时间分辨荧光免疫法检测食品中黄曲霉毒素B1的含量

目的建立高灵敏度的时间分辨免疫法检测黄曲霉毒素B_1的含量。方法通过双功能螯合剂异氰酸苄基二乙烯三胺四乙酸络合稀土元素Eu~(3+)标记抗AFB_1的单克隆抗体,以AFB_1-OVA为固相抗原,优化反应条件,建立直接竞争时间分辨免疫检测方法。结果优化条件后,方法灵敏度为0.02μg/L,IC_(50)为0.73μg/L,线性检测范围为0.01~30μg/L,大米、花生、黄豆和干果样品回收率在91%~104%之间,检测结果准确;与结构类似物AFB_2、AFG_1、AFG_2、AFM_1的交叉反应率分别为17.3%、4.49%、2.37%和0.73%。结论本方法灵敏度高、特异性强、检测快速,可以满足实际样品的检测需求。     

2014年黄曲霉毒素在饲料产品中的污染分布规律

针对1 655个检测样品,通过酶联试剂盒初筛和液相色谱法验证两种形式进行饲料产品中黄曲霉毒素B1污染状况调查。黄曲霉毒素B1在雏禽配合饲料和蛋禽配合饲料中超标最严重,超标率分别为11.0%和14.0%。饲料产品中黄曲霉毒素B1污染在不同地区中存在较大差异,华中地区蛋禽配合饲料超标率达到34.1%,而华北和东北地区的蛋禽配合料超标率为0。不同规模的企业饲料产品受黄曲霉毒素B1污染程度同样存在差异,中小企业的超标率明显高于大企业。     

间接竞争ELISA检测试剂盒测定粮油食品中的 黄曲霉毒素B1

目的建立一套适用于食品中黄曲霉毒素B1的快速、简便、准确的检测方法,同时,进一步验证本公司研制的黄曲霉毒素ELISA检测试剂盒的检测效果。方法用酶联免疫法和高效液相色谱法分别对市售的食用油、花生、谷物样品中AFB1的污染程度进行检测分析。结果用ELISA法和HPLC法测定食用油样品的回收率分别为87.1%~92.7%和85.8%~100.8%;花生样品的回收率分别为76.0%~92.8%和76.3%~92.9%;谷物样品的回收率分别为82.6%~92.7%和82.7%~106.4%,花生加标样品6次平行测定的RSD分别为1.29%~2.46%和5.15%~8.70%,11份阳性样品测定结果表明2种方法具有很好的一致性(r2=0.995)。结论 ELISA法和HPLC法均有很好的线性关系,且测定结果相近。本公司研制的黄曲霉毒素ELISA试剂盒可以快速地检测食品中黄曲霉毒素B1,分析周期短,分析效率高。     

Effect of the inclusion of adsorbents on aflatoxin B1 quantification in animal feedstuffs

The extraction efficiency of aflatoxin B₁ (AFB₁) in cattle feed containing nine adsorbents (ADSs) was investigated using two organic/aqueous solvents composed of methanol/water (80/20 v/v; MeOH) and acetone/water (85/15 v/v; AC). Samples were obtained including a highly AFB₁-contaminated (HC) and a low-level AFB₁-contaminated (LC) feedstuff (15.33 and 7.57 µg kg^–1, respectively), nine ADSs (four clay minerals; one yeast cell wall-based product; one activated carbon and three commercial ADS products) at two different levels of inclusion (10 and 20 g kg^?1). After solvent extraction and immunoaffinity column clean-up, all samples were analysed for AFB₁ by high-performance liquid chromatography (HPLC) with fluorescence detection. For each contamination level (HC and LC), the data obtained were analysed using a factorial arrangement in a completely randomized design. Means were compared with the correspondent controls using the Dunnett's test. No statistical difference was found in AFB₁ levels of feedstuffs not containing ADSs when extracted with AC or MeOH, even if numerically higher values were obtained with AC. A dose-dependent effect (p < 0.01) of ADSs inclusion was observed on AFB₁ recoveries that were lower when the higher ADS level (20 g kg^?1) was included in the HC and LC feedstuffs. Higher AFB₁ recoveries were obtained using AC compared with MeOH, both in HC (75.0% versus 12.0%, respectively) and in LC (84.0% versus 22.8%, respectively) ADSs containing feedstuffs. However, when the activated carbon and the sodium bentonite were included in feeds, lower AFB₁ concentrations with respect to control values (p < 0.001 and <0.05, respectively) were obtained also using AC. The data obtained in this study indicate that routine use of the MeOH solvent for AFB₁ analysis of unknown feedstuffs, can produce misleading results if they contain an ADS.     

