Aflatoxin B1 in betel nuts (Areca catechu L.) imported to Pakistan from different regions of South Asia

Aflatoxin B₁ (AFB₁) levels were evaluated in betel nuts (Areca catechu L.) being imported to Pakistan during 2010–2011. In total, 278 betel nut samples (India = 21, Indonesia = 51, Sri-Lanka = 34 and Thailand = 172) were received from the Department of Customs and were analysed by thin layer chromatography (TLC). All Indian origin betel nuts showed AFB₁ contamination ranging from 11.7–262.0 µg kg^?1 with a mean of 92.5 µg kg^?1. Among Indonesian and Sri Lankan shipments, 80.4% and 73.5% betel nuts were contaminated with AFB₁ ranging between 3.3–39.2 and 6.5–103.4 µg kg^?1 with a mean of 11.6 and 35.0 µg kg^?1, respectively. However, only 30.2% of Thailand origin samples showed AFB₁ contamination ranging 3.3–77.0 µg kg^?1 with a mean of 6.6 µg kg^?1. The widespread occurrence of AFB₁ increases the hazard associated with betel nuts. Thus, strict control is a pre-requisite for the production and import/export of psychoactive substances as betel nuts.     

Aflatoxins in animal feed in Iran

One hundred and forty-six samples of animal feed (barley, n = 60; wheat bran, n = 22; wheat dry pulp, n = 29; and canola meal, n = 35) were collected in 2011 from Mashhad (Khorasan, Iran). Aflatoxins (AFs) were determined in these samples after immunoaffinity column clean-up by high-performance liquid chromatography (HPLC) with fluorescence detection. Aflatoxin B₁ (AFB₁) contamination was found in 28 samples: in five of the barley samples (8.3%) at a mean level of 0.48 µg·kg^?1, in two wheat bran samples (9.0%) at a mean level of 0.88 µg·kg^?1, in 10 wheat dry pulp samples (34.5%) at a mean level of 0.30 µg·kg^?1 and in 11 canola meal samples (31.4%) at a mean level of 0.92 µg·kg^?1. AFB₁ levels were below the maximum levels of Iran regulations (5 µg·kg^?1) and the EU maximum limit (5 µg·kg^?1).     

Relative severity of fumonisin contamination of cereal crops in West Africa

Traditional and improved varieties of maize, pearl millet and sorghum were planted by small-scale farmers under the direction of the International Institute for Tropical Agriculture in two Nigerian agro-ecological zones: the Sudan Savanna and the Northern Guinea Savanna. Samples were collected for the determination of Fusarium infection and fumonisin (B₁, B₂ and B₃) contamination. A previous paper reported Aspergillus infection and aflatoxin contamination of these samples. Fusarium infection levels, measured by per cent kernels infected, were modest with mean levels for the above cereals of 16% ± 11% (SD), 12% ± 7% and 13% ± 16%, respectively. However, the Fusarium species recovered from maize were predominantly the fumonisin producers F. verticillioides and F. proliferatum, together making an infection rate of 15% ± 10%, whereas these species were present to a limited extent only in the other two cereals, 1% ± 1% for pearl millet and 2% ± 6% for sorghum. Fumonisin contamination was variable but reflected the diversity of Fusarium producers in these three cereals. Mean levels were 228 ± 579 µg kg^–1 (range < 5–2860 µg kg^–1) for maize, 18 ± 7 µg kg^–1 (range = 6–29 µg kg^–1) for pearl millet and 131 ± 270 µg kg^–1 (range < 5–1340 µg kg^–1) for sorghum. Together with previous results on aflatoxin, this study confirmed the co-occurrence of aflatoxins and fumonisins in maize as well as in the traditional African cereals, millet and sorghum (89% co-occurrence across all three cereals). The low fumonisin levels may be ascribed to the use of good agricultural practices. Of the Fusarium species present, those in maize consisted mainly of fumonisin producers, the opposite of what was observed in pearl millet and sorghum. It is concluded that replacement of maize by pearl millet and sorghum could improve food safety with regards to aflatoxin B and fumonisin B exposure.     

