Non-destructive SPE-UPLC-based Quantification of Aflatoxins and Stilbenoid Phytoalexins in Single Peanut (Arachis spp.) Seeds.

Aflatoxins are highly carcinogenic secondary metabolites of some fungal species, particularly Aspergillus flavus. Aflatoxins often contaminate economically important agricultural commodities, including peanuts, posing a high risk to human and animal health. Due to the narrow genetic base, peanut cultivars demonstrate limited resistance to fungal pathogens. Therefore, numerous wild peanut species with tolerance to Aspergillus have received substantial consideration by scientists as sources of disease resistance. Exploring plant germplasm for resistance to aflatoxins is difficult since aflatoxin accumulation does not follow a normal distribution, which dictates the need for the analyses of thousands of single peanut seeds. Sufficiently hydrated peanut (Arachis spp.) seeds, when infected by Aspergillus species, are capable of producing biologically active stilbenes (stilbenoids) that are considered defensive phytoalexins. Peanut stilbenes inhibit fungal development and aflatoxin production. Therefore, it is crucial to analyze the same seeds for peanut stilbenoids to explain the nature of seed resistance/susceptibility to the Aspergillus invasion. None of the published methods offer single-seed analyses for aflatoxins and/or stilbene phytoalexins. We attempted to fulfill the demand for such a method that is environment-friendly, uses inexpensive consumables, and is sensitive and selective. In addition, the method is non-destructive since it uses only half of the seed and leaves the other half containing the embryonic axis intact. Such a technique allows germination and growth of the peanut plant to full maturity from the same seed used for the aflatoxin and stilbenoid analysis. The integrated part of this method, the manual challenging of the seeds with Aspergillus, is a limiting step that requires more time and labor compared to other steps in the method. The method has been used for the exploration of wild Arachis germplasm to identify species resistant to Aspergillus and to determine and characterize novel sources of genetic resistance to this fungal pathogen.

Prevalence and concentration of mycotoxins in bovine feed and feed components: A global systematic review and meta-analysis.

Mycotoxins are secondary metabolites produced by fungi and identified as contaminants in animal feed. They have potentially harmful effects, including carcinogenicity, mutagenicity, and repro-toxicity in animals and humans. As a result of climate change, there is the potential for a change in the prevalence and concentration of mycotoxins in animal feed components. This necessitates an assessment of the present and emerging threats to the food supply chain from mycotoxins. This systematic review and meta-analysis study synthesised studies on mycotoxin contamination and prevalence in cattle feed components. The studies were collected from scientific databases Web of Knowledge, Scopus, and Embase between 2011 and 2022. The meta-analysis synthesised 97 studies on the prevalence and the concentration of aflatoxins, ochratoxin A, deoxynivalenol, zearalenone, fumonisin and T-2/HT-2 toxins in feed components. Aflatoxin was highly prevalent (59 %), with a concentration of 2.58-3.92 µg kg-1 in feed components. Ochratoxin A had a global prevalence of 31 % with a concentration of 5.56-12.41 µg kg-1. Deoxynivalenol had a global concentration of 233.17-327.73 µg kg-1 and a prevalence of 74 %. Zearalenone had a prevalence of 70 % and a concentration of 42.47-66.19 µg kg-1. The concentration and prevalence of fumonisins was 232.19-393.07 µg kg-1 and 65 %, respectively. The prevalence and concentration of T-2/HT-2 toxins were 45 % and 23.54-35.12 µg kg-1, respectively. The synthesised concentration of the mycotoxins in the overall feed components was lower than the regulated and guidance values set by the European Union. However, in a few cases, the 95th percentile exceeded these concentration values due to high levels of uncertainty attributed to lower sample size, and thus, need to be considered while conducting risk assessments. The study highlights climates and regions likely to be conducive to the emergence of mycotoxin risk, especially considering the potential influences of climate change.

One-pot green synthesis of ZIF-8/IgG composite for the precise orientation and protection of antibody and its application in purification and detection of aflatoxins in peanut oil.

