免疫亲和柱结合超高效液相色谱-串联质谱法测定牛奶中的16种真菌毒素比较研究

目的 通过比较5种不同商品化多合一免疫亲和柱的可检测目标毒素种类、回收率和稳定性,筛选性能最优的免疫亲和柱,建立免疫亲和前处理-超高效液相色谱-串联质谱法(ultraperformanceliquid chromatography-tandem mass spectrometry, UPLC-MS/MS)测定牛奶中16种真菌毒的方法。方法 牛奶样品分别经5种不同多毒素免疫亲和柱进行净化富集,用优化后的UPLC-MS/MS,在多反应监测模式下测定,同位素内标法定量,筛选覆盖牛奶中真菌毒素污染种类全面、回收率和稳定性最佳的免疫亲和柱,并进行免疫亲和柱性能评价和方法学验证,最终将方法应用于实际样品检测。结果 免疫亲和柱A对牛奶中16种真菌毒素在低、中、高3个添加水平均具有良好的准确度和精密度,加标回收率在83.6%～126.8%之间,相对标准偏差为0.2%～18.4%。柱A内各毒素残留水平低于仪器检出限,柱容量在21.8～1317.5 ng之间。使用免疫亲和柱A富集净化样品,各目标毒素在线性范围内线性良好,相关系数均大于0.99,定量限为0.0010～0.2000ng/g。应用该方法对牛奶质...     

Mycotoxin Biomarkers of Exposure: A Comprehensive Review

To date, the use of biomarkers has become generally accepted. Biomarker‐driven research has been proposed as a successful method to assess the exposure to xenobiotics by using concentrations of the parent compounds and/or metabolites in biological matrices such as urine or blood. However, the identification and validation of biomarkers of exposure remain a challenge. Recent advances in high‐resolution mass spectrometry along with new analytical (post‐acquisition data‐mining) techniques will improve the quality and output of the biomarker identification process. Chronic or even acute exposure to mycotoxins remains a daily fact, and therefore it is crucial that the mycotoxins’ metabolism is unravelled so more knowledge on biomarkers in humans and animals is acquired. This review aims to provide the scientific community with a comprehensive overview of reported in vitro and in vivo mycotoxin metabolism studies in relation to biomarkers of exposure for deoxynivalenol, nivalenol, fusarenon‐X, T‐2 toxin, diacetoxyscirpenol, ochratoxin A, citrinin, fumonisins, zearalenone, aflatoxins, and sterigmatocystin.     

Changes in ochratoxin A and type B trichothecenes contained in wheat flour during dough fermentation and bread-baking

Ochratoxin A (OTA) and type B trichothecenes are mycotoxins that occur frequently in cereals and thus can be found in cereal by-products such as bread. The aim of this work was to study the variation of the levels of OTA, deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON) and nivalenol (NIV) during the bread-making process. This was done by using wheat flour spiked with different levels of toxins. Mycotoxin levels were controlled after fermentation of the dough with yeasts (Saccharomyces cerevisiae) and after further baking at different temperature–time combinations. Analysis of variance (ANOVA) of the results showed a significant reduction in OTA level (p < 0.05) during fermentation of the dough. The reduction ranged between 29.8% and 33.5%, depending on the initial concentration of toxin in the flour. During this period, the level of the other mycotoxins studied was not modified. By contrast, in the baking phase there were significant changes in the levels of the four mycotoxins, although the reduction was similar under all the baking conditions. Considering all the temperature–time conditions tested, it can be concluded that during the baking period the average reduction of OTA, NIV, 3-ADON, and DON was 32.9%, 76.9%, 65.6%, and 47.9%, respectively.     

粮食类饲料资源真菌毒素防制技术探讨

真菌毒素是一类有毒次级代谢物,对人畜健康有严重的影响,粮食饲料中一旦发生真菌毒素的污染,将对畜牧产品造成极大的经济损失.本文介绍了粮食饲料中真菌毒素的危害,并给出了可能的预防和控制措施.在真菌毒素控制方法中,根据它们降解毒素的不同原理,分成物理、化学、生物方法分别做了介绍.这些预防措施已经在一些领域使用.     