Distribution of aflatoxins and fumonisins in dry-milled maize fractions

The aim was to evaluate the distribution of aflatoxins and fumonisins in fractions derived from the dry-milling of contaminated maize. Two maize lots with different contamination levels were processed and sampled: the first (maize 1) had aflatoxin B₁ (AFB₁) and fumonisin B₁ (FB₁) levels of 3.6 and 5379 µg kg^?1, respectively; the second (maize 2) had corresponding levels of 91.1 and 8841 µg kg^?1, respectively. The cleaning step reduced AFB₁ and FB₁ levels by 8 and 11% in maize 1 and by 57 and 34% in maize 2. The subsequent removal of bran and germ led to a further decrease in contamination levels in the products destined for human consumption. In the latter, AFB₁ was uniformly distributed, while FB₁ was concentrated in the finer size fractions. Contamination of raw maize 1 (3.6 µg kg^?1) was below the European Union AFB₁ limit of 5 µg kg^?1 for unprocessed maize, but among the final products only coarse flour (1.7 µg kg^?1) was within the European Union limit of 2 µg kg^?1, while grits and fine flour showed higher levels (2.7 and 2.5 µg kg^?1, respectively). As regards cleaned maize, a different distribution of the two toxins was observed in the kernels: AFB₁ contamination was more superficial and concentrated in germ, while FB₁ contamination affected the inner layers of the kernels.     

Aflatoxins contamination of maize in Serbia: the impact of weather conditions in 2015

ABSTRACT

In recent years climate changes recorded in temperate regions of Europe have led to aflatoxin (AF) contamination of maize. Thus, the aim of this study was to investigate the influence of weather conditions on levels of aflatoxin B₁ (AFB1), aflatoxin B₂ (AFB2), aflatoxin G₁ (AFG1) and aflatoxin G₂ (AFG2) in 180 maize samples collected from the main maize-growing regions (Western Ba?ka, North Banat, South Banat and Central Serbia) in Serbia after harvest in 2015. The concentrations of AFs were determined by a validated HPLC method with post-column derivatisation and fluorescence detection (HPLC-FLD). The presence of AFB1, AFB2, AFG1 and AFG2 was detected in 57.2%, 13.9%, 5.6% and 2.8% of maize samples in the concentration ranges of 1.3–88.8 µg kg^–1, 0.60–2.8 µg kg^–1, 1.8–28.5 µg kg^–1 and 2.1–7.5 µg kg^–1 respectively. The recorded smaller amount of precipitation and especially higher air temperatures during the summer of 2015 were favourable for AF production, which resulted in 32.2% and 21.1% of samples being unsuitable for human consumption, since AFB1 and the sum of AFs concentrations were above 5.0 and 10.0 µg kg^–1 respectively. Furthermore, the findings in this study indicate that the microclimate conditions in the investigated regions had a great influence on the contamination frequency of maize with AFs. The highest percentage of samples unsuitable for human consumption, considering both AFB1 and total AFs content were 72.5% and 51.5% respectively from Central Serbia, whilst the lowest percentages of 15.6% and 6.2% respectively were found in Western Ba?ka. These findings confirmed that maize should be continuously monitored in order to protect human and animal health from the harmful effects caused by AFs contamination.     

