Nutrition-based health: cell-based bioassays for food antioxidant activity evaluation

Food science has progressively evolved and now there are wide evidences that foods have biological activities that are beyond their classical nutritional value. In this field, the antioxidant activity of pure compounds, food, feed, and dietary supplements has been extensively studied and numerous analytical approaches and assay models have been developed, involving various systems from simple chemical assays to animal models and human studies. This article is an overview of different cell-based models that have been used for testing the antioxidant properties of food, feed, and dietary supplements. Advantages, drawbacks, and technical problems to develop and validate suitable, robust, and high-throughput cell-based bioassays for screening food antioxidant activity will be discussed.     

鞣花酸和石榴皮多酚提取物抗氧化活性的比较

对鞣花酸提取物(EA)和石榴皮多酚提取物(PE)的抗氧化活性进行比较研究。通过HPLC法检测EA和PE中鞣花酸的含量,Folin-酚法检测PE中总多酚含量;采用H2O2处理HepG2细胞建立细胞氧化应激损伤模型,MTT法比较不同浓度的EA和PE对HepG2细胞氧化应激损伤的保护作用;采用DPPH法、·OH清除法检测PE和EA体外抗氧化作用。结果表明:与EA相比,PE中鞣花酸含量很低,但PE和EA均以剂量依赖的方式提高氧化损伤的HepG2细胞生存率,有显著的保护作用,且在同一的浓度下,PE的保护作用显著高于EA。PE和EA对DPPH法、·OH清除法的清除率均呈剂量依赖性增强。     

真空提取核桃粕多酚的抗氧化活性研究

目的 探究真空提取的核桃粕(walnut meal)多酚的抗氧化活性。方法 以非真空条件下提取的多酚为对照,采用超声波辅助真空法提取核桃粕多酚,用HPD-100型大孔树脂对多酚进行纯化,福林酚(Folin-Ciocalte)法测定多酚含量,通过试剂盒检测多酚对1,1-二苯基-2-苦基肼自由基(1,1-diphenyl-2-picrylhydrazyl,DPPH)、超氧阴离子自由基的清除率及Fe³⁺还原能力(ferric reducing antioxidant power,FRAP)来评价其体外抗氧化活性,用cck8(cell countingkit-8)法测定不同浓度的核桃粕多酚对HepG2细胞的毒性作用,确定3个无毒性作用浓度,以770μmol/L的H₂O₂诱导HepG2细胞建立氧化应激损伤模型,通过测定HepG2细胞中丙二醛(malonicdialdehyde, MDA)含量、超氧化物歧化酶(superoxide dismutase, SOD)活力、谷胱甘肽(glutathione, GSH)含量探究12.5、25...     

咖啡酸维生素C酯的合成、抑菌活性和抗氧化性研究

研究了以3,4-二羟基苯甲醛、丙二酸和维生素C为原料,经过Knoevenagel缩合和直接酯化一锅法合成咖啡酸维生素C酯的方法,并考察了咖啡酸维生素C酯的抑菌活性和抗氧化性。在催化剂SO42-/ZrO2的作用下,3,4-二羟基苯甲醛与丙二酸首先发生Knoevenagel缩合生成咖啡酸,产物不需要分离,加入维生素C继续进行酯化反应,以85.1%的产率得到了咖啡酸维生素C酯,产物结构用1HNMR和IR进行确证。抑菌活性实验表明,咖啡酸维生素C酯对金黄色葡萄球菌、大肠杆菌、枯草杆菌、酿酒酵母、青霉、黄曲霉和黑曲霉均有较强的抑制作用,对细菌和酵母的抑制作用高于霉菌。抗氧化性实验表明,咖啡酸维生素C酯可以有效清除DPPH自由基和羟基自由基,清除效果明显好于维生素C。      

加工工艺对牡丹籽油脂肪酸成分、理化性质及抗氧化能力的影响

目的 研究加工工艺对牡丹籽油的脂肪酸组成、理化性质及抗氧化特性的影响。方法 采用直接冷榨法、炒制+冷榨法和超临界CO2萃取技术制取牡丹籽油, 以出油率、优质品质、脂肪酸组成和抗氧化能力为指标比较了不同工艺制备的牡丹籽油的品质。结果 直接冷榨的出油率(5.6%)远低于炒制+冷榨(8.3%)和超临界CO2萃取(8.2%), 直接冷榨制备的油脂品质和活性成分及抗氧化活性与超临界CO2萃取无显著性差异(P>0.05), 炒制+冷榨则会造成脂肪酸组成和抗氧化能力的较大差别(P<0.05)。结论 冷榨可以较好地保留牡丹籽油的活性成分和品质。     

