Analysis Of Aflatoxins Research Articles

Microbial analysis and quantitative assessment of aflatoxins from edible dried insects (Palm Weevil, Cricket and Shea Tree Caterpillar) consumed in southwestern and southeastern regions of Nigeria

Currently in Nigeria, a large number of edible insects are known, and they are globally consumed. The aim of this research was to investigate the microbial load and aflatoxin levels in palm weevil, cricket, and shea butter caterpillars. Six different dried edible insect samples were obtained from different States in the country, (5 palm weevils from Ibadan and Owerri State, 5 shea tree caterpillars from Owerri, 10 palm weevils from Ogun, and 10 crickets from Ibadan and Ondo state). All the samples were packaged in sterile zip lock bags, microbial load analysis was carried out using Standard Microbial Technique, and aflatoxin quantification was done using High-Performance Liquid Chromatography (HPLC). Statistical analysis was carried out using the Statistical Package for Social Sciences (SPSS) version. Microbial analysis of cricket from Ibadan and Ondo showed high counts of total aerobic bacterial load (104.0 ×107 cfu/g and 91.0 ×10 7 cfu/g respectively), the highest aflatoxin quantification of fungi present had values of (17.00 μg/kg and 12.00 μg/kg) in cricket from Ondo and Ibadan respectively. The aflatoxin level was above the permissible limits for ready-to-eat edible dried insects (AFB1; 2μg/kg, Total aflatoxins; 4 μg/kg). Microbial Identification of bacteria and fungi colonies isolated from the palm weevils, crickets, and shea tree caterpillar revealed 3 dominant species of bacteria (Staphylococcus aureus, Escherichia coli, and Proteus mirabilis), three mycotoxin-producing fungi were isolated which includes; Aspergillus niger, Aspergillus terreus, and Aspergillus flavus. The presence of E. coli signifies a potential risk to food safety. Also, the presence of Aspergillus flavus in most of the edible insect sample suggests a potential risk for aflatoxin production. The findings of this study indicate an urgent need for strict quality control measures to ensure the safety of edible insects consumed in Nigeria. Additionally, research into effective processing methods to reduce contamination is recommended.

Standardization and HPTLC Fingerprint Studies on the Ethanol Extracts of <i>Vitex negundo</i> Leaves and <i>Opuntia dillenii</i> Cladode

Background: In traditional Indian system of medicine Vitex negundo Linn is referred as Sarvaroganivarani - the remedy for all diseases. The cladodes of Opuntia dillenii possess anti-inflammatory, potent hypotensive and antihyperglycemic effects. Aim: This study aimed to perform comparative standardization and HPTLC fingerprinting on ethanolic leaf extracts of Vitex negundo and cladodes of Opuntia dillenii. Methods: Extracts were prepared using cold maceration. Physicochemical studies of powdered crude drugs, preliminary phytochemical analysis, heavy metal analysis for arsenic, lead, cadmium and mercury, test for aflatoxin B1, G1, B2, G2 and microbial analysis were carried out in both ethanolic plant extracts. The chromatographic fingerprints of ethanolic extracts of plants were recorded using HPTLC and the solvent system used was chloroform and methanol in the ratio 9:1. Results: The outcomes of preliminary phytochemical analysis revealed the presence of alkaloids, flavonoids, phenols, glycosides and steroids in both the plant extracts. Proteins, saponins and tannins were absent. Analysis of heavy metals and aflatoxins were Below Quantification Limits (BQL). Microbes E. coli, Salmonella, Staphylococcus aureus, and Pseudomonas aeruginosa, were absent. Total aerobic bacterial count and yeast mould count were well within the acceptable limits. The results obtained from HPTLC fingerprint lane analysis showed 12 and13 bands at wavelengths 254nm and 366nm respectively. Conclusion: The present study offers valuable information and act as a tool that can applied in standardization of ethanolic plant extracts of Vitex negundo and Opuntia dillenii.