This study presents a novel approach toward the one-pot green synthesis of ZIF-8/IgG composite, focusing on its precise orientation and protection of the anti-aflatoxins antibody. The antibody orientation is achieved through the specific binding of IgG to the Fc region of the antibody, while the antibody protection is accomplished by the structural change restriction of ZIF-8 framework to the antibody. Consequently, the antibody exhibits enhanced target capability and significantly improved tolerance to organic solvents. The ZIF-8/IgG/anti-AFT was employed for the purification and detection of AFTs by coupling with UPLC. Under optimized conditions, the recoveries of spiked AFTs in peanut oils are between 86.1% and 106.4%, with relative standard deviations (RSDs) ranging from 0.8% to 8.8%. The linearity range is 0.5-20.0 ng for AFB1 and AFG1, 0.125-5.0 ng for AFB2 and AFG2, the limit of detection is 0.1 ng for AFB1 and AFG1, 0.03 ng for AFB2 and AFG2.

Aflatoxins and fumonisins in conventional and organic corn: a comprehensive review.

This review analyzes the occurrence and co-exposure of aflatoxins and fumonisins in conventional and organic corn, and compares the vulnerability to contamination of both. The risks of fungal contamination in corn are real, mainly by the genera Aspergillus and Fusarium, producers of aflatoxins and fumonisins, respectively. Aflatoxins, especially AFB1, are related to a high incidence of liver cancer, and the International Agency Research of Cancer (IARC) classified them in group 1A 'carcinogenic to humans'. The occurrence in conventional corn is reported in many countries, including at higher levels than those established by legislation. IARC classified fumonisins in group 2B 'possibly carcinogenic to humans' due to their link with incidence of esophageal cancer. However, comparing corn and organic and conventional by-products from different regions, different results are observed. The co-occurrence of both mycotoxins is a worldwide problem; nevertheless, there is little data on the comparison of the co-exposure of these mycotoxins in corn and derivatives between both systems. It was found that the agricultural system is not a decisive factor in the final contamination, indicating the necessity of effective strategies to reduce contamination and co-exposure at levels that do not pose health risks.

Determination and risk assessment of aflatoxin B1 in the kernel of imported raw hazelnuts from Eastern Azerbaijan Province of Iran.

Aflatoxin B1 (AFB1) is widespread and seriously threatens public health worldwide. This study aimed to investigate AFB1 in imported hazelnut samples in northwest of Iran (Eastern Azerbaijan Province) using High-Performance Liquid Chromatography with a Fluorescent Detector (HPLC-FLD). In all tested samples AFB1 was detected. The mean concentration of AFB1 was 4.20 µg/kg and ranged from 3.145 to 8.13 µg/kg. All samples contained AFB1 levels within the maximum acceptable limit except for one sample. Furthermore, the human health risk assessment of AFB1 from consuming imported hazelnuts by Iranian children and adults was evaluated based on the margin of exposure (MoE) and quantitative liver cancer risk approaches. The MoE mean for children was 2529.76, while for adults, it was 8854.16, indicating a public health concern. The present study found that the risk of developing liver cancer among Iranian children was 0.11100736 per 100,000 people, and in the Iranian adult population was 0.0314496 cancers per 100,000 people. Since environmental conditions potentially affect aflatoxin levels in nuts, countries are advised to monitor aflatoxin contents in imported nuts, especially from countries with a conducive climate for mold growth.

Incidence and health risk assessment of hydrogen cyanide and multi-mycotoxins in Nigerian garri.

