伏马菌素对食品的污染及毒性

本文主要介绍了伏马菌素对人和动物的危害，以及国内外食品中伏马菌素的污染状况，并介绍了几个国家对伏马菌素毒性的风险评价。     

伏马菌素研究进展

伏马菌素是串珠镰刀菌产生一类酶菌毒素,它能污染玉米及其制品,也可污染小麦、稻米等粮食及其制品;由于伏马菌素分布广泛且毒性较大,因此它在食品安全中意义越来越受到人们关注。该文介绍伏马菌素对食品污染状况、伏马菌素毒性及伏马菌素测定方法。     

伏马菌素B1人工抗原的制备及其鉴定

本试验采用戊二醛一步法将小分子半抗原(伏马菌素B1,FB1)与载体蛋白OVA进行偶联,制备人工抗原FB1-OVA。分别采用聚丙烯酰胺凝胶电泳法(SDS-PAGE)、紫外扫描法(UV)、质谱法(MS)和免疫芯片法来检测FB1与OVA的偶联效果。结果证明,成功得到了人工抗原FB1-OVA,而且通过质谱法检测得出FB1-OVA和OVA的分子量分别是51928.612和44265.718,分析其偶联比为10.6:1。     

食品中伏马菌素的污染及污染控制

伏马菌素（Fumonisin）是20世纪80年代末期发现的一类由串珠镰刀菌产生的真菌毒素.伏马菌素分布广泛且毒性较大,因此它在食品安全中的意义越来越受到人们的重视.本文叙述了产伏马菌素的真菌,综述了中国不同地区、不同食品中伏马菌素污染的情况,以及各种降低伏马菌素污染的方法,综述了关于伏马菌素的立法.     

食品伏马菌素及其检测方法研究进展

伏马菌素主要是由串珠镰刀菌产生的一类真菌毒素,主要污染玉米及其制品,也可污染小麦、大米等粮食作物。伏马菌素的检测方法不断发展,有必要对其进行系统分析和研究。     

伏马菌素产生菌的分离及其产毒检测

伏马菌素(fumonisin)是二十世纪八十年代末期发现的一类由串珠镰刀菌产生的真菌毒素。本文从市售玉米中分离出两株产毒镰刀菌,研究其发酵液中伏马菌素B1的提取、纯化方法,利用薄层层析对伏马菌素B1做半定量检测,利用HPLC法对伏马菌素B1做定量检测。同时,对大连市售玉米花生样品进行了伏马菌素B1的检测。     

伏马菌素检测技术的研究进展

综述了伏马菌素的危害与各种检测伏马菌素的方法与机理,指出液相色谱-质谱(LC-MS)联合检测和酶联免疫检测是主要的研究方向。。     

伏马菌素B2人工抗原的制备研究

为制备伏马菌素B2人工抗原,通过戊二醛法和碳二亚胺法将伏马菌素B2偶联到栽体蛋白上,人工抗原免疫BALB/c小鼠后,酶联免疫吸附法测定抗血清的效价,竞争酶联免疫吸附法测定人工抗原的免疫原性.通过氨基化和羧基化的苏丹红作为内参,紫外可见光谱扫描图谱鉴定伏马菌素B2成功偶联到载体蛋白上,免疫原性鉴定证明,碳二亚胺法制备的人工抗原可刺激BALB/c小鼠产生抗伏马菌素B2的抗体.     

伏马菌素检测方法的研究进展

伏马菌素(Fumonisin)主要是由串珠镰刀菌产生的一类真菌毒素,污染情况遍布世界各地,由于伏马菌素能引起马脑白质软化症和猪肺水肿,并且与人的食管癌发病率有关,因此,引起人们越来越多的重视。综合叙述了测定玉米和玉米制品中检测伏马菌素的方法,如气相色谱法(GC)、高效液相色谱法(HPLC)、薄层层析法(TLC)、液相-质谱联用(LC-MS)、酶联免疫(ELISA)等检测方法。     

玉米及其制品中伏马菌素的去除方法

伏马茵素分布广泛且毒性较大,丢弃受伏马菌素污染的玉米将造成巨大的经济损失,因此降解和去除玉米及其制品中伏马菌素的技术和方法日益受到人们的重视.介绍了伏马菌素的性质以及伏马菌素污染食品的情况,对去除伏马菌素的方法,如物理法、化学法、生物法进行总结和阐述,并对去除和降解伏马菌素的前景进行展望.     