Estimation of the daily exposure of Koreans to aflatoxin B1 through food consumption

A survey for aflatoxin B₁ (AFB₁) was conducted on 88 food-grade rice samples randomly collected during July and August 2002 in Seoul, Korea. The presence of AFB₁ was determined by enzyme-linked immunosorbent assay, and the positive samples from enzyme-linked immunosorbent assay were confirmed using high-performance liquid chromatography. Besides this, from the surveying data from the literature published since 1997, the intake of AFB₁ from food in Korea was calculated and compared with the provisional maximum tolerable daily intakes. Naturally occurring AFB₁ was found in 5/88 (6%) samples of rice with an average of 4.8 ng g^-1. A calculated probable daily intake of AFB₁ for Koreans fell into the range 1.19-5.79 ng kg^-1 bw day^-1, hence exceeding the estimated provisional maximum tolerable daily intakes. In conclusion, the exposure of Koreans to AFB₁ could bring about health concerns. This is the first report discovering that rice is the major contributor to the dietary intake of AFB₁ in Korea.     

茶叶中黄曲霉毒素B1的检测方法研究

目的针对茶叶中黄曲霉毒素B1的检测,对比胶体金免疫层析法、高效液相色谱、酶联免疫法的差异。方法以红茶、绿茶、花茶为基质,进行方法验证。并选取具有代表性的红茶、绿茶、乌龙茶、及花茶、萃取茶、药用保健茶分别用不同的方法检测。结果改良后的液相法和胶体金免疫层析法准确度和精密度较高,酶联免疫法存在假阳性。随机检测11种茶叶仅有一种检出黄曲霉毒素B1。     

Determination of aflatoxin B1 levels in peanut butter using an immunoaffinity column clean-up procedure: inter-laboratory study

Ten United Kingdom laboratories participated in an evaluation of an immunoaffinity column sample preparation procedure used to prepare aflatoxin B1 containing extracts obtained from peanut butters contaminated by aflatoxins. Each laboratory was sent seven randomly numbered samples of roasted peanut butter which included two sets of undisclosed triplicates. These two peanut butters were naturally contaminated with aflatoxin B1 at levels of about 12 and 35 micrograms/kg. The other sample was a nominal blank peanut butter containing approximately 2 micrograms/kg of aflatoxin B1 which was also employed by participants for recovery experiments. Participating laboratories were instructed to follow a protocol regarding the use of the immunoaffinity columns for extract preparation, but were allowed a free choice of instrumental technique for quantification of aflatoxin levels. Mean recovery for spikes was 72%. Coefficients of variation for the results from the 10 participants for the two contaminated roasted peanut butters were, respectively, 45% (on a mean of 13.6 micrograms/kg) and 36% (on a mean of 37.2 micrograms/kg).     

高效液相色谱法测定粮谷中黄曲霉毒素B1的研究

建立了用三氯甲烷提取、硅胶小柱净化、三氟乙酸衍生,再以荧光检测器来测定粮谷中黄曲霉毒素B1的高效液相色谱方法.试验证明,该方法安全限量标准最低可达0.4 μg/kg,线性在2～50 μg/kg良好,平均回收率较高,具有准确度高、精密度好、安全限量低的特点.     

Determination of aflatoxin B1 in tiger nut-based soft drinks

An analytical method for the determination of aflatoxin B₁ in a tiger nut-based soft drinks named 'horchata' is described. The method is based on an immunoaffinity clean-up, followed by HPLC separation and fluorescence detection after electrochemical post-column derivatization. The detection limit (S/N = 3) and the quantification limit (S/N = 10) were 0.02 and 0.06 µg kg^-1, respectively. The mean recovery at a level of 2 µg l^-1 was 88% (n = 6) and the coefficient of variation was 9%. The method was applied to conduct a small market survey for a beverage named 'horchata' that is frequently consumed by parts of the population in Southern Europe. Twenty-two samples from Spanish and Belgian supermarkets were analysed. As a result, only one sample was found to contain aflatoxin B₁ at the limit of quantitation of the method.     

Determination of aflatoxin M1 level in milk in the production of baby and children's food using immunoassay]

Using commercial immunokits the concentration of aflatoxin M1 was measured in 376 samples of raw milk from farms in the area of a new dairy plant producing milk baby foods. 87.8% of the samples contained no aflatoxin M1 (detection limit 0.025 micrograms/l) and only 2 samples (0.5%) possessed higher concentration than 0.1 microgram/l, which represents the tolerance limit for aflatoxin M1 in baby milk foods admitted in Czechoslovakia.     

