基于衍生化反应的母乳低聚糖质谱检测方法的建立

对比2-氨基苯甲酸、2-氨基苯甲酰胺和2-氨基吖啶酮这3种衍生剂,选择2-氨基苯甲酸作为最优衍生剂,基于此建立一种超高效液相色谱-串联质谱法定量检测母乳中6种母乳低聚糖(HMOs)含量的方法。样品经除脂、除蛋白、衍生后,采用ACQUITY UPLC BEH Amide色谱柱分离,多反应离子监测(MRM)模式定量测定HMOs含量。方法学验证结果表明,各HMO在设定的浓度范围线性关系良好,相关系数大于0.99。在高、中、低3个水平加标试验中,回收率在83.2%～119.9%之间,日内精密度在1.0%～7.9%之间,日间精密度在4.5%～7.4%之间。2’-岩藻糖基乳糖和3’-岩藻糖基乳糖的定量限为1.5 mg/L,3’-唾液酸乳糖、6’-唾液酸乳糖和乳-N-岩藻五糖-I的定量限为5.0 mg/L,乳-N-岩藻五糖-V的定量限为4.5 mg/L,有较好的灵敏度。方法的精密度与准确度良好,可满足母乳中6种HMO的准确定量需求。     

2′-岩藻糖基乳糖的生理功效以及制备方法研究进展

2′-岩藻糖基乳糖是母乳中含量最高的寡糖,其对婴幼儿的健康发育起到显著促进作用,近年来针对2′-岩藻糖基乳糖的学术研究进展迅速,其生物制备技术已步入产业化实施阶段,未来将成为婴幼儿配方奶粉产业的重要风口。该文就近年来针对2′-岩藻糖基乳糖的生理功效以及制备方法的研究进行综述,总结了最新的研究方向和代表性成果,并对2′-岩藻糖基乳糖生物制备技术的未来发展方向以及进一步提高产量的策略进行了展望。     

基于HPLC-MS/MS的羊乳寡糖GP衍生化条件优化

高效液相色谱-质谱联用仪（HPLC-MS/MS）广泛应用于寡糖分析，但寡糖的复杂结构使寡糖的数据分析准确性存在诸多变数，衍生化效果的好坏对检测灵敏度与准确性至关重要。通过衍生化可以改变寡糖的疏水性或电荷性质，为提高质谱分析中多糖的电离效率和分析灵敏度提供了一种可行的方法。采用吉拉德试剂P(Girard’s Reagent P,GP)对母乳和羊乳寡糖的衍生化效率进行研究，结果表明随着GP溶液质量浓度和体积的增加，寡糖原型显著减少，50 mg 2’-岩藻糖基乳糖（2’-FL）中加入0.1 mol/L GP溶液5 mL时，GP衍生化效率达到99%以上，羊乳寡糖衍生化后6种寡糖检测结果与2’-FL平行一致，表明该种衍生化方法效果良好，此法可为乳寡糖的精准检测分析提供技术参考。     

中国母乳中低聚糖组分及含量变化——以中国广东江门地区为例

密集收取22位中国母亲共163个母乳样品,采用高效阴离子色谱法监测在不同泌乳期中22种母乳低聚糖(human milk oligosaccharides,HMOs)的组成和含量变化。结果表明,15种中性低聚糖含量在泌乳初期(初乳)达到最高值后逐渐减少(成熟乳),但是3-岩藻糖乳糖的含量在整个取样阶段均在持续增加。6种酸性低聚糖在泌乳初期稳定但后期随时间延长逐渐减少,其中唾液酸化-乳糖-N-四糖a只在部分样本中检出。基于Lewis血型类型,可将母亲分为分泌型组和非分泌型组。分泌型母亲的母乳中富含α-1-2-L-岩藻糖基化低聚糖(如2’-岩藻糖乳糖、乳酰-N-岩藻五糖Ⅰ、二岩藻糖基乳糖、乳酰-N-二岩藻六糖Ⅰ)。在分泌型与非分泌型母乳中,α-1-2-L-岩藻糖基化低聚糖的质量浓度范围分别为1 211~7 272 mg/L和100~920 mg/L;然而α-1-3/4-L-岩藻糖基化低聚糖(如乳酰-N-二岩藻六糖Ⅱ、乳糖-N-二岩藻糖基-八糖、3-岩藻糖乳糖、乳酰-N-岩藻五糖Ⅱ)的质量浓度范围则分别为181~2 722 mg/L和476~4 931 mg/L。总体表明,分泌型和非分泌型母亲在泌乳期内的HMOs的组成和含量均在不断变化且差异明显。     