谷物中伏马菌素B1酶联免疫分析法的建立

以匙孔血蓝蛋白(KLH)为载体,采用碳化亚二胺法人工合成伏马菌素B1(FB1)抗原FB1-KLH,免疫大白兔获得特异性良好的抗FB1 多克隆抗体;在多克隆抗体的基础上建立伏马菌素B1 的间接竞争酶联免疫吸附分析方法,该方法IC50 为11.7μg/L,最低检出限1.1μg/L,平均批内和批间变异系数分别为5.8%、10.2%;分析不同溶剂对标准曲线和加标回收率的影响,确定PBS 溶液为样本最佳提取溶剂;在玉米、小麦、大米、高粱谷物样本中添加50～500μg/kg 的FB1 标准品,平均回收率在70.5%～105.6% 之间。将该方法应用于40 份谷物样本中FB1 的检测,检测结果与高效液相色谱的相关系数(R2)为0.9547。该方法简单、灵敏、快速、准确,适用于谷物中FB1 的检测。     

A PCR-Based Assay for the Detection and Differentiation of Potential Fumonisin-Producing Fusarium verticillioides Isolated from Indian Maize Kernels

One hundred and three Fusarium isolates from maize samples collected from different districts of Karnataka state, India, were analyzed with genus-specific, species-specific, and potential fumonisin specific oligonucleotide primers. One set of genus-specific primers ITS F and ITS R based on a highly conserved ITS region of the genus Fusarium were used to differentiate Fusarium species from closely related genera. All the Fusarium species tested scored positive with the ITS pair of primers. Detection and identification of Fusarium verticillioides species was done by using a newly designed reverse primer VERT-R (5′- CGA CTC ACG GCC AGG AAA CC ?3′) based on an intergenic spacer sequence (IGS) combined with an already designed forward primer VERTF-1 (5′-GCG GGA ATT CAA AAG TGG CC -3′) published previously. Out of 103 Fusarium species tested, 83 isolates of F. verticillioides scored positive for VERTF-1/ VERT-R species-specific pair of primers. Further to discriminate potential fumonisin-producing and nonproducing strains of F. verticillioides, the VERTF-1/VERTF-2 set of primers [VERTF-1 (5′-GCG GGA ATT CAA AAG TGG CC -3′) and VERTF-2 (5′-GAG GGC GCG AAA CGG ATC GG -3′)] were used. 64 isolates of F. verticillioides scored positive for VERTF-1/ VERTF-2 pair of primers. In total, three primers, one forward primer VERTF-1 and two reverse primers VERT-R and VERTF-2, were used for the confirmation of F. verticillioides up to the species level and the second pair of primers were used to confirm the potential for fumonisin production. The developed PCR assay should provide a powerful tool for the detection and differentiation of potential fumonisin-producing F. verticillioides strains in a population.     

Fumonisin B1, fumonisin B2, zearalenone and ochratoxin A contamination of maize in Croatia

Mycotoxins are products of moulds that frequently contaminate maize. In this study the presence of mycotoxins fumonisin B₁ (FB₁), fumonisin B₂ (FB₂), zearalenone (ZEA) and ochratoxin A (OTA) was determined in 49 maize grain samples collected in autumn 2002. The most frequent finding was that of FB₁(100%), followed by ZEA (84%) and OTA (39%), while FB₂ was found only in three samples. The co-occurrence of two and three mycotoxins was found in 55 and 37% of samples, respectively. The concentrations (mean ± SD) of FB₁, ZEA and OTA in positive samples were 459.8 ± 310.7, 3.84 ± 6.68 and 1.47 ± 0.38 µg kg^-1, respectively, and the concentrations of FB₂ in three positive samples were 68.4, 109.2 and 3084.0 µg kg^-1. Although such low concentrations of mycotoxins are not a significant source of exposure in countries with a European diet, a few samples with extreme values indicate that thorough control is needed.     

