Occurrence and exposure assessment of aflatoxins in Zhejiang province,China

The purpose of this study was to assess the risk of aflatoxins due to multiple food consumption among the Zhejiang population. Ultra-high-performance liquid chromatography coupled with tandem mass spectrometry method was used to determine aflatoxins in 792 samples. Aflatoxins were detected in 27.1% of the samples at levels between 0.07 and 262.63 μg kg⁻¹, and aflatoxins B₁ was the most frequently detected among different types of samples. 0.8% of peanut oil, 3.39% of nut products as well as 1.1% of condiments contaminated with aflatoxins B₁ exceeded China national tolerance limits. Peanut oil had the highest incidence of aflatoxin, with a range from 0.17 to 22.50 μg kg⁻¹. Using bags conferred limited advantages in reducing aflatoxin contents. Moreover, peanut and rice were the main contributors to dietary exposure to aflatoxins among Zhejiang residents. Finally, the margin of exposure values obtained by rice consumption were far from the safe margin of 10,000, indicating a potential risk to public health. The results pointed out the need for further prioritization of aflatoxins B₁ risk-management actions in Zhejiang.     

Determination of aflatoxin levels in Sudanese edible oils

Fifty-six samples of groundnut, sesame and cottonseed oils form factories, and traditional mills were collected from several localities in Kordofan, Gezira and Khartoum states, Sudan and assessed for aflatoxin B₁ (AFB₁), aflatoxin B₂ (AFB₂), aflatoxin G₁ (AFG₁) and aflatoxin G₂ (AFG₂), using high performance liquid chromatography (HPLC). Aflatoxin B₁ (AFB₁) was detected in eight samples representing 14.3%, the highest incidence of aflatoxin contamination occurred in sesame (7 out of 16 samples, 43.75%) followed by groundnut (1 out of 28 samples, 3.57%) while no aflatoxin contamination was detected in cottonseed oil. Aflatoxin B₁ levels in sesame oil samples ranged from 0.2–0.8 μg/kg and were 0.6 μg/kg in groundnut oil samples. All aflatoxin contaminated samples are unrefined. This paper reports the findings of the first exploratory investigation on presence of aflatoxins in Sudanese edible oils collected from three states.     

Effects of Bacillus subtilis ANSB060 on growth performance,meat quality and aflatoxin residues in broilers fed moldy peanut meal naturally contaminated with aflatoxins

This study was conducted to investigate the toxic effects of aflatoxins and the efficacy of Bacillus subtilis ANSB060 for the amelioration of aflatoxicosis in broiler chickens. Six replicates of ten broilers each were assigned to one of seven dietary treatments, which were labeled C0 (basal diet); M0 (basal diet containing moldy peanut meal); C500 and C1000 (C0 + 500 or 1000 g/t aflatoxin biodegradation preparations, composed mainly of ANSB060); and M500, M1000 and M2000 (M0 + 500, 1000 or 2000 g/t aflatoxin biodegradation preparations). The concentrations of aflatoxin B₁, B₂, G₁ and G₂ in the moldy diets (M0, M500, M100 and M2000) fluctuated around 70.7 ± 1.3, 11.0 ± 1.5, 6.5 ± 0.8 and 2.0 ± 0.3 μg/kg, respectively. The results showed that the M0 diet caused a significant decrease in average daily weight gain and increased feed requirements, with a gain ratio increasing from d 8 to 42, deterioration in meat quality and aflatoxin residues in broilers’ livers as compared with the C0 diet. The addition of ANSB060 to the aflatoxin-contaminated diets offset these negative effects, leading to the conclusion that ANSB060 has a protective effect on growth performance and meat quality while reducing the amount of aflatoxin residues in the livers of broilers fed naturally moldy peanut meal.     

A Survey Using Normal Phase Hplc of Aflatoxins in Domestic and Imported Foods and Dairy Products in the Ussr

