基于斑马鱼幼鱼的塑化剂肝毒性危害识别研究

目的 采用塑化剂邻苯二甲酸二乙基己酯（DEHP）和邻苯二甲酸二丁酯（DBP）作为阳性对照,建立斑马鱼幼鱼脂肪肝毒性风险评估模型进行塑化剂邻苯二甲酸丁酯苯甲酯（BBP）肝毒性危害识别。方法 分析DEHP和DBP暴露后斑马鱼肝脏脂肪信号强度及过氧化物酶体增殖物激活受体（PPAR）靶基因CD36的表达,初步构建斑马鱼幼鱼肝毒性模型;基于此模型对BBP进行肝毒性评价,此外采用肝脏表达绿色荧光转基因斑马鱼幼鱼进行肝密度分析、甘油三酯（TG）定量检测以及成鱼肝脏病理检查进一步验证模型的可靠性,应用基准剂量法（BMD）推导关键效应点。结果 DEHP、DBP表现出极显著诱发脂肪肝风险（均P<0.01）,显著上调CD36相对表达量（P<0.001,P<0.05）,提示造模成功。BBP（0.000 012 5%、0.000 025%、0.000 05%）诱发斑马鱼幼鱼显著的脂质沉积（P<0.05,P<0.001,P<0.001）、显著影响密度（P<0.05,P<0.01,P<0.01）、显著增加TG蓄积（P<0.05,P<0.05,P<0.01）、上调CD36基因表达。此外,BBP（≥0.000 012 5%）诱发斑马鱼成鱼肝脏部位脂质空泡形成、空泡间隙和数量减少,提示BBP具有明显的肝毒性。关键效应为肝脏脂肪信号强度（S）,基准剂量下限（BMDL）值为0.013 mg/L。结论 本研究成功构建了一种通过肝脏脂肪信号强度来识别塑化剂肝毒性的斑马鱼幼鱼模型,结合基准剂量法进一步为塑化剂肝毒性危害识别提供科学依据。     

拟除虫菊酯类农药对斑马鱼幼鱼的神经毒性作用研究

摘要：目的 探究四种拟除虫菊酯类农药（溴氰菊酯、丙烯菊酯、高效氯氰菊酯、顺式氰戊菊酯）对斑马鱼神经毒性的作用,并与大鼠毒性结果进行比较,以期为斑马鱼在食品安全评价中的应用提供参考。方法 对四种农药采用静态暴露方法,分析其在高中低三种剂量下对斑马鱼幼鱼的形态、神经行为的影响;同时以活性氧（Reactive oxygen,ROS）、超氧化物歧化酶（(superoxide dismutase,SOD）、乙酰胆碱酯酶（Acetylcholinesterase,AchE）为指标,研究各农药在高中低剂量下对斑马鱼幼鱼神经化学的影响。结果 四种拟除虫菊酯类农药均对斑马鱼幼鱼造成了体长变短、脊柱弯曲等形态变化;除丙烯菊酯外,其他三种农药均使斑马鱼幼鱼出现显著的运动活性降低趋势;在神经化学方面,与空白组和对照组相比,四种农药随着浓度增加均使斑马鱼幼鱼体内的AchE活性显著下降、SOD活性显著升高;但四种农药对ROS活性变化各有差异：溴氰菊酯与高效氯氰菊酯对斑马鱼幼鱼的ROS影响无显著性差异,而丙烯菊酯和顺式氰戊菊酯在高中剂量下均可导致斑马鱼幼鱼ROS显著增加。结论 拟除虫菊酯农药对斑马鱼幼鱼产生了显著的神经毒性作用,首次发现形态变化和AchE活性的变化与大鼠相关结果体现出较好的一致性,有望替代大鼠用于更加简便快捷的食品安全风险评估;其余指标与大鼠呈现不同程度的差异,可能与两者生物学特性的差别有关。     