离子色谱法测定母乳中的寡聚糖与游离唾液酸

母乳寡聚糖(HMOs)是母乳中第三大营养素,其含量仅次于乳糖与脂肪,是婴儿健康成长的重要营养来源。本研究采用离子色谱法(IC)建立并验证了测定母乳中6种主要母乳寡聚糖含量的方法,包括3-岩藻糖基乳糖(3-FL)、2’-岩藻糖基乳糖(2’-FL)、乳糖-N-新四糖(LNnT)、乳糖-N-四糖(LNT)、6’-唾液酸乳糖(6’-SL)及3'-唾液酸乳糖(3’-SL)。另外,母乳中另一种活性成分唾液酸(Neu5Ac)也可以同时测定。将母乳样用超纯水稀释,微孔滤膜过滤后进样分析。以水-氢氧化钠溶液-乙酸钠溶液为流动相,选用CarboPacTMPA-1分析柱进行色谱分离,用脉冲安培检测器检测。所测7种分析组分在该试验条件下能很好分离,且在较宽的质量浓度范围内与其峰面积呈良好的线性关系(r0.999)。方法的检出限(3 s/n)分别为5.2,0.4,2.8,2.5,1.0,1.8,0.5 mg/L母乳,定量限分别为19.8,9.8,19.9,19.7,9.9,9.9,5.0 mg/L母乳。通过测定分泌型与非分泌型的3段乳得出该方法的线性范围与定量限能够满足各类母乳样品的需求。以母乳作为基质,测得各分析组分的加标回收率(n=6)在91.8%～106.1%之间,母乳基质与加标样测定值的相对标准偏差(n=6)小于3.7%。本方法样品处理简单快速,具有较低的检出限与定量限,较宽的线性范围,良好的重现性与回收率,适用于母乳样品中6种主要寡聚糖与游离唾液酸的准确测定。     

重组大肠杆菌补救途径合成2′-岩藻糖基乳糖的分子克隆和产物合成分析

2′-岩藻糖基乳糖作为人乳寡糖中含量最多的一种，对婴幼儿健康生长发育有重要作用，是极具应用价值的母乳概念配方奶粉添加剂。该研究通过比较不同来源的α-1,2-岩藻糖基转移酶，选定幽门螺杆菌来源的α-1,2-岩藻糖基转移酶基因fucT2与脆弱拟杆菌来源的L-岩藻糖激酶基因fkp共表达，构建了2′-FL的补救合成途径。对乳糖转运蛋白LacY以及2′-FL运出蛋白伯氏耶尔森氏菌来源的糖转运蛋白Tpyb或大肠杆菌来源的糖转运蛋白SetA适量过表达，大幅增加了胞外产物浓度。比较BL21(DE3)、C41(DE3)、JM109(DE3)这3种菌株的2′-FL合成能力，得到了基于JM109(DE3)的2′-FL最终合成菌株J2-V8。通过发酵条件优化，在摇瓶中48 h得到2′-FL的胞外质量浓度为2.67 g/L,较初始菌株提高了5倍，转化率为0.82 mol/mol岩藻糖，以上结果为2′-FL的生物制备技术开发提供了借鉴。     

柱前衍生-高效液相色谱荧光法测定婴幼儿配方乳粉中低聚半乳糖含量

建立了柱前衍生测定婴幼儿配方乳粉中低聚半乳糖的高效液相色谱法。样品经乙酸调节pH到4.5,使用淀粉葡萄糖苷酶和β-半乳糖苷酶酶解、2-氨基苯甲酰胺衍生。经色谱柱分离,荧光检测器检测。通过响应曲面设计优化衍生关键因素水平：衍生剂浓度、衍生温度与衍生时间。定量化合物3"-半乳糖基乳糖在10.0~100 mg/L范围内线性良好(R²为0.9993)。在低聚半乳糖添加水平为2.5、10.0、40.0 g/kg时,回收率在92.5~102.7%之间,RSD(n=6)在3.69~6.69%之间,方法检测限为0.8 g/kg,定量限为2.5 g/kg。本方法操作简单、回收率高和重现性好,可使用3"-半乳糖基乳糖对婴幼儿配方乳粉中的低聚半乳糖进行定量分析。     

2′-岩藻糖基乳糖的酶法合成研究进展和展望

人乳寡糖（human milk oligosaccharide,HMOs）是母乳的关键成分,它对婴幼儿健康成长发挥着重要的作用。2′-岩藻糖基乳糖（2′-fucosyllactose,2′-FL）和3-岩藻糖基乳糖（3-fucosyllactose,3-FL）是HMOs的主要组分,具有抑制病原体感染、调节肠道菌群和提高免疫力等功能,其合成方法成为近年内的研究热点。然而,利用化学法或酶法合成2′-FL和3-FL具有反应过程繁琐、成本高以及产物收率低等缺点,无法满足市场的需求,因此,高效的微生物法已被开发应用于2′-FL和3-FL的商业化生产。本文总结2′-FL和3-FL的制备方法,系统阐述微生物法合成2′-FL和3-FL的工程化策略,为后续2′-FL和3-FL的微生物生产提供一定的研究思路。

     