Abstract

An improved normal phase HPLC method has been developed to study the occurrence of aflatoxins B₁, B₂, G₁, G₂ and M₁ in domestic and imported foods. Ether-methanol-water (95:5:1) mobile phase and fluorometric detector with silica gel packed flow cell were used. The detection limit of the method was 0.1 ug/kg for aflatoxin B₁, coefficients of variation were 11% and 8.5% at contamination levels 10 and 100 ug/kg of aflatoxin B₁ respectively. Recoveries of added aflatoxins B₁, B₂, G₁, G₂ for corn ranged from 78% to 88%. This method allows the determination of aflatoxins B₁, B₂, G₁, G₂, and M₁, B_2a, M_2a as well as other aflatoxin metabolites. The method is used in monitoring aflatoxin contamination of foods, the first stage of which is a preliminary screening of samples by TLC method (the detection limit is 1.0 ug/kg for aflatoxin B₁). A survey of the occurrence of aflatoxins B₁, B₂, G₁, G₂, in Soviet domestic and imported cereals, nuts, beans, and oil seeds harvested in 1985-87 (over 4300 samples) as well as aflatoxin M₁ in domestic dairy products (over 250 samples) was carried out using HPLC and TLC methods. It was shown that 26.9% of imported peanuts, 2.2% of corn and 6.2% of barley were contaminated by aflatoxins at levels exceeding the maximum tolerated level established in the USSR (5 ug/kg for aflatoxin B₁ in foods of all kinds excluding baby foods). Maximum concentrations were 3600, 155 and 8.0 ug/kg respectively. As much as 28.3% of domestic cotton seed samples, which were chosen for the analysis due to toxic effects on animals, were also contaminated by aflatoxins. The USSR aflatoxin monitoring system was demonstrated to be effective during this study.     

Protective Effects of Bacillus subtilis ANSB060 on Serum Biochemistry,Histopathological Changes and Antioxidant Enzyme Activities of Broilers Fed Moldy Peanut Meal Naturally Contaminated with Aflatoxins

The aim of this study was to investigate the toxic effects of aflatoxins and evaluate the effectiveness of Bacillus subtilis ANSB060 in detoxifying aflatoxicosis in broilers. A total of 360 one-week-old male broilers (Ross 308) were assigned to six dietary treatments for five weeks. The treatment diets were: C0 (basal diet); C1.0 (C0 + 1.0 g B. subtilis ANSB060/kg diet); M0 (basal diet formulated with moldy peanut meal); M0.5, M1.0 and M2.0 (M0 + 0.5, 1.0 and 2.0 g B. subtilis ANSB060/kg diet, respectively). The contents of aflatoxin B₁, B₂, G₁ and G₂ in the diets formulated with moldy peanut meal were 70.7 ± 1.3, 11.0 ± 1.5, 6.5 ± 0.8 and 2.0 ± 0.3 µg/kg, respectively. The results showed that aflatoxins increased (p < 0.05) serum aspartate transaminase activity, decreased (p < 0.05) serum glutathione peroxidase activity, and enhanced (p < 0.05) malondialdehyde contents in both the serum and liver. Aflatoxins also caused gross and histological changes in liver tissues, such as bile duct epithelium hyperplasia, vacuolar degeneration and lymphocyte infiltration. The supplementation of ANSB060 reduced aflatoxin levels in the duodenum and counteracted the negative effects of aflatoxins, leading to the conclusion that ANSB060 has a protective effect against aflatoxicosis and this protection is dose-related.     

Aflatoxin distribution and total microbial counts in an edible oil extracting plant. I. Preliminary observations

Groundnut (peanut) kernels, groundnut and cotton-seed pellets and groundnut and cottonseed oils (crude and refined) were screened for aflatoxins. The groundnut kernels and groundnut and cotton-seed pellets were additionally examined for total microbial counts as well as for certain types of micro-organism. All samples contained aflatoxins (B₁, B₂, G₁ and G₂) while a quantitative estimation of aflatoxin B₁ revealed that all samples contained this aflatoxin in varying amounts. The study identified in substantial numbers several types of micro-organisms that have been associated with industrial and health hazards.     

Toxicological study of gamma-irradiated aflatoxins using the chicken embryo

Aflatoxins B₁ and G₁ and mixtures of B₁ and B₂ and G₁ + G₂ were irradiated at doses of 1.0, 5.0, 10.0, or 15.0 Mrad in a ⁶⁰Co γ irradiator. Fertile chick embryos were injected with these aflatoxins or corresponding nonirradiated control aflatoxin (0.05 μg/egg). The lethality of the aflatoxins decreased with increased γ-irradiation dose. The order of decreasing potency of irradiated aflatoxins was B₁, B₁ + B₂, G₁, and G₁ + G₂. Histopathological examination of the livers showed fatty infiltration, hyperemia, and necrosis.     

