海蜇生殖腺酶解肽的抗氧化活性和ACE抑制活性研究

为实现海蜇加工副产物生殖腺的高值化利用,以海蜇性腺脱脂粉为原料,选用中性蛋白酶、木瓜蛋白酶、胰蛋白酶和碱性蛋白酶分别对海蜇生殖腺进行水解,以酶解产物(JGHP)的水解度、DPPH清除活性和ACE抑制活性为指标,选择制备海蜇生殖腺活性肽的工具酶。经超滤膜将酶解产物分成JGPH-P1(大于3000 u)、P2(1000~3000 u)、P3(小于1000 u)三个部分,分别测定三部分的DPPH清除活性和ACE抑制活性。结果表明,中性蛋白酶水解所得产物的水解度、DPPH清除率和ACE抑制活性均优于其他三种蛋白酶;JGHP超滤后,其中组分JGHP-P3的活性较高。利用电子自旋共振(ESR)技术检测JGHP-P3对DPPH·、·OH、O-2·的清除活性。其ACE抑制活性IC50为1.06 mg/m L,DPPH·、·OH、O-2·的清除活性IC50分别为0.38、0.63、0.59 mg/m L。由此可见,经中性蛋白酶水解得到的JGPH中小于1000 u的组分具有较高的抗氧化和ACE抑制活性。     

鳕鱼骨胶原蛋白肽的抗氧化活性

以鳕鱼骨为原料,采用热水提取法提取胶原蛋白,并采用中性蛋白酶、复合蛋白酶、酸性蛋白酶和胃蛋白酶分别进行酶解制备鳕鱼骨胶原蛋白肽。以对.OH、DPPH.和O2-.自由基的清除率作为评价指标,比较4种蛋白酶酶解制备的鳕鱼骨胶原蛋白肽的抗氧化活性。结果表明:4种蛋白酶制备的鳕鱼骨胶原蛋白肽对3种自由基均有一定的清除能力,且其清除能力随样品质量浓度的增大而增强;4种蛋白酶酶解制备的鳕鱼骨胶原蛋白肽对3种自由基的清除能力各不相同,其中对.OH的清除能力最强且有较好的量-效关系;当多肽质量浓度在4～10 mg/mL时,4种蛋白酶酶解胶原蛋白肽对DPPH.的清除能力优于对O2-.的清除能力。     

鸡骨酶解物的抗氧化活性研究

分别选用复合风味蛋白酶、Protamex复合蛋白酶、Alcalase蛋白酶、胰酶、胰蛋白酶、AS.1398中性蛋白酶和木瓜蛋白酶水解鸡骨泥,研究鸡骨酶解物(蛋白质含量3 mg/mL)对DPPH、羟自由基(OH·)的清除能力及其还原性.试验结果表明:Protamex复合蛋白酶酶解鸡骨产物的DPPH清除率最强,即81.44%;复合风味蛋白酶酶解物OH·清除率达30.32%,还原性测定的吸光值为0.3.总之,鸡骨酶解物的DPPH清除能力强于羟自由基清除能力和还原力.在研究不同浓度和水解度对鸡骨酶解物清除DPPH活性的影响时发现,随着酶解物蛋白质质量浓度的增大,酶解物的DPPH清除率升高.复合风味蛋白酶、Protamex复合蛋白酶的鸡骨酶解物蛋白质质量浓度分别为4、6mg/mL时,其DPPH清除率分别达91.49%、97.3%.但DPPH清除率与水解度不呈正相关性,只有在特定水解度下,使抗氧化肽含量最高时,水解物才表现出最强的抗氧化能力.例如复合风味蛋白酶、AS.1398中性蛋白酶、Protamex复合蛋白酶、胰酶的最佳水解度分别为12.59%、23.78%、14.45%、20.04%.     

蜂王浆蛋白肽的制备、鉴定及抗氧化活性研究

以蜂王浆为原料，以DPPH自由基清除率和OH自由基清除率为评价指标，对蜂王浆的酶解工艺进行了系统研究，并对蜂王浆酶解液中的蛋白肽进行了分子量分布分析、凝胶过滤层析纯化、LC-MS/MS分离鉴定、肽的合成及活性验证。结果表明，在酸性蛋白酶、料液比1:4、初始pH 5、酶解温度45℃、加酶量9000 U/g及酶解时间5 h的条件下酶解蜂王浆，所得产物的DPPH自由基清除率和OH自由基清除率最佳。对最优工艺下得到的酶解液层析分离，得到活性最高的组分为F4。对该组分进行质谱鉴定，并筛选出15种肽段进行进一步验证，结果表明，得到序列为FDRIW、YPDWSW、WHDKIF的三条肽DPPH自由基清除效果较好，IC50分别为3.63、3.59、1.00 mg/mL，具有在功能食品开发中的应用潜力。     

