食品包装材料中丙烯酰胺在大鼠肝微粒和S9中的体外代谢研究

目的研究食品包装材料中丙烯酰胺(acrylamide)的体外代谢情况,并确证其代谢产物。方法采用肝微粒体及肝S9体外温孵法,优化代谢条件,对丙烯酰胺进行了体外代谢研究,并用液相色谱-串联质谱法(liquid chromatography-tandem mass spectrometry,LC-MS/MS)检测并确证丙烯酰胺的代谢物。结果通过检测结果发现,丙烯酰胺分别在肝S9和肝微粒体作用下发生显著的代谢反应,其代谢产物之一为环氧丙酰胺。结论丙烯酰胺能通过代谢转化成毒性更强的环氧丙酰胺,因此,监控食品、饮水和周围环境中的丙烯酰胺的含量对于维护人们的健康具有重要的意义。     

食品接触材料中芘的体外代谢模拟分析

目的模拟体内代谢条件,分析芘的体外基本代谢情况。方法采用肝微粒体及肝S9成分2种代谢试剂,体外模拟了肝脏代谢条件,对代谢试剂浓度及代谢时间条件进行优化,采用气相色谱-质谱(GC-MS)法检测芘和羟基芘,比较分析2种代谢试剂对PYR的代谢情况。结果体外代谢最佳孵化时间为60 min,最佳体外代谢试剂浓度为0.5 mg/m L。结论 PYR在肝微粒体代谢试剂中无明显代谢反应发生,在肝S9成分的作用下,代谢生成羟基芘,推断参与PYR代谢的主要为二相酶类。     

两色金鸡菊中马里苷的肠道菌群代谢转化

该研究探讨了两色金鸡菊中马里苷的肠道菌群代谢情况，考察了马里苷代谢规律并探索其原型成分与代谢产物的相关性。采集健康SD大鼠新鲜粪便制备肠菌孵育液，建立大鼠肠道菌群体外孵育模型，与马里苷单体在厌氧环境下共孵育。采用超高效液相色谱-四极杆串联飞行时间高分辨质谱（Ultrahigh Performance Liquid Chromatography Tandem Quadrupole Time-of-flight Mass Spectrometry，UHPLC-Q-TOF-MS/MS）技术对0、6、12、24、36 h的代谢产物进行定性分析。利用外标法对马里苷及其4种主要代谢产物黄诺马苷、异奥卡宁、异绿原酸B、奥卡宁不同时间段的含量进行测定，观察马里苷在肠道菌培养液中的代谢规律。最终在大鼠肠道菌群孵育液中共鉴定出11个代谢产物，发现马里苷在肠道菌群代谢过程中主要发生了异构化、葡萄糖醛酸化、环裂解、羟基化、乙酰化等反应。马里苷在肠道菌的作用下，6 h就已检测不到原型成分，12~24 h主要的代谢产物为异奥卡宁、奥卡宁、7,3'',5''-三羟基黄酮、槲皮素等由马里苷代谢产生的苷元。综上，肠道菌加速了马里苷的水解脱糖，在代谢后期产生的代谢产物均为苷元。     

食品包装材料中有害物质双酚A二缩水甘油醚 体外代谢研究

目的通过模拟体内代谢,对双酚A二缩水甘油醚(BADGE)的体外基本代谢情况进行研究。方法采用肝微粒体、肝S9 2种体外代谢试剂,通过模拟体内肝脏代谢,对BADGE的代谢行为及其代谢产物进行研究。通过代谢试剂浓度及代谢时间条件的优化,采用高效液相色谱-串联质谱(HPLC-MS/MS)作为检测手段对BADGE的体外代谢产物进行分析确证。结果体外代谢最佳孵化时间为60 min,最佳体外代谢试剂浓度为0.5mg/m L,在肝S9及肝微粒体2种体外代谢试剂的作用下,BADGE发生显著的代谢反应。结论本研究与传统的动物试验相比,节约了时间、精力,对食品包装材料的毒理学研究和安全性评价有重要的推动作用。     

基于肝微粒体和肝S9成分的食品接触材料中 五氯苯酚体外代谢分析

目的通过体外代谢方法,研究五氯苯酚(pentachlorophenol,PCP)的代谢情况。方法采用肝微粒体和肝S9成分2种体外代谢试剂模拟体内代谢模式,通过优化代谢反应条件,建立PCP、四氯苯醌(Cl4BQ)2种目标物的气相色谱-质谱联用(GC-MS)检测方法,对PCP进行体外代谢研究分析。结果体外代谢分析的最佳反应条件为代谢试剂浓度0.75 mg/m L、代谢反应时间4 h。结论在最优反应条件下分析比较2种试剂对PCP的代谢情况,验证了PCP的代谢产物为Cl4BQ,且在肝微粒体和肝S9成分中的代谢没有显著差异。     

红薯叶不同溶剂提取物抗氧化性及活性成分鉴定

研究不同极性溶剂对红薯叶中酚类化合物的提取以及提取物抗氧化性的影响,并鉴定提取物中的主要抗氧化成分组成。分别采用极性不同的7 种溶剂（蒸馏水、甲醇、无水乙醇、丙酮、正丁醇、乙酸乙酯和氯仿）从红薯叶中提取多酚,并评价提取物中总酚、总黄酮和花青素的含量,以及对1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基清除能力和还原能力,最后运用高效液相色谱-串联质谱（high performance liquidchromatography tandem mass spectroscopy,HPLC-MS/MS）技术分析抗氧化活性最好的提取物中多酚的主要组成成分。结果表明：提取溶剂的极性对红薯叶中多酚类化合物的提取效率和提取物抗氧化活性有很大的影响,水提物具有最高的粗提物得率（（37.13±1.60）%）,而甲醇提取物中总酚含量（13.80 mg GAE/g）和总黄酮含量（（5.68±0.35）mg QE/g）最高,且具有最好的DPPH自由基清除能力（IC50为0.32 mg/mL）与还原能力（ρ0.5为0.95 mg/mL）。采用HPLC-MS/MS从红薯叶甲醇提取物中鉴定9 种、初步鉴定3 种酚类化合物,鉴定的化合物为咖啡酸、对羟基苯甲酸、1-咖啡酸奎宁酸、3-咖啡酸奎宁酸、异绿原酸A、异绿原酸B、异绿原酸C、3,4,5-三咖啡酰奎尼酸和金丝桃苷。     