Aflatoxin induced inhibition of protein synthesis

The effect of aflatoxins B₁, G₁ and M₁ on protein synthesis in rat liver was studied. Wistar rats (80–100 g body weight) were injected i.p. with aflatoxins B₁, G₁ and M₁ (6, 12 and 6 mg/kg) respectively dissolved in dimethyl sulfoxide and were sacrificed 24 hr after dosing. Aflatoxins B₁ and M₁ but not G₁, inhibited in vivo protein synthesis. Aflatoxins M₁ G₁, but not B₁ inhibited in vitro mitochondrial protein synthesis, whereas both B₁ and M₁ inhibited cytoplasmic protein synthesis.     

Centrifugation-Assisted Solid-Phase Extraction Coupled with UPLC-MS/MS for the Determination of Mycotoxins in ARECAE Semen and Its Processed Products

Mycotoxins can occur naturally in a variety of agriculture products, including cereals, feeds, and Chinese herbal medicines (TCMs), via pre- and post-harvest contamination and are regulated worldwide. However, risk mitigation by monitoring for multiple mycotoxins remains a challenge using existing methods due to their complex matrices. A multi-toxin method for 22 mycotoxins (aflatoxin B₁, B₂, G₁, G₂, M₁, M₂; ochratoxin A, B, C; Fumonisin B₁, B₂, B₃; 15-acetyldeoxynivalenol, 3-acetyldeoxynivalenol, diace-toxyscirpenol, HT-2, T-2, deepoxy-deoxynivalenol, deoxynivalenol, neosolaniol, zearalenone, and sterigmatocystin) using centrifugation-assisted solid-phase extraction (SPE) clean-up prior to ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis for Arecae Semen and its processed products was developed and validated. Several experimental parameters affecting the extraction and clean-up efficiency were systematically optimized. The results indicated good linearity in the range of 0.1–1000 μg/kg (r² > 0.99), low limits of detection (ranging from 0.04 μg/kg to 1.5 μg/kg), acceptable precisions, and satisfactory recoveries for the selected mycotoxins. The validated method was then applied to investigate mycotoxin contamination levels in Areca catechu and its processed products. The mycotoxins frequently contaminating Areca catechu were aflatoxins (AFs), and the average contamination level and number of co-occurring mycotoxins in the Arecae Semen slices (Binlangpian) were higher than those in commercially whole Arecae Semen and Arecae Semen Tostum (Jiaobinlang). Sterigmatocystin was detected in 5 out of 30 Arecae Semen slices. None of the investigated mycotoxins were detected in Arecae pericarpium (Dafupi). The results demonstrated that centrifugation-assisted SPE coupled with UHPLC-MS/MS can be a useful tool for the analysis of multiple mycotoxins in Areca catechu and its processed products.     

Development of an Enzyme-Linked Immunosorbent Assay Method Specific for the Detection of G-Group Aflatoxins

To detect and monitor G-group aflatoxins in agricultural products, we generated class-specific monoclonal antibodies that specifically recognized aflatoxins G₁ and G₂. Of the final three positive and stable hybridomas obtained, clone 2G6 produced a monoclonal antibody that had equal sensitivity to aflatoxins G₁ and G₂, and did not cross-react with aflatoxins B₁, B₂, or M₁. Its IC₅₀ values for aflatoxins G₁ and G₂ were 17.18 ng·mL⁻¹ and 19.75 ng·mL⁻¹, respectively. Using this new monoclonal antibody, we developed a competitive indirect enzyme-linked immunosorbent assay (CI-ELISA); the method had a limit of detection of 0.06 ng·mL⁻¹. To validate this CI-ELISA, we spiked uncontaminated peanut samples with various amounts of aflatoxins G₁ and G₂ and compared recovery rates with those determined by a standard HPLC method. The recovery rates of the CI-ELISA ranging from 94% to 103% were comparable to those of the HPLC (92% to 102%). We also used both methods to determine the amounts of G-group aflatoxins in five peanut samples contaminated by aflatoxin B₁-positive, and their relative standard deviations ranged from 8.4% to 17.7% (under 20%), which demonstrates a good correlation between the two methods. We further used this CI-ELISA to assess the ability of 126 fungal strains isolated from peanuts or field soils to produce G-group aflatoxins. Among these, seven stains producing different amounts of G-group aflatoxins were identified. Our results showed that the monoclonal antibody 2 G6-based CI-ELISA was suitable for the detection of G-group aflatoxins present in peanuts and also those produced by fungi.     

