Aflatoxins in sorghum,sorghum malt and traditional opaque beer in southern Malawi

Samples of sorghum grain and malt, traditional opaque sweet beverage (thobwa) and beer prepared from sorghum malts, were collected from the southern region of Malawi during the humid month of January. The samples were analyzed for total aflatoxins using aflatest VICAM fluorometry procedure. All malt and beer samples, 15% and 43% of the sorghum and thobwa samples, respectively, were contaminated with aflatoxins. The sorghum malt prepared for beer brewing, had a significantly (p < 0.01) higher total aflatoxin content (average 408 ± 68 μg/kg [SEM]) than any other type of sample. The average aflatoxin content in the beer was 22.32 μg/l, which is higher than the permissible maximum level in ready to eat foods set by Codex Alimentarius Commission (10 μg/kg). Thus consumption of opaque sorghum-based traditional beer poses a risk of aflatoxin exposure.     

Studies on contamination level of aflatoxins in some cereals and beans of Pakistan

The occurrence of aflatoxins in some cereals and beans was surveyed in this study. One hundred and sixty samples of cereals and beans were purchased from retail shops and local markets of different locations in Pakistan. The samples were analyzed for total aflatoxins to using immune affinity (IAC) clean up with liquid chromatography and fluorescence detection. The results showed the percentage of contamination for total aflatoxins in the samples such as in; rice (25%), broken rice (15%), wheat (20%), maize (40%) barley (20%) and sorghum (30%), while in red kidney beans (20%), split peas (27%), chick pea (10%), cow pea (20%), and soybean (15%). The highest contamination levels of aflatoxins were found in one wheat sample (15.5 μg/kg), one maize sample (13.0 μg/kg) and one barley sample (12.6 μg/kg). Among all the contaminated samples, two rice, one rice broken, two wheat, three maize, two barley, one sorghum, one red kidney beans, one split peas and two soybeans were above the suggested limit for total aflatoxins (4 μg/kg), set by EU regulations.     

Investigations on the occurrence of mycotoxins in beer

The occurrence of several mycotoxins, including ergot alkaloids, alternariol (AOH), deoxynivalenol (DON), and zearalenone (ZEA) in beer (n = 44) from the German market was studied by using enzyme immunoassay methods. Detection limits in beer were 2.1 μg/L (DON), 0.14 μg/L (ZEA), 0.06 μg/L (ergometrine equivalents), and 0.18 μg/L (AOH). DON was detected in 75% of the samples (2.2–20 μg/L, median 3.7 μg/L). All samples were positive for ZEA (0.35–2.0 μg/L, median 0.88 μg/L) and AOH (0.23–1.6 μg/L, median 0.45 μg/L). Most samples (93%) were positive for ergot alkaloids (0.07–0.47 μg/L, median 0.15 μg/L). Correlating toxin levels in beer with European Union tolerable daily intake (TDI) levels for DON (1 μg/kg b.w.), ZEA (0.25 μg/kg b.w.), and ergot alkaloids (0.6 μg/kg b.w.), beer does not represent a major source of intake of these toxins. No TDI data are available for AOH, but considering toxin levels in other foods, beer does not seem to be a major source of intake of this toxin either. Nevertheless, the frequency of their occurrence warrants further study of ergot alkaloids and AOH in raw materials used for beer brewing.     

Seasonal prevalence level of aflatoxin M1 and its estimated daily intake in Pakistan

Aflatoxin M₁ level was measured in 520 milk samples during autumn, winter, spring and summer seasons of 2013–14 in five districts of Southern Punjab-Pakistan. Analyses were performed by using enzyme linked immuno sorbent assay (ELISA) method. Aflatoxin M₁ was found positive in 93% milk samples in the range of 0.001–0.26 μg/l while 53% samples were found to exceed the European Union maximum Limit for aflatoxins M₁ i.e. 0.05 μg/l. The seasonal prevalence level of AFM₁ was found in the order of winter > spring > autumn > summer. Comparing the aflatoxin M₁ level during different day times, morning milk was found 37–50% more contaminated than evening milk. The estimated daily intake (EDI) of aflatoxins M₁ during different seasons of year for various age groups was found in the range of 0.22–5.45 ng/kg/day. Infants were found in the highest risk group while adults were in the lowest. The results of the study indicate that people of Pakistan are at high risk of health issues related with aflatoxins M₁.     