Aflatoxin B1 and total fumonisin contamination and their producing fungi in fresh and stored sorghum grain in East Hararghe,Ethiopia

Natural contamination of sorghum grains by aflatoxin B₁ and total fumonisin and their producing toxigenic fungi has been studied. A total of 90 sorghum grain samples were collected from small-scale farmers’ threshing floors and 5–6 months later from underground pits during 2013 harvest from three districts of East Hararghe, Ethiopia. Mycotoxin analysis was done using enzyme-linked immunosorbent assay (ELISA). The limits of detection were in the range 0.01–0.03 μg kg^–1. The results revealed that all sorghum grain samples were contaminated with both Aspergillus and Fusarium species. Aflatoxin B₁ was detected at levels ranging from ?1 grain. There were marked variations in aflatoxin B₁ concentrations between fresh and stored samples, with much higher levels in the latter. Total fumonisin levels varied between 907 and 2041 µg kg^?1 grain across the samples. Lowest total fumonisin was recorded in freshly harvested sorghum grain samples. Sorghum is a main staple cereal in the studied districts and its consumption per day per person is high. Daily intake of low doses of mycotoxin-contaminated food stuff over a period of time could lead to chronic mycotoxicosis.     

Survey of 11 mycotoxins in wheat flour in Hebei province,China

A survey of 11 mycotoxins in 348 wheat flour samples marketed in Hebei province of China were analysed by liquid chromatography-tandem mass spectrometry, was carried out. The selected mycotoxins consisted of four aflatoxins (AFs: AFB₁, AFB₂, AFG₁ and AFG₂) and seven Fusarium toxins, i.e. deoxynivalenol (DON), nivalenol, 3-acetyldeoxynivalenol and 15-acetyldeoxynivalenol, zearalenone, Fusarenon-X and deoxynivalenol-3-glucoside. Results indicated that most of the wheat samples analysed were contaminated with mycotoxins. Wheat was most susceptible to DON (91.4% contamination), with a mean level of 240 μg kg^?1. On average the probable daily intake (PDI, expressed as µg kg^?1 body weight day^?1) of mycotoxins was within the provisional maximum tolerable daily intake (PMTDI, 2.0 µg kg^?1 of body weight day^?1) as set by the Joint FAO/WHO Expert Committee on Food Additives. Nevertheless, exposure assessment revealed that the maximum PDI of mycotoxins was 4.06 µg kg^?1 body weight day^?1, which was twice the PMTDI value. Thus, consistent monitoring is recommended, as to keep the contamination level under control.     

Aflatoxins in composite spices collected from local markets of Karachi,Pakistan

This survey was carried out to evaluate the occurrence of total aflatoxins (AFs; B₁+B₂+G₁+G₂) in unpacked composite spices. A total of 75 samples of composite spices such as biryani, karhai, tikka, nihari and korma masalas were collected from local markets of Karachi, Pakistan, and analysed using HPLC technique. The results indicated that AFs were detected in 77% (n = 58) samples ranging from 0.68 to 25.74 µg kg^?1 with a mean of 4.63 ± 0.95 µg kg^?1. In 88% (n = 66) samples, AFs level was below the maximum limits (ML = 10 µg kg^?1) as imposed by EU. Furthermore, 61% (n = 46) tested samples contained AFs level between 1 and 10 µg kg^?1, 9% (n = 7) exhibited AFs contamination ranged 10?20 µg kg^?1 and only 3% (n = 2) of the investigated samples contained AFs levels higher than the ML of 20 µg kg^?1 for total aflatoxins as set by the USA. It was concluded that there is need to establish a strict and continuous national monitoring plan to improve safety and quality of spices in Pakistan.     

Co-occurrence of aflatoxins,ochratoxin A and citrinin in “egusi” melon (Colocynthis citrullus L.) seeds consumed in Ireland and the United Kingdom

The natural co-occurrence of aflatoxins (AFs), ochratoxin A (OTA) and citrinin (CIT) in melon seed samples obtained from retailers and households in Ireland and the United Kingdom (UK) was evaluated. AFs and OTA were determined by HPLC with fluorescence detection while CIT was analysed by HPLC-MS/MS. AFB₁ was detected in all (100%) samples (mean = 9.7 μg kg^?1; range = 0.2–66.5 μg kg^?1). Mean total AFs was 12.0 μg kg^?1 (range = 0.3–82 μg kg^?1). Commercially retailed samples showed a significantly higher AFB₁ contamination (p < 0.05) than the household samples. OTA occurred in 3 (13.6%) samples, while 4 (18.2%) were contaminated with CIT at very low levels. In this study, 68% of the melon seed samples were contaminated above the 2 μg kg^?1 EU limit for AFB₁ in oilseeds. These results highlight the need for the development of strategies to reduce AF contamination in “egusi” for human consumption.     