Aflatoxins B1, B2 and G1 modulate cytokine secretion and cell surface marker expression in J774A.1 murine macrophages

Aflatoxins are fungal products which occur in food and feed. They are potent hepatocarcinogens, and are known to cause immunosuppression. We investigated the effect of aflatoxin B₁ (AFB₁), aflatoxin B₂ (AFB₂) and aflatoxin G₁ (AFG₁) exposure, alone and in combination, on the secretion of key pro- and anti-inflammatory cytokines from the murine macrophage cell line, J774A.1. Exposure of macrophages to low doses of aflatoxin (0.01 or 0.1 ng/mL) resulted in a statistically significant change in the secretion of a number of cytokines following stimulation with lipopolysaccharide (LPS), a component of Gram-negative bacterial cell walls. Specifically, treatment with AFB₁ or AFB₂ alone significantly decreased (P < 0.01) the secretion of the anti-inflammatory cytokine interleukin (IL) 10 (IL-10), while the secretion of the pro-inflammatory cytokine IL-6 was significantly increased (P < 0.01). In addition, aflatoxin exposure affected expression levels of key cell surface markers involved in the inflammatory response. Toll-like receptor 2 (TLR2) and Cluster of Differentiation 14 (CD14) expression levels decreased significantly (P < 0.01), but Toll-like receptor 4 (TLR4) expression was unaffected. This data provides further insight into the mechanisms by which aflatoxins modulate the host immune response to exert their immunosuppressive activity.     

Dietary Risk Assessment and Consumer Awareness of Mycotoxins among Household Consumers of Cereals,Nuts and Legumes in North-Central Nigeria

This study characterized the health risks due to the consumption of mycotoxin-contaminated foods and assessed the consumer awareness level of mycotoxins in households in two north-central Nigerian states during the harvest and storage seasons of 2018. Twenty-six mycotoxins and 121 other microbial and plant metabolites were quantified by LC-MS/MS in 250 samples of cereals, nuts and legumes. Aflatoxins were detected in all food types (cowpea, maize, peanut and sorghum) except in millet. Aflatoxin B₁ was the most prevalent mycotoxin in peanut (64%) and rice (57%), while fumonisin B₁ occurred most in maize (93%) and beauvericin in sorghum (71%). The total aflatoxin concentration was highest in peanut (max: 8422 µg/kg; mean: 1281 µg/kg) and rice (max: 955 µg/kg; mean: 94 µg/kg), whereas the totals of the B-type fumonisins and citrinin were highest in maize (max: 68,204 µg/kg; mean: 2988 µg/kg) and sorghum (max: 1335 µg/kg; mean: 186 µg/kg), respectively. Citrinin levels also reached 51,195 µg/kg (mean: 2343 µg/kg) in maize. Aflatoxin and citrinin concentrations in maize were significantly (p < 0.05) higher during storage than at harvest. The estimated chronic exposures to aflatoxins, citrinin and fumonisins were high, resulting in as much as 247 new liver cancer cases/year/100,000 population and risks of nephrotoxicity and esophageal cancer, respectively. Children who consumed the foods were the most vulnerable. Mycotoxin co-occurrence was evident, which could increase the health risk of the outcomes. Awareness of mycotoxin issues was generally low among the households.     

Comparative effects of curcumin and resveratrol on aflatoxin B1-induced liver injury in rats

Aflatoxin B₁ is a potent hepatotoxic and hepatocarcinogenic mycotoxin. Lipid peroxidation and oxidative DNA damage are the principal manifestations of aflatoxin B₁-induced toxicity that could be counteracted by antioxidants. Many plant constituents have been reported to prevent liver damage associated with lipid peroxidation. In this study, curcumin (polyphenolic antioxidant purified from turmeric) and resveratrol (polyphenol obtained from grapes) were evaluated for possible protection against liver injury induced by aflatoxin B₁ in rats. Adult male Fischer rats were divided into six groups including untreated control, curcumin control (200 mg/kg BW), resveratrol control (10 mg/kg BW) and aflatoxin B₁ (25 μg/kg BW). Other two groups were administered either curcumin or resveratrol along with aflatoxin B₁. The study was carried out for 90 days. At the end of the experiment period, blood and tissue samples were collected from the animals before they were killed. Livers were collected for histopathologic studies and fixed in 10% buffered formalin solution. Serum was used for estimation of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and γ-glutamyl transferase (γ-GT) enzymes. The lipid peroxidation, reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were estimated in liver homogenates. The results revealed that aflatoxin B₁ administration caused liver damage as indicated by statistically significant (P < 0.05) increase in serum ALT, AST and γ-GT levels. In addition, there were general statistically significant reductions in the activities of GSH, SOD, CAT, GSH-Px, and an increase in lipid peroxidation in the liver of aflatoxin B₁-treated group compared to the untreated control group. Curcumin showed a significant hepatoprotective activity by lowering the levels of serum marker enzymes, lipid peroxidation and elevating the levels of GSH, SOD, CAT and GSH-Px. However, resveratrol failed to protect from the aflatoxin B₁-induced liver injury. These findings suggest that curcumin but not resveratrol has a hepatoprotective effect against aflatoxin B₁-induced liver injury.     