The Effective Sorting Technologies for Reducing Aflatoxins in Nuts and Dried Figs in Turkey

Background: RASFF notifications serve to determine the hazards responsible for food and feed. In case of determining risks to human health, the information provided to member countries via the rapid alarm system at the same time warned us to take the necessary measures. Objectives: In this research, the RASFF portal alerts over the last three years related to aflatoxins in hazelnut, fig, and peanut products were examined. The impact of image processing technologies on lowering the danger of aflatoxins in shelled hazelnuts and dried figs was also investigated, as well as Critical Control Points (CCPs) on aflatoxin hazard. Methods: The RASFF database was used in the RASFF Window portal and analyzed for aflatoxin alerts. ISO 22000:2018 was used for hazard and risk analysis for raw hazelnut, peanut, and fig products. Aflatoxin analyses were performed according to the AOAC 2000/991.31 method. Prior to and after hazelnut sorting methods, levels of aflatoxin were measured. Besides, aflatoxin contamination in peanuts harvesting in 2023 years from three local producers in Turkey was studied. Results: The number of RASFF notifications for hazelnuts, peanuts/groundnuts, dried figs, and their products for the period of 03/01/2020 to 01/06/2023 were 84, 283 and 173, respectively. As a result of HACCP implementation, especially for aflatoxin hazard management, optical-laser-manual sorting and aflatoxin detection using UV light stages are accepted as CCPs. When nuts were sorted using optical and laser systems were detected decreasing total aflatoxin levels. The mean value of Aflatoxin B1(AFB1) and total aflatoxins were reduced from 7.80±1.08 and 13.56±1.06 to 3.63±0.39 and 7.21±0.71, respectively. 79.16% of all sorted samples were found to comply with legal limits. Investigated peanut samples for aflatoxin B1 contents were found between 5.63 and 27.75 µg kg-1 in 19.5%. Conclusion: The alerts based on aflatoxin contamination above legal limits have been carried on for nuts and dried figs. The physical sorting methods using optical selection and high-resolution laser sensor systems aflatoxin-contaminated nuts from healthy nuts applied for in-shell Turkish nut sorting were able to assess the quality of the batch studied, and the systems were assigned significantly effective to reduce aflatoxin contents. Food business operators in Turkey, which is among the countries exporting agricultural and food products to the European Union (EU), have tended to prefer the most advanced technologies. Efficacies of reduction of aflatoxin by different physical and other (such as imaging, X-ray and acoustic, audio signals and hyperspectral images of moldy natural nuts etc.) methods will be developed in the near future.

Development and validation of extraction and clean-up procedures for UPLC-MS/MS analysis of aflatoxins in spices.

UPLC-MS/MS analytical conditions for the analysis of aflatoxins in spices were optimized and validated in this study. Liquid-liquid partition-based protocols for the cleaning up of extracts using common organic solvents such as acetonitrile, hexane, and ethyl acetate were developed and validated. The developed liquid-liquid partition methods were compared with immuno-affinity column and QuEChERS clean-up methods for the UPLC-MS/MS analysis of aflatoxins in 8 spices. The reduction of lipophilic components using the partition with hexane is particularly useful in spices like red pepper that have higher levels of fatty acids, carotenoids, sterols, triterpenoids, etc. The subsequent partitioning with ethyl acetate considerably reduced the matrix interference from the polar components and increased the sensitivity. The cleaning up of spice extracts using liquid-liquid partition techniques resulted in limits of quantification (LOQ) of 2-5 µgL-1 in UPLC-MS/MS analysis. Trueness, repeatability, and reproducibility of the methods were in acceptable ranges. The accuracy of the developed methods was further verified by analyzing aflatoxins in naturally incurred samples of spices and comparing the results with those obtained from the immuno-affinity column cleanup-HPLC-FD method.

