Antioxidant activity of half <Emphasis Type="Italic">N</Emphasis>-acetylated water-soluble chitosan in vitro

Different molecular weight water-soluble chitosan (half N-acetylated chitosan) was prepared and the structure of water-soluble chitosan was characterized by FT-IR. The pH dependence of water solubility of water-soluble chitosan was evaluated from turbidity. Total antioxidant activity, reducing power, superoxide anion radical and hydroxyl radical quenching assay, metal chelating activity, and H₂O₂ scavenging activity were used for the evaluation of different molecular weight half N-acetylated chitosan in vitro. Low-molecular weight water-soluble chitosan (WSC4) exhibited high reductive capacity and expressed good inhibition of linoleic acid peroxidation in the linoleic acid model system. WSC4 (0.25 mg/mL) scavenged 78.8% of superoxide radical. At 5 mg/mL, scavenging percentage of WSC1, WSC2, WSC3, and WSC4 against hydroxyl radical was 49.3%, 66.8%, 77.1%, and 83.7%, respectively. These results indicate that water-soluble chitosan is an ideally natural antioxidant, and its antioxidant activity depends on its molecular weight.     

复合蛋白酶水解玉米谷蛋白产物的抗氧化活性研究

以玉米谷蛋白为原料,利用复合蛋白酶对其进行限制水解,探讨不同水解时间所获得的谷蛋白酶解物的分子量分布和抗氧化活性。结果表明:不同水解时间获得的酶解物分子量分布差异较大。水解时间为120min的酶解物中,分子量分布为6 511.51~307.32Da的肽段占94.36%,此时获得的酶解物的抗氧化活性最高,其对DPPH自由基、O-2·、·OH的清除率分别为58.86%,82.64%,37.21%;还原力为0.236;与亚铁离子的螯合能力为29.92%。     

Chitosan glucose complex – A novel food preservative

Chitosan glucose complex (CGC), a modified form of chitosan was prepared by heating chitosan with glucose. Fluorescence and the browning reaction of CGC indicated the presence of the Maillard reaction product (MRP). CGC showed excellent antioxidant activity while chitosan or glucose alone did not have any significant activity (p < 0.05). The IC₅₀ value of CGC for DPPH radical scavenging was 51.1 μg/ml. The efficacy of CGC in scavenging hydroxyl and superoxide anion radicals was also very high but it showed a low reducing power. The antimicrobial activity of CGC was similar to chitosan against E. coli, Pseudomonas, Staphylococcus aureus and Bacillus cereus, the common food spoilage and pathogenic bacteria. The minimum inhibitory concentration of CGC or chitosan was 0.05%. Addition of CGC to lamb meat increased its shelf life by more than 2 weeks during chilled storage. It also enhanced the shelf life of pork cocktail salami to 28 days. CGC is endowed with both antioxidant and antimicrobial activity and thus is a promising novel preservative for various food formulations.     

Antioxidant and free radical scavenging activities of pigments extracted from molasses alcohol wastewater

The pigments from molasses alcohol wastewater were extracted by the macroporous resin adsorption method. The antioxidant and free radical scavenging activities of these pigments were also investigated. The adsorptive characteristics of five macroporous resins including HPD-600, HPD-500, D301-R, NKA-II and D296-R were studied and the results showed that the macroporous resin HPD-600 was most appropriate for extracting the pigments from molasses alcohol wastewater. The antioxidant and free radical scavenging activities of pigments extracted from alcohol wastewater were evaluated using nitrate, hydroxyl radical, superoxide anion radical and 1,1-diphenyl-2-picrylhydrazyl (DPPH) in vitro model systems. The pigment extract exhibited a concentration-dependent radical scavenging activity in all the systems. Meanwhile, scavenging activity of pigment extract in the DPPH system was found to be significantly (P < 0.05) higher than that in other systems and the 50% inhibitory concentration (IC₅₀ value) was about 0.07 mg/ml. The scavenging effect of pigment extract on superoxide anion radical was very weak with IC₅₀ value greater than 10 mg/ml.     

