油料和食用油中真菌毒素快速测定方法的研究

本研究对油料（大豆、油菜籽）以及食用植物油中黄曲霉毒素、赭曲霉毒素A和玉米赤霉烯酮的快速检测方法进行了研究,建立了快速检测的筛选方法（免疫亲和柱层析净化荧光光度法）和确证方法（免疫亲和柱层析净化高效液相色谱法）.     

出入境农产品中真菌毒素的污染、检测及控制

<正>真菌毒素(Mycotoxin)是一类由真菌产生的毒性二次代谢产物,广泛污染农作物、食品及饲料等植物源性产品。目前已知的有200多种,按其主要产毒菌种可分为曲霉菌毒素(如黄曲霉毒     

免疫亲和柱的制备及在真菌毒素检测中应用的研究进展

真菌毒素是真菌的次生代谢产物,是引起谷物和饲料污染的重要元凶,其检测手段日趋完善。目前免疫亲和柱在真菌毒素检测分析方面已经被广泛使用,本文综述了免疫亲和柱的原理及其制备方式,及其在几种常见真菌毒素检测方面的应用。      

高效液相色谱法测定粮食中真菌毒素的效果评价

本实验依据国家标准GB/T 5009.111,GB/T 5009.96和GB/T 5009.209中高效液相色谱法分别测定小麦样品中脱氧雪腐镰刀菌烯醇(DON)、赭曲霉毒素A(OTA)以及玉米赤霉烯酮(ZEN)这3种毒素的含量.按照检疫检验相关标准和文件要求,对该方法实践操作后结果的不确定性展开评定.结果表明:本次试验...     

高效液相法同时检测谷物中玉米赤霉烯酮和赭曲霉毒素A

该研究开发一种快速、灵敏同时检测玉米、小麦和稻米中玉米赤霉烯酮(zearalenone,ZEN)和赭曲霉毒素A(ochratoxin A,OTA)的高效液相色谱检测方法。将样品采用乙腈/水(80/20,体积比),200 r/min,30℃振荡提取30 min后,经Oasis PRiME HLB固相萃取柱净化后,上样检测。该方法ZEN和OTA检测限(limit of detection,LOD)为3.7μg/kg和0.11μg/kg,定量限(quantification Limit,LOQ)为12.25μg/kg和0.38μg/kg,线性范围分别为10μg/kg^2000μg/kg和0.2μg/kg^200μg/kg,加标样品中不同浓度的ZEN和OTA回收率为83.0%~101.3%,日内精密度和日间精密度分别为3.12%~7.03%和3.57%~9.3%。该方法适用于玉米、小麦和稻米中ZEN和OTA的同时检测。     

真菌毒素性质及免疫原制备方法的研究进展

真菌毒素是一些真菌在生长过程中所产生的毒性很强的次级代谢产物,通过食品或饲料危害动物和人类的健康及安全。能充分认识真菌毒素性质,准确快速的检测真菌毒素是控制其危害的最有效手段,现阶段酶联免疫(ELISA)检测方法已经成为真菌毒素快速检测的主要趋势,本文对一些主要真菌毒素的性质、种类、危害以及真菌毒素的ELISA 检测方法中重要环节--免疫原制备方法的研究做一综述。     

HPLC测定玉米中玉米赤霉烯酮和赭曲霉毒素A

建立玉米中玉米赤霉烯酮(Zearalenone,ZEN)和赭曲霉毒素A(Ochratoxin A,OTA)的超高效液相色谱检测方法。样品用乙腈/水(90∶10,体积比)振荡提取,过滤后过真菌毒素净化柱,取上清液氮气吹干,用流动相溶解残渣,过0.22μm有机滤膜于进样瓶,采用高效液相色谱检测法(High Performance Liquid Chromatography,HPLC)分析。在ZORBAX SB-C18反相柱上分离后,ZEN以甲醇/水(60∶40,体积比)作为流动相,OTA以乙腈/水/乙酸(43∶56∶1,体积比)为流动相,荧光检测器检测。ZEN的线性范围为0.5μg/mL~6.0μg/mL,相关系数大于0.99;OTA的线性范围为0.05μg/mL~0.8μg/mL,相关系数大于0.99。3个不同水平的加标平均回收率为86.4%~114.1%,相对标准偏差不大于10%。ZEN和OTA的检测限分别为5.0μg/kg和1.0μg/kg。应用该方法对收集的不同玉米样品进行测定,发现ZEN和OTA污染较严重,超标率均为71.4%。该方法具有操作简单、灵敏度高、重现性好等特点,能够用于玉米样品中真菌毒素的测定。     

不同质量玉米中3种真菌毒素含量的研究

采用酶联免疫吸附试验(ELISA)对不同质量玉米中黄曲霉毒素B₁、呕吐毒素和玉米赤霉烯酮3种真菌毒素含量进行了研究。试验结果表明,3种真菌毒素含量与生霉粒含量和杂质含量之间没有明显的相关性;玉米中3种真菌毒素含量并没有呈现同时增加或降低的趋势;新玉米中3种真菌毒素远低于储藏3个月后的陈玉米;东北玉米中3种真菌毒素含量均明显低于山东玉米。     

