苦丁茶提取物的分离纯化、鉴定及不同萃取部位活性分析

以苦丁茶为原料，对其乙醇提取物和不同溶剂萃取部位的抗氧化活性、抗补体活性、抑制癌细胞活性进行探究，并对活性最佳的萃取部位进一步分离纯化，分析鉴定出单体化合物。抗氧化活性以总还原力、清除1,1-二苯基-2-三硝基苯肼自由基和超氧阴离子自由基的能力为指标，通过溶血实验评价抗补体活性，并采用MTT法测定对人乳腺癌细胞MCF-7的存活率，结合多种分离技术获得不同单体化合物，通过对质谱、核磁共振波谱的数据分析鉴定其化学结构。结果表明，苦丁茶乙醇提取物及各部位均具有良好的抗氧化效果，其中乙酸乙酯部位不仅在实验质量浓度范围内有较强的抗氧化活性（0.8 mg/mL均达到90%以上），还具有良好的抗补体活性（79.67±0.99）%，能很好抑制人乳腺癌MCF-7细胞增殖活性（54.8±0.26）%，对乙酸乙酯部位进行分离纯化，总共分离得到11 个单体化合物。由此可见，苦丁茶乙醇提取物和各部位均具有抗氧化、抗补体活性和人乳腺癌MCF-7细胞抑制活性，乙酸乙酯作为萃取剂时活性最佳。分离的单体化合物鉴定后多为酚类、黄酮类物质。     

铁皮石斛抗肿瘤活性成分的探究

为了探究铁皮石斛中的抗肿瘤活性成分,本研究采用乙酸乙酯、95%乙醇、水三种不同极性的溶剂依次对铁皮石斛茎的干燥粉末进行了提取,初步得到了三种提取物;初提取物在MTT细胞增殖检测结果的指引下,结合硅胶柱层析、TLC薄层分析和凝胶柱层析进行分离纯化,最终筛选出具有强抗肿瘤活性的组分。结果显示乙酸乙酯提取物经分离纯化后得到了最强抗肿瘤活性成分,其对肝癌细胞HepG-2、胃癌细胞SGC-7901和乳腺癌细胞MCF-7的增殖都有明显的抑制效果,半抑制浓度IC50依次为:0.45μg/mL、0.36μg/mL、0.34μg/mL,经波谱学数据鉴定,该化合物为金钗石斛菲醌。金钗石斛菲醌首次分离自金钗石斛,其抗肿瘤功效已经得到证实,并引起了广泛的关注,本研究成果对以铁皮石斛为原料的药品和保健食品的开发具有重要的指导意义。     

桦褐孔菌提取物抗乳腺癌活性部位筛选及成分分析

以人乳腺癌MDA-MB-231细胞筛选桦褐孔菌抗乳腺癌活性部位,并对其活性部位的成分进行初步分析。采用MTT法检测发酵液不同萃取部位抗乳腺癌活性;通过细胞形态观察进一步了解药物对细胞的影响;划痕试验检测桦褐孔菌活性部位对细胞迁移的影响;采用LC-MS/MS对活性部位的成分进行分析。结果表明,桦褐孔菌石油醚萃取部位对MDA-MB-231细胞增殖的抑制作用最强,且呈量效和时效关系,作用24 h的IC50值为76.17μg/mL;细胞形态观察显示石油醚萃取部位对乳腺癌MDA-MB-231细胞的凋亡作用随着药物质量浓度的升高而加强;划痕实验显示,药物对细胞迁移有较强的抑制作用;LC-MS/MS数据初步表征了其中的14个化学成分,初步鉴定了其中的10种成分,这10种成分首次从桦褐孔菌中得到鉴定,虽然其他4个物质未能鉴定,但亦对其相对分子质量和分子式进行了初步表征。证实了石油醚部位是桦褐孔菌抗乳腺癌MDA-MB-231细胞的主要活性部位,能抑制乳腺癌细胞增殖并诱导细胞凋亡,为进一步开发利用桦褐孔菌食药用资源提供科学依据。     