Garri is a granular, starchy food prepared by the fermentation of mashed cassava. Hydrogen cyanide (HCN) and mycotoxins are contaminants in certain foods at different points along the food value chain. The incidence and contamination levels of HCN and multi-mycotoxins in garri from five agroecological zones of Nigeria were determined using a spectrophotometric method and ultra-high-performance liquid chromatography-tandem mass spectroscopy (UHPLC-MS/MS), respectively. The health risk associated with the consumption of contaminated garri was assessed. The health risk assessment model was used to calculate the dietary exposure of humans to the mycotoxins in garri. This was done by estimating the daily intake (EDI), the percentage tolerable daily intake (%TDI), the annual hepatocellular carcinoma (HCC) cases attributable to exposure to aflatoxins (AFs) in garri, as well as the HCC risk. The average intake of garri was estimated at 0.303 kg/day for a Nigerian adult. The incidence of HCN was 98.3% (0.056-2.463 mg/kg), and fermentation reduced the HCN level in garri more than other processing steps. The twenty-one mycotoxins identified and quantified were all within maximum levels, as applicable to those that are regulated by the EU. The %TDI for the other mycotoxins, with the exception of AFs, showed no alarming health risk with garri consumption. Annual HCC cases resulting from AF in garri were estimated at 10-60 cases for HBsAg + ve individuals and 4-23 cases for HBsAg - ve individuals based on 8.1% hepatitis B virus (HBV) incidence. Results further revealed no interdependence between HCN levels and mycotoxin content. This work suggests an unlikely chance of acute toxicity from HCN and major mycotoxins from a garri-based diet in Nigeria. Hence, it is recommended that concerned regulatory bodies maintain the existing permissible limits for HCN in Garri.

Association of aflatoxin with gallbladder cancer in a case-control study nested within a Chinese cohort.

We evaluated whether aflatoxin B1 (AFB1 ) exposure was associated with later risk of developing gallbladder cancer (GBC). We measured AFB1 -lysine albumin adducts in baseline samples from the Shanghai Cohort Study of 18 244 men aged 45 to 64 years (recruited 1986-1989). We included 84 GBC cases with sufficient serum and 168 controls matched on age at sample collection, date of blood draw and residence. We calculated adjusted odds ratios (ORs) and 95% confidence intervals (95% CIs) for detectable vs non-detectable AFB1 -lysine albumin adducts and gallbladder cancer. AFB1 -lysine albumin adducts were detected in 50.0% of GBC cases, and risk of GBC was twice as high in those with detectable vs undetectable levels (OR = 2.0, 95% CI = 1.0-3.9). ORs ranged from 1.8 (95% CI = 0.75-4.3) for 0.5 to <1.75 pg/mg vs undetectable adduct levels to 2.2 (95% CI = 0.91-5.6) for >3.36 pg/mg vs undetectable, suggesting a dose-response (Ptrend = .05). When restricted to cases diagnosed before the median time to diagnosis after blood draw (18.4 years), results were similar (OR = 2.2, 95% CI = 0.80-5.8) to those for the entire follow-up duration. The OR was 9.4 (95% CI = 1.7-51.1) for individuals with detectable AFB1 -lysine albumin adducts and self-reported gallstones compared to individuals with neither. Participants with detectable AFB1 -lysine albumin adducts at baseline had increased risk of developing GBC, replicating the previously observed association between AFB1 exposure and providing the first evidence of temporality.

Total aflatoxin and ochratoxin A levels, dietary exposure and cancer risk assessment in dried fruits in Türkiye.

This study aimed to measure total aflatoxin (AF) (AFB1, AFB2, AFG1, and AFG2) and ochratoxin A (OTA) levels in dried fruit samples and to evaluate the potential dietary exposure and cancer risk to these mycotoxins in Kayseri/Türkiye. Dried fruit samples were collected between April-May 2021. A total of 11 dried grapes and apricot samples, 7 dried fig and plum samples were collected. Total aflatoxins and OTA in dried fruits were determined by ELISA method. Then, the margin of exposure (MOE) and cancer risk were calculated. Total AF was detected in dried fruit samples between 42.86%, and 100%. Between 18.18% and 57.14% of samples exceeded the European Commission (EC) limits for total AF. Moreover, OTA was detected in all samples. Between 71.43% and 100% of samples exceeded the EC limits for OTA. Cancer risk due to OTA exposure was higher than total AF and it was determined that OTA exposure could pose a risk for public health (MOE < 10,000). Although mycotoxin exposure seems to be low due to the low consumption of dried fruit in Türkiye, the risk of exposure and cancer may increase because of complying with the recommendations of the dietary guidelines. The findings provide new insights into exposure to total AF and OTA through the consumption of dried fruit.