免疫亲和层析及其在真菌毒素检测中应用

该文简介免疫亲和层析工作原理,载体要求与选择,免疫亲和柱制备与使用,重点综述免疫亲和层析在黄曲霉毒素,赭曲霉毒素,玉米赤霉烯酮,脱氧雪腐镰刀菌烯醇,伏马毒素,T-2毒素等真菌毒素检测中应用最新进展。     

乳酸菌抗真菌活性及其抑制真菌毒素的效果

讨论了乳酸菌抗真菌的活性及其清除真菌毒素的3种可能机制，即①乳酸菌通过对霉菌生长的直接抑制，从而抑制真菌毒素的形成；②通过产生代谢产物抑制真菌菌株或降解真菌毒素到无毒或毒性较低的化合物，以此降低真菌毒素的危害；③借助乳酸菌细胞壁与真菌毒素(黄曲霉毒素B1和玉米赤霉烯酮)的物理结合来清除介质中的毒素危害。乳酸菌的抗真菌活性和清除霉菌毒素的能力显然受到菌株、细胞浓度、细胞处理方式和环境条件的影响。新型发酵乳菌株筛选过程中应该考虑乳酸菌菌株的抗真菌活性及其清除真菌毒素的能力。     

Dietary exposure to mycotoxins of the Hong Kong adult population from a Total Diet Study

Dietary exposure of Hong Kong adults to mycotoxins and their metabolites including aflatoxins (AFs), ochratoxin A (OTA), fumonisins (FNs), deoxynivalenol (DON), acetyldeoxynivalenols (AcDONs) and zearalenone (ZEA) was estimated using the Total Diet Study (TDS) approach to assess the associated health risk to the local people. Sixty commonly consumed food items, collected in four seasons, were sampled and prepared as consumed. These mycotoxins were primarily found at low levels. The highest mean levels (upper bound) were: AFs, 1.50 µg kg^–1 in legumes, nuts and seed; OTA, 0.22 µg kg^–1 in sugars and confectionery; FNs, 9.76 µg kg^–1 in cereals and their products; DON and AcDONs, 33.1 µg kg^–1 in cereals and their products; and ZEA, 53.8 µg kg^–1 in fats and oils. The estimated dietary exposures of Hong Kong adults to the mycotoxins analysed were well below the respective health-based guidance values, where available. For AFs, the upper-bound exposure for high consumers is 0.0049 µg kg bw^–1 day^–1, which was estimated to contribute to about 7.7 (< 1%) of liver cancer cases when compared with 1222 liver cancer cases per year in Hong Kong. The percentage contributions of the estimated 95th percentile dietary exposures (lower and upper bound) to the health-based guidance values of individual mycotoxins were: ochratoxin A, 3.6–9.2%; fumonisins, 0.04–8.5%; deoxynivalenol and acetyldeoxynivalenols, 21.7–28.2%; and zearalenone 3.3–34.5%. The findings indicate that dietary exposures to all the mycotoxins analysed in this study were unlikely to pose an unacceptable health risk to the Hong Kong population.     

小麦粉中溴酸钾含量快速测定方法研究

根据溴酸钾(KBrO3)可在酸性条件下氧化3,3',5,5'-四甲基联苯胺(TMB)而显色的原理,建立了小麦粉中溴酸钾定性检测的目视比色法和定量检测的分光光度法。研究了体系酸度、显色液用量、反应时间对显色的影响,并确立了最优化的实验条件。结果表明:在最优化实验条件下,显色体系的颜色随溴酸钾含量的升高而逐渐加深,溴酸钾浓度在0~4 mg/L范围内符合比尔定律,线性回归方程为ΔA=0.2437C-0.005 2,R2=0.999 3。可用于实际样品的测定,目视比色法可快速确定溴酸钾的含量范围,分光光度法的回收率为96.51%~101.70%,RSD为1.79%~3.12%,能够满足市场监管的需求。     

Validation of the procedure for the simultaneous determination of aflatoxins ochratoxin A and zearalenone in cereals using HPLC-FLD