基于氧化石墨烯的荧光适配体传感器检测食品中真菌毒素

目的：利用核酸适配体（aptamer,APT）和氧化石墨烯（graphene oxide,GO）组装一种基于荧光共振能量转移原理的荧光适体传感器,用于两种真菌毒素的同时检测。方法：GO通过π-π堆积作用将荧光标记的黄曲霉毒素B1（aflatoxin B1,AFB1）APT1和伏马毒素B1（fumonisin B1,FB1）APT2吸附在其表面。由于荧光共振能量转移效应,APT上荧光基团的荧光被GO猝灭;当向溶液中加入靶标AFB1和FB1时,APT1和APT2分别与AFB1和FB1结合,从GO中游离出来,恢复荧光。结果：该传感器为AFB1和FB1的荧光检测提供快速、灵敏的方法,AFB1的检出限为0.15?ng/mL,FB1的检出限为0.12?ng/mL。同时对白酒样品进行加标回收实验,回收率分别为92.00%～100.81%（AFB1）和89.00%～99.50%（FB1）。结论：本研究构建的荧光APT传感器用于两种真菌毒素的同时检测具有高灵敏度和特异性,同时该APT传感器同样适用于其他真菌毒素的多重检测。     

分级制备法评估粉碎粒径对玉米中黄曲霉毒素B1测定的影响

评估粉碎粒径对黄曲霉毒素B1（AFB1）测定的影响,改进样品制备方法,提高现行国标检测方法对玉米原粮样品AFB1测定结果的重复性。以天然AFB1阳性玉米为实验材料,采用分级制备方式得到不同粒径范围的四个级别样品,使各级样品的粒径分布差异最大化,通过各级别样品中AFB1测定值及变异系数评估样品制备对AFB1测定值的影响,得到最佳样品制备要求和称样量,进行验证。样品粉碎后的不同粒径样品与该部分的AFB1含量成反比;当称样量为5 g时,应将样品粉碎至全部通过0.6 mm网筛;当称样量为10 g时,应将样品粉碎至全部通过0.9 mm网筛,以使检测结果重复性符合GB/T 32465—2015《化学分析方法验证确认和内部质量控制要求》的要求。研究结果兼顾AFB1测定准确性与操作效率,可扩大应用于其他真菌毒素的检测,对完善食品中真菌毒素检测方法具有参考意义。     

Use of a bio-wipe kit to detect fumonisin B1 in faecal materials

Fusarium toxins with reference to fumonisin B1 (FB1) have long been regarded as contaminants of maize and maize-based related products. However, when consumed they can cause intoxication, especially in humans. Therefore, effective quantitative methods for assessing the dietary exposure of this toxic fungal metabolite are required. The objective of this investigation was to evaluate the effect on the use of a bio-wipe kit, which is a faecal material collection kit, to detect the presence of FB1. Faecal materials were collected from a rural farming community in Gauteng Province, South Africa. In total, 200 samples of faecal material were analysed for Fusarium species using a serial dilution method, while FB1 was further analysed and quantified by reversed-phase TLC and HPLC. The study showed the presence of 11 different Fusarium species grown on potato dextrose agar culture medium of which F. verticillioides and F. proliferatum, producers of FB1, and F. oxysporum were the dominant species. Fumonisin B1 was recorded at an incidence rate of 65% of the total using TLC. Results from HPLC showed that 84% were positive at different ranges of concentration for FB1. This study supports the use of a bio-wipe as a rapid method to determine human exposure to FB1.     