Antioxidant and melanogenesis‐inhibitory activities of collagen peptide from jellyfish (Rhopilema esculentum)

BACKGROUND: Jellyfish collagen was hydrolysed with trypsin and properase E, and jellyfish collagen peptide (JCP) was purified from the enzymatic hydrolysate using ion exchange chromatography and gel filtration. The antioxidant activity of JCP in a linoleic acid emulsion system, its superoxide anion‐ and hydroxyl radical‐scavenging activities and its copper‐chelating ability were evaluated in vitro. Initial investigations of JCP's ability to inhibit melanogenesis were carried out using cultured B16 melanoma cells. RESULTS: The molecular weight distribution of JCP was from 400 to 1200 Da. Amino acid analysis showed that JCP was rich in Gly, Pro, Ser, Ala, Glu and Asp and had a total hydrophobic amino acid content of 384.2 g kg^?1. JCP showed high antioxidant activity (IC₅₀147.8 µg mL^?1), superoxide anion‐scavenging activity (IC₅₀21.9 µg mL^?1), hydroxyl radical‐scavenging activity (IC₅₀16.7 µg mL^?1) and copper‐chelating ability (IC₅₀88.7 µg mL^?1) in vitro. It also significantly inhibited intracellular tyrosinase activity, decreased melanin content and enhanced glutathione synthesis (P < 0.05). Furthermore, JCP decreased intracellular cAMP levels and suppressed tyrosinase mRNA expression. CONCLUSION: Based on the results of this study, JCP exerts anti‐melanogenic actions via its antioxidant properties and copper‐chelating ability. JCP could be used as a natural skin‐lightening agent in the medicine and food industries. Copyright © 2009 Society of Chemical Industry     

Purification and identification of antioxidant peptides from corn gluten meal

Corn gluten meal was hydrolyzed by alkaline protease and Flavourzyme to obtain the antioxidant peptides. The antioxidant activities of the hydrolysates or peptides were evaluated by free radical scavenging capacity (1,1-diphenyl-2-picrylhydrazyl/2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt/hydroxyl radical/superoxide radical anion), metal ion (Fe²⁺/Cu²⁺) chelating activity and lipid peroxidation inhibitory capacity. The hydrolysates were separated by ultrafiltration, and those with molecular weight <10 kDa exhibited highest antioxidant activity in all relevant assays. The hydrolysates were subsequently purified by gel filtration chromatography, and fraction F3 showed the highest antioxidant activity. Three peptides were identified from fraction F3 using LC–ESI–Q–TOF MS/MS as Leu-Pro-Phe (375.46 Da), Leu-Leu-Pro-Phe (488.64 Da) and Phe-Leu-Pro-Phe (522.64 Da). These peptides exhibited good free radical scavenging activity and lipid peroxidation inhibitory effect. Thus, corn gluten meal may be used as a potential source of antioxidant peptides for food and nutraceutical applications.     

取代度相同的N-酰化低聚壳聚糖清除自由基作用的研究

低聚壳聚糖经N-酰化得到取代度相同的N-马来酰低聚壳聚糖和N-邻苯二甲酰低聚壳聚糖,其取代度均为0.25。用红外光谱对其结构进行表征,凝胶色谱测定其相对分子质量大小。并考察了其对羟基自由基(.OH)和1,1-二苯基苦基苯肼自由基(DPPH)的清除能力。结果表明:取代度相同,N-邻苯二甲酰低聚壳聚糖对.OH、DPPH的清除能力优于N-马来酰低聚壳聚糖。这说明取代度相同时,取代基的结构会影响N-酰化低聚壳聚糖对自由基的清除活性。     

Antioxidant activities of red pepper (Capsicum annuum) pericarp and seed extracts

