ThermoFisher Scientific检测食品中五种黄曲霉毒素的完整解决方案

黄曲霉毒素因具有致癌性及强急性毒性,所以在食品和农产品贸易中为必检项目,尤其经过蒙牛乳制品中黄曲霉毒素M1超标事件后,人们对黄曲霉毒素的关注愈来愈热.黄曲霉毒素是霉菌的二级代谢产物,目前主要关注的黄曲霉毒素有黄曲霉毒素B1、B2、G1、G2、M1,其中黄曲霉毒素B1的毒性和致癌性最强,而黄曲霉毒素M1是黄曲霉毒素B1的代谢物.我国规定,乳品及乳制品中黄曲霉毒素M1限量为0.5μg/kg,粮食中黄曲霉毒素B1为10μg/kg.     

真菌毒素在欧洲国家酿造啤酒中的感染情况

本文对欧洲国家生产的106种啤酒样品中赭曲霉毒素A、单端孢霉烯、伏马毒素和黄曲霉毒素的感染情况进行了调查。采用配有荧光检测器的高效液相色谱仪对赭曲霉毒素A和黄曲霉毒素进行分析．同时采用气质联用技术和液质联用技术分别检测单端孢霉烯和伏马毒素。任一样品中均未检出黄曲霉毒素,但在相当多的样品中检出赭曲霉毒素A、单端孢霉烯及伏马毒素,但其在所有样品中的含量都处于较低水平,欧洲不同国家之间的啤酒存在着显著差异。就赭曲霉毒素A而言,南欧的啤酒样品中含量均低于O．040μg／L,而其他欧洲国家啤酒样品中的含量明显较高．最高达到0．189μg／L（P〈O．001）,意大利啤酒中伏马毒素的含量明显更高（P=O．006）。     

高效液相色谱串联质谱法测定花生及制品中的五种真菌霉素

建立了高效液相色谱-串联质谱(HPLC-MS/MS)测定花生及制品中黄曲霉毒素B1、黄曲霉毒素M1、脱氧雪腐镰刀菌烯醇、赭曲霉毒素A、玉米赤霉烯酮五种真菌霉素的快速分析方法。用甲醇-水(55:45,V/V)对样品进行提取,采用真菌毒素免疫亲和柱萃取,在ESI+模式下采用多反应监测(MRM)模式进行检测。目标物在C18色谱柱上实现了有效分离,在6 min内完成一个样品的分析,相关系数(r2,n=6)大于0.999,检测结果稳定、灵敏。黄曲霉毒素B1、黄曲霉毒素M1、脱氧雪腐镰刀菌烯醇线性范围0.5~50.0μg/L,检出限为0.05μg/kg(LOD,S/N=3),赭曲霉毒素A、玉米赤霉烯酮线性范围5.0~100.0μg/L,检出限为0.5μg/kg(LOD,S/N=3),方法回收率为86.8~102.7%,精密度RSD为0.36~4.79%。该方法快速、灵敏,适用于花生及制品中黄曲霉毒素B1、黄曲霉毒素M1、脱氧雪腐镰刀菌烯醇、赭曲霉毒素A、玉米赤霉烯酮五种真菌霉素的检测与确证。     

免疫亲和层析荧光光度法测定枸杞中黄曲霉毒素

利用免疫亲和层析荧光光度法测定枸杞中的黄曲霉毒素(AFT),样品由甲醇-水提取,提取液经过滤、稀释,用黄曲霉毒素免疫亲和柱净化。以甲醇洗脱,溴溶液显色,荧光光度计测定黄曲霉毒素(B1+B2+G1+G2)含量。枸杞的最低检出限为1μg/kg,5μg/kg的加标回收率为99.6%,10μg/kg的加标回收率为114.0%。该方法准确、快速、安全。     

细菌降解黄曲霉毒素B1的研究进展

黄曲霉毒素B1是目前已发现的黄曲霉毒素中毒性最强的一种,其具有强肝毒性、高致突变性和高致畸性。广泛存在于农产品及饲料食品中,对人类健康存在严重威胁,同时对粮食和畜牧业造成严重的经济损失。细菌降解黄曲霉毒素B1是一种有效、安全和环保的解毒方法,该文通过对黄曲霉毒素B1降解的影响因素、细菌胞外酶和胞内酶等对黄曲霉毒素B1的降解机理以及黄曲霉毒素B1降解菌活性产物的应用研究等方面对细菌降解黄曲霉毒素B1进行了论述,并对细菌降解黄曲霉毒素B1应用前景进行展望,为以后更进一步研究提供较为全面的资料。     

