Comparison of the 2,2'-azinobis-3-ethylbenzotiazo-line-6-sulfonic acid (ABTS) and ferric reducing anti-oxidant power (FRAP) methods to asses the total antioxidant capacity in extracts of fruit and vegetables

A comparison was made on the use of two spectrophotometric methods, the ferric reducing antioxidant power (FRAP) method and the 2,2'-azinobis-3-ethylbenzotiazoline-6-sulfonic acid (ABTS) method, for the measurement of the total antioxidant capacity (TAC) of plant foods. The correlations of TAC measured by the two methods were highly significant in both water-soluble (r2= 0.90) and water-insoluble extracts (r2= 0.98) from 13 strawberry samples. Also a corresponding comparison of TAC in extracts from 14 plant species showed high correlation coefficients, r2= 0.98 for water-soluble extracts and r2= 0.88 for water-insoluble extracts. The ratio of TAC values obtained with the two methods (ABTS/FRAP) varied between 0.7 and 3.3 for different plant extracts indicating that they contained antioxidants with varying reactivity in the two methods. TACs in six pure antioxidant substances were ranked in the following order by both methods: quercetin > ferulic acid > catechin > rutin > caffeic acid > Trolox = chlorogenic acid. The two methods showed similar TAC values for quercetin, rutin, caffeic acid and chlorogenic acid while ferulic acid and catechin gave higher results with the ABTS method than with the FRAP method, and such differences probably explain the varying ratios of ABTS/FRAP obtained in foods. Regarding storage TAC in water-soluble strawberry extracts stored at -20 or -80 degrees C was stable for at least five months while storage at 4 degrees C decreased the TAC value with 40% during five weeks of storage. The study showed that both the ABTS and FRAP methods can be used for convenient monitoring of the antioxidant capacities in fruit and vegetables, and that different antioxidants had varying reactivity in the two methods.     

凤头姜醇溶和水溶黄酮的抗氧化作用

比较凤头姜水溶和醇溶黄酮对超氧阴离子自由基、1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基、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)自由基的清除能力和总抗氧化能力(铁离子还原能力法),同时用VC和2,6-二叔丁基对甲酚(butylated hydroxytoluene,BHT)作对照。结果表明:在DPPH自由基清除能力测定中,水溶黄酮的抗氧化效果强于醇溶黄酮和BHT,但弱于VC,水溶和醇溶黄酮的半抑制浓度(half maximal inhibitory concentration,IC_(50))分别为585.75μg/m L和1 013.45μg/m L;在ABTS+·、超氧阴离子自由基清除能力和总抗氧化能力测定中,醇溶黄酮的抗氧化效果均强于水溶黄酮、VC和BHT,醇溶和水溶黄酮对ABTS+·的IC_(50)分别为21.90μg/m L和87.54μg/m L;在超氧阴离子自由基清除能力测定中,水溶和醇溶黄酮的IC_(50)分别为26.56μg/m L和22.29μg/m L;在铁离子还原能力测定中,水溶黄酮和醇溶黄酮的TEAC1 000分别为45.78μg/m L和36.42μg/m L。综合研究发现,凤头姜水溶黄酮和醇溶黄酮在不同抗氧化体系中的抗氧化效果存在差异。     

Total antioxidant capacity and content of flavonoids and other phenolic compounds in canihua (Chenopodium pallidicaule): an Andean pseudocereal