QuEChERS-表面增强拉曼光谱法测定花生中黄曲霉毒素B1

目的 建立基于QuEChERS净化和表面增强拉曼光谱法(surface-enhanced Raman spectroscopy,SERS)测定花生中黄曲霉毒素B1(aflatoxin B1,AFB1)的分析方法.方法 以纳米银(nano silver,AgNPs)作为SERS活性基底,结合QuEChERS样品预处理技术...     

Co-contamination of aflatoxin B1 and fumonisin B1 in food and human dietary exposure in three areas of China

Aflatoxins and fumonisins are ubiquitous foodborne toxicants and the co-occurrence of these mycotoxins in human foods represents a significant public health concern, which has been strongly associated with human aflatoxicosis, neural tube defects, as well as many types of primary cancers. In this study the co-contamination of aflatoxin B(1) (AFB(1)) and fumonisin B(1) (FB(1)) in food and human dietary exposure was investigated in residents of three different areas of China. A total of 209 food samples were measured for AFB(1) and FB(1). The median AFB(1) levels were 13.5, 2.3 and 1.3 μg kg(-1) and the median FB(1) levels were 2.6, 0.4 and 0.3 mg kg(-1) in corn samples collected from Huaian (a high-risk area for oesophageal cancer), Fusui (a high-risk area for liver cancer) and Huantai (a low-risk area for both oesophageal and liver cancers), respectively. The median level of AFB(1) in plant oil of Fusui was the highest (52.3 μg kg(-1)) among all food samples analysed. Co-contamination of these two mycotoxins was found in corn, rice and wheat flour. Based on measured food consumption data, the averaged daily dietary intake of AFB(1) was 0.397 μg (range = 0.269-1.218 μg) in residents of Huantai, 1.723 μg (0.224-49.772 μg) in Huaian, and 2.685 μg (1.006-14.534 μg) in Fusui. The averaged FB(1) daily dietary intake was 92.4 μg (range = 55.0-362.1 μg) for residents of Huantai, 460.0 μg (83.2-2894.5 μg) in Huaian, and 138.6 μg (30.0-10,541.6 μg) in Fusui. These data suggest that the co-exposure to AFB(1) and FB(1) in residents of rural China may contribute to the aetiology of human chronic diseases in high-risk areas.     

Co-contamination of aflatoxin B1 and fumonisin B1 in food and human dietary exposure in three areas of China

Aflatoxins and fumonisins are ubiquitous foodborne toxicants and the co-occurrence of these mycotoxins in human foods represents a significant public health concern, which has been strongly associated with human aflatoxicosis, neural tube defects, as well as many types of primary cancers. In this study the co-contamination of aflatoxin B₁ (AFB₁) and fumonisin B₁ (FB₁) in food and human dietary exposure was investigated in residents of three different areas of China. A total of 209 food samples were measured for AFB₁ and FB₁. The median AFB₁ levels were 13.5, 2.3 and 1.3?µg?kg^?1 and the median FB₁ levels were 2.6, 0.4 and 0.3?mg?kg^?1 in corn samples collected from Huaian (a high-risk area for oesophageal cancer), Fusui (a high-risk area for liver cancer) and Huantai (a low-risk area for both oesophageal and liver cancers), respectively. The median level of AFB₁ in plant oil of Fusui was the highest (52.3?µg?kg^?1) among all food samples analysed. Co-contamination of these two mycotoxins was found in corn, rice and wheat flour. Based on measured food consumption data, the averaged daily dietary intake of AFB₁ was 0.397?µg (range?=?0.269–1.218?µg) in residents of Huantai, 1.723?µg (0.224–49.772?µg) in Huaian, and 2.685?µg (1.006–14.534?µg) in Fusui. The averaged FB₁ daily dietary intake was 92.4?µg (range?=?55.0–362.1?µg) for residents of Huantai, 460.0?µg (83.2–2894.5?µg) in Huaian, and 138.6?µg (30.0–10,541.6?µg) in Fusui. These data suggest that the co-exposure to AFB₁ and FB₁ in residents of rural China may contribute to the aetiology of human chronic diseases in high-risk areas.     