Survey of aflatoxins in maize tortillas from Mexico City

In Mexico, maize tortillas are consumed on a daily basis, leading to possible aflatoxin exposure. In a survey of 396 2-kg samples, taken over four sampling days in 2006 and 2007 from tortilla shops and supermarkets in Mexico City, aflatoxin levels were quantified by HPLC. In Mexico, the regulatory limit is 12?µg?kg^?1 total aflatoxins for maize tortillas. In this survey, 17% of tortillas contained aflatoxins at levels of 3–385?µg?kg^?1 or values below the limit of quantification (<LOQ) and, of these, 13% were >12?µg?kg^?1 and 87% were below the regulatory limit. Average aflatoxin concentrations in 56 contaminated samples were: AFB₁ (12.1?µg?kg^?1); AFB₂ (2.7?µg?kg^?1); AFG₁ (64.1?µg?kg^?1) and AFG₂ (3.7?µg?kg^?1), and total AF (20.3?µg?kg^?1).     

Transfer of aflatoxin B1 and fumonisin B1 from naturally contaminated raw materials to beer during an industrial brewing process

The aim of this research was to determine the fate of aflatoxins (AFs) and fumonisins (FBs) naturally occurring in raw materials (maize grit and malted barley) during four industrial brewing processes. The aflatoxin B₁ (AFB₁) level in raw materials varied from 0.31 to 14.85 µg kg^?1, while the fumonisin B₁ (FB₁) level (only in maize grit) varied from 1146 to 3194 µg kg^?1. The concentration in finished beer ranged from 0.0015 to 0.022 µg l^?1 for AFB₁ and from 37 to 89 µg l^?1 for FB₁; the other aflatoxins and fumonisin B₂ were not found in beer samples. The average percentage of toxins recovered in finished beer, referring to the amounts contained in raw materials, were 1.5% ± 0.8% for AFB₁ and 50.7% ± 4.7% for FB₁. These results were mainly due to the different solubility of the two mycotoxins during the mashing process. If raw materials comply with the limits fixed by European Commission Regulations, the contribution of a moderate daily consumption of beer to AFB₁ and FB₁ intake does not contribute significantly to the exposure of the consumer.     

Fumonisin B1 in cereal mixtures marketed in Brazil

The aim of this study was to detect and quantify fumonisin B₁ (FB₁) in cereal mixtures marketed in Brazil. Fifteen samples from different lots were acquired each month by internet from supermarkets during seven months, adding up to 105 analysed samples. The unit sample constituted of an original package with a minimum of 250 g. Extraction and clean-up of samples for FB₁ determination were carried out using immunoaffinity columns. Identification and quantification of FB₁ were performed by high performance liquid chromatography. Eighty-eight (83.8%) samples were contaminated with FB₁ and four (3.8%) presented levels above 500 µg kg^?1 (634, 703, 1269 and 1876 µg kg^?1). Maximum FB₁ + FB₂ levels allowed by Brazilian regulations will reach 1500 µg kg^?1 for corn flour in 2016 and 1000 µg kg^?1 for others corn products. This study showed that even at levels below the legislative limits, human exposure to this toxin can occur constantly.     