Spherical Fe2O3 nanoparticles inhibit the production of aflatoxins (B1 and B2) and regulate total soluble solids and titratable acidity of peach fruit.

Aflatoxin is a group I carcinogen and causes significant public health and food safety risks, throughout the world. This study was carried out to assess the levels of aflatoxin contamination in diseased peach (Prunus persica L.) fruit and their control using myco-synthesized iron oxide nanoparticles (Fe2O3 NPs). Diseased peach fruit were diagnosed to be infected with Aspergillus flavus. The isolated pathogen was cultured under UV light (365 nm) and exposed to ammonium hydroxide (31 %) vapors, which confirmed its ability to produce aflatoxin. For the control of this disease, Fe2O3 NPs were synthesized in the filtrate of a biocontrol fungus (Trichoderma harzianum) and characterized before analyzing their potential in disease control. FTIR spectrum described the presence of capping and reducing agents (secondary amines, alcohol, alkyne and aromatic compounds) on the surface of Fe2O3 NPs. X-ray Diffraction (XRD) described the crystalline size (7.78), while the spherical shape of Fe2O3 NPs was described by the SEM analysis. The EDX spectrum indicated the successful formation of Fe2O3 NPs by showing strong signals of iron (74.38 %). All concentrations displayed mycelial growth inhibition, in vitro and the greatest growth reduction (65.4 %) was observed at 1 mg/ml concentration of NPs. At the same concentration of Fe2O3 NPs, significant control of fruit rot of peach was also observed, in vivo. Treatment of Fe2O3 NPs maintained higher soluble solids, sucrose, total sugar, ascorbic acid, titratable acidity and firmness of peach fruit. Diseased fruit were further investigated for the presence and detection of aflatoxins. All three methods viz. thin layer chromatography (TLC), enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography (HPLC) confirmed a higher production of aflatoxins in control plants, while this production was significantly reduced in Fe2O3 NPs-treated peach fruit.

Synthesis of an ordered macroporous metal-organic framework for efficient solid-phase extraction of aflatoxins from milk products.

Aflatoxins (AFs) exhibit hepatotoxicity, immunotoxicity, and carcinogenicity, and their detection in food has attracted widespread concern. An ordered macroporous metal-organic framework (OM-ZIF-8) based on solid-phase extraction (SPE) was used to extract six AFs from milk products. The SPE conditions, including eluting solvent, eluting volume, amounts of OM-ZIF-8, pH of loading solution, loading solvent, ionic strength, loading flow rate, and elution flow rate, were exhaustively optimized. Under optimal parameters, the six AFs were detected by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The OM-ZIF-8 exhibited satisfactory AFs extraction performance through ordered macropore structure, π-π interaction, coordination interaction, and electrostatic interaction. Furthermore, linearity in the range of 0.01-100 ng mL-1 with low detection limits of 0.002-0.0150 ng mL-1 was obtained, and the relative recoveries of AFs were 80.3-110 % with relative standard deviation ≤8.7 %. Thus, this research provides a promising platform for the analysis of trace AFs in complex foods.

Assessment of dietary exposure and levels of mycotoxins in sorghum from Niger State of Nigeria.

This study reports levels of mycotoxins in sorghum from Niger State, Nigeria, and provides a comprehensive assessment of their potential health risks by combining mycotoxin levels and dietary exposure assessment. A total of 240 samples of red and white sorghum were collected from both stores and markets across four microclimatic zones. Fungal species were identified using a dilution plate method. Aflatoxins (AFs), deoxynivalenol, nivalenol, and ochratoxin (OTA) were quantified using HPLC, whereas cyclopiazonic acid, fumonisins (FUMs) and zearalenone were quantified using ELISA. A. flavus and A. fumigatus were dominant fungal species followed by F. verticilloides, A. oryzae and P. verrucosum. Aflatoxins (mean: 29.97 µg/kg) were detected in all samples, whereas OTA (mean: 37.5 µg/kg) and FUMs (mean: 3269.8 µg/kg) were detected in 72% and 50% of the samples, respectively. Mycotoxins frequently co-occurred in binary mixtures of AFs + OTA and AFs + FUMs. Dietary exposure estimates were highest for FUMs at 230% of TDI and margin of exposures (MOEs) for both AFs and OTA (<10,000) indicating a potential risk associated with combined exposure to AFs and OTA. The Risk of hepatocellular carcinoma cases (HCC/year) attributable to AFs and OTA exposure from sorghum was estimated to be 5.99 × 105 and 0.24 × 105 cases for HBsAg + individuals based on 13.6% HBV incidence. Similarly, the HCC/year for AFs and OTA were assessed to be 3.59 × 105 and 0.14 × 105 at an 8.1% prevalence rate. Therefore, the results of this study demonstrate the high prevalence and dietary exposure to mycotoxins through sorghum consumption, raising public health and trade concerns.