Comparative Oxidative Metabolism of Aflatoxin B1 and Palmotoxins B0 and G0 by Rat Liver Microsomal Fractions

Abstract

1. The enzymic transformations of aflatoxin B₁ and palmotoxins B₀ and G₀ by cell fractions of rat liver have been studied. Metabolism involves NADPH-dependent enzymes with max. activity in the presence of nicotinamide, MgCl₂, glucose-6-phosphate and NADP.

2. The substances all undergo demethylation with production of formaldehyde. With both aflatoxin B₁ and palmotoxin B₀, two metabolites were detected by t.l.c. The two metabolites of aflatoxin B₁ correspond to aflatoxin M₁ and possibly aflatoxin B_2a. The two derivatives of palmotoxin B₀ have not been characterized but, like those of aflatoxin B₁, are probably hydroxylation products, as they are more polar than the parent compound. Only one derivative of palmotoxin G₀ was detected.

3. Aflatoxin B₁ showed a higher rate of metabolism than the two other substances.     

Sensitive Quantification of Aflatoxin B1 in Animal Feeds,Corn Feed Grain,and Yellow Corn Meal Using Immunomagnetic Bead-Based Recovery and Real-Time Immunoquantitative-PCR

Aflatoxins are considered unavoidable natural mycotoxins encountered in foods, animal feeds, and feed grains. In this study, we demonstrate the application of our recently developed real-time immunoquantitative PCR (RT iq-PCR) assay for sensitive detection and quantification of aflatoxins in poultry feed, two types of dairy feed (1 and 2), horse feed, whole kernel corn feed grains, and retail yellow ground corn meal. Upon testing methanol/water (60:40) extractions of the above samples using competitive direct enzyme linked immunosorbent assay, the aflatoxin content was found to be <20 μg/kg. The RT iq-PCR assay exhibited high antigen hook effect in samples containing aflatoxin levels higher than the quantification limits (0.1–10 μg/kg), addressed by comparing the quantification results of undiluted and diluted extracts. In testing the reliability of the immuno-PCR assay, samples were spiked with 200 μg/kg of aflatoxin B₁, but the recovery of spiked aflatoxin was found to be poor. Considering the significance of determining trace levels of aflatoxins and their serious implications for animal and human health, the RT iq-PCR method described in this study can be useful for quantifying low natural aflatoxin levels in complex matrices of food or animal feed samples without the requirement of extra sample cleanup.     

用流式细胞光度术(FCM)研究争光霉素对肿瘤及正常细胞周期的影响

用流式细胞光度术发现七种争光霉素组分(A₂+B₂混合品,A₂,A₂,A₅,A₆,A₅₀₃₃和B₂)都使中国仓鼠卵巢细胞(CHO)阻断在G₂期,而对G₁→S→G₂的进行无影响。这种G₂期阻断呈剂量依赖性。不同组分的G₂阻断效应不同,在40 μg/ml时,以A₄和A₅最强,其次为A₆,A₂,A₂+B₂混合物和B₂,而A₅₀₃₃无影响。用等毒性剂量,所有组分都使小鼠艾氏腹水癌产生多倍化细胞。由于争光霉素的G₂期阻断效应与抑瘤存在着平行关系,我们认为争光霉素的抑瘤作用与癌细胞被阻断在G₂期有关。     

Reduction of Aflatoxins in Apricot Kernels by Electronic and Manual Color Sorting