桑叶蛋白活性多肽的酶解制备

该研究旨在探讨蛋白血管紧张素转换酶（angiotensin I-convening enzyme,ACE）抑制肽及抗氧化肽的酶解制备方法。选取3种蛋白酶（碱性蛋白酶、中性蛋白酶及芽孢杆菌蛋白酶）酶解制备桑叶功能活性多肽,以ACE抑制活性为主要指标并辅以水解度（degree of hydrolysis,DH）评价ACE抑制活性,以DPPH自由基清除能力和总抗氧化能力（total antioxidant capacity,T-AOC）为指标评价其抗氧化性能。运用单因素逐级优化法对酶解反应的酶解pH值、底物浓度、加酶量、酶解温度和酶解时间进行参数优化。结果表明,3种蛋白酶的酶解产物均具有ACE抑制活性,中性蛋白酶酶解产物效果最佳,酶解工艺条件为底物浓度20 g/L、加酶量7.5%、酶解时间50 min、酶解温度55℃、pH7.0时,酶解产物的ACE抑制活性为81.23%,DH为21.41%。在不同蛋白酶优化条件下测定3种酶解产物的抗氧化能力,中性蛋白酶DPPH自由基清除率和总抗氧化能力均为最高,分别为80.702%和4.717 mmol/g。     

鲨鱼肉酶解物的抗氧化作用及其稳定性研究

以还原能力和金属离子螯合能力、清除DPPH·、O2-·、OH·自由基能力为指标,考察鲨鱼肉酶解物的抗氧化活性,同时以清除自由基DPPH·的活性为指标,分析温度和pH值对其抗氧化作用稳定性的影响。结果表明:鲨鱼肉酶解液产物具有一定的还原能力和金属离子螯合能力,其清除DPPH·、O2-·、OH·自由基的IC50值分别为10.16、23.71、19.82mg/mL。酶解产物耐热性强;在酸性环境中,鲨鱼肉酶解液能较好地保持其抗氧化活性,但在碱性环境中抗氧化活性丧失较快。     

蛋白酶酶解核桃饼粕的抗氧化活性

为研究碱性蛋白酶酶解核桃饼粕制得的酶解液的抗氧化活性,运用碱溶酸沉提取法提取核桃饼粕蛋白,利用碱性蛋白酶酶解制得核桃饼粕酶解液及核桃饼粕分离蛋白酶解液.通过对·OH、O2-、DPPH自由基清除能力及总还原力的测定,对比核桃饼粕与其分离蛋白酶解产物的抗氧化活性.结果 显示,·OH清除率分别为17.32％和18.11％,O...     

油莎豆粕抗氧化肽的制备及其稳定性研究

采用碱性蛋白酶和木瓜蛋白酶对油莎豆粕蛋白质进行复合酶解制备油莎豆粕抗氧化肽,以DPPH自由基清除率为指标,确定制备油莎豆粕抗氧化肽的最佳条件。此外,评价了油莎豆粕抗氧化肽的体外抗氧化活性和稳定性。结果表明,制备油莎豆粕抗氧化肽的最适条件为碱性蛋白酶：木瓜蛋白酶=1∶1,酶的质量分数5%、底物质量浓度46 mg/mL、酶解温度61.7℃、酶解pH 6.7、酶解时间2.5 h,在此条件下,DPPH自由基清除率为88.45%。油莎豆粕抗氧化肽的体外抗氧化活性优良,当油莎豆粕抗氧化肽为5 mg/mL时,DPPH·清除率达88.69%、·OH清除率为42.57%、O■清除率为39.17%、ABTS自由基清除率达87.98%,还原能力为0.2627。油莎豆粕抗氧化肽具有热稳定性及冻融稳定性,在酸性和中性条件下稳定,但不耐受碱性条件,经胃肠消化后,活性下降比较明显。     

抗氧化活性林蛙胎卵多肽酶解酶的筛选

以东北林蛙胎卵为原料,以酶解液对DPPH自由基和羟自由基(.OH)清除率为指标,分别研究木瓜蛋白酶、碱性蛋白酶、中性蛋白酶、复合蛋白酶和胰蛋白酶在最佳酶作用条件下酶解时间对酶解液清除DPPH自由基、.OH的影响,筛选出适合生产抗氧化活性林蛙胎卵蛋白肽的碱性蛋白酶、木瓜蛋白酶、中性蛋白酶,再经过复合双酶酶解林蛙胎卵实验,结果表明,以中性蛋白酶与碱性蛋白酶复合酶解林蛙胎卵3h得到肽液的.OH清除率与DPPH自由基清除率最高,达到80.91%和70.45%。     

马氏珠母贝肉酶解产物清除自由基活性的研究

对马氏珠母贝肉的风味酶酶解产物对超氧阴离子自由基、羟自由基及DPPH·的清除效果进行了研究.结果表明,马氏珠母贝肉酶解产物对三种自由基均有一定的清除活性,量效关系明显;其清除O2-·、·OH、DPPH·三种自由基的IC50分别为1.513、1.423、2.618mg/mL,清除自由基能力总体高于牡蛎和海湾扇贝提取物.     