野生苦菜体外抗氧化及抑制肿瘤活性研究

以野生苦菜为原料,采用1,1-二苯基-2-三硝基苯肼(DPPH)清除能力、2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)清除能力、羟基自由基清除能力和铁离子还原能力法评定其抗氧化能力,采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法评定苦菜提取物对雌激素依赖性人类乳腺癌细胞MCF-7和雌激素非依赖性人乳腺癌细胞MDA-MB-231的抑制活性。结果表明,苦菜甲醇提取物对ABTS、DPPH都具有较强的清除能力,且具有较强的铁离子还原能力,对雌激素依赖性人类乳腺癌细胞MCF-7和雌激素非依赖性人乳腺癌细胞MDA-MB-231均有显著的抑制活性。苦菜的甲醇提取物对MCF-7和MDA-MB-231半抑制率分别为99、107μg/mL。     

朝鲜族大酱中异黄酮的提取工艺优化及其抗肿瘤活性研究

为优化朝鲜族大酱异黄酮的提取工艺,并研究朝鲜族大酱中异黄酮的抗肿瘤活性,本实验用超声波法提取朝鲜族大酱中的异黄酮,通过单因素实验和正交实验优化其提取工艺,并通过CCK-8法对正常乳腺细胞、MDA-MB-231和MCF-7两种乳腺癌细胞的增殖抑制情况进行了测定,用Western Blot法检测了朝鲜族大酱中异黄酮处理后的两种乳腺癌细胞凋亡相关蛋白的表达情况。通过单因素和正交试验确定其最佳提取条件为提取时间80 min、料液比1:24、温度50 ℃,此时得率最高,为0.802%。CCK-8结果显示,不同浓度的朝鲜族大酱异黄酮对正常细胞无毒副作用,对MDA-MB-231和MCF-7两种乳腺癌细胞的增殖抑制情况均呈剂量-时间依赖性,且均有显著性差异(p<0.05)。对不同浓度朝鲜族大酱异黄酮处理过的两种细胞凋亡相关蛋白的检测结果显示,两种细胞随着异黄酮浓度的增加,其促凋亡蛋白Bax,caspase-3的表达量与空白对照组相比均显著上升(p<0.05),而抗凋亡蛋白Bcl-2的表达量显著下降(p<0.05),且呈现剂量依赖性。表明朝鲜族大酱异黄酮对乳腺癌细胞的生长能够起到抑制作用。     

北虫草不同极性部位体外抗肿瘤活性研究

目的研究山东北虫草不同极性部位体外抗肿瘤活性。方法采用系统溶剂法,分别用石油醚、乙酸乙酯、正丁醇和蒸馏水依次提取北虫草不同极性部位。以乳腺癌MCF-7细胞、食管癌ECA-109细胞和肺癌A549细胞为研究对象,用MTT法检测北虫草不同极性部位对MCF-7细胞、ECA-109细胞和A549细胞的抑制作用。结果石油醚部位在100μg/ml浓度时,对MCF-7细胞、ECA-109细胞和A549细胞的抑制率较强,乙酸乙酯部位在50,100μg/ml浓度对ECA-109细胞的抑制率较强。结论北虫草石油醚部位和乙酸乙酯部位体外抗肿瘤活性较强,值得进一步研究其化学成分。     

Delphinidin抗乳腺癌EMT体外效应研究

研究飞燕草素(Delphinidin,Dp)抑制HER-2~+乳腺癌细胞(MDA-MB-453)和HER-2~-乳腺癌细胞(MCF-7)上皮细胞-间充质转化(EMT)效应。将不同浓度Dp分别处理MDA-MB-453和MCF-7细胞48 h,于倒置显微镜观察细胞形态及数量变化后,用蛋白质免疫印迹法和逆转录聚合酶链反应检测EMT标志性蛋白E-cadherin、N-cadherin、Vimentin、Fibronectin及关键调控因子锌指转录因子(Snail 1)表达。80μmol/L和100μmol/L的Dp能使MDA-MB-453和MCF-7乳腺癌细胞的E-cadherin标志蛋白表达上调,而降低与细胞迁移和侵袭能力有关的N-cadherin、Vimentin、Fibronectin及Snail因子表达水平。Dp体外具有抗MDA-MB-453和MCF-7乳腺癌EMT的效应,研究结果可为植物黄酮类化合物抗乳腺癌转移及辅助治疗提供新支撑。     