Total antioxidant capacity (TAC), total phenolic compounds (TPH), total flavonoids (TF) and individual phenolic compounds were determined in canihua collected at approx. 3850 m altitude. The TAC values varied among samples from 2.7 to 44.7 by the ferric reducing antioxidant power (FRAP) method and from 1.8 to 41 by the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) method expressed as micromol of Trolox equivalents/g dw. The content of TPH was 12.4-71.2 micromol gallic acid equivalents/g dw and that of the TF ranged between 2.2 and 11.4 micromol of catechin equivalents/g dw. The data obtained by the four methods showed several significant correlations. Prior to analysis by HPLC, the samples were subjected to acid hydrolysis and in the water-soluble extracts this led to an up to 20-fold increase in the TAC values in comparison with the values of the nonhydrolysed samples. HPLC analysis showed the presence of eight major compounds identified as catechin gallate, catechin, vanillic acid, kaempferol, ferulic acid, quercetin, resorcinol and 4-methylresorcinol. Their estimated contribution to the TAC value (FRAP method) indicated that resorcinols contributed most of the antioxidant capacity of the water-soluble extract. The results show that canihua is a potential source of natural antioxidant compounds and other bioactive compounds which can be important for human health.     

洋甘菊各萃取相抗氧化活性及其有效成分分析

目的:筛选洋甘菊醇提物抗氧化活性最强的萃取组分,并对该组分抗氧化活性的有效成分进行研究。方法:采用DPPH·、ABTS+·、O₂-·自由基清除能力和FRAP抗氧化功能测试方法,对比研究了洋甘菊醇提物不同极性萃取组分的抗氧化活性,并运用HPLC等方法对抗氧化能力最强萃取组分进行有效成分分析。结果:洋甘菊乙酸乙酯萃取组分的抗氧化能力最强,对DPPH·、ABTS+·和O₂-·的IC₅₀分别为25.6、30.6和83.3 mg/mL,总抗氧化能力(以trolox计)为16.7 mg/mL。成分分析表明,乙酸乙酯萃取相中总酚和总黄酮含量分别为25.84%和14.93%,在检测到的10种多酚或黄酮类化合物中,木犀草素、蒙花苷和木犀草素-7-葡萄糖含量较丰富。结论:洋甘菊醇提物的乙酸乙酯萃取组分具有良好的抗氧化能力可能与其富含的多酚和黄酮成分有关。     

Characteristics of peptide preparations obtained during enzymatic hydrolysis and ultrafiltration of milk proteins for use in specialized nutrition products

+Ultrafiltrated bovine milk proteins with whey protein/casein ratios 20/80, 60/40 and 97/3 were subjected to pancreatin hydrolysis, and the hydrolysates obtained were fractionated to high- and low-molecular weight (LMW) fractions by means of ultrafiltration. The preparations obtained were characterized according to routine physico-chemical indices, amino-nitrogen content, amino acids score, molecular weight distribution, and concentration of precipitating milk antigens. The capacity of LMW fractions for inducing oral anaphylactic sensitization in guinea pig and for changing its susceptibility to histamine LD50 was studied. The conclusion has been made that LMW preparations with whey/casein ratios 20/80 and 60/40 could be effectively used in specialized formulae intended for nutrition of allergic children and adults, nursing women, and also for enteral nutrition.     

A physiologic approach to test the global antioxidant response of foods. The GAR method

Several methods have been applied to measure antioxidants in foods. Extraction methods have previously relied on chemical methods which are non-physiological or based on enzymatic hydrolysis. Whatever the method used, the insoluble fraction is systematically excluded. The global antioxidant response (GAR) method use an in vitro approach with enzymatic digestion, designed to mimic digestion through the gastrointestinal tract, aimed to release antioxidants from foods. A total of 27 samples were analysed using the ABTS and FRAP assays applied to the soluble and insoluble fractions. The GAR method showed a higher antioxidant activity compared with the usual chemical extraction and the Quencher (direct) method. The soluble fraction was more antioxidant than the chemical extracts due to the release of compounds from the hydrolysis of carbohydrates and proteins. In addition, the GAR method allowed the measurement of the antioxidant activity in the insoluble fraction, which was important in fibre-rich samples.     