Reliable identification of grapevine rootstock varieties using RAPD PCR on woody samples

Since grapevine ( Vitis spp .) rootstock material is being traded increasingly as disbudded woody material a lack of distinctive morphological features on such material necessitates an alternative and reliable means of identification. Methods described here were developed for rapid and efficient extraction of DNA from woody samples rich in phenolic compounds and polysaccharides, and for subsequent identification of varieties by RAPD PCR. Using these methods, and with the application of only one selected RAPD primer, we were able to differentiate sixteen rootstock varieties, including the seven varieties most commonly used in Germany. Problems commonly encountered with reproducibility of RAPD patterns were avoided by choosing primers with a dinucleotide sequence and a high G/C content that allowed a rather high annealing temperature of 45°C. Methods described here should also be useful for other horticultural crops, especially those with woody tissues rich in phenolic compounds and polysaccharides.     

An internet compendium of analytical methods and spectroscopic information for monomers and additives used in food packaging plastics

An internet website (http://cpf.jrc.it/smt/) has been produced as a means of dissemination of methods of analysis and supporting spectroscopic information on monomers and additives used for food contact materials (principally packaging). The site which is aimed primarily at assisting food control laboratories in the European Union contains analytical information on monomers, starting substances and additives used in the manufacture of plastics materials. A searchable index is provided giving PM and CAS numbers for each of 255 substances. For each substance a data sheet gives regulatory information, chemical structures, physico-chemical information and background information on the use of the substance in particular plastics, and the food packaging applications. For monomers and starting substances (155 compounds) the infra-red and mass spectra are provided, and for additives (100 compounds); additionally proton NMR are available for about 50% of the entries. Where analytical methods have been developed for determining these substances as residual amounts in plastics or as trace amounts in food simulants these methods are also on the website. All information is provided in portable document file (PDF) format which means that high quality copies can be readily printed, using freely available Adobe Acrobat Reader software. The website will in future be maintained and up-dated by the European Commission's Joint Research Centre (JRC) as new substances are authorized for use by the European Commission (DG-ENTR formerly DGIII). Where analytical laboratories (food control or other) require reference substances these can be obtained free-ofcharge from a reference collection housed at the JRC and maintained in conjunction with this website compendium.     

Chlorohydrins of bisphenol A diglycidyl ether (BADGE) and of bisphenol F diglycidyl ether (BFDGE) in canned foods and ready-to-drink coffees from the Japanese market

BADGE.2HCl and BFDGE.2HCl were determined in 28 samples of ready-to-drink canned coffee and 18 samples of canned vegetables (10 corn, 5 tomatoes and 3 others), all from the Japanese market. HPLC was used as the principal analytical method and GCMS for confirmation of relevant LC fractions. BADGE.2HCl was found to be present in one canned coffee and five samples of corn, BFDGE.2HCl in four samples of canned tomatoes and in one canned corn. No sample was found which exceeded the 1mg/kg limit of the EU for the BADGE chlorohydrins. However the highest concentration was found for the sum of BFDGE.2HCl and BFDGE.HCl.H₂O at a level of 1.5mg/kg. A Beilstein test confirmed that all cans containing foods contaminated with BADGE.2HCl or BFDGE.2HCl had at lest one part coated with a PVC organosol.     

European priorities for research to support legislation in the area of food contact materials and articles

A strong science base is required to underpin the planning and decision-making process involved in determining future European community legislation on materials and articles in contact with food. Significant progress has been made in the past 5 years in European funded work in this area, with many developments contributing to a much better understanding of the migration process, and better and simpler approaches to food control. In this paper this progress is reviewed against previously identified work-areas (identified in 1994) and conclusions are reached about future requirements for R&D to support legislation on food contact materials and articles over the next 5 or so years.     

Aroma characterization of various apricot varieties using headspace–solid phase microextraction combined with gas chromatography–mass spectrometry and gas chromatography–olfactometry

The characterization of the aromatic profile of several apricot cultivars with molecular tracers in order to obtain objective data concerning the aromatic quality of this fruit was undertaken using headspace–solid phase microextraction (HS–SPME). Six apricot cultivars were selected according to their organoleptic characteristics: Iranien, Orangered, Goldrich, Hargrand, Rouge du Roussillon and A4025. The aromatic intensity of these varieties measured by HS–SPME–Olfactometry were defined and classified according to the presence and the intensity of grassy, fruity and apricot like notes. In the six varieties, 23 common volatile compounds were identified by HS–SPME–GC–MS. Finally, 10 compounds, ethyl acetate, hexyl acetate, limonene, β-cyclocitral, γ-decalactone, 6-methyl-5-hepten-2-one, linalool, β-ionone, menthone and (E)-hexen-2-al were recognized by HS–SPME–GC–O as responsible of the aromatic notes involved in apricot aroma and considered as molecular tracers of apricot aromatic quality which could be utilized to discriminate apricot varieties.     