柠檬酸对花生B族黄曲霉毒素脱毒效果的研究

研究分析不同浓度的柠檬酸溶液在不同的液料比和不同作用时间下花生中B族黄曲霉毒素的含量变化。结果表明,随着柠檬酸溶液浓度的提高,液料比的增加,作用时间的延长,花生中B 族黄曲霉毒素的含量降低,脱毒效果越好。用80g/L 的柠檬酸溶液,液料比(V/m)为5:1 对霉变后染有B 族黄曲霉毒素(AFB1 和AFB2)的花生颗粒(98.60μg/kg)浸泡30min 具有很好的脱毒效果,处理后,黄曲霉毒素含量小于20μg/kg,达到国标GB2761 - 2005 对花生中黄曲霉毒素限量的要求。     

柱后衍生-高效液相色谱法检测核桃粉中黄曲霉毒素B1含量的研究

建立了一种免疫亲和层析净化-高效液相色谱法测定核桃粉中黄曲霉毒素B．含量的办法。将核佻粉样品用甲醇-水提取、稀释后直接过免疫亲和枉净化,经甲醇洗脱后用HPLCNIJ定。该方法检测黄曲霉毒素B．的回收率在84．0～92．7％之间,相对标准偏差在1．51～4．38％之间,最低检出限为0．5μg／kg,且检测结果准确可靠,重复性好,可进行核挑粉中黄曲霉毒素B1含量的检测。     

免疫亲和层析净化荧光光度法快速测定酱油及醋中黄曲霉毒素

为检测酱油和醋中黄由霉毒素的含量，建立了免疫亲和层析净化荧光光度法。试样由甲醇一水提取，提取液被过滤、稀释后，上交联着黄曲霉毒素B1、B2、G1、G2特异抗体的免疫亲和层析柱净化。以甲醇通过免疫亲和层析柱洗脱，溴溶液衍生，荧光光度计测定衍生物的黄曲霉毒素(B1 B2 G1 G2)含量。酱油和醋中检出限分别为2．5μgkg和1μg／kg。添加回收率在85％以上。该方法准确、简单、快速、安全。     

免疫亲和柱净化-大体积流通池高效液相色谱法同时检测食品中6种黄曲霉毒素

建立了免疫亲和柱净化-大体积流通池无需衍生同时测定食品中6种黄曲霉毒素(Aflatoxins)的检测方法。样品采用乙腈-水(84:16,V/V)超声辅助提取,用免疫亲和柱净化,经过XBridge TM C18柱(150 mm×4.6 mm,5μm)分离。采用乙腈-甲醇-水(15:25:60,V/V)为流动相,大体积流通池荧光检测器检测,无需衍生,外标法定量。6种黄曲霉毒素在9.5 min内有效分离,从样品前处理到结果分析整个过程小于50 min。黄曲霉毒素B1、B2、G1、G2、M1、M2的检出限(LOD,S/N=3)分别为0.05μg/kg、0.02μg/kg、0.05μg/kg、0.02μg/kg、0.04μg/kg、0.03μg/kg,能够满足我国对食品中黄曲霉毒素限量的要求。6种黄曲霉毒素标准曲线线性良好,线性相关系数r2大于0.999;样品加标回收率为77.6%~90.5%,精密度RSD为2.42%~6.08%。本方法简便、准确、灵敏度高,无需衍生即可同时检测食品中6种黄曲霉毒素,适用于食品中6种黄曲霉毒素的快速定量测定。     

徐州市市售酱油、食用醋及啤酒中黄曲霉毒素的检测

利用酶联免疫法测定徐州市市售酱油、食用醋、啤酒中的黄曲霉毒素B1。从徐州市场所售食品中选择不同品牌的酱油、食用醋、啤酒共59个样品。在59个样品中,58个样品中的黄曲霉毒素B1的含量低于国家标准中黄曲霉毒素B1允许量（≤5μg/kg）,1个样品中的黄曲霉毒素B1的含量超标,该样品中黄曲霉毒素B1的含量为8.530μg/kg,总合格率为98.30%。其中20个酱油样品的合格率100%,18个啤酒样品的合格率100%,21个食用醋样品的合格率为95.24%。     

Efficiency of different extraction solvent mixtures used in analyses of aflatoxins from a certified peanut meal reference material