Assessment of Quality parameters and Safety in Indigofera tinctoria L. (Avuri) Rootbark extract

Avuri (Indigofera tinctoria Linn) has widely been used for the treatment of poisonous bites, epilepsy, nervous disorders, bronchitis, dermatological disorders, and liver ailments in Indian traditional medicine. Extensive research has been made into the aerial extracts of this plant for medicine preparation. Despite being used in medicinal formulations, especially in the treatment of poisonous bites the root bark has been less studied. This study is the physiochemical and safety of methanolic extract of Avuri verpattai. Roots were excised from mature plants collected from the Hills of Palani, then the root bark was peeled and air-dried. Extraction was done in a Soxhlet apparatus using a methanolic extraction procedure. Further, the extracts were used for preliminary phytochemical, physiochemical analysis, and safety analysis like aflatoxin, pesticide residue, Microbial contamination, and heavy metals. The phytochemical analysis showed the presence of alkaloids, flavanoids, steroids, triterpenoids, coumarin, phenol, tannin, saponin, sugar and betacyanin. The physiochemical tests conducted on the root bark extract unveiled specific parameters: pH(4.73), Total ash (3.59% w/w), acid insoluble ash (0.4751% w/w), loss on drying at 105°C(3.45% w/w), water-soluble extractive (8.74%w/w), and alcohol soluble extractive (1.91% w/w). The bulk density was 0.2496g/ml, while the tapped density was determined as 0.4940g/ml for the extracts. Safety assessment conducted via aflatoxin analysis, microbial contamination, and pesticide residues, indicated the absence of such toxins in the extract. Additionally, trace amounts of heavy metals mercury and arsenic were detected at levels of 0.334 and 1.970ppm, respectively. The study reveals that the Methanolic extract of Avuri verpattai has significant quality and safety. Consequently, these parameters can serve in the study of Avuri verpattai Extract, as an important Siddha medicinal preparation.

Silver amplified immunosensor via effective fluorogenic Ag+-imidazole aggregation for detection of AFB1

BackgroundCereals are susceptible to aflatoxin contamination during storage and transportation, which is highly carcinogenic and teratogenic, and seriously threaten human health. The accurate and rapid detection of total aflatoxin (including aflatoxin B1, B2, G1, and G2) is of great importance for food safety. Conventional fluorescence immunoassays have the advantage of being sensitive and fast; however, these methods can be affected by strong background and matrix interference. Therefore, the development of ultrasensitive, cost-effective, and interference rejection sensors for detecting aflatoxins in moldy grains is vital for food safety and human health. ResultsIn this paper, a broad-spectrum aflatoxin monoclonal antibody was prepared by using hybridoma cell fusion technology. An aggregation-induced emission (AIE) based immunosensor via silver amplification coupled with a fluorogenic Ag+ probe was established for AFB1 analysis. Silver nanoparticles are decomposed into numerous Ag+ by H2O2, and then Ag+ further specifically binds with imidazole-modified AIE molecules, improving the sensitivity and anti-interference ability of the method. The IC50 and IC15 of AIE-based immunosensor for AFB1 were 0.019 and 0.0014 μg/L, respectively, 2.3-fold and 5.8-fold higher than those of icELISA. The AIE-based immunosensor was also used to analyze AFB1 from actual cereal samples, with spiked recoveries ranging from 72.91 to 115.92 %. In addition, the method was used to detect total aflatoxins in moldy grains. SignificanceBased on the advantages of broad-spectrum aflatoxin monoclonal antibody, high-efficiency metal signal amplification, and functional AIE molecule, a sensitive, accurate, cost-effective, and time-saving method was developed for the analysis of total aflatoxins in cereals. Moreover, the proposed signal amplification strategy shows great potential for analyzing other trace-level small molecular pollutants.

Aflatoxins Levels in Concentrate Feeds Collected from Specialized Dairy Farms and Local Markets in Selected Urban Centers of Eastern Ethiopia.