黄芩叶毒理学安全性研究

目的 对黄芩叶进行系统毒理学研究得到安全性基础数据。方法 以黄芩叶提取物为受试物,采用食品安全国家标准方法,对黄芩叶开展急性经口毒性试验、细菌回复突变试验、哺乳动物红细胞微核试验、小鼠精母细胞染色体畸变试验以及90 d经口毒性试验,进行系统的毒理学安全性评价。结果 黄芩叶提取物急性经口毒性试验雌雄小鼠LD₅₀>20.0 g/kg?BW,雌雄大鼠LD₅₀>15.0 g/kg?BW,属于实际无毒级;细菌回复突变试验、哺乳动物红细胞微核试验及小鼠精母细胞染色体畸变试验3项遗传毒性检测结果均为阴性;90 d经口毒性试验中,2.0、4.0、8.0 g/kg?BW 3个剂量组大鼠的体质量、进食量、食物利用率、血液学、血生化等指标,与对照组比较无显著性差异。结论 在本试验剂量条件下,未发现黄芩叶提取物摄入对实验动物产生毒性作用,换算后黄芩叶NOAEL值为24.0 g/kg?BW。     

液相色谱—柱后衍生法测定苹果中5种氨基甲酸酯类农药残留量的不确定性评定

目的:对苹果中5种氨基甲酸酯类农药检测结果的准确度进行分析。方法:依据NY/T 761—2008对苹果中5种氨基甲酸酯类农药残留量进行检测,并对其测定过程的不确定度进行评定和分析。结果:苹果中克百威、灭多威、3-羟基克百威、涕灭威、异丙威含量以其扩展不确定度形式分别表示为(0.105 7±0.007 8),(0.107 7±0.006 2),(0.109 2±0.005 8),(0.103 9±0.008 4),(0.106 1±0.007 6) mg/kg (k=2)。 结论:试验中不确定度的主要来源为标准物质、回收率、样品前处理和检测仪器,其中整体表现为U_rel(标准)＞U_rel(回收率)＞U_rel(前处理)＞U_rel(仪器)。     

玉米全粉中黄曲霉毒素B1标准物质的研制

目的 研制均匀性和稳定性满足计量要求的玉米全粉中黄曲霉毒素B₁成分分析标准物质,用于粮食及其制品中黄曲霉毒素B₁的质量控制。方法 采用高效液相色谱法对玉米全粉中黄曲霉毒素B₁候选物进行均匀性检验、稳定性考察,选取8家具有较高检测水平的实验室开展多家实验室联合定值。结果 均匀性检验通过方差分析可知,F_统计值＜F_临界值 。稳定性监测结果通过分析可知,候选物中黄曲霉毒素B₁未观测到不稳定性。确定了玉米全粉中黄曲霉毒素B₁标准值和不确定度。结论 该标准物质具有良好的均匀性,可在常温 (<25℃)下保存,在低于60℃的条件下运输。玉米全粉中黄曲霉毒素B₁标准物质定值结果为27±3μg/kg,k=2(95%置信区间),已被批准发布为国家二级标准物质。     

高峰淀粉酶活性对饮料中维生素B2测定结果的影响

目的 调查分析高峰淀粉酶来源和酶解参数对饮料中维生素B₂（核黄素-5''-磷酸钠来源）测定结果的影响。方法 按GB 5009.85—2016对受试样品中维生素B₂（核黄素-5''-磷酸钠来源）的含量进行检测,考察高峰淀粉酶及其工作条件对检测结果的影响。结果 不同来源的高峰淀粉酶对维生素B₂（核黄素-5''-磷酸钠来源）的实测结果有明显的影响,实测值与理论值最大相差10倍以上;前处理过程中建议选择酶解时间12 h,酶解pH值6.0~6.5。结论 国标方法检测饮料中维生素B₂（核黄素-5''-磷酸钠来源）时,预先检测高峰淀粉酶的酶解活性是十分必要的。     

源于中高温大曲的米根霉制备米曲工艺优化及应用

目的:提高大曲白酒的出酒率。方法:从中高温大曲中筛选出高产糖化酶的菌株,将该菌株制作成米曲并应用至大曲白酒的酿造中,测定出酒率。结果:分离得到一株高产糖化酶菌株M-1,鉴定结果为米根霉(Rhizopus oryzae)。用该菌株制备成米曲的最优工艺条件为m_麸皮∶m_米粉为2∶8、米曲含水量45%、培养时间72 h、干燥温度40 ℃,此时米曲糖化酶活力高达864.50 U/g。当M-1米曲添加量为粮食质量的2%时,白酒出酒率提高至43.17%。结论:建立了一种将米根霉制备成米曲来提高中高温大曲糖化酶活力的方法。     