Method validation for quantitative analysis of aflatoxins (AFs), ochratoxin A (OTA) and zearalenone (ZEA) in cereals using HPLC with fluorescence detector (FLD) is described. Mycotoxins were extracted with methanol?:?water (80?:?20) and purified with a multifunctional AOZ immunoaffinity column before HPLC analysis. The validation of the analytical method was performed to establish the following parameters: specificity, selectivity, linearity, limits of detection (LOD) and quantification (LOQ), accuracy, precision (within- and between-day variability), stability, robustness, measurement of performance, and measurement of uncertainty. Calibration curves were linear (r?>?0.999) over the concentration range, from the LOQ to 26, 40 and 400?ng/g for AFs, OTA and ZEA, respectively. LOD and LOQ were 0.0125 and 0.05?ng/g for aflatoxin B1 (AFB1) and G1 (AFG1), 0.0037 and 0.015?ng/g for aflatoxin B2 (AFB2) and G2 (AFG2), as well as 0.05 and 0.2?ng/g for OTA and 0.5 and 2?ng/g for ZEA, respectively. The mean recovery values were 77–104% for different concentrations of AFs, OTA and ZEA in spiked cereal samples. Both intra- and inter-day accuracy and precision were within acceptable limits. This method was successfully applied for the simultaneous determination of mycotoxins for 60 cereal samples collected from Malaysian markets. Fifty per cent of the cereal samples were contaminated with at least one of these mycotoxins, at a level greater than the LOD. Only one wheat sample and two rice samples were contaminated with levels greater than the European Union regulatory limits for AFs and OTA (4 and 5?ng/g). The means and ranges of mycotoxins obtained for the cereal samples were 0.4?ng/g and 0.01–5.9?ng/g for total AFs; 0.18?ng/g and 0.03–5.3?ng/g for OTA; and 2.8?ng/g and 2.4–73.1?ng/g for ZEA, respectively. The results indicate that the method is suitable for the simultaneous determination of AFs, OTA and ZEA in cereals and is suitable for routine analysis.     

Aflatoxins,ochratoxin A and zearalenone in Brazilian corn cultivars

Past surveys indicated that the occurrence of aflatoxins, zearalenone and ochratoxin A was not a problem in corn and corn products in the state of São Paulo, Brazil. However, according to recent studies, a change in pattern has been detected. To obtain a better overview, these toxins were searched for in 110 samples of freshly harvested corn, corresponding to 48 commercial cultivars planted at three different locations in the state. Aflatoxin contamination was found in 60 (54.5%) of the samples, in levels ranging from 6 to 1600 µg kg^?1 aflatoxin B₁. Insect control was exercised, so this was not the main route of corn infection. Endosperm type, germplasm type, number of days to flowering, and length of time the mature corn remained in the field had no effect on aflatoxin contamination. Ochratoxin A was found in two samples (206 and 128 µg kg^?1) and zearalenone in one sample (4640 µg kg^?1). Possible causes of the increase in aflatoxin levels may lie in the changing nature of the commercial cultivars employed, associated with the forsaking of the original landraces, and in a change in the toxigenicity pattern of the corn mycoflora Aspergillus flavus/Aspergillus parasiticus prevailing strains. © 2001 Society of Chemical Industry     

胶体金免疫层析法快速测定玉米和小麦中玉米赤霉烯酮

采用高效液相色谱法对玉米赤霉烯酮阳性样品进行定值,将阳性样品制备为不同毒素含量的验证样品,对胶体金免疫层析法的准确性、重复性、稳定性、最低检出限、误差范围进行了验证试验,并且完成了对实际样品的检测,同时采用酶联免疫法（ELISA）进行对比,试验结果为：最低检出限为60μg/kg,其检测结果与酶联免疫法基本一致,方法具有...     

Optimization and validation of a HPLC method for simultaneous determination of aflatoxin B1, B2, G1, G2, ochratoxin A and zearalenone using an experimental design

A reversed-phase HPLC optimization strategy is presented for investigating the separation and retention behavior of aflatoxin B₁, B₂, G₁, G_2, ochratoxin A and zearalenone, simultaneously. A fractional factorial design (FFD) was used to screen the significance effect of seven independent variables on chromatographic responses. The independent variables used were: (X1) column oven temperature (20–40°C), (X2) flow rate (0.8–1.2?ml/min), (X3) acid concentration in aqueous phase (0–2%), (X4) organic solvent percentage at the beginning (40–50%), and (X5) at the end (50–60%) of the gradient mobile phase, as well as (X6) ratio of methanol/acetonitrile at the beginning (1–4) and (X7) at the end (0–1) of gradient mobile phase. Responses of chromatographic analysis were resolution of mycotoxin peaks and HPLC run time. A central composite design (CCD) using response surface methodology (RSM) was then carried out for optimization of the most significant factors by multiple regression models for response variables. The proposed optimal method using 40°C oven temperature, 1?ml/min flow rate, 0.1% acetic acid concentration in aqueous phase, 41% organic phase (beginning), 60% organic phase (end), 1.92 ratio of methanol to acetonitrile (beginning) and 0.2 ratio (end) for X1–X7, respectively, showed good prediction ability between the experimental data and predictive values throughout the studied parameter space. Finally, the optimized method was validated by measuring the linearity, sensitivity, accuracy and precision parameters, and has been applied successfully to the analysis of spiked cereal samples.     

免疫亲和柱同时净化-HPLC法测定植物油中黄曲霉毒素和玉米赤霉烯酮

建立了免疫亲和柱同时净化-高效液相色谱法检测植物油中的黄曲霉毒素B1、B2、G1、G2和玉米赤霉烯酮的分析方法。黄曲霉毒素B1、B2、G1、G2的方法的定量限均为0.1μg/kg,玉米赤霉烯酮的方法的定量限为5.0μg/kg,相对标准偏差低于10.4%,回收率为82.0%～95.5%。该方法简便快速、灵敏准确、重现性好,可以用于对植物油中黄曲霉毒素B1、B2、G1、G2和玉米赤霉烯酮进行快速定量检测。