Aflatoxin constitutes a significant concern for food and feed safety, posing detrimental health risks to both animals and humans. This study aimed to examine the prevalence and concentration of aflatoxins in maize feed, total mixed ration, and wheat bran collected from specialized dairy farms and local markets in three major urban centers in eastern Ethiopia. A total of 180 feed samples were collected from September 2021 to January 2022 in Chiro town, Dire Dawa city, and Harar city. These samples underwent thorough extraction and immunoaffinity clean-up before aflatoxin analysis using HPLC/FLD. The results revealed that AFB1, AFB2, AFG1, AFG2, and TAF contamination was detected in 72.2%, 66.1%, 71.1%, 68.7%, and 82.8% of the feed samples, respectively. The corresponding mean levels of each aflatoxin were 28.15 ± 3.50, 3.3 ± 0.40, 19.87 ± 1.87, 2.7 ± 0.32, and 54.01 ± 4.72 µg/kg, respectively. The occurrence and levels of aflatoxin varied across different study sites and feed types. Notably, feeds from Dire Dawa city exhibited significantly higher mean levels of AFB1 (43.98 ± 5.3 µg/kg), AFB2 (5.69 ± 0.6 µg/kg), AFG1 (32.25 ± 2.7 µg/kg), and AFG2 (5.01 ± 0.5 µg/kg) than feeds from other urban centers did. Additionally, a significantly higher occurrence of AFB1 (29.4%) and AFG1 (28.3%) was detected in feed from Dire Dawa city. Similarly, the total mixed ration (TMR) displayed significantly higher levels of AFB1 (50.67 ± 5.2 µg/kg), AFB2 (4.74 ± 0.6 µg/kg), AFG1 (32.87 ± 2.6 µg/kg), and AFG2 (3.86 ± 0.5 µg/kg) compared to the other feed types. Moreover, a significantly higher occurrence of AFB1 (30.7%) and AFG1 (28.7%) was detected in the TMR. Furthermore, a moderate correlation was observed between the count of aflatoxigenic Aspergillus species and the levels of TAF in the feed samples. Overall, this study underscores the widespread presence of aflatoxin contamination in dairy feeds in eastern Ethiopia, highlighting the urgent need for stringent monitoring and mitigation measures to ensure food and feed safety, as well as public health.

Assessment of the Traditional Processing Methods in the Reduction of Aflatoxin Levels in Maize and Maize Products in Kenya

Aflatoxins are fungal secondary toxic metabolites produced predominantly by certain strains of the Aspergillus molds. The objective of this study was to investigate the effectiveness of Kenyan traditional processing methods in reduction of aflatoxin in maize. The traditional methods used included; alkaline cooking using Magadi soda and maize cob ash, decortication by pounding wet maize with mortar and pestle, dehulling in mechanical mill and exposure to solar radiation. Standardization of alkali cooking was done before actual treatment of the contaminated samples. Naturally contaminated samples of maize were sampled and used in the experiment. Aflatoxin analysis before and after each of the treatments was done using ELISA method and total aflatoxins expressed in parts per billion (ppb) (dry weight basis). The results were subjected to statistical analysis at 5% significance levels (p≤0.05). The reduction of aflatoxin was found to be highest when maize was boiled in maize cob ash solution which resulted in loss of aflatoxin from 83.1±0.3 ppb to 7.0±3.9 ppb. Dry decortication reduced aflatoxin from 51.3±15.3 to 9.6±0.8 ppb, boiling in Magadi soda led to a drop from 59.5±3.818 ppb to 13.4±0.424 ppb, solar irradiation caused a drop from 60.8±1.8 ppb to 13.7±0.1 ppb and pounding in Magadi soda resulted in least loss of aflatoxin from 81.5±0.3 to 72.7±0.2ppb. Aflatoxin loss in all the treatments was significant except for dry decortication. Only dry decortication and boiling in maize cob ash solution brought down the aflatoxin levels to below the maximum allowable limit as set by the Kenya Bureau of Standards (10ppb)

Molecularly imprinted polymer based on covalent organic framework coated steel substrate as the mass spectrometric ionization source for the direct detect of aflatoxins in complex food matrices