Incidence of zearalenone and fumonisins in Bulgarian cereal production

Ninty-one small-grain cereals (wheat, barley, maize) collected during the 2007 harvest in Bulgaria were tested for zearalenone (ZON) and fumonisins contamination. Analytical methods based on immunoaffinity clean-up and detection by liquid chromatography was used after validation. Limits of detection for ZON in different matrices were below 4.0 μg/kg in barley and wheat, and slightly higher for maize (17.6 μg/kg). The limit of quantification for ZON was 12 μg/kg in barley and wheat, and 58.8 μg/kg in maize. Recovery values ranged between 84% and 105%. The occurrence of ZON in cereals were rather low and only single incidences was found – up to 148 μg/kg for maize and 36.6 μg/kg for other cereals. Fumonisins in maize have showed a widespread distribution (in 94.7% of tested samples). One of the tested samples was contaminated above the established maximum limits for unprocessed maize.     

Natural occurrence of mycotoxins in cereals and spices commercialized in Morocco

Sixty samples of cereals (20 of corn, 20 of barley, and 20 of wheat) and 55 samples of spices (14 of paprika, 12 of ginger, 14 of cumin, and 15 of pepper) purchased from popular markets of Rabat and Salé in Morocco were analyzed for mycotoxins.Cereals samples were all analyzed for ochratoxin A (OTA). The average levels of contamination were 1.08, 0.42, and 0.17 μg/kg for corn, wheat, and barley, respectively. Samples of corn were also analyzed for zearalenone (ZEA) and fumonisin B₁ (FB₁) the average contaminations were 14 and 1930 μg/kg, respectively. Co-occurrence of OTA, FB₁, and ZEA was also checked. Spices samples were analyzed only for aflatoxins (AFs) and the average contaminations found for aflatoxin B₁ (AFB₁) were 0.09, 0.63, 2.88 and 0.03 μg/kg for black pepper, ginger, red paprika and cumin, respectively. The higher level of contamination was found in red paprika (9.68 μg/kg).The present report is the first one ever drafted on the natural co-occurrence of OTA, FB₁ and ZEA in cereals and on the occurrence of AFs in spices from Morocco.     

Fast and sensitive aflatoxin B1 and total aflatoxins ELISAs for analysis of peanuts,maize and feed ingredients

Aflatoxins are a group of carcinogenic compounds produced by Aspergillus fungi that can grow on different agricultural crops. Both acute and chronic exposure to these mycotoxins can cause serious illness. Due to the high occurrence of aflatoxins in crops worldwide fast and cost-effective analytical methods are required for the identification of contaminated agricultural commodities before they are processed into final products and placed on the market. In order to provide new tools for aflatoxin screening two prototype fast ELISA methods: one for the detection of aflatoxin B₁ and the other for total aflatoxins were developed. Seven monoclonal antibodies with unique high sensitivity and at the same time good cross-reactivity profiles were produced. The monoclonal antibodies were characterized and two antibodies showing IC₅₀ of 0.037 ng/mL and 0.031 ng/mL for aflatoxin B₁ were applied in simple and fast direct competitive ELISA tests. The methods were validated for peanut matrix as this crop is one of the most affected by aflatoxin contamination. The detection capabilities of aflatoxin B₁ and total aflatoxins ELISAs were 0.4 μg/kg and 0.3 μg/kg for aflatoxin B₁, respectively, which are one of the lowest reported values. Total aflatoxins ELISA was also validated for the detection of aflatoxins B₂, G₁ and G₂. The application of the developed tests was demonstrated by screening 32 peanut samples collected from the UK retailers. Total aflatoxins ELISA was further applied to analyse naturally contaminated maize porridge and distiller's dried grain with solubles samples and the results were correlated with these obtained by UHPLC-MS/MS method.     

A survey of the incidence and level of aflatoxin contamination in a range of locally and imported processed foods on Malawian retail market