Household dietary exposure to aflatoxins from maize and maize products in Kenya

Aflatoxicosis has repeatedly affected Kenyans, particularly in the eastern region, due to consumption of contaminated maize. However, save for the cases of acute toxicity, the levels of sub-lethal exposure have not been adequately assessed. It is believed that this type of exposure does exist even during the seasons when acute toxicity does not occur. This study, therefore, was designed to assess the exposure of households to aflatoxins through consumption of maize and maize products. Twenty samples each of maize kernels, muthokoi and maize meal were randomly sampled from households in Kibwezi District of Makueni County in Eastern Kenya and analysed for aflatoxin contamination. The samples were quantitatively analysed for aflatoxin contamination using HPLC. The uncertainty and variability in dietary exposure was quantitatively modelled in Ms Excel using Monte Carlo simulation in @Risk software. Aflatoxins were found in 45% of maize kernels at between 18 and 480 μg kg^–1, 20% of muthokoi at between 12 and 123 μg kg^–1, and 35% of maize meal at between 6 and 30 μg kg^–1. The mean dietary exposure to aflatoxin in maize kernels was 292 ± 1567 ng kg^?1 body weight day^?1, while the mean dietary exposure to aflatoxin in maize meal and muthokoi were 59 ± 62 and 27 ± 154 ng kg^?1 body weight day^?1 respectively. The results showed that the amount and frequency of consumption of the three foods is the more important contributing factor than the mean aflatoxin concentration levels, to the risk of dietary exposure to aflatoxins.     

Aflatoxin B1 in eggs and chicken livers by dispersive liquid–liquid microextraction and HPLC

A rapid, low-cost and simple technique has been developed for the determination of aflatoxin B₁ (AFB₁) in eggs and livers using high-performance liquid chromatography (HPLC) with UV detection. In this study, the presence of AFB₁ was investigated in 150 eggs and 50 chicken livers from the local market of Tabriz, Iran. AFB₁ was extracted with a mixture of acetonitrile:water (80:20) and cleaned up by dispersive liquid–liquid microextraction which is a very economical, fast and sensitive method. AFB₁ was quantified by HPLC-UV without need for any complex derivatisation in samples to enhance the detection. The results showed that 72% of the liver and 58% of the egg samples were contaminated with AFB₁ ranging from 0.30 to 16.36 µg kg ^?1. limit of detection and limit of quantification for AFB₁ were 0.08 and 0.28 µg kg ^{? 1}, respectively. The proposed method is suitable for fast analysing of AFB₁ in egg and liver samples.     

Aflatoxin M1 in raw milk from different regions of São Paulo state – Brazil

A total of 635 raw milk samples from 45 dairy farms, from three regions of São Paulo state – Brazil, were evaluated during 15 months for aflatoxin M₁ (AFM₁). AFM₁ was determined by high performance liquid chromatograph with fluorescence detection. AFM₁ was detected (>0.003 µg kg^?1) in 72.9%, 56.3% and 27.5% of the samples from Bauru, Araçatuba and Vale do Paraíba regions, respectively. The mean AFM₁ contamination considering all the samples was 0.021 µg kg^?1. Furthermore, the concentration of AFM₁ was quite different among Bauru (0.038 µg kg^?1), Araçatuba (0.017 µg kg^?1) and Vale do Paraíba (<0.01 µg kg^?1) regions. Only three samples (0.5%) had higher contamination than the tolerated limit in Brazil (0.50 µg kg^?1) and 64 samples (10.1%) had a higher contamination than the maximum limit as set by the European Union (0.050 µg kg^?1). The estimated AFM₁ daily intake was 0.358 and 0.120 ng kg^?1 body weight per day for children and adults, respectively.     