大高良姜的化学成分及对人乳腺癌细胞MCF-7的细胞毒作用研究

目的研究大高良姜的化学成分及所得部分苯丙素类化合物对人乳腺癌细胞MCF-7的细胞毒活性。方法用柱色谱技术分离、纯化,通过理化常数和波谱分析鉴定化合物结构;细胞培养MTT染色法测定化合物对肿瘤细胞的毒性。结果分离并鉴定出10个化合物,分别为:二乙酸-反式-对香豆醇酯(Ⅰ),对乙酰氧基苯内烯醇(Ⅱ),对乙酰氧基苯丙烯醚(Ⅲ),1-乙酰氧基胡椒酚乙酸酯(Ⅳ),反式对羟基桂皮醛(Ⅴ),反式对羟基桂皮酸(Ⅵ),反式-对羟基桂皮醛乙酯(Ⅶ),二对乙酰氧基苯乙烯甲烷(Ⅷ),对羟基苯甲醛(Ⅸ),2,5-二羟基苯甲酸(Ⅹ);化合物Ⅰ对人乳腺癌细胞MCF-7生长抑制作用较强,化合物Ⅱ～Ⅵ无明显细胞毒作用。结论化合物Ⅱ,Ⅹ系首次从该植物中分离到;化合物Ⅰ对人乳腺癌细胞MCF-7有一定的细胞毒作用。     

酸浆五种部位醇提物的活性比较研究

目的:探讨酸浆根、茎、叶、宿萼和浆果醇提物的抑菌、抗氧化、抗肿瘤和抑炎活性。方法:采用牛津杯法检测酸浆五种部位醇提物对金黄色葡萄球菌、大肠杆菌和枯草芽孢杆菌的抑菌活性;DPPH清除法检测其抗氧化活性;Alamar Blue法检测其对人乳腺癌细胞株MCF-7、MDA-MB-231和肝癌细胞株Bel7402的抑制效果;一氧化氮检测试剂盒测定其抗炎活性。结果:浆果对金黄色葡萄球菌和大肠杆菌的抑菌效果最好,叶对枯草芽胞杆菌抑菌活性较明显,抑菌圈分别为(16.63±0.49)、(14.97±0.37)、(15.83±0.29)mm。宿萼的抗氧化活性最高,浓度为5 mg/m L时对DPPH的清除率为96.79%±2.09%,基本和同剂量的VC效果相当。酸浆各部位醇提物对肿瘤细胞的抑制作用呈现浓度依赖性,其中根对MCF-7的抑制活性较强,宿萼对MDA-MB-231和Bel7402的活性较强,IC50分别为(31.46±1.49)、(48.84±1.69)、(65.48±1.82)μg/m L。叶的抗炎效果最佳,茎抗炎效果较弱,10μg/m L时NO产生量分别为11.01、27.73μmol/L。结论:酸浆具有一定的抑菌、抗氧化、抗肿瘤和抗炎活性,为酸浆的功能开发及活性成分分离提供一定的理论依据。     

三种菊科植物抗癌抗炎组分分离与功能研究

本研究以甜叶菊、蒲公英、野菊花三种菊科植物为材料,采用水、乙醇/乙酸乙酯、甲醇和正己烷4种溶剂提取,硅胶柱层析、TLC分离纯化,以小鼠肝癌细胞Hepa 1c1c7和小鼠巨噬细胞Raw 264.7为模型,通过测定醌还原酶活性和一氧化氮抑制率,筛选具有抗癌和抗炎的活性组分。结果:甜叶菊、蒲公英、野菊花的乙醇/乙酸乙酯提取物经分离纯化后得到最强活性组分,其诱导醌还原酶倍增的浓度分别为0.26-1.59μg/mL、0.45-3.73μg/mL和0.60-0.92μg/mL;甜叶菊、蒲公英组分一氧化氮抑制率达到50%的浓度分别为14.02-19.04μg/mL、48.90-86.05μg/mL。这一研究结果为深入开展菊科植物抗癌抗炎功能成分的分离鉴定及其作用机理打下了前期研究基础。     

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.     

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.     

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.     

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.     

Analysis on Economic Indicators from 62 Enterprises in Printing Machinery Industry in Q1 2014

Operation of printing machine industry was still unsatisfactory in the first quarter of 2014.Analysis on operation of printing machine industry.a.Market demand was not strong;sales of product undulated and declined.According to the statistics,the total industrial output value fell by 19.28% in the first quarter of 2014 than the average quarter value in 2013; industrial added value decreased by 4.16%; sales revenue dropped by 22.83%. h. Business operation of enterprises was in poor condition. c. R＆D of new products is an important transformation guarantee for enterprises. d. To take self explore new ways upgrading advantages,and explore new ways.