Deoxynivalenol in wheat,maize, wheat flour and pasta: surveys in Hungary in 2008–2015

Among Fusarium mycotoxins, deoxynivalenol (DON) is the most common contaminant in case of cereals and cereal-based foods in Hungary. In this study, Hungarian wheat (n = 305), maize (n = 108), wheat flour (n = 179) and pasta (n = 226) samples were analysed (N = 818). The samples were collected during 2008–2015 in Hungary. Applied methods of analysis were enzyme-linked immunosorbent assay and liquid-chromatography coupled with a mass spectrometer. Results were compared and evaluated with Hungarian weather data. Among cereal samples, in 2011, wheat was contaminated with DON (overall average ± standard deviation was 2159 ± 2818 µg kg^?1), which was above the maximum limit (ML). In case of wheat flour and pasta, no average values above ML were found during 2008–2015, but higher DON contamination could be observed in 2011 as well (wheat flour: 537 ± 573 µg kg^?1; pasta: 511 ± 175 µg kg^?1).     

Aflatoxin M1 and ochratoxin A in raw bulk milk from French dairy herds

Mycotoxins in milk are a public health concern and have to be regularly monitored. A survey on the presence of aflatoxin M₁ (AFM₁) and ochratoxin A (OTA) in raw bulk milk was conducted in 2003 in the northwest of France, the main French milk-producing basin. Randomly selected farms (n = 132) were characterized by a diet based on corn silage and containing a large proportion of on-farm produced cereals, feeding sources that are frequently contaminated by mycotoxins. Farms were surveyed twice in winter and in summer. At each sampling time, a trained surveyor completed a questionnaire recording farm management procedures and production traits. The AFM₁ was found in 3 out of 264 samples but at levels (26 ng/L or less) that are below the European legislation limit of 50 ng/L. Traces of AFM₁ (less than 8 ng/L) were also found in 6 other samples. The OTA was detected in 3 samples also at low levels, 5 to 8 ng/L. Farms that tested positive to the presence of mycotoxins, 12 in total including 6 farms that had traces of AFM₁, differed from negative farms by a more extensive use of total mixed rations, 58 vs. 27%. In addition, the positive farms tended to have lower milk yields. Although the incidence of milk contamination with AFM₁ and OTA at the farm level was low during the period studied, production and management data from the surveyed farms suggest a link between feeding management practices and mycotoxin contamination.     

超高效液相色谱-串联质谱法检测小麦中19种真菌毒素

目的 建立超高效液相色谱-串联质谱法用于检测小麦中19种真菌毒素。方法 样品经乙腈?水?乙酸(70:29:1, V:V:V)振荡提取并低温高速离心后,直接用水稀释提取液并再次低温高速离心,上清液过膜后经C18柱分离,最后用三重四极杆质谱在多反应监测模式下外标法定量测定。结果 在最佳分析条件下,各真菌毒素靶标均获得良好线性范围,相关系数(r)均>0.999;检出限和定量限分别为0.001~5 μg/kg和0.003~15 μg/kg;在3个不同浓度下的加标回收率位于77.08%~119.77%之间,相对标准偏差为0.59%~9.99% (n=6)。结论 该方法具有操作简单、稳定可靠、检测通量高、检测成本低的优点,有望用于大批量小麦产品中真菌毒素质量安全风险评估筛查及验证分析,为政府部门粮食质量安全监管提供技术支撑。     

霉菌毒素检测与脱毒技术研究进展

霉菌毒素是某些霉菌在谷物或饲料上生长繁殖过程中产生的有毒代谢物,威胁动物乃至人类的健康。有关毒素的脱毒技术一直是国内外的一个研究热点,其中生物法脱毒是现在最有前景的一种脱毒方法,主要通过微生物的吸附和降解作用来达到脱毒的效果。本文结合最新的研究成果,详细介绍了霉菌毒素的毒性、污染现状及主要的检测方法,并对霉菌毒素物理、化学、生物脱毒技术进行了概述,并对未来的研究方向加以展望。     

畜产品中霉菌毒素残留危害、防控及检测技术

霉菌毒素广泛存在于饲料中,我国饲料中霉菌毒素检出率高达90%,其中对畜牧业危害最大的霉菌毒素主要有6种。畜禽食用了这样的饲料,霉菌毒素会引起畜禽的慢性中毒、疾病甚至死亡,饲料霉变产生的霉菌毒素被动物采食后,可能被机体的肠肝循环中被反复重吸收。因此,霉菌毒素会在动物机体的各个组织、内脏、鸡蛋、牛奶中残留形成残留。人食用了这样的畜产品,会对人类的健康产生很大的潜在威胁,甚至导致畸、致突变、致癌、肝中毒、肾中毒等危害。本研究简要介绍了目前霉菌毒素的防控措施、脱毒方法、检测技术等方法的研究进展,以期对霉菌毒素的防治起到一定的参考作用。