Comparison of the effects of different types of soya milk on the total antioxidant capacity of black tea infusions

Experimental evidence has shown that the polyphenols in black tea exhibit strong antioxidant potential in vitro and in vivo. The objective of the present study was to analyse and compare the total antioxidant capacity (TAC) of five brands of tea and to evaluate whether the addition of different varieties of soya milk decreased the TAC compared to semi-skimmed bovine milk. Each of the teas analysed was a significant source of antioxidants (7796–10,434 μmol/l FRAP). The addition of milk lowered the TAC of each of the teas analysed. When compared to tea with semi-skimmed bovine milk, each of the five teas analysed had either significantly higher antioxidant values or no change after the addition of soya milk. The addition of soya milk to black tea may be a useful alternative to semi-skimmed bovine milk if wishing to maintain the overall antioxidant potential of the tea infusion.     

Effect of Extraction Techniques on Phenolic Content, Antioxidant and Antimicrobial Activity of Bauhinia purpurea: HPTLC Determination of Antioxidants

Bauhinia purpurea leaf was extracted by Soxhlet, ultrasonication and maceration extraction methods using ethanol (99.5%, v/v) to obtain Soxhlet (SBE), ultrasonicated (UBE) and macerated (MBE) B. purpurea leaf extract. The effects of different extracting methods on the polyphenolic content and antioxidant activities using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), ferric-reducing antioxidant power (FRAP) and total antioxidant capacity (TAC) were investigated. Disc diffusion and broth dilution methods were also carried out to find the antibacterial activity of these extracts. Findings of this study showed that UBE possessed significant (P < 0.05) polyphenolic constituents followed by MBE and SBE. All the extracts exhibited good DPPH and ABTS radical scavenging as well as potential reducing ability in TAC and FRAP methods. UBE possessed significant (P < 0.05) radical scavenging activity and reducing ability followed by MBE and SBE. Even the results of antibacterial activity were similar to antioxidant activity, with UBE inhibiting most of the bacteria followed by MBE and SBE. All the extracts were subjected to thin layer chromatography (TLC) bioautography followed by high-performance TLC densitometric determination, and the results show that extraction using ultrasonication method yields the highest amount of antioxidant compounds among the three methods mentioned earlier. This study confirms ultrasonic extraction to be an ideal, simple and rapid method to obtain antioxidant- as well as antibacterial-enriched B. purpurea leaf extract. The HPTLC fingerprint profile can be used as a reference data for the standardisation of B. purpurea leaf.     

Antioxidant capacity of cow milk,whey and deproteinized milk

The total antioxidant capacity (hydrophilic plus lipophilic) of sixteen different commercial samples of pasteurized and ultra high temperature (UHT) treated milk was determined using the oxygen radical absorbance capacity assay using fluorescein as a fluorescent probe. A significant correlation between the percentage of fat and the value of the total antioxidant capacity was found in milk samples obtained from the same batch of raw milk. Analyses of the whole milk, whey and deproteinized milk showed that the major contributor to the total antioxidant capacity of whole milk was the casein fractions, while albumin was the major contributor to the total antioxidant capacity of whey protein. Hydrophilic antioxidant compounds, such as vitamin C and uric acid, were the main contributors to the total antioxidant capacity of the deproteinized milk. Significant differences in total antioxidant capacities were found between whey and deproteinized samples obtained from pasteurized and UHT-treated milk, the values being higher for the former.     

枇杷花提取物抗氧化活性研究

采用DPPH、ABTS、FRAP和福林酚(F-C)四种方法测定枇杷花酚类化合物的抗氧化活性,比较最优条件下蒸馏水、甲醇、乙醇三种溶剂的提取效率。结果表明,DPPH、ABTS、F-C法所测抗氧化活性顺序一致,即为水提物>甲醇提取物>乙醇提取物,但当量大小有所差异,其中F-C实验结果最大;FRAP实验中,甲醇提取物抗氧化活性略大于水提物,乙醇提取物活性最低。其原因可能是由于提取溶剂极性不同,反应机理不同,FRAP法基于单电子转移原理。因此,枇杷花酚类物质在乙醇中的溶解性较差。     