干燥方式、储存温度和品种对玉米中伏马毒素B1的影响

伏马毒素B_1广泛存在于玉米中,是玉米及其制品质量安全的重要危害因子。选取23个玉米品种为研究对象,对不同干燥方式(烘干、晒干和晾干)和不同储藏温度(4、15、25℃、常温)对其中伏马毒素B_1的影响进行研究,结果表明:玉米进行烘干、晒干和晾干后放置3个月后,伏马毒素B_1含量为烘干晒干晾干,玉米水分含量与伏马毒素B_1含量显著相关(r=0.678);不同温度中放置3个月后,伏马毒素B_1含量由小到大为4℃15℃25℃室温;烘干、4℃储存玉米中伏马毒素B_1含量分别为0～1.39 mg/kg(平均值0.40 mg/kg)、0.08～1.60 mg/kg(平均值0.78 mg/kg),均低于美国FDA规定的最高限量(2 mg/kg);品种对玉米中伏马毒素B_1含量影响显著。烘干、4℃储存、选取高抗镰刀菌玉米品种可有效控制在储藏过程中伏马毒素B_1对玉米的污染。     

Accumulation of Fumonisins in Maize Hybrids Inoculated under Field Conditions with Fusarium moniliforme Sheldon

The accumulation of fumonisin B₁ (FB₁) and fumonisin B₂ (FB₂) after inoculation of 14 maize hybrids with Fusarium moniliforme has been examined in Poland during the seasons 1992–1995. Fumonisins have been detected in all 14 hybrids in amounts ranging from 0·09 mg kg^-1 FB₁+0·02 mg kg^-1 FB₂ to 93·2 mg kg^-1 FB₁+15·8 mg kg^-1 FB₂. Inoculated ears contained from <1% up to 60% of Fusarium damaged (=mouldy) kernels (FDK). Fumonisins' levels in FDK ranged from 5·1 mg kg^-1 FB₁+1·4 mg kg^-1 FB₂ up to 196·0 mg kg^-1 FB₁+62·0 mg kg^-1 FB₂. Significant influence of the year of inoculation on ear infection and fumonisins accumulation was observed. Only one hybrid (Mona) exhibited low disease score, low percentage of FDK (1·1–2·6%) and low levels of fumonisins (FB₁ from 0·09 to 1·66 mg kg^-1 and FB₂ from 0·02 to 0·42 mg kg^-1) during 3 years of experiments. © 1997 SCI.     

玉米中黄曲霉毒素B1测定的不确定度评定

采用超高效液相色谱法对玉米中黄曲霉毒素B1的不确定度进行评估。根据CNAS-GL006-2018《化学分析中不确定度的评估指南》和JJF1059.1-2012《测量不确定度评定与表示》中的有关规定,建立测定玉米中黄曲霉毒素B1不确定度的数学模型,逐层对不确定度来源进行分析。通过对不确定度分量进行量化和合成,得出当玉米中黄曲霉毒素B1为3.60μg/kg时,其相对扩展不确定度为0.14μg/kg(k=2)。结果表明,影响检测结果的主要因素为样品非均匀性、回收率、校准过程及样品处理液定容;其量化结果分别为0.0140、0.009、0.007、0.0058。     

环介导等温扩增法检测转基因玉米MON89034

根据MON89034外源插入片段与植物基因组序列设计特异性引物,筛选最佳引物并对反应体系和反应条件进行优化,最终建立转基因玉米MON89034转化体特异性LAMP检测方法。对该方法进行了特异性、灵敏度、稳定性和重复性测试。结果表明:该方法能够特异性检测出MON89034玉米;检测其灵敏度达到1 pg;以转基因玉米MON89034 DNA标准品质量分数为1.00%,0.10%,0.05%的样品为模板,其稳定性好、重复性高,假阴性率为0。本试验设计的LAMP方法适用于特异性检测转基因玉米MON89034。