Samples of locally (Malawian) processed and imported maize- and groundnut-based food products (peanut butters, roasted groundnuts, peanut based therapeutic foods, instant baby cereals, maize puffs and de-hulled maize flour) were collected from popular markets of Lilongwe City, Malawi. The samples were analysed in order to determine the frequency and extent of aflatoxin contamination, using immuno-affinity column and reversed-phase liquid chromatography with post-column photochemical derivatization and fluorescence detection. No aflatoxins were detected in all samples of imported baby cereal and locally processed de-hulled maize flour. However, all locally processed maize based baby foods had aflatoxins above EU maximum tolerable level of 0.1 μg/kg. In 75% of locally processed maize puffs, aflatoxins were detected at levels of up to 2 μg/kg. Peanut based therapeutic foods had aflatoxin level between 1.6 and 2.9 μg/kg, exceeding the EU tolerable maximum level (0.1 μg/kg) set for food for health purposes. Locally processed peanut butters had aflatoxins levels in the range of 34.2–115.6 μg/kg, which was significantly higher than their imported counterparts (<0.2–4.3 μg/kg). Samples of locally processed skinned and de-skinned roasted groundnuts had aflatoxins in range of 0.5–2.5 μg/kg and 0.6–36.9 μg/kg, respectively. These results highlight the need for rigorous monitoring of aflatoxins in commercially available processed products in order to reduce likely health risks associated with dietary aflatoxin intake.     

An initial characterization of aflatoxin B1 contamination of maize sold in the principal retail markets of Kigali,Rwanda

Food security considerations have shifted in recent years, with the recognition that available food should also be nutritious and safe. There is a growing evidence base for contamination of maize and other crops by fungal toxins in the tropics and sub-tropics. As an initial snapshot of contamination by one of these toxins in Rwanda, Aflatoxin B1 (AFB1) was analyzed in 684 samples of maize flour collected in seven principal retail markets of Kigali and in 21 samples of animal feed from seven feed vendors. Two rounds of sample collections were carried out, the first in September 2014 and the second in January 2015. A questionnaire given to vendors was used to determine if gender and education level of vendors, origin of maize and awareness of aflatoxins had any significant effect on AFB1 level in collected samples. Enzyme-Linked Immunosorbent Assay (ELISA) and Immuno-affinity fluorimetery were used to analyze samples. Only markets had a significant effect on AFB1 level; for the two collections, differences were inconsistent among markets. In the first round, market means of AFB1 varied between 8.0 ± 5.57 μg/kg and 24.7 ± 23.74 μg/kg and for the second round, between 10.4 ± 8.4 μg/kg and 25.7 ± 25.85 μg/kg. In most animal feed samples AFB1 was >100 μg/kg. None of the vendors interviewed was aware of the risk of mycotoxin contamination in their maize-based flours and feed. Limits set by the United States Food and Drug Administration (20 μg/kg) for total aflatoxins and European Commission (2 μg/kg) for AFB1 for maize flour imports, were varied between 2–35% and 66–100% of samples, respectively. The implications of this study for human and animal health in Rwanda suggest that expanded surveys are needed to understand the scope of contamination, given the influence of environment and other factors on aflatoxin accumulation. Available options to mitigate and monitor aflatoxin contamination can be further deployed to reduce contamination.     

The occurrence of the selected fusarium mycotoxins in Czech malting barley

In 2008–2011, the occurrence of deoxynivalenol (DON), zearalenone (ZON), T-2 toxin (T-2), and HT-2 toxin (HT-2) was studied in 325 malting barley samples collected from various regions of the Czech Republic. The highest occurrence of fusarium mycotoxins was recorded in crop 2009 (2213.5, 59.4 and 145.0 μg/kg for DON, ZON and ∑T-2, HT-2, respectively). Only one measured value exceeded the maximum allowable limit for DON set by the European Union.The validated method of high-performance liquid chromatography coupled with mass spectrometry was used for the analysis of the above mentioned toxins. Limits of detection were 1.5 μg/kg for DON, ZON and HT-2 and 0.3 μg/kg for T-2, the limits of quantification were 5.0 μg/kg for DON, ZON and HT-2 and 1.0 μg/kg for T-2.     

A survey of ethyl carbamate in beer from Chinese market

Result of a survey of ethyl carbamate (EC) in beer was carried out by gas chromatography-mass spectrometry (GC/MS). 217 samples were purchased from supermarkets in five cities of China. The concentrations of EC ranged from n.d. to 19.6 μg/kg with an average level of 2.2 μg/kg, and positive rate was 95.4%. Mean in domestic beer (N = 142, incidence 97.3%) was 1.8 μg/kg, lower than the 2.9 μg/kg in imported beer (N = 75, incidence 94.4%). The result of domestic beer showed that EC level in canned beer (N = 73, mean = 1.5 μg/kg) was lower than that in bottled beer (N = 69, mean = 2.1 μg/kg). Additionally, EC level of beer from different countries was varied. Furthermore, a strong positive correlation of EC content in beer with alcohol content, storage time, temperature and light exposure was observed.     