血浆中85种有毒生物碱快速筛查方法的建立及其应用

目的 利用超高效液相色谱-四级杆/静电场轨道阱高分辨质谱建立血浆中85种有毒生物碱快速筛查方法。方法 构建85种生物碱高分辨数据库；血浆经乙腈沉淀蛋白进行预处理，采用Acquity Waters BEH C₁₈色谱柱（100 mm×2.1 mm，1.7 μm），以5 mmol/L甲酸铵（含0.1%甲酸）和乙腈为流动相进行梯度洗脱，对比了3种数据采集方式下（Full mass/dd-MS²、Full mass/AIF、Full mass/DIA）加标血浆中85种有毒生物碱筛查情况。结果 血浆在5、50和250 ng/mL 3个浓度添加水平下，Full mass/DIA模式下的筛选准确率均为最高。利用生物碱数据库，选择Full mass/DIA模式采集样品数据信息，建立了血浆中85种有毒生物碱快速筛查方法。80%以上生物碱在血浆中的检出限低于10 ng/mL。利用该方法成功在滇乌头碱毒物代谢动力学实验获得的3份Wistar大鼠血浆中筛选到了目标物滇乌头碱及其可能的代谢产物脱氧乌头碱及印乌头碱。结论 该方法快速、高效，可以在无标准品情况下实现突发性有毒生物碱中毒血浆的快速筛查。     

高效液相色谱法同时测定保健食品中6种黄酮类化合物

目的:建立同时测定保健食品中汉黄岑苷、4’-羟基汉黄岑素、去甲汉黄岑素、黄岑素、汉黄岑素、白杨素的分析方法。方法:样品经丙酮提取,C₁₈固相萃取柱净化,采用高效液相色谱仪,以样品基质匹配的标准曲线法进行定量分析。色谱柱为Phenomenex Luna^?C₁₈(250 mm×4.6 mm,5 μm),通过流动相(乙腈—乙酸溶液)进行梯度洗脱,流速为1.0 mL/min,柱温为30 ℃,检测波长为280 nm。结果:保健食品基质中6种目标物分离情况良好,相应的浓度与峰面积呈良好的线性关系,相关系数(R²)均＞0.99,精密度和稳定性试验RSD≤3.08%,加标平均回收率为83%~100%(RSD＜4.63%)。方法检出限为0.02~0.07 mg/kg,方法定量限为0.08~0.25 mg/kg。结论:该方法重现性好、有机溶剂消耗少,适用于保健食品中6种黄酮类化合物的同时测定。     

液相色谱—串联三重四极杆复合线性离子阱质谱法测定水生蔬菜中45种抗生素

目的:建立超高效液相色谱—三重四极杆复合线性离子阱质谱法同时测定水生蔬菜中多种抗生素类化合物。方法:样品以含0.1%甲酸的乙腈溶液为溶剂,加入N-丙基乙二胺(PSA)、无水Na₂SO₄和C₁₈净化处理,质谱采用分时段多反应检测—信息关联—增强子离子(SMRM-IDA-EPI)扫描模式及谱库检索技术,通过对化合物保留时间、离子对及EPI谱库检索对比,外标法定量。结果:45种抗生素在1.0~100.0 μg/L质量浓度范围内线性关系良好(R²＞0.99),检出限为0.3~1.0 μg/kg,定量限为1.0~3.0 μg/kg,在3个浓度添加水平下,45种抗生素的回收率为60.4%~121.7%,相对标准偏差为0.9%~10.2%。结论:该方法快速简便、定性能力强,适用于水生蔬菜中45种抗生素类化合物的筛查确证和定量检测。     

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.     

Influence des ions sur le pouvoir hydratant de l'ure e: e tude sur peau de porc ex vivo

This study deals with the influence of ions (NaCl and MgSO4) in a W/O emulsion containing 10% urea. Moisturization kinetics are assessed by corneometry on pig skin ex vivo. The formula's influence on urea penetration is measured by infrared spectrometry with an ATR device and the stripping method. Corneometry and spectroscopy were chosen to record simultaneously the hydratation levels and urea localization into superficial cell layers. Urea crystallization after evaporation of emulsions and aqueous solutions is described. Results show that urea does not hydrate nor penetrate when applied to the skin through an aqueous gel. In a W/O emulsion, sodium chloride increases the ability of urea to moisturize without improving penetration. In vitro urea crystallization is disturbed by sodium chloride or magnesium sulphate for solutions and emulsions. This stabilization by ions is correlated with good moisturization values. The stabilization of urea in the solute state provided by ions increases its water epidermal binding capacity without enhancing penetration.