Ambient mass spectrometry allows direct analysis of various sample types with minimal or no pretreatment. However, due to the influence of matrix effects, there are sensitivity and issues in analyzing trace analytes in complex food samples. In this work, we developed a spray mass spectrometry platform based on SSS@TPBD-TPA@MIPs (Stainless steel substrate (SSS), terephthalaldehyde (TPA), N, N, N′, N′-tetrakis(p-aminophenyl)-p-phenylenediamine (TPBD), molecularly imprinted polymer (MIP)), for rapid, in situ, high-throughput, highly enrichment efficiency and highly selective trace analysis of aflatoxins. By simplifying the sample pretreatment and directly applying high voltage for ESI-MS, the analysis can be completed within 1 min. The established method base on SSS@TPBD-TPA@MIPs exhibited high sensitivity and accuracy when determine trace level AFs in maize and peanuts. The results demonstrated a good linear relationship within the range of 0.01–10 μg/L, with the determination coefficient (R2) ≥ 0.9956. The limits of detection (LODs) was 0.035–0.3 ng/mL and limits of quantitation (LOQs) was 0.12–0.99 ng/mL, with acceptable recovery rate of 82.09–115.66 % and good repeatability represented by the relative standard deviation (RSD) less than 17.43 %. Furthermore, SSS@TPBD-TPA@MIPs exhibited excellent reusability, with more than 8 repeated uses, and showed good adsorption performance.

Analysis of mold and aflatoxins in processed fish and their control with electrolyzed water

Analysis of mold and aflatoxins in processed fish and their control with electrolyzed water

Aflatoxins in Wheat Grains: Detection and Detoxification through Chemical, Physical, and Biological Means.

Wheat (Triticum aestivum L.) is an essential food crop in terms of consumption as well as production. Aflatoxin exposure has a widespread public health impact in economically developing nations, so there is a need to establish preventive techniques for these high-risk populations. Pre-harvest and post-harvest practices are the two strategies used to control aflatoxin contamination, which include the use of genetically modified crops that show resistance against Aspergillus infection, the use of pesticides, changing the planting and harvesting time of crops, and physical, chemical, and biological methods. In this research, aflatoxin detection and quantification were performed in different wheat varieties to determine quantitative differences in comparison to the European Commission's limit of 4 ppb aflatoxins in wheat. TLC for qualitative and the ELISA kit method for quantitative analysis of aflatoxins were used. Out of 56 samples, 35 were found contaminated with aflatoxins, while the remaining 21 samples did not show any presence of aflatoxins. Out of the 35 contaminated samples, 20 samples showed aflatoxin contamination within the permissible limit, while the remaining 15 samples showed aflatoxin concentration beyond the permissible level, ranging from 0.49 to 20.56 ppb. After quantification, the nine highly contaminated wheat samples were detoxified using physical, chemical, and biological methods. The efficiency of these methods was assessed, and they showed a significant reduction in aflatoxins of 53-72%, 79-88%, and 80-88%, respectively. In conclusion, the difference in aflatoxin concentration in different wheat varieties could be due to genetic variations. Furthermore, biological treatment could be the method of choice for detoxification of aflatoxins in wheat as it greatly reduced the aflatoxin concentration with no harmful effect on the quality of the grains.

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.

Phytochemicals analysis and aflatoxin B1 detoxification potential of leaves extract of Moringa oleifera and Calotropis procera

The present study aimed to identify the presence of certain classes of phytochemicals in the leaf extract of medicinal herbs viz. Moringa oleifera and Calotropis procera, using qualitative detection tests and explored the potential of aqueous and ethanolic extract to inhibit aflatoxin production by thin layer chromatography at 25 °C and pH (7) of different incubation times i.e. 0-, 1-, 3-, 6- and 24-h. Qualitative phytochemical analysis reported that the aqueous leave extracts of M. oleifera and C. procera contained tannins, phlobatannins, quinones, steroids, sugar, betacyanins, fatty acids, phenols, and volatile oils. Aflatoxin analysis reported that the ethanolic extract of M. oleifera was found more effective than detoxifying 100% of AFB1 after 24 h of incubation. In the case of C. procera, the aqueous extract reduced 96.5% of AFB1 and ethanolic extract reduced 96% of AFB1 after 24-h of incubation. The results revealed that natural plant products have a high potential to reduce AFB1 and could contribute to mitigation plans for AFB1. There is a need for further characterisation using techniques such as GC-MS, LC-MS, or NMR which would provide valuable information on the chemical composition of the extracts.