鲢鱼肽美拉德反应产物超滤组分的抗氧化活性研究

鲢鱼肽美拉德反应产物(MRPs)经5 k Da、1 k Da超滤膜分离、然后再经溶剂分级后,获得了不同组分,分析其抗氧化活性与理化特性。通过对不同组分的DPPH自由基清除活性、金属离子螯合活性和总抗氧化能力的测定可以看出,水溶性组分中的高分子量化合物比低分子量化合物具有较高抗氧化活性,而醇溶性组分中高分子量的抗氧化活性比低分子量的低。MRPs中水溶性组分F-3(5 k Da)对DPPH自由基清除活性、对金属离子的螯合活性和总抗氧化力最高;水溶性组分中F-3和F-2(1 k Da~5 k Da)的褐变程度较高,F-3的荧光强度最高;FI-IR分析发现,F-2和F-3在860.01 cm-1和948.61 cm-1处有强烈的吸收峰,这表明肽因美拉德反应发生了结构的变化。因此,鲢鱼肽MRPs中起主要抗氧化作用的为水溶性物质,且其中相对高分子量的有色水溶性产物抗氧化活性较强。     

Bioactive composition and antioxidant potential of different commonly consumed coffee brews affected by their preparation technique and milk addition

Coffee is one of the most popular beverages in the world, prepared and consumed in many different ways. Taste, aroma and composition of the coffee brew vary depending on the preparation method. Therefore, this study investigates the effect of different brewing methods on the polyphenol and methylxanthine composition and antioxidant capacity of thirteen different coffee brews. The content of total phenols and flavonoids was determined spectrophotometrically and the content of chlorogenic acid derivates (3-CQA, 4-CQA and 5-CQA) and caffeine using the high performance liquid chromatography (HPLC-PDA). Antioxidant capacity of coffee brews was evaluated by using the ABTS (2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)) and FRAP (ferric-reducing antioxidant power) assays. Instant coffee brews showed the highest values in content of total phenols, chlorogenic acid derivates, caffeine and antioxidant capacity, which significantly decreased by milk addition. The antioxidant capacity of coffee brews was in compliance with the total phenol content and content of chlorogenic acid derivates.     

Comparison of antioxidant capacity of cow and ewe milk kefirs

This research aimed to evaluate the effects of using either grain or commercial starter culture on the antioxidative capacity of cow and ewe milk kefirs. The antioxidant capacity of kefir samples during fermentation and 21 d of storage was assessed by using 3 assays: 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation decolorization; 2,2-diphenyl-1-picrylhydrazyl (DPPH^?) radical scavenging activity assay; and Fe⁺³-reducing power (ferric reducing antioxidant power assay, FRAP). Vitamin E and β-carotene contents were also quantified. All kefir samples exhibited varying values for DPPH, ABTS, and FRAP assays depending on the starter culture and milk type. Vitamin E and β-carotene contents were similar in all kefir samples during storage. The results of this study suggest that milk type (cow or ewe) and culture type (kefir grains or commercial starter) were the significant parameters for the antioxidative activity of kefir.     

COMPOSITION OF MAJOR FLAVANONE GLYCOSIDES AND ANTIOXIDANT CAPACITY OF THREE CITRUS VARIETIES

This paper reported on the composition of major flavanone glycosides (FGs) and antioxidant capacity of three Chinese citrus varieties during maturity, namely, Ponkan (P; citrus poonensis Hort. ex Tanaka), Satsuma mandarin (S; citrus unshiu Marc.) and Huyou (H; citrus paradisi Macf. cv. Changshanhuyou). The results showed that citrus peel is a rich source of FGs. Total penolics contents (TPC) of methanol extract of P, S and H were determined. The total antioxidant capacity (TAC) of P, S and H extracts was compared based on vitamin C equivalent antioxidant capacity by 2, 2-diphenyl-1-picrylhydrazyl radical (DPPH·), 2, 2 ' -azinobis (3-ethylbenzothiazoline-6-sulfonate) (ABTS ^·+ ), reducing power and ferric reducing antioxidant power (FRAP) assay. Phenolic compounds may contribute a lot to TAC of fresh fruits, especially for unripe fruits. The correlations among TPC, DPPH·, ABTS ^·+ , reducing power and FRAP assay were rather high ( P <  0.05).