Textural Properties of Cold-set Gels Induced from Heat-denatured Whey Protein Isolates

A 9% whey protein (WP) isolate solution at pH 7.0 was heat-denatured at 80°C for 30 min. Size-exclusion HPLC showed that native WP formed soluble aggregates after heat-treatment. Additions of CaCl2 (10–40 mM), NaCl (50–400 mM) or glucono-delta-lactone (GDL, 0.4–2.0%, w/v) or hydrolysis by a protease from Bacillus licheniformis caused gelation of the denatured solution at 45°C. Textural parameters, hardness, adhesiveness, and cohesiveness of the gels so formed changed markedly with concentration of added salts or pH by added GDL. Maximum gel hardness occurred at 200 mM NaCl or pH 4.7. Increasing CaCl2 concentration continuously increased gel hardness. Generally, GDL-induced gels were harder than salt-induced gels, and much harder than the protease-induced gel.     

Tests of potential functional barriers for laminated multilayer food packages. Part I: Low molecular weight permeants

The advent of the functional barrier concept in food packaging has brought with it a requirement for fast tests of permeation through potential barrier materials. In such tests it would be convenient for both foodstuffs and materials below the functional barrier (sub-barrier materials) to be represented by standard simulants. By means of inverse gas chromatography, liquid paraffin spiked with appropriate permeants was considered as a potential simulant of sub-barrier materials based on polypropylene (PP) or similar polyolefins. Experiments were performed to characterize the kinetics of the permeation of low molecular weight model permeants (octene, toluene and isopropanol) from liquid paraffin, through a surrogate potential functional barrier (25 μm-thick oriented PP) into the food simulants olive oil and 3% (w/v) acetic acid. These permeation results were interpreted in terms of three permeation kinetic models regarding the solubility of a particular model permeant in the post-barrier medium (i.e. the food simulant). The results obtained justify the development and evaluation of liquid sub-barrier simulants that would allow flexible yet rigorous testing of new laminated multilayer packaging materials.     

Occurrence of bisphenol-F-diglycidyl ether (BFDGE) in fish canned in oil

The levels of bisphenol-F-diglycidyl ether (BFDGE) were quantified as part of a European survey on the migration of residues of epoxy resins into oil from canned fish. The contents of BFDGE in cans, lids and fish collected from all 15 Member States of the European Union and Switzerland were analysed in 382 samples. Cans and lids were separately extracted with acetonitrile. The extraction from fish was carried out with hexane followed by re-extraction with acetonitrile. The analysis was performed by reverse phase HPL C with fluorescence detection. BFDGE could be detected in 12% of the fish, 24% of the cans and 18% of the lids. Only 3% of the fish contained BFDGE in concentrations considerably above 1mg/kg. In addition to the presented data, a comparison was made with the levels of BADGE (bisphenol-A-diglycidyl ether)analysed in the same products in the context of a previous study.     

Contribution of European research to risk analysis

The European Commission's, Quality of Life Research Programme, Key Action 1—Health, Food & Nutrition is mission-oriented and aims, amongst other things, at providing a healthy, safe and high-quality food supply leading to reinforced consumer confidence in the safety of European food. Its objectives also include the enhancing of the competitiveness of the European food supply. Key Action 1 is currently supporting a number of different types of European collaborative projects in the area of risk analysis. The objectives of these projects range from the development and validation of prevention strategies including the reduction of consumers risks; development and validation of new modelling approaches; harmonization of risk assessment principles, methodologies, and terminology; standardization of methods and systems used for the safety evaluation of transgenic food; providing of tools for the evaluation of human viral contamination of shellfish and quality control; new methodologies for assessing the potential of unintended effects of genetically modified (genetically modified) foods; development of a risk assessment model for Cryptosporidium parvum related to the food and water industries; to the development of a communication platform for genetically modified organism, producers, retailers, regulatory authorities and consumer groups to improve safety assessment procedures, risk management strategies and risk communication; development and validation of new methods for safety testing of transgenic food; evaluation of the safety and efficacy of iron supplementation in pregnant women; evaluation of the potential cancer-preventing activity of pro- and pre-biotic ('synbiotic') combinations in human volunteers. An overview of these projects is presented here.