In this study, we examined the antioxidant activities of red pepper (Capsicum annuum, L.) pericarp and red pepper seed extracts. The extracts were evaluated by various antioxidant assays, including 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical scavenging, superoxide anion radical scavenging, hydroxyl radical scavenging, [2,2′‐azino‐bis(3‐ethylbenzthiazoline‐6‐sulphonic acid)] (ABTS) radical scavenging, ferrous chelating activity, superoxide dismutase (SOD) activity and reducing power, along with the determination of total phenolic and flavonoid contents. All the extracts showed strong antioxidant activity by the testing methods. The red pepper pericarp extract exhibited strong ferrous chelating activity and high scavenging activity against free radicals, including both the hydroxyl and DPPH radicals, but it exhibited weaker scavenging activity for the superoxide anion radical and for SOD. In contrast, the red pepper seed extract exhibited strong SOD activity and high scavenging activity against the superoxide anion radical, but showed weaker ferrous chelating activity, hydroxyl radical scavenging, and DPPH radical scavenging. We observed that the reducing power level and ABTS radical scavenging activity of the red pepper seed were higher than those of the red pepper pericarp at the highest tested concentration. Most of the test results for the red pepper seed and red pepper pericarp extracts increased markedly with increasing concentration; however, the metal chelating, SOD and ABTS radical scavenging activities did not increase with the concentration. Highest total phenolic and flavonoid contents were obtained from the red pepper pericarp extracts. Overall, the red pepper seed and red pepper pericarp extracts were highly effective for the antioxidant properties assayed, with the exceptions of ferrous chelating activity, hydroxyl radical scavenging and SOD activity.     

In vitro determination of antioxidant activity of proteins from jellyfish Rhopilema esculentum

In this study, the antioxidant activity of proteins isolated from jellyfish, Rhopilema esculentum Kishinouye (R. esculentum), was determined by various antioxidant assays, including superoxide anion radical-scavenging, hydroxyl radical-scavenging, total antioxidant activity, reducing power and metal chelating activity. Butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), α-tocopherol, vitamin C and mannitol were used as standards in those various antioxidant activities. The crude protein (CP) and the protein fractions isolated by Sephadex chromatography, first peak (FP) and second peak (SP), had very low reductive power and metal chelating abilities compared to EDTA, but they showed strong scavenging effects on the superoxide anion radical, hydroxyl radical and varying total antioxidant activity. FP and SP exhibited stronger scavenging effects on the superoxide anion radical than BHA, BHT or α-tocopherol. The EC₅₀ values of FP and SP were 6.12 and 0.88 μg/ml, respectively, while values EC₅₀ of BHA, BHT and α-tocopherol were 31, 61 and 88 μg/ml, respectively. CP, FP and SP showed far higher hydroxyl radical-scavenging activities than did vitamin C or mannitol. The EC₅₀ values of CP, FP and SP were 48.76, 45.42 and 1.52 μg/ml, but EC₅₀ values of vitamin C and mannitol were 1907 and 4536 μg/ml, respectively. In a β-carotene–linoleate system, SP and CP showed antioxidant activity, but lower than BHA. Of the three samples, SP had the strongest antioxidant activity. So, SP may have a use as a possible supplement in the food and pharmaceutical industries.     

酶解缫丝蚕蛹蛋白抗氧化肽的分离与稳定性研究

目的：研究酶解缫丝蚕蛹蛋白抗氧化肽的分离和稳定性,为其开发应用提供基础数据。方法：超滤分离酶解缫丝蚕蛹蛋白抗氧化肽,比色法测定其抗氧化能力。结果：超滤分级后得到分子质量为200～3 000 D的酶解缫丝蚕蛹蛋白抗氧化肽对O2－•、•OH、1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基的IC50分别为18.85 mg/mL、3.13 mg/mL、35.23 μg/mL。该抗氧化肽经过4 h胃蛋白酶+胰蛋白酶处理前后对DPPH自由基清除率分别为81.05%、82.26%;在pH 4、8条件下处理1 h,对DPPH自由基清除率分别为98.02%、11.80%（100 μg/mL）;在温度95 ℃条件下处理1 h,对DPPH自由基清除率为87.11%（100 μg/mL）;用浓度为1.0 mol/L的NaCl处理6 h,相比对照组,对DPPH自由基清除率由51.58%下降到22.68%（60 μg/mL）。结论：超滤后得到的分子质量为200～3 000 D的酶解缫丝蚕蛹蛋白抗氧化肽的抗氧化能力比酶解原液有一定提高;且在酸性、高温、脱盐处理后对DPPH自由基的清除能力保持较好;胃肠道消化酶对其DPPH自由基清除能力的影响不显著（P＞0.05）。     