PRACTICAL APPLICATION

Citrus flavonoids are abundant in citrus fruits, especially FGs, such as hesperidin and naringin; and they are attracting more attentions for their biological properties and health benefits. The sun-dried citrus peels ( chenpi ) are usually used in the traditional Chinese medicine for high content of hesperidin. Investigation on the composition of FGs in peel and edible part of citrus fruits during maturity is necessary to establish a database for the future use in health foods and ingredients. Meanwhile, four methods were employed simultaneously to achieve a comprehensive evaluation on the antioxidant capacity of the three citrus varieties. The high antioxidant potential of the three selected citrus varieties made them desirable for dietary prevention of cardiovascular and other diseases.     

蓝莓花、茎、叶酚类物质含量及抗氧化活性比较

比较蓝莓花、嫩茎、嫩叶及老叶总多酚、总黄酮、原花青素的含量,并研究其抗氧化性能差异。结果表明:嫩叶中总多酚含量最高,为43.77 mg GAE/g(以干质量计,下同),分别是花、嫩茎和老叶的3.45、1.42、1.12倍;老叶中总黄酮、原花青素含量最高,为68.01 mg RE/g、23.29 mg CAE/g,分别是花的2.57倍和4.61倍、嫩茎的1.79倍和1.23倍、嫩叶的1.03倍和1.98倍。抗氧化性能中,老叶的铁还原能力最大,其次是嫩茎和花,嫩叶较差;清除·OH的能力为嫩茎嫩叶花老叶;清除DPPH自由基能力和总抗氧化能力均为嫩茎嫩叶老叶花甲醇提取物,均高于对照品;蓝莓各部分提取物对DPPH自由基的清除能力最强,总抗氧化能力次之,清除·OH能力和铁还原能力较差。     

响应面法优化黄浆水发酵液制备工艺及其抗氧化活性研究

采用植物乳杆菌(Lactobacillus plantarum)、嗜酸乳杆菌(Lactobacillus acidophilus)和清酒乳杆菌(Lactobacillus sakei)3种乳酸菌对黄浆水进行组合发酵,以发酵液DPPH自由基清除率为评价指标,研究发酵温度、发酵时间、脱脂乳粉添加量、葡萄糖添加量和接种量对发酵液的影响。在单因素试验基础上,采用响应面法优化制备高抗氧化活性黄浆水发酵液的工艺条件。结果表明最佳发酵参数为:接种量1%、发酵温度37℃、发酵时间37.50 h、脱脂乳粉添加量8%、葡萄糖添加量5%。在此条件下制备的黄浆水发酵液DPPH自由基清除率为82.36%。抗氧化活性试验表明:黄浆水发酵液提取物抗氧化能力得到显著提升,其清除DPPH自由基、羟自由基和ABTS自由基的半抑制质量浓度(IC50)分别为2.03 mg/mL、1.12 mg/mL和0.30 mg/mL。     

白穗醋栗多酚鉴定及其抗氧化活性分析

采用东北农业大学小浆果资源圃内白穗醋栗果实,利用固相萃取技术进行分离纯化得到多糖、花色苷、非花色苷等组分,测定各组分的化学成分,并进一步测定各组分的总酚以及总花色苷含量,使用1,1-二苯基-2-三硝基苯肼自由基、2,2’-联氮双(3-乙基苯并噻唑啉-6-磺酸)阳离子自由基、铁离子还原能力等方法测定其各组分抗氧化能力。结果表明：白穗醋栗果实中含有丰富的非花色苷多酚为78 种,花色苷多酚只有一种为矢车菊素-3-葡萄糖苷,可以作为一种潜在的花青素提取原料;白穗醋栗果实多酚粗提液总酚含量为（34.93±0.47）mg/g;总花青素含量为（2.99±0.98）mg/g;果实表现出了较强的抗氧化活性。     