Four analytical methods with different extraction solvent mixtures were used to compare the efficiency of the extraction step in the analysis of aflatoxins from a reference material with a certified value of 206 ± 13 µg aflatoxin B₁ kg^-1. The extraction mixtures used were chloroform-water (100 + 10), acetonitrile-water (60 + 40), acetone-water (85 + 15) and methanol-water (80 + 20). The analytical values determined from the extraction mixtures after correction for recovery were 211, 204, 163 and 120 µg kg^-1, respectively. The small amounts of aflatoxin B₂ in the reference material followed the same pattern. The selection of extraction solvent mixture is very important for achieving the true value and that correction for recovery does not always fully compensate for getting the correct analytical value.     

Pilot survey of aflatoxin–albumin adducts in sera from Egypt

Aflatoxins are potent liver carcinogens that frequently contaminate cereals in developing countries. Aflatoxin exposure has been predicted in Egypt but, to date, no studies have measured the level of aflatoxin–albumin (AF–alb) adducts as a validated biomarker to assess exposure. In this pilot survey, a limited number of sera samples, available from a hepatocellular carcinoma (HCC) case-control study in Egypt, were analysed. AF–alb was detected in 24/24 samples from HCC-negative individuals (geometric mean 9.0 pg mg^?1; range 3.5–25.8pg mg^?1), while 7/22 samples from HCC-positive cases had detectable AF–alb (geometric mean 2.6 pg mg^?1; range: non-detectable–32.8 pg mg^?1). These AF–alb data do not represent a case-control comparison due to inherent difficulties in comparing markers of dietary intake between controls and patients with disease. Although these data are limited, the potential health consequences of aflatoxin exposure in this region merit further investigation.     

超高效液相色谱-串联质谱法测定食品中黄曲霉毒素B1和M1的方法研究

应用超高效液相色谱-串联质谱联用技术（UPLC-MS/MS）,在多反应离子检测（MRM）方式下对食品中的黄曲霉毒素B1（AFB1）和M1（AFM1）污染量进行检测.样品经乙腈-水混合试剂提取,中性氧化铝柱净化,经Agilem ZORBAX Eclipse plus C18色谱柱（100 mm×4.6 mm,1.8 μm）,以甲醇和10 mmol/L NH4Ac溶液（含0.1％的甲酸）为流动相洗脱分离,以MRM方式进行定量分析.AFB1和AFM1分别在0.12～6.12 μg/L和0.11～2.28 μg/L质量浓度范围内线性关系良好,相对标准偏差（RSD）＜5％,加标回收率为81.3％～97.2％.该方法具有低成本、准确、快速、简便、灵敏度高等优点,可满足我国对AFB1和AFM1检测限要求.     

Aflatoxins and ochratoxin A and their Aspergillus causal species in Tunisian cereals

ABSTRACT

Occurrence of aflatoxins (AFs) AFB1, AFB2, AFG1, AFG2 and ochra toxin A (OTA) was studied in 65 samples of stored and freshly harvested wheat, barley and maize collected in Tunisia. The mycotoxins were simultaneously extracted and quantified by high performance liquid chromatography. Determination of AF-producing (section Flavi) and OTA-producing Aspergillus species (sections Nigri and Circumdati) was conducted in these samples by species-specific polymerase chain reaction (PCR). Results showed that most of maize samples were contaminated with AFs, data after storage showing lower values than those collected at harvest. All contaminated maize samples contained AFG1 and AFG2, among which 27.78% also had AFB1 and AFB2. This AFs pattern was consistent with the A. parasiticus toxin profile. A. flavus however showed the highest frequency in maize but was also found in barley and wheat where no AFs were detected. In contrast, OTA was neither found in maize nor in barley and only one wheat sample contained OTA. A. niger was the only OTA-producing species detected.     