Fluorine-functionalized covalent organic framework as efficient solid phase extraction sorbent for adsorption of aflatoxins in nuts.

In this study, a new fluorine-functionalized covalent organic framework (F-COF) was designed and fabricated by the direct polycondensation of tris(4-aminophenyl)amine and 2,3,5,6-tetra-fluoroterephthaldehyde for the first time. F-COF exhibited a remarkably enhanced adsorption capability compared with that of the fluorine-free COF. The favorable adsorption of aflatoxins was attributed to multiple interactions including pseudo hydrogen bond, F-O, π-π, F-π interactions and hydrophobic interactions between F-COF and aflatoxins. By coupling F-COF based solid phase extraction with high-performance liquid chromatography equipped with fluorescence detector, a rapid and sensitive method for determining aflatoxins (aflatoxin B1, B2, G1 and G2) in nuts (peanuts and pistachios) was established. Under optimal conditions (35 mg F-COF, 100 mL sample solution, 3 mL min-1 as sample loading rate, pH<7, 0.2 mL acetonitrile as desorption solvent), the limits of detection for aflatoxins were 0.02-0.30 ng g-1. The linear range was 0.08-16.0 ng g-1 and the recoveries of the F-COF-based method were 83.5-114 % with relative standard deviations less than 8.0 %.

Assessment of the Adverse Health Effects of Aflatoxin Exposure from Unpackaged Peanut Oil in Guangdong, China.

Aflatoxins are liver carcinogens and are common contaminants in unpackaged peanut (UPP) oil. However, the health risks associated with consuming aflatoxins in UPP oil remain unclear. In this study, aflatoxin contamination in 143 UPP oil samples from Guangdong Province were assessed via liquid chromatography-tandem mass spectrometry (LC-MS). We also recruited 168 human subjects, who consumed this oil, to measure their liver functions and lipid metabolism status. Aflatoxin B1 (AFB1) was detected in 79.72% of the UPP oil samples, with levels ranging from 0.02 to 174.13 µg/kg. The average daily human intake of AFB1 from UPP oil was 3.14 ng/kg·bw/day; therefore, the incidence of liver cancer, caused by intake of 1 ng/kg·bw/day AFB1, was estimated to be 5.32 cases out of every 100,000 persons per year. Meanwhile, Hepatitis B virus (HBV) infection and AFB1 exposure exerted a synergistic effect to cause liver dysfunction. In addition, the triglycerides (TG) abnormal rate was statistically significant when using AFB1 to estimate daily intake (EDI) quartile spacing grouping (p = 0.011). In conclusion, high aflatoxin exposure may exacerbate the harmful effects of HBV infection on liver function. Contamination of UPP oil with aflatoxins in Guangdong urgently requires more attention, and public health management of the consumer population is urgently required.

Evaluating aflatoxins and Sudan dyes contamination in red chili and turmeric and its health impacts on consumer safety of Lahore, Pakistan.