Co-exposures of aflatoxins with deoxynivalenol and fumonisins from maize based complementary foods in Rombo,Northern Tanzania

Children consuming maize based foods in Tanzania may be exposed to multiple mycotoxins. We estimated co-exposures of aflatoxins with Deoxynivalenol (DON) and fumonisins for children in rural Tanzania. Food consumption by the children was estimated by twice administering a 24 h dietary recall questionnaire to mothers of 18–24 months old children in Kikelelwa village. Each mother also, provided a sample of maize based flour used for feeding her child in the previous day. Each child's body weight (bw) was measured by following standard procedures. Aflatoxins, DON and fumonisins were determined in each sample using validated HPLC methods. Exposures for a mycotoxin were estimated by multiplying flour consumption (g/child/kgbw/day) by its contamination (μg/kg). Complete data were obtained for 41 children. Maize flour consumption ranged from 16 to 254 g/child/day. Thirteen (32%) of the 41 children consumed flour with detectable aflatoxin levels (range, 0.11–386 μg/kg), resulting in exposures from 1 to 786 ng/kg bw/day. All these children exceeded the aflatoxins exposure of concern (0.017 ng/kg bw/day). Eighteen (44%) of the children consumed flour with detectable DON levels (57–825 μg/kg) and 34 (83%), detectable fumonisins levels (63–2284 μg/kg), resulting in respective exposure ranges of 0.38–8.87 μg/kg bw/day and 0.19–26.37 μg/kg bw/day. Twelve (66%) of the DON exposed children and 56% of the fumonisins exposed children exceeded the respective provisional tolerable daily intakes of 1 μg/kg bw and 2 μg/kg bw. Co-exposures for aflatoxins with both DON and fumonsins were determined in 10% of the 41 children. Co-exposures of aflatoxins with fumonisins alone were found in 29% and of fumonisins with DON alone in 41% of the children. The study showed that children consuming maize based complementary foods in Northern Tanzania are at a risk of exposure to multiple mycotoxins. We recommend adoption of appropriate measures to minimize exposures of multiple mycotoxins in Tanzania.     

Simultaneous determination of mycotoxin in commercial coffee

Mycotoxins are secondary metabolites produced by filamentous fungi that usually contaminate food products. Coffee is a natural product susceptible to mycotoxin contamination. The present study evaluates the presence of nivalenol, deoxynivalenol, T-2 and HT-2 Toxin, diacetoxyscirpenol, aflatoxin B₁, aflatoxin B₂, aflatoxin G₁, aflatoxin G₂, fumonisin B₁, fumonisin B₂, ochratoxin A, zearalenone, enniatin A, enniatin A₁, enniatin B, enniatin B₁, and beauvericin in coffee samples, using liquid chromatography tandem mass spectrometry (LC-MS/MS). The results show that zearalenone was not present in any sample. In the positive samples the contents of fumonisins ranged from 58.62 to 537.45 μg/kg, emerging mycotoxins ranged from 0.10 to 3569.92 μg/kg, aflatoxins ranged from 0.25 to 13.12 μg/kg, and trichothecenes, excepting nivalenol, ranged from 5.70 to 325.68 μg/kg. Nivalenol presented the highest concentrations, from 0.40 to 25.86 mg/kg. Ochratoxin A ranged from 1.56 to 32.40 μg/kg, and five samples exceeded the maximum limit established by the European Commission.     

Aflatoxins in dates and dates products

A total of 153 samples of dates and dates products (date cookies, date cake and date halva) from Punjab and Khyber Pakhtunkhwa, Pakistan were evaluated for aflatoxins (AFs) using HPLC equipped with fluorescence detection. Thirty eight out of 96 samples (39.6%) of different date varieties and 18 out of 57 (31.6%) samples of date products contained AFs. The total mean level of AFs ranged between 2.90 to 4.96 μg/kg and 2.76 to 4.80 μg/kg in dates and dates products, respectively. About 16 and 20 samples of dates were found above the permissible level for AFB₁ and total AFs, respectively (i.e. 2 μg/kg, 4 μg/kg). Furthermore, two samples of date's cookies and one sample of date cake were found above the level of AFB₁ and total AFs and three and five samples of date halva were found above the recommended limit for AFB₁ and total AFs, respectively. The high occurrence of AFs may cause health hazards and limit exports.     