小麦粉污染霉菌的分离鉴定及产黄曲霉毒素 能力的研究

目的对污染小麦粉中所含霉菌进行分离和菌株鉴定,并对所分离菌株的产黄曲霉毒素能力进行评价。方法使用马铃薯-葡萄糖琼脂培养基和麦汁琼脂培养基对小麦粉污染的霉菌进行分离和纯化,根据菌落形态、显微形态观察和ITS序列分析结果对分离菌株进行鉴定,采用PCR技术检测黄曲毒霉合成路径的关键基因来判断菌株的潜在产毒能力,最后用高效液相色谱法确认菌株是否产毒。结果共分离出5株菌株,分别鉴定为链格孢霉(NHF1)、橘灰青霉(NHF2)、黑曲霉(NHF3)和米曲霉(NHF4、NHF5),其中2株米曲霉具有潜在的产黄曲霉素的能力,在一定条件下会产生黄曲霉毒素。结论需要加强小麦粉微生物检测,尤其是霉菌污染的检测、管理和控制,全面制定小麦粉中污染微生物的限量标准,尤其是霉菌的限量值。     

Aflatoxin contamination in food commodities in Bangladesh

During September 2009, we performed a rapid cross-sectional study to investigate the extent of aflatoxin contamination among common Bangladeshi foods. We collected eight common human food commodities (rice, lentils, wheat flour, dates, betelnut, red chili powder, ginger and groundnuts) and poultry feed samples from two large markets in each of three cities in Bangladesh. We quantified aflatoxin levels from pooled subsamples using fluorescence high-performance liquid chromatography. Aflatoxin levels were highest in dates and groundnuts (maximum 623 and 423?ng/g), respectively. Samples of betelnut (mean 30.6?ng/g), lentils (mean 21.2?ng/g) and red chili powder (>20?ng/g) also had elevated levels. The mean aflatoxin level among poultry feed samples was 73.0?ng/g. Aflatoxin levels were above the US maximum regulatory levels of 20?ng/g in five of eight commonly ingested human food commodities tested.     

加热对减少花生中黄曲霉毒素水平的作用

将花生在各种不同的加热温度和加热时间下进行实验,观察黄曲霉毒素在不同温度及不同加热时间下的减少水平。研究结果表明,加热可以减少花生中黄曲霉毒素的水平,并随着加热温度和加热时间的增加,毒素减少量也增加;黄曲霉毒素减少速率与样品最初被污染的水平有关;AFTG1的热稳定性比AFTB1差。     

Aflatoxin Intake from Maize-Based Diets in a Rural Population in Southern India

The aflatoxin intake through the consumption of maize was assessed in 12 households in a rural population in southern India. This was based on the measurement of aflatoxin levels in cooked maize and the quantity of consumption of the maize. The average aflatoxin intakes in nine out of 12 household ranged from 0·33 to 1·5 μg day^-1. Individual intakes varied from 0·08 to 2·22 μg day^-1. When assessed on a body weight basis, aflatoxin intakes in the younger age groups were observed to be higher. The average intakes in different age groups 1–5, 5–12, 12–18 and above 18 years were observed to be 47, 35, 33 and 22 ng kg^-1 body weight day^-1, respectively. Aflatoxin intakes calculated from aflatoxin levels in maize before cooking tended to be higher by 36% when compared with intakes calculated from cooked maize. This significant difference (P<0·001) was attributed to the effect of cooking on aflatoxin levels in maize. © 1997 SCI.     

黄曲霉毒素的检测及其降解方法进展

黄曲霉毒素主要是由真菌寄生曲霉和黄曲霉产生的次生代谢产物,具有极强的毒性、致癌性,在自然界中普遍存在。文中论述了不同物质中黄曲霉毒素的检测方法及应用,同时介绍了黄曲霉毒素的降解方法及成果,并对黄曲霉毒素检测及降解技术的发展趋势进行了展望。     

牛奶及奶粉中黄曲霉毒素M1的快速测定

采用免疫亲和柱－荧光光度法快速测定了牛乳及乳制品中黄曲霉毒素M1。试样经过离心、脱脂、过滤后滤液经过键合有黄曲霉毒素M1特殊抗体的免疫亲和柱净化，此抗体对黄曲霉毒素M1具有专一的识别能力，黄曲霉毒素M1键合在分离柱中的抗体上，用甲醇与水之比为10：90的混合液将免疫亲和柱上杂质除去；以甲醇与水之比为80：20的混合液通过分离柱洗脱；加入溴溶液衍生，以提高测定灵敏度，衍生化后的洗脱液于荧光光度计中测定黄曲霉毒素M1，检测低限为0．1μg/kg；在0．1－1．0μg/kg范围内，回收率为89．6％－96．1％，变异系数为0．52％－5．8％，分析一个样品的时间小于30min，分析过程中不使用黄曲霉毒素M1标准物质，结果表明，本方法具有准确、简单、快速、安全等优点，可以满足少量和批量样品的检测需要。