11种植物油的脂肪酸组成与抗氧化活性比较

对4种新资源食品油、2种高端食用油和5种普通植物油的脂肪酸组成和抗氧化活性进行比较研究,以评价不同类型植物油的质量。采用气-质联用色谱法测定了11种植物油的脂肪酸组成,应用DPPH、ABTS、FRAP法对不同植物油的全油以及极性部位的抗氧化活性进行测定。结果显示:新资源食品油(茶叶籽油除外)的多不饱和脂肪酸含量均大于65%,其中α-亚麻酸高达38%以上,而普通植物油以及牛油果油和橄榄油中α-亚麻酸含量仅有0.11%～8.67%; 11种植物油对DPPH·清除能力从大到小依次为杜仲籽油美藤果油玉米油大豆油葵花籽油牡丹籽油菜籽油橄榄油牛油果油花生油茶叶籽油;对ABTS+·的清除能力从大到小依次为橄榄油菜籽油杜仲籽油大豆油玉米油美藤果油花生油牛油果油茶叶籽油牡丹籽油葵花籽油;对FRAP Fe3+-TPTZ还原能力从大到小依次为杜仲籽油美藤果油菜籽油橄榄油玉米油花生油大豆油牡丹籽油葵花籽油牛油果油茶叶籽油。在脂肪酸组成方面,新资源食品油有较大优势,DPPH、ABTS、FRAP 3种方法所得的每种植物油的抗氧化活性不同,但杜仲籽油、美藤果油、菜籽油和玉米油在3种方法中均表现出较好的抗氧化活性。     

Effect of dietary antioxidant and fatty acid supply on the oxidative stability of fresh and cooked pork

The effect of dietary oil (linseed or soybean oil) and antioxidant treatment (α-tocopheryl acetate (AT; 40ppm) versus a cocktail (AOC; 200ppm): α-tocopheryl acetate+rosemary+citric acid+gallic acid) on colour, lipid and protein oxidation of fresh and processed pork was investigated. No effect of oil source on different parameters of oxidation was seen. No effect of antioxidant treatment on colour stability of fresh longissimus thoracis (LT) or cooked cured ham (CCH) was observed. For both antioxidant treatments, lipid oxidation in fresh LT and CCH was well controlled during display. However, lipid oxidation increased significantly in pre-frozen uncured cooked meat under aerobic conditions. No unambiguous effect of antioxidant treatment on protein oxidation was observed. There seemed to be no clear link between colour, protein and lipid oxidation. At the dose used in this study, no additional or synergistic effects of the extra components of the AOC on the different oxidation parameters was found.     

西瓜籽油脂肪酸成分分析及抗氧化活性研究

目的：研究西瓜籽油脂肪酸成分及抗氧化活性。方法：采用索氏回流法对西瓜籽进行提油,对西瓜籽油进行酯化处理,用气相色谱—质谱联机（GC-MS）对其脂肪酸的成分、含量进行分析,通过其对铁离子的还原能力和对DPPH自由基、羟基自由基及超氧阴离子自由基清除能力的测定。结果：西瓜籽油中含有6种脂肪酸,其中不饱和脂肪酸相对含量达到50%以上;西瓜籽油对铁离子具有还原能力,对DPPH自由基的IC₅₀值为3.7 mg/mL,对超氧阴离子自由基的IC₅₀值为0.031 mg/mL,对羟基自由基的IC₅₀值为12.04 mg/mL。结论：西瓜籽油富含不饱和脂肪酸,有较好的抗氧化活性。     

A systematic survey of antioxidant activity of 30 Chinese medicinal plants using the ferric reducing antioxidant power assay

The antioxidant activities and total phenolic contents of 30 Chinese medicinal plants were evaluated using the ferric reducing antioxidant power assay and the Folin–Ciocalteu method, respectively. The Chinese medicinal plants were extracted by the traditional method, boiling in water and also in 80% methanol. A significant and linear correlation coefficient between the antioxidant activity and the total phenolic content was found in both aqueous (R² = 0.7917) and methanol (R² = 0.7584) extracts. Phenolic compounds are thus a major contributor of antioxidant activity. Comparing the extraction efficiency of the two methods, the boiling water method extracted phenolic compounds more efficiently, and antioxidant activity of the extract was higher. It was found that the Chinese medicinal plants Rhodiola sacra Fu, the stem of Polygonum multiflorum Thunb. and the root of P. multiflorum Thunb. possessed the highest antioxidant activities and thus could be potential rich sources of natural antioxidants.     