Physicochemical characterization, Phytochemical analysis and Anti-microbial activity of Elathy mathirai

Elathy Mathirai is a Siddha herbomineral formulation indicated to cure sori(itching), sirangu(Scabies), suram(fever), vaayu, maandham(gastro intestinal disorder) . The aim of the study is to standardize Elathy Mathirai to ensure its quality, purity and safety through its physicochemical, chromatographic, phytochemical, heavy metal, pesticide residue and aflatoxins analysis and the secondary objective is to evaluate its antimicrobial property. This study determines the physicochemical paramaters that are loss on drying, total ash, acid insoluble ash, water soluble extractive, alcohol soluble extractive and pH as 5.667% , 5%, 0.014 %, 14.93 %, 8.3% and 5.6 respectively denotes purity and quality of the drug. From the study results it is ensured that the test drug Elathy mathirai is free from pesticide toxicity and aflatoxins. Qualitative preliminary phytochemical analysis shows the presence of alkaloids, Flavanoids,steroids, triterpenoids, saponins, coumarins, phenol, tannin, sugar and betacyanin. HPTLC finger printing analysis reveals the presence of seven prominent peaks Rf value ranges from 0.04 to 0.39 corresponds to the presence of seven components present within it. It was observed from the results of In vitro anti microbial assay that the formulation EM reveals significant level of activity against bacteria Klebsiella pneumonia and observed moderate activity against Staphylococcal aureus, Pseudomonas aeruginosa and posess consistent anti fungal activity against Candida albicans and there is no significant effect on Aspergillus niger. The obtained results shows that the EM complies with the regulatory standard and also possess significant anti microbial property against the tested microbes.

Assessment of mycotoxin contamination in Rwanda: A comparison of agro-ecological zones

Aflatoxin contamination of maize in Rwanda and underlying causes are poorly documented. This study aimed to address this gap where maize samples for aflatoxins analysis were collected from two different agro-ecological zones and the temperature and relative humidity conditions during the time preceding sample collection were monitored to determine the conditions conducive to this contamination. Maize was analysed for moisture content and aflatoxin contamination and the temperature and humidity data were used to estimate the saturation deficit of the ambient air as an indicator of risk of contamination, which is high when the saturation deficit is low. Results showed an average lower saturation deficit of 0.0018 kg H₂O kg−1 in the high-altitude Volcanic region compared to the 0.00095 kg H₂O kg−1 recorded in the lower altitude region of the Eastern Savannah. Given the high moisture content in most of the analysed samples, this meant that it is more challenging to dry maize to the safe storage moisture content in the Volcanic region compared to the Eastern Savannah. The mean aflatoxin contamination was 40.45 μg kg−1 in the Volcanic region significantly higher than 4.66 μg kg−1 in the Eastern Savannah. Aflatoxin G1 was predominant among the contaminated samples. Affordable drying technologies that use renewable sources of energy should be developed and made accessible to farmer cooperatives.

Investigation of total aflatoxin in nuts and dried fruits and consumption habits during pregnancy in Türkiye

