纤维素酶法水解大豆异黄酮糖苷研究

为了提高大豆异黄酮的生物活性作用,获得更多的大豆异黄酮苷元成分,对纤维素酶水解大豆异黄酮进行了研究,通过单因素试验和正交试验,酶水解的影响因素主要为水解温度、水解时间、酶-底物质量比和酶水解pH值。得到纤维素酶制备大豆异黄酮苷元的较优工艺条件:纤维素酶水解时间17 h,水解温度48.5℃,水解pH值5.0,酶-底物质量比为1.05%。通过小试试验,以脱脂豆粕原料进行提取大豆异黄酮,纤维素酶水解得到苷元产品收率为0.1%,总苷元质量分数为30.79%;以30%大豆异黄酮粉为原料进行水解,获得产品收率为10.2%,总苷元质量分数为47.81%。     

酶法催化大豆异黄酮糖苷水解苷元的工艺研究

试验主要研究在单因素基础上,通过正交试验,考察酶的用量、反应温度和反应时间对大豆异黄酮糖苷水解苷元工艺的影响,确立了水解反应的最佳工艺条件。试验结果表明,当反应温度为52℃,酶用量为14 mg,反应时间2.8 h,大豆异黄酮总的水解产率可达到82.91%以上。用高效液相色谱对水解产物进行定量分析。超声波协同酶催化大豆异黄酮糖苷水解苷元的工艺研究具有操作简单、安全、成本低廉等特点,适用于大豆异黄酮糖苷转化苷元的水解工艺。     

酶水解对大豆异黄酮粗提物中苷元含量的影响

采用β-葡萄糖苷酶水解的方法将大豆异黄酮糖苷转化为苷元,以染料木素和大豆苷元含量为指标,通过单因素试验对水解过程中的不同影响因素进行了考察。以染料木素含量为指标,运用正交试验优化了β-葡萄糖苷酶水解大豆异黄酮的工艺条件为反应温度40℃、水解时间1.5h、水解介质pH4.5、水解底物浓度10mg/mL,在此条件下,水解得到的大豆异黄酮苷元中染料木素的含量可达到22.91%。     

纤维素酶催化大豆异黄酮的水解

为充分发挥大豆异黄酮生物价值,采用纤维素酶催化糖苷型大豆异黄酮水解制备游离苷元型大豆异黄酮。通过考察10种纤维素酶对糖苷型大豆异黄酮总水解率和苷元型大豆异黄酮总转化率的影响,筛选得到一种成本较低且水解效果较好的纤维素酶,用于催化水解糖苷型大豆异黄酮,优化了该纤维素酶在水解工艺中底物质量浓度、酶添加量、反应体系pH、酶解温度、酶解时间等参数。结果表明：选择来源于Trichoderma viride的纤维素酶作为大豆异黄酮水解用酶;底物质量浓度0.8～2.0 mg/mL、酶添加量7%～11%、反应体系pH 5.0、酶解温度55℃、酶解时间5～6 h是较经济有效的水解工艺参数,实验优化过程中,大豆异黄酮总水解率超过90%,总转化率接近60%。因此,采用纤维素酶催化水解大豆异黄酮可显著增加游离苷元含量,提高大豆异黄酮利用价值。     

糖化酶水解大豆异黄酮

研究了糖化酶水解大豆异黄酮的技术工艺,利用糖化酶水解大豆异黄酮粉得到大豆异黄酮苷元。通过单因素试验对水解过程中的不同影响因素进行考察,运用正交试验优化了糖化酶水解大豆异黄酮的反应条件,优化结果为:水解温度55℃,pH值4.8,时间7 h,水解率可达98.18%。     

高效液相色谱法快速测定大豆异黄酮制品中有效成分的研究

采用高效液相色谱法测定大豆异黄酮制品中染料木苷、黄豆苷、染料木黄酮和黄豆苷元含量.固定相为Agilent-1100 C18柱(3.9 x 150mm);以甲醇:水=35%～45%为流动相,进行梯度洗脱,流速为1.0～2.0mL/min;柱温为室温;检测波长为260nm.实验结果表明,市售30%大豆异黄酮制品中,含大豆异黄酮糖苷近28%,其中染料木苷为4%、黄豆苷为8%,苷元形式异黄酮未检出;采用酶水解法制备的大豆异黄酮水解物中含染料木黄酮14.3%、黄豆苷元9.8%,其大豆异黄酮苷元占大豆异黄酮总含量的96%.     

大豆异黄酮糖苷水解研究进展

该文在介绍大豆异黄酮的组成、化学结构、特性及生理功能基础上,详细介绍水解法制备大豆异黄酮苷元原理、工艺流程、大豆异黄酮苷元分离纯化及检测方法。     

纤维素酶水解大豆异黄酮糖苷混合物的抗氧化活性

大豆异黄酮主要以糖苷类型的分子形式存在于大豆中,但是其具有生物活性的部分主要是苷元.为了建立一种体外生物转化大豆异黄酮糖苷为大豆异黄酮苷元的新方法,采用纤维素酶水解70%乙醇提取脱脂大豆粕得到的异黄酮粗提物,对比了水解前后两样品的异黄酮组成差异,并测定了其清除DPPH自由基的能力.结果表明:纤维素酶能完全水解大豆苷和染料木苷为大豆素和染料木素;而不能水解丙二酰基染料木苷和丙二酰基大豆苷.纤维素酶处理以后的大豆异黄酮糖苷混合物达到DPPH自由基50%清除率所需要的异黄酮浓度是未处理的大豆异黄酮糖苷混合物的1/2.56.     

乳酸催化大豆异黄酮糖苷水解的工艺研究

研究乳酸催化水解大豆异黄酮糖苷的最佳工艺条件。以大豆异黄酮糖苷的水解率为评价指标,通过单因素试验对水解过程中的不同影响因素进行考察,运用正交试验优化了乳酸水解大豆异黄酮糖苷的反应条件,结果为,水解温度163℃,水解时间2.5 h,乳酸水溶液浓度2.0 mol/L,水解率可达100%。优选所得的大豆异黄酮糖苷水解生成苷元的工艺简单易行,实用性高。     

大豆异黄酮水解物制备及放大实验

用发酵黑曲霉得到的β-葡萄糖苷酶水解40%的大豆异黄酮粉,通过正交实验确定最佳水解条件为:加酶量100u,底物浓度20mg/mL,50℃,水解1h。将水解液降温至4℃,4000r/min离心分离30min,离心后沉淀物在-18℃下预冻,于-40℃冷冻干燥,得到固态大豆异黄酮苷元,经检测苷元转化率90.12%、大豆皂苷3.87%、大豆异黄酮94.36%、大豆苷元43.45%、染料木素46.26%。放大实验结果:苷元转化率78.56%、大豆皂苷4.93%、大豆异黄酮88.97%、大豆苷元36.47%、染料木素38.81%。     

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.     

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.     

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.     

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.