低分子量透明质酸的制备及其抗氧化活性的研究

首先研究了Fe2+-S2O2-8氧化体系硫酸根自由基(SO-4·)对透明质酸(HA)的氧化降解作用,初步确定了SO-4·氧化降解HA制备低分子量透明质酸(LMWHA)的实验条件。接着HA降解产物经过乙醇沉淀以及凝胶柱层析纯化,采用高效凝胶过滤色谱法(HPGFC)测得纯化后LMWHA分子量为1.45×105Da。最后采用不同的体外抗氧化实验,并以HA为对照,研究了LMWHA的抗氧化活性,发现SO-4·降解后生成的LMWHA的抗氧化活性高于降解前的。本实验说明用SO-4·降解HA制备具有抗氧化活性LMWHA是可行的,为LMWHA抗氧化药物的开发提供了理论依据。     

金银花中绿原酸的双水相萃取及抗氧化能力研究

为合理利用大别山金银花资源,以罗田县的金银花及其茎、叶为试验材料,利用双水相体系萃取绿原酸,并研究了其抗氧化活性。结果表明：50 mL丙酮/磷酸氢二钾(K2HPO4)双水相体系萃取绿原酸的最佳条件为酮水比8.5∶1.5(V/V),磷酸氢二钾添加量3 g,水提液绿原酸质量浓度22.9 g/L。在此条件下,绿原酸含量为56.27%,收率为98.46%,且其绿原酸的羟基自由基清除能力较强,从而为大别山金银花资源的开发利用提供了理论依据。     

芝麻粕植酸的超声波辅助提取工艺优化及其抗氧化活性研究

为探索芝麻粕植酸的超声波辅助醋酸法提取最佳工艺条件,采用单因素试验及响应面法对植酸提取工艺进行优化,并分析芝麻粕植酸的抗氧化活性。结果表明:芝麻粕植酸提取的最佳工艺条件为超声波功率270 W、超声波时间20 min、醋酸质量分数15%、料液比1∶16、提取时间115 min、提取温度51℃,在最佳工艺条件下芝麻粕植酸的提取率为73.24%。体外抗氧化试验表明,芝麻粕植酸具有一定的抗氧化活性,其还原力、DPPH自由基清除率和羟自由基清除率的较适质量浓度分别为80、100μg/mL和120μg/mL。     

红枣可溶性膳食纤维的抗氧化活性研究

为研究红枣可溶性膳食纤维（Soluble Dietary Fiber,SDF）的体外抗氧化活性,采用Na NO₂-Al（NO₃）₃-Na OH法和Folin-Ciocalteu法分别测定红枣SDF中总黄酮和总多酚含量,并通过DPPH·、ABTS⁺·、·OH、O₂^-·和H₂O₂自由基清除体系和还原能力评价红枣SDF的抗氧化活性。结果表明:红枣SDF中总黄酮和总多酚含量分别为（1.78±0.04）mg/g和（4.12±0.09）mg/g;红枣SDF具有较强的体外抗氧化活性,对DPPH·和H₂O₂清除能力较强,2.0 mg/m L红枣SDF的清除率分别可达到79.81%和50.07%,尤其对DPPH·的清除能力最强,其半数抑制浓度(IC₅₀)为0.725 mg/m L,对ABTS⁺·、·OH、O₂^-·的清除能力较弱,并具有一定还原能力。红枣SDF具有一定的体外抗氧化活性,可作为一种有效的天然食品抗氧化剂或用于开发红枣膳食纤维功能性食品。      

Effect of dietary antioxidant and increasing corn oil inclusion on milk fat yield and fatty acid composition in dairy cattle

The objective of this study was to examine the effect of a dietary synthetic antioxidant on feed intake, yields of milk and milk components and milk fatty acids (FA), in combination with increasing concentrations of dietary corn oil to provide increasing rumen unsaturated fatty acid load (RUFAL) challenges. Twenty-six Holstein cows (177 ± 57 d in milk; mean ± standard deviation) were assigned to treatment in a randomized complete block design. Treatments were a control diet (CON; n = 13 cows) or the same diet supplemented with a synthetic antioxidant (AOX; 6.1 g/d; dry blend of ethoxyquin and propyl gallate, Novus International Inc., St. Charles, MO; n = 13 cows). In period 1 (21 d), no supplemental corn oil was fed; in periods 2, 3, and 4 (14 d each), corn oil was supplemented at 0.7, 1.4, and 2.8% of the diet [dry matter (DM) basis] to incrementally increase RUFAL. For all variables measured, no significant interactions were detected between treatment and period, indicating no differences between the CON and AOX treatments at all levels of oil inclusion. Intake of DM was lower for AOX compared with CON but AOX had no effect on milk yield or milk fat concentration and yield. Milk protein yield and feed efficiency (energy-corrected milk/DM intake) tended to be greater for AOX compared with CON. Increasing dietary corn oil concentration (RUFAL) decreased DM intake, milk yield, milk fat concentration and yield, and feed efficiency. The AOX treatment increased the concentration and yield of 16-carbon milk FA, with no effect on de novo (<16 carbon) or preformed (>16 carbon) milk FA. Milk FA concentration of trans-10 C18:1, trans-10,cis-12 C18:2, and trans-9,cis-11 C18:2 were unaffected by AOX but increased with increasing RUFAL. In conclusion, supplementation with AOX did not overcome the dietary-induced milk fat depression caused by increased RUFAL.     