葛根抗氧化肽的分离及清除自由基活性研究

目的：分离纯化葛根抗氧化肽并对其进行体外清除自由基活性的研究。方法：以野生葛根为实验材料,经30%的乙醇浸提、透析、冷冻干燥,进一步采用DEAE阴离子交换层析,反相高效液相色谱的方法,对葛根抗氧化肽进行纯化,对不同组分进行还原能力的测定,还原能力最强的命名为PE1。对PE1进行茚三酮反应和分子质量测定。采用邻苯三酚自氧化法、DPPH法、Fenton反应法检测PE1体外对超氧阴离子自由基、DPPH自由基、羟自由基的清除作用。结果：反相高效液相色谱检测PE1纯度,显示为单一峰,PE1与茚三酮反应呈阳性,分子质量约为746D,PE1在体外可以有效清除以上3种自由基。在质量浓度为0.652mg/mL时,羟自由基清除率可达到80.01%;在5mg/mL质量浓度条件下,对超氧阴离子自由基的清除率高达97.35%;在3.5mg/mL质量浓度条件下,对DPPH自由基的清除率达到75.02%,其IC50为2.83mg/mL。结论：PE1具有较强的清除自由基活性。     

不同蛋白酶对小麦蛋白酶解物抗氧化活性的影响

小麦蛋白是小麦淀粉加工的副产物,酶解是提高小麦蛋白溶解性和功能性的有效方式,而酶解用酶种类可能对酶解产物的功能性如抗氧化活性有一定影响。采用碱性蛋白酶、中性蛋白酶、胃蛋白酶、风味蛋白酶、胰蛋白酶、木瓜蛋白酶6种常用的蛋白酶分别对小麦蛋白进行酶解,并对酶解4 h后酶解物的多肽得率、分子质量分布、1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除率、超氧阴离子自由基(O_2~-·)清除率、羟自由基(·OH)清除率等反映水解程度和抗氧化能力的主要指标进行评价。结果表明,风味蛋白酶酶解物中多肽得率最高,达91.44%,且分子质量小于3 000 D的多肽含量达76.9%;酶解物质量浓度为3 mg/m L时,木瓜蛋白酶酶解物对DPPH自由基清除作用最好,清除率为65.12%(P0.01),其次是风味蛋白酶(58.43%)和碱性蛋白酶(55.29%);碱性蛋白酶酶解物对O_2~-·清除率效果最好,清除率为58.68%(P0.01),其次是风味蛋白酶(49.25%);碱性蛋白酶和木瓜蛋白酶酶解物对·OH清除效果最佳,清除率分别为59.23%和58.16%。结果说明,蛋白酶种类对小麦蛋白酶解物抗氧化活性影响显著,风味蛋白酶对提高蛋白水解程度和生成小分子质量多肽的作用明显,而碱性蛋白酶、木瓜蛋白酶和风味蛋白酶对提高酶解产物抗氧化活性效果较好。     

Isolation and characterization of three antioxidant pentapeptides from protein hydrolysate of monkfish (Lophius litulon) muscle