基于模块优化强化大肠杆菌合成乳糖-N-新四糖的研究

乳糖-N-新四糖(lacto-N-neotetraose,LNnT)作为人乳寡糖核心组分之一,在婴幼儿成长发育过程中发挥着重要作用。为寻找高效的LNnT生物合成方法,探究模块优化对大肠杆菌合成LNnT产量的影响,以大肠杆菌BL21(DE3)ΔlacZ为出发菌株,根据合成路径中的关键代谢物质将LNnT合成路径划分为:外源酶所在路径的模块A,UDP-半乳糖合成路径的模块B和UDP-N-乙酰氨基葡萄糖合成路径的模块C。利用不同拷贝数质粒初步优化模块A、B和C的表达强度,当大肠杆菌BL21(DE3)ΔlacZ共表达重组质粒pRSF-lgtA-A.act和pET-galE时,LNnT产量最高,达0.87g/L。通过CRISPR/Cas9技术敲除setA和ugd强化模块A和模块B,获得的重组菌株E20合成LNnT产量达1.16g/L。摇瓶发酵条件优化后,重组菌株E20合成LNnT产量达1.28g/L。在5L发酵罐中,LNnT分批补料发酵产量达15.53g/L,发酵过程最高生产强度为0.43g/(L·h)。模块优化强化大肠杆菌高效合成LNnT有望为人乳寡糖的高效生物合成提供理论基础,进而推进婴幼儿配方食品产业的革新。     

人乳中低聚糖的含量及其常用分析方法的研究进展

人乳中常见的低聚糖包括3-岩藻糖基乳糖(3FL)(0.07~0.86 g/L)、乳糖-N-四糖(LNT)(0.18~1.18 g/L)、乳糖-N-新四糖(LNn T)(0.10~2.04 g/L)、2'-岩藻糖基乳糖(2'FL)(0.45~4.04 g/L)、乳酰-N-岩藻五糖I~III(LNFP I~III)、乳糖-N-二岩藻糖基-六糖I~II(LNDFH I~II)等,它们在促进婴儿生长发育、增强免疫力、调节肠道菌群方面有重要意义。目前分析人乳中低聚糖常用的方法有高效液相色谱法(HPLC)和高效阴离子交换色谱-脉冲电化学检测法(HPAEC-PED)。本文综述了人乳中主要低聚糖的含量及常用分析方法。     

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.     

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.     

Analysis of benzo[a]pyrene in spiked fatty foods by second derivative synchronous spectrofluorimetry after microwave-assisted treatment of samples

A simple, rapid and inexpensive method has been developed for the determination of benzo[a     

The development of reference materials for paralytic shellfish poisoning toxins in lyophilized mussel. II: Certification study

This paper describes the second part of a project undertaken to develop certified mussel reference materials for paralytic shellfish poisoning toxins. In the first part two interlaboratory studies were undertaken to investigate the performance of the analytical methodology for several PSP toxins, in particular saxitoxin and decarbamoyl-saxitoxin in lyophilized mussels, and to set criteria for the acceptance of results to be applied during the certification exercise. Fifteen laboratories participated in this certification study and were asked to measure saxitoxin and decarbamoyl-saxitoxin in rehydrated lyophilized mussel material and in a saxitoxin-enriched mussel material. The participants were allowed to use a method of their choice but with an extraction procedure to be strictly followed. The study included extra experiments to verify the detection limits for both saxitoxin and decarbamoyl-saxitoxin. Most participants (13 of 15) were able to meet all the criteria set for the certification study. Results for saxitoxin.2HCl yielded a certified mass fraction of <0.07 mg/kg in the rehydrated lyophilized mussels. Results obtained for decarbamoyl-saxitoxin.2HCl yielded a certified mass fraction of 1.59+/-0.20 mg/kg. The results for saxitoxin.2HCl in enriched blank mussel yielded a certified mass fraction of 0.48 +/- 0.06 mg/kg. These certified reference materials for paralytic shellfish poisoning toxins in lyophilized mussel material are the first available for laboratories to test their method for accuracy and performance.     

Announcing a new international peer-reviewed journal:Food Science and Human Wellness now accepting manuscript submission in English

<正>We are pleased to announce the launch of a new international peer-reviewed journal-Food Science and Human Wellness,ISSN 2213-4530,which is an open access journal,produced and hosted by Elsevier B.V.on behalf of Beijing Academy of Food Sciences.Food Science and Human Wellness is an international peer-reviewed English journal that provides a forum for the dissemination of the     

Chinese CTP Market

According to Printing and Printing Equipment Industries Association of China（PEIAC）＇s statistics to the plate manufucturer in China, in 2013, the actual offset plate production has reached 346 million square meters in China. Among them, the CTP production volume was 245 million square meters, up by 11% than that of last year; the total sales of the CTP plate was 239 million square meters, up by 13%.     

Preparatory Work of Print China 2015 is Going Smoothly

正Held at Guangdong Modern International Exhibition Center,Print China 2015 will cover 7exhibition halls,besides the original Hall No.3,4,5,6,7,the newly built F zone of Hall 3 will be used too.The total area will be140,000 square meters.Hall 3:Offset and large printing equipment,package printing equipment,post press