showed promising antioxidant and hypoglycemic activity via the inhibition of carbohydrate digestive enzymes. These activities may be of interest from functional point of view and for the revalorization of the spice blend in gastronomy also outside the African country.     

The antioxidant and radical scavenging activities of black pepper (Piper nigrum) seeds

Water and ethanol crude extracts from black pepper (Piper nigrum) were investigated for their antioxidant and radical scavenging activities in six different assay, namely, total antioxidant activity, reducing power, 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, and metal chelating activities. Both water extract (WEBP) and ethanol extract (EEBP) of black pepper exhibited strong total antioxidant activity. The 75 microg/ml concentration of WEBP and EEBP showed 95.5% and 93.3% inhibition on peroxidation of linoleic acid emulsion, respectively. On the other hand, at the same concentration, standard antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and alpha-tocopherol exhibited 92.1%, 95.0%, and 70.4% inhibition on peroxidation of linoleic acid emulsion, respectively. Also, total phenolic content in both WEBP and EEBP were determined as gallic acid equivalents. The total phenolics content of water and ethanol extracts were determined by the Folin-Ciocalteu procedure and 54.3 and 42.8 microg gallic acid equivalent of phenols was detected in 1 mg WEBP and EEBP.     

液质联用法同时测定保健食品中维生素E、维生素E乙酸酯和辅酶Q10

目的采用高效液相色谱-串联质谱法(HPLC-MS/MS),建立保健食品中维生素E、维生素E乙酸酯和辅酶Q10的检测方法。方法样品经过乙腈超声提取(50℃),采用HPLC-MS/MS法进行检测。色谱条件:采用Agilent Eclipse-plus C18(2.1 mm×50 mm,1.8μm)色谱柱,流动相为乙醇-甲醇(含0.1%甲酸)溶液,梯度洗脱,流速为0.2 ml/min,柱温为35℃。质谱条件:电喷雾离子源(ESI+),干燥气温度为250℃,鞘气流速为11 L/min,源电压为4 kV,检测方式为多重反应监测(MRM)模式。结果维生素E、维生素E乙酸酯和辅酶Q10在相应的浓度范围内与峰面积都呈良好线性关系(r>0.99),加样回收率为78.1%~96.3%,精密度RSD<6%,方法检出限在0.008μg/g^0.15μg/g。结论此方法测定快速准确、灵敏度高,专属性强,可用于保健食品中维生素E、维生素E乙酸酯和辅酶Q10的检测。     

In vitro antioxidant and hypoglycemic activities of Ethiopian spice blend Berbere

The metal chelating activity, antioxidant properties, and the effect on carbohydrate-hydrolyzing enzymes of Ethiopian spice blend Berbere have been investigated. Berbere contains a total amount of phenols corresponding to 71.3 mg chlorogenic acid equivalent per gram of extract and a total flavonoid content of 32.5 mg quercetin equivalent per gram of extract. An increase of the resistance towards forced oxidation was obtained when Berbere was added to sunflower oil. In order to evaluate the bioactivity of the non-polar constituents, an n-hexane extract was obtained from Berbere. The gas chromatography–mass spectrometry analysis revealed the presence of 19 fatty acids constituents (98.1% of the total oil content). Among them, linoleic acid was the major component (72.0% of the total lipids). The ethanolic extract had the highest ferric-reducing ability power (35.4 μM Fe(II)/g) and DPPH scavenging activity with a concentration giving 50% inhibition (IC₅₀) value of 34.8 μg/ml. Moreover, this extract exhibited good hypoglycemic activity against α-amylase (IC₅₀ = 78.3 μg/ml). In conclusion, Ethiopian spice blend Berbere showed promising antioxidant and hypoglycemic activity via the inhibition of carbohydrate digestive enzymes. These activities may be of interest from functional point of view and for the revalorization of the spice blend in gastronomy also outside the African country.     

In vitro neuroprotective effects of the leaf and fruit extracts of Juglans regia L. (walnut) through enzymes linked to Alzheimer's disease and antioxidant activity

Several extracts of the leaves and fruits of Juglans regia L. were assessed for their neuroprotective effects through antioxidant and anti-cholinesterase methods. Anticholinesterase activity was determined against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), the enzymes vital for Alzheimer's disease, at 50, 100 and 200 μg ml(-1). Antioxidant activity was tested using radical scavenging activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH), N,N-dimethyl-p-phenylenediamine (DMPD), superoxide (SO), nitric oxide (NO) and hydrogen peroxide (H(2)O(2)) radicals, as well as ferric ion-chelating capacity, ferric- and phosphomolybdenum-reducing antioxidant power at 500, 1000 and 2000 μg ml(-1). Total phenol and flavonoid quantification of the extracts was calculated. The extracts scavenged DPPH radical in varying degrees; however, they did not scavenge DPMD and H(2)O(2). Only the dichloromethane and water extracts were able to quench SO (10.09 ± 1.38%) and NO (24.09 ± 2.19%) radicals, respectively, at low level. The extracts showed either low or no BChE inhibition and no AChE inhibition.     

Vitamin E in foods from high and low linoleic acid diets

As part of a human diet study, vitamin E activity was estimated in foods used in seven daily menus. Each menu was designed to contain 35% fat calories with either 10 or 30 gm/day of linoleic acid (18:2) and 500 mg/day of cholesterol. To estimate vitamin E activity, each food used in the menus was analyzed for alpha and gamma tocopherol content by high-pressure liquid chromatography with fluorescence detection. This article reports the alpha and gamma tocopherol contents of those foods, tocopherol contributions from each food in one sample 2,400-kcal menu, and the mean daily vitamin E activity (milligram alpha tocopherol equivalents) of all seven menus at five caloric levels. Major sources of alpha tocopherol (greater than 10% of the RDA) common to both diets (10 and 30 gm linoleic acid) were olive oil and a few fruits and vegetables. Additional major sources in the 30-gm linoleic acid diets were polyunsaturated fatty acid (PUFA) vegetable oils and margarine. Contrary to a common assumption, increasing the level of PUFA in the menus did not necessarily result in higher milligram equivalents of alpha tocopherol because soybean oil, with a tocopherol composition that is predominantly gamma tocopherol, was the major source of linoleic acid in the diets. Thus, vitamin E activity was not necessarily increased when soybean oil was substituted for a less saturated fat such as olive oil, which has mostly alpha tocopherol.