Spices are contaminated with aflatoxins (AFs) and Sudan dyes which are classified as class Group 1 and Group 3 human carcinogens by the International Agency for Research on Cancer (IARC) respectively and their prolonged exposure may raise a human health concern. A total of 474 samples of red chili and turmeric were collected from Lahore city and were subjected to quantitative and qualitative AFs and Sudan dyes analysis by thin layer chromatography (TLC) respectively. The number of red chili and turmeric samples with ≥10 µg/kg of total AFs (European Union standard limit) were 70% and 33% respectively and considered unfit for human consumption. The presence of Sudan dyes in red chili and turmeric samples was 67% and 27% respectively. The mean estimated daily intake (EDI) among females and males was 0.0019 µg/kg bw/day, 0.0012 µg/kg bw/day for red chili, and 0.0008 µg/kg bw/day, 0.0006 µg/kg bw/day for turmeric respectively. The mean value of margin of exposure (MOE) among females and males for ingestion of AFs-contaminated red chili and turmeric was 210.25, 332.13, 501.02, and 699.31 respectively. Therefore, the current study demands a continuous monitoring plan and the implementation of novel techniques to enhance the product's quality and protect public health.

Endocrine Effect of Some Mycotoxins on Humans: A Clinical Review of the Ways to Mitigate the Action of Mycotoxins.

Fungi such as Aspergillus spp. and Fusarium spp., which are commonly found in the environment, pose a serious global health problem. This study aims to present the results of epidemiological studies, including clinical cases, on the relationship between human exposure to some mycotoxins, especially zearalenone and aflatoxin, and the occurrence of reproductive disorders. In addition, examples of methods to reduce human exposure to mycotoxins are presented. In March 2023, various databases (PubMed, Google Scholar, EMBASE and Web of Science) were systematically searched using Google Chrome to identify studies evaluating the association between exposure to mycotoxins and the occurrence of complications related to impaired fertility or cancer incidence. The analysed data indicate that exposure to the evaluated mycotoxins is widespread and correlates strongly with precocious puberty, reduced fertility and increased cancer incidence in women and men worldwide. There is evidence to suggest that exposure to the Aspergillus mycotoxin aflatoxin (AF) during pregnancy can impair intrauterine foetal growth, promote neonatal jaundice and cause perinatal death and preterm birth. In contrast, exposure to the Fusarium mycotoxin zearalenone (ZEA) leads to precocious sexual development, infertility, the development of malformations and the development of breast cancer. Unfortunately, the development of methods (biological, chemical or physical) to completely eliminate exposure to mycotoxins has limited practical application. The threat to human health from mycotoxins is real and further research is needed to improve our knowledge and specific public health interventions.

Research on highly sensitive quantitative detection of aflatoxin B2 solution based on THz metamaterial enhancement.

Food such as cereal crops, oil crops and dairy products are very easy to produce highly toxic and carcinogenic aflatoxins during inappropriate storage. Therefore, it is of great significance to achieve rapid, non-destructive and highly sensitive detection of aflatoxin. A terahertz metamaterial sensor with "X" compound double-peak structure is designed based on electromagnetic theory to realize highly sensitive detection of aflatoxin B2 solution. It is found that the amplitude of the transmission peak of the terahertz transmission spectrum of aflatoxin B2 (AFB2) solution around 1.2 THz and 2.0 THz gradually decreased with the increase of the concentration of aflatoxin B2 solution, and the frequency of the transmission peak gradually shifted to high frequency with the increase of the concentration of aflatoxin B2 solution, hence a full concentration model was established. And a strategy of first classifying concentration intervals and then building a grouped quantitative model was proposed. The Limit of Detection (LOD) of the interval sub-model of low and medium concentration of aflatoxin B2 solution has been greatly improved with the LOD of the optimal grouping model was 7.28 × 10-11 mg/ml, 4.19 × 10-9 mg/ml and 1.22 × 10-7 mg/ml, respectively. This research verifies the feasibility of terahertz metamaterial sensor based on "X" composite double-peak structure combined with THz-TDS technology for highly sensitive detection of aflatoxin B2 solution. And it provides a new rapid, non-destructive and highly sensitive detection of aflatoxin in food.

[Determination of four aflatoxins in feeds by high throughput automated immunoaffinity magnetic beads purification-ultra performance liquid chromatography].