In the current study, an efficient method had been developed to acquire the antioxidant hydrolysate of monkfish muscle protein (MPH) using trypsin by an orthogonal (L₉(3)⁴) test. Under the optimum conditions of enzymolysis time 4 h, enzyme-to-substrate ratio (E/S) 2%, enzymolysis temperature 40 °C and pH 8.0, the DH (Degree of hydrolysis) and hydroxyl radical scavenging activity of MPH reached 19.83 ± 0.82% and 58.05 ± 3.01%, respectively. By using ultrafiltration, gel filtration chromatography and reversed phase high performance liquid chromatography (RP-HPLC), three antioxidant pentapeptides were isolated from MPH, and their amino acid sequences were identified as Glu-Trp-Pro-Ala-Gln (MPH-P1), Phe-Leu-His-Arg-Pro (MPH-P2), and Leu-Met-Gly-Gln-Trp (MPH-P3) with molecular weights of 629.68 Da, 668.80 Da, and 633.77 Da, respectively. MPH-P1, MPH-P2, and MPH-P3 exhibited good scavenging activities on hydroxyl radical (EC₅₀ 0.269, 0.114 and 0.040 mg/ml), DPPH radical (EC₅₀ 2.408, 3.751, and 1.399 mg/ml), and superoxide anion radical (EC₅₀ 0.624, 0.101, and 0.042 mg/ml) in a dose-dependent manner. MPH-P3 was also effective against lipid peroxidation in the model system. The antioxidant activities of MPH-P1, MPH-P2, and MPH-P3 were due to their small sizes and the presence of antioxidant and hydrophobic amino acid residues within their sequences. The results of this study suggested that the protein hydrolysate and/or its isolated peptides might be effectively used as food additives for retarding lipid peroxidation occurring in foodstuffs.     

不同处理条件对猴头菇抗氧化活性的影响

研究不同加工、烹制条件对猴头菇抗氧化活性的影响。在不同处理条件下处理猴头菇子实体,然后从清除羟自由基的能力、还原力、清除超氧阴离子自由基和清除DPPH自由基的能力4个方面测定不同处理条件对猴头菇抗氧化活性的影响。结果显示,随烘烤温度升高,清除羟自由基的能力与还原力显著升高,清除超氧阴离子自由基的能力和清除DPPH自由基的能力显著降低。微波处理对清除羟自由基基本没有影响,但能显著升高还原力、清除超氧阴离子自由基和清除DPPH自由基的能力。煮处理中,清除超氧阴离子自由基的能力、还原力和清除DPPH自由基的能力显著高于炒处理;而对于清除羟自由基,炒处理则显著高于煮处理。研究结果为猴头菇加工、烹制条件的改善提供理论依据。     

羧甲基壳聚糖的降解及其抗氧化性能的研究

羧甲基壳聚糖是壳聚糖的一种重要衍生物。本研究采用过氧化氢氧化法对羧甲基壳聚糖进行降解,并考察了降解时间及过氧化氢用量对降解的影响。用粘度法和改进的Schales’方法测定了降解产物的分子量。并用流动注射化学发光分析法检测了降解产物的抗氧化能力--对羟基自由基的清除活性。结果表明:随着过氧化氢在降解体系中的浓度的增大,降解产物的分子量将趋于一固定值;当H2O2在总反应体系中浓度大于1.3mol/L,降解时间大于16h,将得到分子量低至1100左右的羧甲基壳聚糖;经降解得到的羧甲基壳聚糖均具有一定的羟基自由基清除活性,且分子量越小,其抗氧化性能越强。     

不同乳酸菌胞外分泌物抗氧化活性的研究

以1,1-二苯基-2-三硝基苯肼（DPPH）自由基、羟自由基、超氧阴离子自由基清除能力和还原能力为评价指标,以维生素C作阳性对照,分析不同来源的28株乳酸菌的胞外分泌物体外抗氧化活性,并研究不同抗氧化活性测定方法间的相关性。结果表明,菌株GB8、WC5和GC2抗氧化能力较强,其中菌株GB8对超氧阴离子自由基清除率（98.26%）和还原能力最高（0.964）,菌株WC5对DPPH自由基的清除率最高,为74.90%;菌株GC2对羟自由基、DPPH自由基及超氧阴离子自由清除率分别为58.67%、73.09%、87.50%,还原能力为0.806,表现出最强的综合抗氧化能力。4种体外抗氧化活性分析方法的相关性结果表明,羟自由基与DPPH自由基（R2=0.633）、超氧阴离子与还原能力（R2=0.488）方法间达到了极显著相关（P＜0.01）;羟自由基与还原能力（R2=0.295）方法间达到了显著相关（P＜0.05）。     

Antioxidant activities of squid protein hydrolysates prepared with papain using response surface methodology