Background: During pregnancy, adequate energy, nutrient intake, and food safety are important. Aims: This study aimed to examine the presence of aflatoxin, which has toxigenic properties, in nuts and dried fruits consumed during pregnancy. Subjects and Methods: Forty-five pregnant women living in Istanbul and Balikesir cities provinces, who were selected using the convenience sampling method, were applied face-to-face interview method on their attitudes and behaviors regarding their preferences for consuming nuts and dried fruits and storage conditions between February and April 2022. For aflatoxin analysis, the samples were taken from the nuts and dried fruits consumed by the pregnant women. The total aflatoxin content in the samples was analyzed by the ELISA method with a quantitative aflatoxin high-sensitivity test kit. Results: The most commonly consumed nuts by pregnant women during pregnancy; were walnuts and hazelnuts, respectively. It was determined that pregnant women buy nuts and dried fruits first from the nut shop and second from the market. Regarding storage preferences, it was seen that the participants mostly stored the nuts in the closed kitchen cabinet or refrigerator cabinet. For the storage material preferences, glass, porcelain, and packaging were preferred. For aflatoxin results, no detectable level of aflatoxin was found in 35 samples, and the presence of aflatoxin in 3 samples (1.43 ppb, 1.523 ppb, and 1.804 ppb, respectively) was detected. Conclusion: Nuts and dried fruit consumption preferences of pregnant women differed according to the products. No aflatoxin was found in the nuts and dried fruits at a level that could threaten the health of pregnant women. Keywords: Aflatoxin, Nuts, Dried Fruits, Pregnancy, ELISA method

Evidence of the effect of pre and postharvest practices on aflatoxin contamination in the Forest and Savannah ecozones of Ghana

Food contamination by aflatoxins is a global concern due to their adverse effects on food security, trade, and health. This study contributes to the evidence on the connection between pre and postharvest practices of maize farmers and the levels of aflatoxin contamination, based on a survey of 400 farmers across Forest and Savannah zones in Ghana. The survey involving a semi-structured questionnaire was conducted between September 2019 and March 2020. Information on preharvest practices such as weed control, fertilizer application, and irrigation; and postharvest practices such as drying, shelling, storage, and pest control among others were explored by the questionnaire. A multistage sampling process was employed to determine the sample size of four hundred respondents. One kg of maize was randomly sampled from the farmers for aflatoxin analysis. The results showed that the least aflatoxin type found in the maize sample was Aflatoxin G2 (0.207 μg/kg), while the highest was Afltoxin B1 (9.618 μg/kg). The Savannah zone recorded a lower contamination level of 11.63 μg/kg than the Forest zone of 23.53 μg/kg. Aflatoxin G1 and Aflatoxin G2 were less prevalent among the samples than Aflatoxins B1 and B2. The contamination levels in both zones exceeded the maximum limits, set by Codex, and EU at 10 μg/kg, and 4 μg/kg respectively. Only the Savannah zone contamination level met the maximum limit of contamination of Ghana which is set at 15 μg/kg. About 73% of farmers who demonstrated awareness of aflatoxin contamination had their produce within the maximum limit of Ghana, CODEX, and the EU. Preharvest practices such as fertilizer application, hoeing, and chemical weed control lowered aflatoxin contamination in both zones. The results showed that awareness creation and adoption of pre and postharvest interventions are vital in reducing aflatoxin contamination in the two ecozones.

Simultaneous evaluation and monitoring ochratoxin A, aflatoxins, and zearalenone contamination levels of cereals from Iranian retail market using HPLC-FLD: A 3-year survey

Mycotoxins are important for farmers and public health. Due to the widespread use of cereals around the world, their monitoring is of great importance. In this study, the simultaneous determination and analysis of aflatoxins (AFs), ochratoxin A (OTA), and zearalenone (ZEN) in Iranian cereal samples, including 150 rice, 150 wheat, and 150 corn samples, were carried out using an immunoaffinity column and analyzed using HPLC-FLD. Under the optimal conditions, the linear range was 0.01–30, 0.03–40 and 0.2–350 ng g-1 for AFs, OTA, and ZEN, respectively. The detection of limit was between 0.0028–0.0035, 0.015 and 0.061 ng g−1, and the qualitative of limit was between 0.0093–0.015, 0.052 and 0.203 for AF, OTA, and ZEN, respectively. In conclusion from the 450 cereal samples, only 7 % contained mycotoxins, and the maximum residue limit (MRL) were less than 1 %. These results show that grain products in Iran are well controlled by the Health and Food and Drug Organization.