Occurrence of aflatoxin contamination in maize throughout the supply chain in the Democratic Republic of Congo

Aflatoxin occurs ubiquitously in maize and is both an economic and a public health concern due to its carcinogenic effects. Nevertheless, in developing countries such as the Democratic Republic of Congo, there is still a dearth of information regarding its natural occurrence, exposure to both humans and animals and strategies for controlling its proliferation or contamination of food crops such as maize. The present study comparatively assessed the aflatoxin occurrence in maize samples collected in the DRC throughout the food supply chain. The analytical method used during this experiment was found to be accurate with a recovery ranging between 70.65 and 98.20%, and also precise (RSD<15%) except for AFG₁, which showed an RSD of 18.05%. Moreover, the limits of detection (LOD) and quantification (LOQ) for aflatoxin ranged from 0.31 to 0.69 μg/kg and from 1 to 2 μg/kg, respectively. The results also revealed that in the pre-harvest period, 32% of maize samples (out of a total of 50) were positive, with aflatoxins at a range of 1.5–51.23 μg/kg for AFB1 and 3.1–103.89 μg/kg for total aflatoxin. As the supply chain progresses, the contamination of maize samples also increases, with 100% of the collected maize samples found to contain aflatoxins at 300 times higher than the maximum limit of 10 μg/kg for total aflatoxin as set by the WHO. The aflatoxin content of maize samples was found to increase drastically at up to 500 times between the city store and the distribution system at market level. This was confirmed with a statistical analysis showing a significant difference (p < 0.01) between city store/market samples and pre-harvest, harvest, farm store and after-transportation samples.     

Aflatoxigenic fungi and aflatoxins occurrence in sultanas and dried figs commercialized in Brazil

The presence of aflatoxins, Aspergillus flavus and Aspergillus parasiticus in dried fruits was investigated. A total of 62 dried fruit samples were analyzed (24 black sultanas, 19 white sultanas and 19 dried figs). A total of 10 A. flavus isolates were found, nine in one white sultana sample (corresponding to 18% infection) and one isolate in dried figs (2%), and all of them were aflatoxin B₁ and B₂ producers. A. parasiticus was not found. Aflatoxins were detected in 3 of 19 (16%) white sultana samples analyzed and, the limits were not higher than 2.0 μg/kg. In dried figs 11 of 19 (58%) samples were contaminated with aflatoxins and, with exception of one sample that was contaminated with 1500 μg/kg of B₁ aflatoxin, the others had less than 2.0 μg/kg. Neither aflatoxigenic or aflatoxins contaminated black sultanas.     

Incidence of aflatoxins in commercial nuts in the holy city of Mekkah

The incidence of aflatoxins contamination in different nuts has been studied. Two hundred and sixty four samples of nuts were collected randomly from the main outlets across seven municipalities of the holy city of Mekkah. The samples were first screened for total aflatoxins using Aflatest immunoaffinity column (IAC) technique. The IAC results showed that 70 samples (26.5%) were contaminated with AFT in levels ranged between 1.0 and 110 μg/kg. The percentages of contamination of AFT in groundnuts were (18.4%), Pistachio (34%), walnuts (50%), cashew (15%), hazelnut (43%) and almond (17%). Only 23 samples (8.7%) had levels equal or exceeded the maximum tolerable limit for AFT in EU (4 μg/kg), and only 7 samples (2.65%) had exceeded the MTL in Saudi Arabia (20 μg/kg). The samples that exceeded MTL of Saudi Arabia were five pistachio sample (110, 45, 43, 34 and 30 μg/kg) and two peanut samples (100 and 28 μg/kg). Quantitative testing of total and individual AFs was conducted in selected samples from the IAC results with contamination levels exceeded the MTL of EU (23 samples) by using HPLC and fluorescence detection. AFB1 was detected in 22 out of 23 samples with contamination level in most of the positive samples exceeded the MTL set by EU regulations (2 μg/kg). The highest levels of AFB1 were detected in two sample of pistachio (411 and 126 μg/kg) and one sample of peanut (73.9 μg/kg). Aflatoxin G2 is detected in 5 out of 23 samples, pistachio (2), walnut (2) and hazelnut (1). While, B2 and G1 were detected in 17 samples in levels ranged between ND–10.7 μg/kg and ND–12.9 μg/kg, respectively.     