Aflatoxin (AFT) is an extremely toxic and highly toxic carcinogenic substance. This is particularly problematic due to the risk of aflatoxin contamination in raw feed materials and products during production, transportation, and storage. In this study, immunoaffinity magnetic beads (IMBs) were prepared for the purification of four aflatoxins (aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1) and aflatoxin G2 (AFG2)). The aflatoxin contents were then determined rapidly and accurately using ultra performance liquid chromatography (UPLC). More specifically, the coupling ratio of magnetic beads (MBs) to the aflatoxin monoclonal antibody was initially optimized, wherein an MB volume of 1 mL and an antibody content of 2.0 mg was found to meet the purification requirements of this method. The magnetic properties of the MBs and the IMBs were then investigated using a vibrating sample magnetometer (VSM) at room temperature. As a result, the maximum saturation super magnetizations of the MBs and the IMBs were determined to be 28.61 and 23.22 emu/g, respectively, indicating that the saturation magnetization intensity of the IMBs was reduced by coupling with a non-magnetic antibody. However, the saturation magnetization intensity remained sufficiently high to permit magnetic separation from the solution. In addition, the appearance of the IMBs was examined using a biomicroscope, and it was clear that the magnetic cores were wrapped in agarose gel. Furthermore, the reaction time between the IMBs and the aflatoxins was investigated, and the optimal reaction time for meeting the purification requirements was determined to be 2 min. The stability of the IMBs was then evaluated under refrigerated storage conditions at 4 ℃. It was found that the prepared IMBs maintained a high aflatoxin enrichment capacity for at least eight months. Through the examination of three different extraction solutions, a mixture of acetonitrile and water (70∶30, v/v) was found to be optimal for the extraction of aflatoxins from the feed samples. Moreover, five sample dilutions and purification effects were also examined, and phosphate-buffered saline (containing 0.5% Tween-20) was selected as the preferred sample dilutant. With the optimized conditions, the effectiveness of using IMB for the purification of different feed samples was investigated. The resulting UPLC chromatogram showed no spurious peaks close to the target peaks, demonstrating a good purification performance. Following matrix spiking (5, 20, and 40 µg/kg, calculated based on AFB1) of the four feed samples (i. e., soybean meal, distillers dried grains with solubles, pig feed, and chicken feed), the spiked recoveries of the four aflatoxins ranged from 91.1% to 119.4% with a relative standard deviation (RSD) of <6.9%. In addition, the inter-day precision was 4.5% to 7.5%, and the method exhibited a good reproducibility. Subsequently, the developed method was used to detect AFB1 using reference materials. The test value was 18.6 µg/kg with an accuracy of 110.3%, thereby constituting satisfactory results. Upon testing 21 randomly purchased feed samples using this method, four of these samples contained AFB1, and the test results obtained using the developed method and stable isotope dilution LC-MS/MS were comparable. It was therefore apparent that the IMB purification method combined with UPLC analysis exhibited a good accuracy for aflatoxin determination. Thus, an automatic purification system was established to facilitate the operation and use of IMBs. This system was able to purify 24 samples simultaneously in 30 min. An IMB purification kit for was also designed and produced for aflatoxin detection in feed samples. The kit contained the sample dilutant, IMBs, the washing solution, and the eluent. After extraction of the feed sample, the extraction solution was added to the sample wells provided in the kit, and the purification system automatically completed the steps of aflatoxin enrichment, impurity washing, and elution of the target toxin. It should be noted that the purification process does not require the operator to manually add the solution, thereby simplifying operation. Overall, the purification method established in this study achieved the high-throughput and automatic purification of the four aflatoxins in feed samples.

Food aflatoxin exposure assessment in Sichuan Province, China.