Effect of thermal processing and digestive protease on the antioxidant capacity of fruit juice–milk beverage model systems under simulated gastrointestinal digestion

The effects of thermal processing and digestive protease on the antioxidant capacity of fruit juice–milk beverage (FJMB) models containing whey protein (WP) and chlorogenic acid or catechin (CAT) under in vitro digestion were investigated. After gastric digestion, the FJMB showed a significant (P < 0.05) increase in ABTS, but little change in ferric reducing antioxidant power (FRAP). Intestinal digestion decreased the FRAP and increased ABTS except in the model containing 0.1% CAT. There were different effects (none, masking, synergistic effect) in the antioxidant activity of the WP and phenolics under digestion. Pasteurisation (63 °C/30 min) had no significant effect on the FJMB's antioxidant capacity, while sterilisation (121 °C/10 min) significantly (P < 0.05) increased, decreased or had no significant effect on ABTS and FRAP under digestion, depending on the model. Pepsin and pancreatin significantly (P < 0.05) decreased the fruit phenolics' ABTS. Ferric reducing antioxidant power was not significantly influenced by pepsin during gastric digestion, but was significantly (P < 0.05) decreased by pancreatin during intestinal digestion. These results would be helpful to improve the efficacy of fruit phenolics in FJMB subjected to thermal processing and to maintain the health benefits of this kind of functional products in the food industry.     

A Comparative Study of the Analysis of Antioxidant Activities of Liquid Foods Employing Spectrophotometric, Fluorometric, and Chemiluminescent Methods

The antioxidant profile of liquid foods is complex and includes different lipid and water-soluble compounds. These should be considered when assessing total antioxidant capacity (TAC) of these beverages, since it may act synergistically rather than individually. This study describes and compares the use of spectrophotometric methods (Trolox equivalent antioxidant capacity, TEAC and 2,2-diphenyl-1-picrylhydrazyl, DPPH), fluorometric (oxygen radical antioxidant capacity, ORAC), and photochemiluminescence (PCL) for the measurement of the TAC of different liquid foods (fruit juice mixed with milk and vegetables beverage). An evaluation was also made for the influence of certain compounds (ascorbic acid, phenolic compounds, total carotenoids, and tocopherols) with antioxidant capacity that was present in the samples studied. The different methods studied allow the determination of the TAC of the analyzed foods in a precise and accurate way. The TAC values in the studied samples differ from the applied method. An overall antioxidant potency composite index was calculated by assigning each test's equal weight. When an index score was applied, ORAC method had the higher antioxidant capacity values in the analyzed liquid foods in comparison with the other methods. The correlations among the different methods used for the determination of the antioxidant capacity depend on food, that is, mainly due to compounds (lipid and water soluble) of the different food matrix. In addition, ascorbic acid was the main contributor to antioxidant capacity of fruit juice mixed with milk beverages measured with the different methods. However, in vegetables beverages, phenolic compounds were found to correlate more significantly with antioxidant capacity values.     

ORAC and TEAC assays comparison to measure the antioxidant capacity of food products

Oxygen radical antioxidant capacity (ORAC) and trolox equivalent antioxidant capacity (TEAC) assays were compared to estimate the total antioxidant capacity (TAC) of orange juice, milk, and an orange juice-milk beverage. When the TEAC method was used with this beverage, an increase in the concentration of orange juice corresponded to an increase in TAC, but increasing the percentage of milk did not increase the TAC value. When the ORAC method was applied, it was seen that increased concentrations of juice or milk corresponded to greater antioxidant capacity. An evaluation was also made of the influence of certain compounds (ascorbic acid, gallic acid, β-carotene, lutein, zeaxanthin and albumin) with antioxidant capacity that were present in the samples studied.