几种传统食品中黄曲霉毒素B1的检测与安全评价

采用酶联免疫法检测月饼、花生油、花生糖和豌豆酱等传统食品中的黄曲霉毒素B1（简称AFT B1）含量，分析AFT B1超标的原因及其安全性，结果表明：大部分传统食品中AFT B1在安全限量之内，少数几类产品存在安全隐患。     

低温法检测菜籽油黄曲霉毒素B1的研究

按照GB 5009. 22-2016《食品安全国家标准食品中黄曲霉毒素B族和G族的测定》第二法检测菜籽油中黄曲霉毒素B1时存在杂峰较多、无法准确定性及定量等问题,本法在GB 5009. 22-2016第二法的基础上优化了萃取溶剂,并通过降低萃取溶剂温度来降低菜籽油中杂质溶解度的方法,达到了理想的净化效果。在样品中加入已放入4℃冰箱冷藏30 min的甲醇-乙腈(体积比70∶30),振荡后取上清液氮吹至近干并衍生,衍生液氮吹至近干后利用初始流动相溶解并注入高效液相色谱仪检测。采用Waters XBridge BEH C18色谱柱(4. 6 mm×100 mm,2. 5μm),以蒸馏水和乙腈为流动相进行梯度洗脱,在流速0. 8 m L/min、柱温40℃、进样量10μL条件下,进行定性和定量分析。结果表明:方法线性范围为0. 1~4. 0 ng/m L,定量限为0. 1μg/kg,回收率为78. 0%~90. 4%,相对标准偏差为1. 61%~4. 15%。与国标方法相比,本方法可有效去除菜籽油中杂质,能准确地进行定性和定量,回收率和准确度均较好,可应用于菜籽油中黄曲霉毒素B1的日常检测。     

酶联免疫法检测混合植物油中黄曲霉毒素B1的含量

目的建立酶联免疫法测定混合植物油中黄曲霉毒素B_1含量的分析方法。方法采用直接竞争酶联免疫吸附法对样品进行测定。样品经甲醇溶液提取,提取的抗原及黄曲霉毒素B_1标记物与微孔板上的黄曲霉毒素B_1特异性单克隆抗体竞争结合,洗涤除去未结合杂质。将底物加入孵育,产生蓝色产物,加入终止液终止反应蓝色产物变成黄色产物,450nm处测量吸光度。结果本方法在2h内完成混合植物油中黄曲霉毒素B_1含量的测定。黄曲霉毒素B_1的加标浓度在1.0～20.0μg/kg之间时的回收率为76.5%～120.2%,方法定量限为1.00μg/kg。结论该方法快速、准确、灵敏,适合测定混合植物油中黄曲霉毒素B_1。     

我国大陆地区食品中柄曲霉素和黄曲霉毒素B1的污染调查

为了解我国食品中黄曲霉毒素B1（Aflatoxin B1,AFB1）和柄曲霉素（Sterigmatocystin,STC）污染情况,采集自我国大陆地区1137份的食品样品以及室温储存1-2年的米份样品315份,采用高效液相色谱-串联质谱方法分析其中AFB1和STC的含量。检测结果显示食品中AFB1检出率范围为0%-40%,谷物类和谷物基加工品检出率分别为0.2%和7.8%;坚果检出率为8.7%,其中有两份样品中AFB1含量超标,啤酒中AFB1的检出率为0%。STC检出率为12.9%,平均含量为0.39 μg/kg,谷物类、谷物基加工品、坚果和啤酒中检出率分别为2.6%,28.3%,4.3%和0%。相比于所采集的超市和农贸市场样品,室温储存1-2年的米粉样品中AFB1和STC的阳性率均较高,分别为4.4%和84.8%,平均含量为0.84和2.38 μg/kg。     

棉籽及棉籽相关产品中黄曲霉毒素B1含量的测定及迁移规律分析

建立了棉籽及棉籽相关产品中黄曲霉毒素B_1含量的测定方法。将样品混合均匀粉碎,称样量5 g,提取溶剂为86%乙腈溶液,固液比1∶10,超声提取10 min,提取液经免疫亲和柱层析净化后,利用高效液相色谱法检测其中的黄曲霉毒素B_1,并对黄曲霉毒素B_1的迁移规律进行分析。结果表明,该方法的日内精密度为1. 82%～4. 41%,日间精密度为6. 00%,样品加标回收率为97%～108%,可满足日常实际检测及质量控制要求。在棉籽加工过程中,黄曲霉毒素B_1绝大部分迁移到棉籽蛋白中,棉籽油中几乎没有。这一结果说明生产过程中黄曲霉毒素B_1污染主要来源于棉籽原料且易迁移到棉籽蛋白中。