Squid protein hydrolysates (SPH) were prepared from the Indian squid Loligo duvauceli using papain. Response surface methodology (RSM) was used for optimization of hydrolysis conditions, including temperature, time, and the enzyme-substrate ratio using DPPH radical scavenging activity as a response. The amino acid composition of SPH was compared with raw squid muscle. In vitro antioxidant activities were evaluated based on reducing power, metal chelation, ABTS, hydroxyl radical, and superoxide anion radical scavenging assays. SPH exhibited good ABTS radical scavenging activities of 96.50±0.90%, superoxide anion radical scavenging activities of 96.4±0.89%, reducing powers of 0.71±0.02, moderate hydroxyl radical scavenging activities of 64.03±2.11%, and metal chelating activities of 52.04±1.02%. In vivo antioxidant activities determined using a sardine minced model system showed 42% reduction in formation of secondary oxidative products as thiobarbituric acid reactive substances (TBARS), almost equivalent to reduction by ascorbic acid of 41.42% at 400 ppm.     

橄榄酚类提取物的抗氧化性能

为进一步开发利用橄榄中的酚类物质,文中通过测定橄榄酚类提取物的铁离子还原能力、亚铁离子螯合能力,及对2,2'-氨基-二(3-乙基-苯并噻唑啉-6-磺酸)自由基(ABTS+.)、1,1-二苯-2-苦基苯肼自由基(DPPH.)、羟基自由基(.OH)和超氧阴离子自由基(O2-.)的清除能力,以考察橄榄酚类提取物的体外抗氧化性能,并与食品抗氧化剂丁基羟基茴香醚(BHA)和Vc进行对比。结果表明:橄榄酚类提取物具有较好的体外抗氧化性能,其ABTS+.、DPPH.清除率分别高达99%和95%以上,.OH、O2-.清除率可达50%左右,铁离子还原力约1 700μmol/L Trolox,亚铁离子螯合率最高为14.9%。且提取物ABTS+.、DPPH.和.OH清除能力及铁离子还原能力均强于BHA和Vc,而O2-.清除能力强于BHA但弱于Vc。另外,橄榄酚类粗提物和纯化物相同浓度下的ABTS+.、DPPH.、.OH清除能力和铁离子还原能力相当(P>0.05),而其O2-.清除能力和亚铁离子螯合能力表现有所差异。     

首乌叶黄酮抗氧化性能及清除自由基能力的研究

测定了首乌叶黄酮的总体抗氧化能力、清除超氧阴离子自由基能力、清除羟基自由基能力、清除DPPH自由基能力和对卵黄脂蛋白脂质抗氧化能力,并与合成抗氧化剂维生素C(VC)和没食子酸丙酯(PG)进行了比较。结果表明:首乌叶黄酮溶液具有良好的抗氧化效果,在一定浓度范围内,首乌叶黄酮溶液对卵黄脂蛋白脂质抗氧化能力优于VC和PG,其总体抗氧化能力、清除超氧阴离子自由基能力优于PG略低于VC,清除羟自由基能力优于VC低于PG,但清除DPPH自由基能力比VC和PG略弱。     

Effects of extraction methods on antioxidant activities of polysaccharides from camellia seed cake

Four purified polysaccharides (COP-H, COP-U, COP-E, and COP-A) were obtained from seed cake of Camellia oleifera Abel. by hot water extraction, ultrasonic-assisted extraction, enzyme extraction, and alkali extraction, respectively. Their structural characterizations were determined, and antioxidant activities were investigated. The results of FT-IR demonstrated that the characteristic absorption bands exhibited slight difference among four polysaccharides. HLPC analysis presented that the molecular weights were calculated to be 394, 297, 462, and 429 kDa for COP-H, COP-U, COP-E, and COP-A, respectively. GC–MS analysis showed that the monosaccharide compositions were identical to four polysaccharides, while the molecular ratios exhibited significant difference. Four polysaccharides showed antioxidant activities in a concentration-dependent manner. Among four polysaccharides, COP-E showed a higher scavenging activity on DPPH, hydroxyl radical, and superoxide anion radical. These findings suggest camellia seed cake polysaccharides have potential as a natural antioxidant.