野生柳叶蜡梅籽油理化性质、脂肪酸组成及抗氧化活性研究

以成熟的野生柳叶蜡梅种子为材料,采用超声波辅助法提取籽油,测定籽油的理化性质、维生素E含量和脂肪酸组成,并以ABTS自由基清除作用和金属离子螯合作用为指标评价籽油的抗氧化活性。结果表明:野生柳叶蜡梅籽油酸价(KOH)为2. 28 mg/g、碘值(I)为109. 47 g/100 g、过氧化值为2. 03 mmol/kg、皂化值(KOH)为185. 81 mg/g、水分及挥发物为0. 82%、相对密度(20℃)为0. 927 5、折光指数(20℃)为1. 467 3、维生素E含量为109. 27 mg/kg;从野生柳叶蜡梅籽油中共鉴定出22种脂肪酸,包括8种饱和脂肪酸(24. 28%)和14种不饱和脂肪酸(75. 37%),含量较高的脂肪酸有亚油酸(55. 60%)、棕榈酸(20. 26%)、油酸(8. 08%)、10,13-十八碳二烯酸(8. 02%);野生柳叶蜡梅籽油具有良好的抗氧化活性,对ABTS自由基清除效果优于葡萄籽油,对金属离子螯合作用弱于葡萄籽油。     

鱼腐乳发酵过程中游离氨基酸含量及抗氧化活性的变化

利用总状毛霉（Mucor racemosus）和红曲霉（Monascus）F1对鱼豆腐进行发酵研制鱼腐乳,对鱼腐乳发酵过程中的游离氨基酸及抗氧化活性的变化规律进行分析。结果表明：在发酵过程中,鲜味氨基酸、必需氨基酸、疏水氨基酸、总氨基酸的含量都呈逐渐升高的趋势,在后熟第28天时分别达到了138.57 mg/100 g、491.68 mg/100 g、340.06 mg/100 g、915.77 mg/100 g。在发酵过程中DPPH自由基清除率和羟自由基清除率也呈逐渐升高的趋势,在后熟第28天时分别达到70.30%和63.11%,分别比发酵开始时提高了53.09%和24.14%。     

魔芋葡甘聚糖及其油酸酯化物的体外抗氧化初步研究

对魔芋葡甘聚糖(KGM)和魔芋葡甘聚糖油酸酯(KGM酯)体外抗氧化性进行了研究。通过氧化诱导试验、清除DPPH自由基、清除ABTS自由基、对Fe~(2+)螯合能力、还原能力试验发现:KGM和KGM酯均有一定的抗氧化性,酯化度越高,抗氧化能力越强;当菜籽油中KGM和酯化度最高的KGM酯1添加量为0.8%时,与空白对照相比其氧化诱导时间分别延长5.94 h和2.56 h;在KGM酯与茶多酚、V_C、TBHQ协同试验中,KGM酯也表现出一定抗氧化性;质量浓度为0.5 mg/m L时,KGM和KGM酯1对DPPH自由基清除率分别为61.68%和36.93%;质量浓度为2.0 mg/m L时,KGM和KGM酯1对ABTS自由基清除率分别为20.31%和17.26%,对Fe~(2+)螯合率分别为17.58%和9.79%。虽然KGM酯化后其抗氧化能力稍有减弱,但仍具有抗氧化性,且与酯化程度呈正相关。     

核桃花中绿原酸、新绿原酸和隐绿原酸的提取工艺响应面优化及其抗氧化活性研究

目的 探讨干燥核桃花中绿原酸、隐绿原酸和新绿原酸的最佳提取工艺及其抗氧化活性。方法 采用超声波辅助乙醇法提取核桃花中的绿原酸、新绿原酸和隐绿原酸, 以提取温度、乙醇体积分数、提取时间和料液比为自变量, 以得率的总评归一值(overall desirability, OD)为因变量, 应用响应面对提取工艺进行优化, 并通过体外实验评价了其抗氧化活性。结果 最佳提取工艺为：温度77℃, 乙醇体积分数67%, 提取时间28 min, 料液比1: 23(g/mL)。此时,绿原酸、新绿原酸和隐绿原酸的总提取率最佳, OD值可达0.91, 对DPPH自由基、ABTS自由基和羟自由基的半数抑制浓度(Half-maximal Inhibitory concentrations, IC50)值分别为41.63、41.68和34.43 μg/mL。结论 优选出的工艺稳定可行, 且提取液具有较好的体外抗氧化活性, 为核桃花中绿原酸类成分的进一步开发利用提供了理论基础。