Aflatoxins (AFs) are frequent contaminants in crops worldwide and can cause adverse health effects in exposed humans. Since foods AFs (AFB1, AFB2, AFG1, AFG2) contamination in Sichuan Province are unexplored, we conducted a study to assess AFs exposure in the population. In total, 318 samples, including grains, red chilli, red chilli powder, and vegetable protein beverages, were collected from 13 cities of Sichuan Province, China, in 2022. AFs were detected in all types of foods except for wheat flour, the highest incidence was found in red chilli powder (75.0%). The concentrations of AFtot (the total aflatoxins) ranged between ND (not detected) and 54.20 µg kg-1. It was observed that the AFs profile was dominated by AFB1. The AFB1 content ranged from ND to 52.60 µg kg-1 across food types. According to EU maximum limits (ML) of AFs, 2.8% of samples exceeded the AFtot limits. For AFB1, 0.4% and 4.3% of samples exceeded the China and EU limits, respectively. In this study, packaging types and sampling sites were selected as parameters influence food aflatoxin contamination. Nevertheless, there was no significant difference between different samples. According to exposure assessment and risk characterization, AFtot daily exposure was shown to be 0.263 and 283.936 ng kg-1 bw for the lower and upper exposure. The MOE value derived from consumption grains and red chilli pepper products were generally bellow 10 000, and liver cancer cases based on these two foods consumption could range from < 0.001 to 0.16 cases per year/10 000 persons.

Handheld Fluorescence Spectrometer Enabling Sensitive Aflatoxin Detection in Maize.

Aflatoxins are among the main carcinogens threatening food and feed safety while imposing major detection challenges to the agrifood industry. Today, aflatoxins are typically detected using destructive and sample-based chemical analysis that are not optimally suited to sense their local presence in the food chain. Therefore, we pursued the development of a non-destructive optical sensing technique based on fluorescence spectroscopy. We present a novel compact fluorescence sensing unit, comprising both ultraviolet excitation and fluorescence detection in a single handheld device. First, the sensing unit was benchmarked against a validated research-grade fluorescence setup and demonstrated high sensitivity by spectrally separating contaminated maize powder samples with aflatoxin concentrations of 6.6 µg/kg and 11.6 µg/kg. Next, we successfully classified a batch of naturally contaminated maize kernels within three subsamples showing a total aflatoxin concentration of 0 µg/kg, 0.6 µg/kg and 1647.8 µg/kg. Consequently, our novel sensing methodology presents good sensitivity and high potential for integration along the food chain, paving the way toward improved food safety.

Survey of knowledge, and attitudes to storage practices preempting the occurrence of filamentous fungi and mycotoxins in some Ghanaian staple foods and processed products.

Mycotoxigenic fungi can infect and produce potent mycotoxins in foodstuffs prior to harvest, during harvest (field fungi), and in storage after harvest (storage fungi), which when ingested, can result in adverse health effects. This study was aimed at assessing the knowledge, attitudes, and practices adopted by the Ghanaian populace to help mitigate the occurrence of molds and mycotoxins in foods. A cross-sectional survey involving a structured questionnaire was conducted with 642 respondents from twelve regions of Ghana. Descriptive statistics and analyses of variance were calculated. Correct Classification Rate (CCR) was measured to assess the utility of a logistic regression model. The results of the study showed that the majority of 299 (46.6%) of the respondents were between the ages of 18-25. Age and educational level were related to knowledge about the occurrence of fungi and mycotoxins in foods (p < 0.05). More than half the respondents, 50% indicated that they knew of aflatoxins as a major mycotoxin present in food. Higher education directly influenced on the knowledge of mycotoxicosis and the management of stored food to present intoxication by fungal metabolites. 502 (32.9%) knew that consuming foods with toxins could cause stomach aches. The most commonly consumed food commodity despite the presence of visible growth of fungi was bread (35.3%). The average KAP score for knowledge showed that, out of 100%, there was adequate knowledge (63.8%) among the members of the Ghanaian populace. Favorable environmental conditions of high humidity (> 85% ERH) and temperature (> 28-32 °C) enhance the proliferation of fungi in most foods and the attendant production of mycotoxins such as aflatoxins, ochratoxins, and fumonisins are associated with several severe human and animal health conditions; mycotoxicosis was associated with high fever, pain, vomiting, suppression of immunity, cancer, etc. when these foods are consumed on regular basis for a prolonged length of time. Future examination of the food items used for the School Feeding Programme in Ghana will offer opportunities to examine the risks of feeding youth with fungal-contaminated food preparations from providers.