Reduced graphene oxide-zinc iron oxide nanomaterial as selective dispersive solid-phase extraction sorbent for extraction and enrichment of aflatoxins from milk combined with UHPLC-MS/MS analysis

Fungal species capable of producing aflatoxins frequently contaminate the feed provided to dairy animals, leading to the subsequent occurrence of aflatoxin contamination in milk. Milk is renowned for its exceptional nutritional composition, and the consumption of milk contaminated with aflatoxins by humans can engender a multitude of health complications. In this study, reduced graphene oxide zinc iron oxide (rGO-Zn/Fe2O4) nanomaterial was utilized as a dispersive solid phase extraction adsorbent to separate and enrich aflatoxins (aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, and aflatoxin M1) from milk. The rGO-Zn/Fe2O4 nanomaterial was morphologically characterized by scanning electron microscopy, transmission electron microscopy, and X-ray diffraction, indicating that Zn/Fe2O4 nanoparticles were embedded on the surface of rGO. The acetonitrile/formic acid (99/1, v/v) combined with ultrasonic treatment for a duration of 40 min expressed an efficient extraction solvent. Employing the matrix-spiked technique, different parameters of the dispersive solid-phase extraction (d-SPE) method were meticulously optimized, yielding the following conditions: 2% acetonitrile as loading solvent, 15 mg of rGO-Zn/Fe2O4 as the adsorbent quantity, 15 min of ultrasonication as adsorption time, and n-hexane as the washing solvent. The desorption reagent was 5 mL acetonitrile, with a desorption time of 5 min. The combination of the dispersive solid-phase extraction (d-SPE) technique with validated UHPLC-MS/MS quantitative analysis demonstrated excellent selectivity, a robust linear relationship (R2 ≥ 0.994), remarkable sensitivity (limit of quantification ranging from 0.006 to 0.013 µg/L), and satisfactory recovery rates of 78%–104% at three different matrix-spiked levels. Moreover, the method exhibited precision with relative standard deviations (RSD) ranging from 2.4% to 10.8%. Following appropriate pretreatment procedures, the milk samples underwent rigorous aflatoxin analysis, leading to the identification of AFM1 presence in all of the examined milk samples (n = 9). However, the contamination levels detected were found to be within the permissible limits as defined by regulatory standards.

Analysis of Heavy metal, Aflatoxin, Pesticide Residue and Microbial Contamination of Siddha Herbal Formulation Muppirandai Chooranam

Aim: The aim of the study was to evaluate the presence of Heavy metal, Aflatoxin, Pesiticide residue and Microbial contamination of Siddha herbal formulation Muppirandai Chooranam (MRC).
 Place of study: Heavy metal analysis, Aflatoxin assay, Pesiticide residue, Microbial contamination analysis were conducted at Noble Research Solutions, Kolathur, Chennai -99.
 Methodology: The Siddha formulation Muppirandai Chooranam was prepared as per Good Manufacturing Practices(GMP) guidelines and the Heavy metal analysis, Aflatoxin assay, Pesiticide residue, Microbial contamination analysis were conducted at Noble Research Solutions, Kolathur, Chennai -99. 
 Results: The results of Heavy metal analysis of Muppirandai Chooranam (MRC) shown the presence of Lead at 3.54 PPM and Arsenic, Cadmium, Mercury at Below Detection Limit (BDL). Aflatoxin assay of MRC shown the absence of Aflatoxin B1, Aflatoxin B2, Aflatoxin G1, Aflatoxin G2. Pesticide residue analysis showed that there were no traces of Pesticide residues such as Organo chlorine, Organo phosphorus, Organo carbamates and Pyrethroids. In Microbial contamination analysis, Test for Specific Pathogen shown the absence of Organisms E-coli, Salmonella, Staphylococcus Aureus, Pseudomonas Aeruginosa and No Bacterial and fungal growth or colonies were observed in the Sterility test of MRC as per the methods of AYUSH specifications.
 Conclusion: From the results, it is concluded that the study medicine MRC has Heavy metal content below the permissible limit as per PLIM guidelines of AYUSH, and the sample were free from Aflatoxins, Pesiticides, Microbes, and Specific Pathogens which ensures that the study medicine Muppirandai Chooranam was safe therapeutically.