The efficacy of color sorting on reducing aflatoxin levels in shelled apricot kernels was assessed. Naturally-contaminated kernels were submitted to an electronic optical sorter or blanched, peeled, and manually sorted to visually identify and sort discolored kernels (dark and spotted) from healthy ones. The samples obtained from the two sorting approaches were ground, homogenized, and analysed by HPLC-FLD for their aflatoxin content. A mass balance approach was used to measure the distribution of aflatoxins in the collected fractions. Aflatoxin B₁ and B₂ were identified and quantitated in all collected fractions at levels ranging from 1.7 to 22,451.5 µg/kg of AFB₁ + AFB₂, whereas AFG₁ and AFG₂ were not detected. Excellent results were obtained by manual sorting of peeled kernels since the removal of discolored kernels (2.6%–19.9% of total peeled kernels) removed 97.3%–99.5% of total aflatoxins. The combination of peeling and visual/manual separation of discolored kernels is a feasible strategy to remove 97%–99% of aflatoxins accumulated in naturally-contaminated samples. Electronic optical sorter gave highly variable results since the amount of AFB₁ + AFB₂ measured in rejected fractions (15%–18% of total kernels) ranged from 13% to 59% of total aflatoxins. An improved immunoaffinity-based HPLC-FLD method having low limits of detection for the four aflatoxins (0.01–0.05 µg/kg) was developed and used to monitor the occurrence of aflatoxins in 47 commercial products containing apricot kernels and/or almonds commercialized in Italy. Low aflatoxin levels were found in 38% of the tested samples and ranged from 0.06 to 1.50 μg/kg for AFB₁ and from 0.06 to 1.79 μg/kg for total aflatoxins.     

Simplified Synthesis and Stability Assessment of Aflatoxin B1-Lysine and Aflatoxin G1-Lysine

Aflatoxins B₁ (AFB₁) and G₁ (AFG₁) are carcinogenic mycotoxins that contaminate crops such as maize and groundnuts worldwide. The broadly accepted method to assess chronic human aflatoxin exposure is by quantifying the amount of aflatoxin adducted to human serum albumin. This has been reported using ELISA, HPLC, or LC-MS/MS to measure the amount of AFB₁-lysine released after proteolysis of serum albumin. LC-MS/MS is the most accurate method but requires both isotopically labelled and unlabelled AFB₁-lysine standards, which are not commercially available. In this work, we report a simplified synthetic route to produce unlabelled, deuterated and ¹³C₆ ¹⁵N₂ labelled aflatoxin B₁-lysine and for the first-time aflatoxin G₁-lysine. Additionally, we report on the stability of these compounds during storage. This simplified synthetic approach will make the production of these important standards more feasible for laboratories performing aflatoxin exposure studies.     

Determination of aflatoxin B1 levels in Iranian rice by ELISA method

This study was carried out to detect the presence of aflatoxin B₁ (AFB₁) in 40 samples of Tarom rice from Iran. Enzyme-linked immunosorbent assay (ELISA) was applied to analyze AFB₁ in the samples. All the analyses were conducted twice. Aflatoxin B₁ was found in all rice samples, the concentration of AFB₁ ranged from 0.29 to 2.92?µg/kg. The AFB₁ concentration mean in the rice samples produced in 2013 was higher (P < 0.05) than the findings in rice in 2012.. However, 25 of the 40 samples exceeded the maximum prescribed limit, i.e. 2?µg/kg of European Union Regulations and also none of the samples reached the maximum prescribed limit 5?µg/kg of the Institute of Standards and Industrial Research of Iran (ISIRI) for aflatoxin B₁. Although, rice is ranked the second among cereal staples consumed food in Iran and many countries, it can make a serious health problem for people even for a small amount of aflatoxin.     

Development of a sensitive enzyme-linked immunosorbent assay for the detection of fumonisin B1 in maize

Fumonisin B₁ (FB₁) is a mycotoxin, mainly produced by Fusarium fungi and present in food and feed. It causes harmful effects on human and animal health. Therefore, it is necessary to develop sensitive and reliable screening methods. In this study, a highly sensitive monoclonal antibody (MAb) against FB₁, clone 2D7, was produced, and the 50% inhibition concentration (IC₅₀) of the MAb was 2.2 ng/mL in buffer. The MAb showed high cross-reactivity with fumonisin B₂ (FB₂), and negligible cross-reactivity with other mycotoxins. A sensitive indirect competitive enzyme-linked immunosorbent assay (icELISA) based on this MAb was developed for the determination of FB₁ in maize. In spiked samples (100, 200 and 500 μg/kg), the average recoveries ranged from 78 ± 11 to 107 ± 4%, and the coefficient of variation ranged from 3 to 15%. The limit of detection of the icELISA was 5.4 μg/kg. This method was compared to liquid chromatography tandem mass spectrometry (LC-MS/MS) using naturally contaminated samples, and the correlation coefficient was above 0.82. These results show the reliability of the icELISA method for the determination of FB₁ in maize.