Mycological quality and mycotoxin contamination of Sri Lankan peppers (Piper nigrum L.) and subsequent exposure assessment

The aim of the study was to characterize the toxigenic moulds and to screen different mycotoxins in peppers (Piper nigrum L.) of Sri Lankan origin. Aspergillus flavus, Aspergillus parasiticus, Aspergillus niger and Penicillium spp. were found to be the most dominant fungi. Characterization of the moulds was carried out in A. flavus and parasiticus agar (AFPA) and malt extract agar (MEA) in 77 black pepper (BP) and 11 white pepper (WP) samples. In total, 73% of the BP and 64% of the WP samples were contaminated with A. flavus and/or A. parasiticus (AfAp). A BP sample with water activity (a_w) 0.70 recorded the highest count of AfAp (4.3*10⁴ CFU/g). Moreover, 75% of the BP samples exceeded the safe a_w limit (0.65) set by the European Spice Association (ESA). The frequency of occurrence of A. niger in BP was 62% with counts up to 1.3*10³ CFU/g. Penicillium spp. were found in 61% and 55% of the BP and WP samples, respectively. In BP 94% of the samples had a Penicillium contamination below 10³ CFU/g. Other Aspergillus spp, found in peppers included, Aspergillus terrus, Aspergillus tamarii, Aspergillus candidus, Aspergillus penicilloides, Aspergillus sydowii and Aspergillus fumigatus. Mould counts in BP (10²–10⁴ CFU/g) were significantly higher than that of WP (<10² CFU/g). Apart from the occurrence of “classical mycotoxins” of spices, aflatoxins (<LOQ-18 μg/kg) and ochratoxin A (<LOQ-79 μg/kg), other toxins including fumonisin B1 (<LOQ-135 μg/kg), sterigmatocystin (<LOQ-49 μg/kg) and citrinin (<LOQ-112 μg/kg) were detected in peppers. In total, 63% of the BP samples were contaminated with at least one mycotoxin. Mycotoxin contamination in WP was significantly less compared to BP. The exposure to aflatoxins and ochratoxin A by consuming pepper remains harmless considering the existing pepper dietary intake data of the Sri Lankan population.     

Survey of aflatoxins and ochratoxin A in retail market chilies and chili sauce samples

The aim of this study was to determine the levels of the aflatoxins (AFs) and ochratoxin A (OTA) in 312 samples of whole chili, chili powder, crushed chili and chili sauce samples from suburbs, open market and food restaurants of Punjab, Pakistan. The analysis was carried out using HPLC with fluorescence detector, after immunoaffinity column clean-up. The results have shown that 176 out of 312 (56.4%) samples were to be positive with AFs and 126 out of 312 (40.4%) sample of chilies were found to be contaminated with OTA. Total mean level of AFB₁ and total AFs in chilies were 12.50 ± 1.91, 15.16 ± 2.22 μg/kg, respectively. The total mean level of OTA in chilies was found 16.68 ± 2.58 μg/kg, ranged from LOD to 120.9 μg/kg. Sample 39.5, 26.3 and 32.7% of chilies were found containing level of AFB₁, total AFs and OTA, higher than the recommended limits for EU, respectively. The dietary levels of 3.26, 3.52, and 3.84 μg/kg were determined for AFB₁, total AFs and OTA in chilies. The incidence and levels of AFs and OTA in chilies are higher and could pose serious health hazards for consumers.     

Development of a rapid magnetic bead-based immunoassay for sensitive detection of zearalenone

Here we demonstrate a novel magnetic bead-based enzyme-linked immunosorbent assay (MB-ELISA) for zearalenone (ZEN) detection. Firstly, an anti-ZEN monoclonal antibody (mAb) was prepared by hybridoma technique, and immobilized on carboxyl modified MBs to obtain mAb-MBs. In addition, the biotinylated ZEN-BSA was labelled by streptavidin-HRP for use as competitor. Based on the mAb-MBs and streptavidin-HRP labelled ZEN-BSA, a MB-ELISA which contains only one 20 min antigen-antibody reaction step and takes no more than 45 min for dozens of samples analysis was developed. The half maximal inhibitory concentration (IC₅₀) and limit of detection (LOD) of the MB-ELISA are 1.78 ng/mL and 0.13 ng/mL, respectively. And the MB-ELISA working range for corn samples analysis is from 5.0 μg/kg to 255.2 μg/kg. The recoveries for ZEN spiked corn samples ranged from 82.3 to 110.5% with coefficient of variation (CV) under 8.9%. For natural corn samples analysis, the results of MB-ELISA showed good agreement with the results of conventional direct competitive ELISA (R² = 0.9742).