超高效液相色谱法测定清咽糖中6种绿原酸类化合物的含量

目的建立超高效液相色谱法同时测定清咽糖中新绿原酸、绿原酸、隐绿原酸、3,4-O-二咖啡酰基奎宁酸、3,5-O-二咖啡酰基奎宁酸和4,5-O-二咖啡酰基奎宁酸6种绿原酸类化合物的含量。方法采用Agilent Eclipse Plus C18 RRHD色谱柱(2.1 mm×150 mm, 1.8μm),以乙腈、0.4%磷酸水溶液为流动相,梯度洗脱,流速0.8 mL/min,检测波长327 nm,柱温35℃。结果 6种成分在相应的线性范围内呈良好的线性关系。精密度、重复性、稳定性实验中6种成分峰面积的相对标准偏差均小于3.0%,平均加样回收率为93.77%～104.37%,相对标准偏差为0.28%～1.86%。结论该方法具有良好的专属性和重现性,加样回收率实验符合要求,结果准确、稳定,适用于同时测定清咽糖中新绿原酸、绿原酸、隐绿原酸、3,4-O-二咖啡酰基奎宁酸、3,5-O-二咖啡酰基奎宁酸和4,5-O-二咖啡酰基奎宁酸的含量。     

高效液相色谱-串联质谱法测定饮料中7种绿原酸类物质

目的：建立了一种高效液相色谱-串联质谱法(High performance liquid chromatography-tandem mass spectrometry, HPLC-MS/MS)同时快速、准确测定饮料中7种绿原酸类物质含量的方法。方法：研究不同柱温、流速、色谱柱、提取溶剂、超声时间等对7种绿原酸类物质测定的影响,最终确定试样经90%甲醇水溶液超声提取,采用RP18(2.1 mm×100 mm,1.7μm)色谱柱分离,HPLC-MS/MS同时测定新绿原酸、绿原酸、隐绿原酸、异绿原酸A、异绿原酸B、异绿原酸C、1,5-二咖啡酰奎宁酸成分含量。结果：对HPLC-MS/MS检测7种成分的检测条件优化和方法学考察,包括色谱条件。结论：该检测方法简便、准确、可靠,线性关系、精密度、重复性、稳定性、加样回收率均符合要求,可用于饮料中7种绿原酸类成分的定量分析。     

HPLC法同时测定甘薯叶中5种酚酸和异槲皮苷

咖啡酸、绿原酸、3,4-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、4,5-二咖啡酰奎宁酸和异槲皮苷是甘薯叶中的主要活性成分,本研究建立高效液相色谱（high performance liquid chromatography, HPLC）同时测定这6种成分的方法。通过对检测波长、色谱柱、洗脱溶剂、洗脱流速等参数优化得到的最佳分析条件为:ZORBAX SB-C₁₈（4.6 mm×250 mm,5 μm）为分析柱,以0.1%甲酸水溶液（A）和乙腈（B）为流动相,采用梯度洗脱:0～12 min,10%~25% B;12～18 min,25% B,流速0.8 mL/min,检测波长326 nm,柱温30 ℃。咖啡酸、绿原酸、3,4-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、4,5-二咖啡酰奎宁酸和异槲皮苷的线性范围分别为0.25~25.0、0.5~50.0、5.0~100.0、10.0~200.0、5.0~100.0和0.5~50.0 μg/mL。该方法分析时间短（18 min）,线性度（R2≥0.998）、稳定性（RSD≤1.97%）、精密度（RSD≤1.56%）和加标回收率（95.14%~103.22%）好。对江西10个不同产地甘薯叶目标成分分析发现,宜春丰城的甘薯叶具有最高的总酚含量。     

LC-MS/MS方法同时检测超声提取迷迭香(Rosmarnus officinalis L.)叶片中的4种主要成分

目的:建立一种同时测定迷迭香中迷迭香酸、鼠尾草酸、绿原酸和咖啡酸含量的LC-MS/MS分析方法,并优化超声辅助提取这4种物质的方法。方法:液相条件:采用Agilent Eclipse XDB-C18色谱柱(150mm×4.6mm,5μm),流动相为乙腈和水(含0.1%甲酸)进行梯度洗脱。质谱条件:ESI离子源,MRM负离子扫描方式,检测在不同提取溶剂、料液比、超声时间、pH值及酸种类的超声波辅助提取条件下4种物质的含量。结果:咖啡酸、绿原酸、迷迭香酸和鼠尾草酸在质量浓度范围分别为0.0005～0.5、0.0005～0.5、0.002～0.2、0.01～1mg/mL的条件下线性关系良好,并符合方法学考察要求;在最优提取条件下迷迭香酸含量为4.07mg/g,鼠尾草酸含量为14.50mg/g,绿原酸含量为8.49mg/g,咖啡酸含量为2.92mg/g。结论:该分析方法可快速灵敏地实现同时检测,并优化了这4种化合物的最佳提取条件。     

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.     

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.     

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.     

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.