Natural co-occurrence of ochratoxin A,ochratoxin B and aflatoxins in Sicilian red wines

The natural occurrence of ochratoxin A, ochratoxin B, aflatoxin B₁, aflatoxin B₂, aflatoxin G₁ and aflatoxin G₂ (OTA, OTB, AFB₁, AFB₂, AFG₁, AFG₂) in red wines was investigated by HPLC/FLD after immunoaffinity column clean-up in 57 market samples produced in Sicily (Italy). The results showed a very low incidence of these mycotoxins in analysed samples, confirming the high degree of quality and safety of Sicilian red wines. The results indicated 71.9% and 64.9% positive samples for OTA and OTB respectively, with an average level of 0.13 μg l^–1, well below the European maximum permitted levels (MLs). The aflatoxin most frequently detected in the samples was AFG₁, present in 57.9% of samples, while the other aflatoxins were rarely present. Recovery experiments were carried out on eight mycotoxin-free red wines spiked with OTA, OTB, AFB₁, AFB₂, AFG₁ and AFG₂ at two different levels. The limits of detection (LODs) in wines were 0.02 µg l^–1 for OTA, 0.04 µg l^–1 for OTB, 0.03 µg l^–1 for AFG₁, AFG₂ and AFB₂, and 0.05 µg l^–1 for AFB₁. A good correlation was found, with good performances in term of precision for the method.     

Assessment of dietary exposure to aflatoxin B1 from corn arepas in Colombia

ABSTRACT

The aim of this research was to evaluate dietary exposure to aflatoxin B1 (AFB₁) in corn arepas in Colombia. In addition, in this study an assessment considering compliance to the maximum level of AFB₁ (4 µg kg^?1) for this food was conducted. AFB₁ concentration data in corn arepas were obtained from 168 samples. The samples were collected from factories in 16 departments in Colombia. AFB₁ was quantified by High-Performance Liquid Chromatography with a fluorescence detector. Consumption data and body weight (b.w.) were measured from the 2005 Colombian National Survey of Nutritional Status. Probabilistic estimates were made by Monte Carlo simulation of dietary exposure and margin of exposure (MOE) segmented by age group. The results showed that 27% of corn arepa samples were contaminated with AFB₁, with an average concentration of 15.1 µg kg^?1 and a maximum value of 111.1 µg kg^?1. The stochastic dietary exposure assessment showed that the age group most exposed was children between 4 and 8 years old (10.014 ng (kg b.w. day)^?1). In addition, the MOE values for all age groups were lower than 10,000, indicating a potential risk for consumers. However, in the scenario where AFB₁ concentration level complies with the maximum limit of 4 µg kg^?1 AFB₁, the level of concern could be reduced for the adult population between 14 and 64 years old because the MOE value is above 10,000.     

Quantitation of aflatoxins in walnut kernels by high-performance liquid chromatography with fluorescence detection

A total of 85 walnut samples collected between October 2012 and April 2013 in different provinces of Turkey were analysed for the presence of aflatoxins (AFs). The method involved methanol–water extraction, clean-up with immunoaffinity columns and a sensitive high-performance liquid chromatography coupled with fluorescence detection after post-column derivatisation. The method was validated for selectivity, linearity, trueness, precision, limit of detection and limit of quantification (LOQ), which met the performance criteria as set by EC regulation No. 401/2006. LOQs were 0.07, 0.04, 0.09 and 0.05 µg kg^–1 for AFB₁, AFB₂, AFG₁ and AFG₂, respectively. AFs were present in 9.4% of walnut samples (8/85) at total AFs levels ranging from 0.09 to 15.4 µg kg^–1. Only one of eight walnut samples exceeded the European Union limit of 2 and 4 µg kg^–1 for AFB₁ and total AFs, respectively.     

Aflatoxin B1 in peanut oil from Western Guangdong,China, during 2016–2017

Aflatoxin B₁ (AFB₁) occurrence in peanut oil samples randomly collected from family workshops in western Guangdong during 2016–2017 (n = 427) was surveyed. AFB₁ content was screened by enzyme-linked immunosorbent assay (ELISA) protocol and analytically confirmed with an UPLC-MS/MS method. The limit of detection of the ELISA method was 1.08 μg kg^?1. The recovery values ranged from 84.4 to 92.6% with relative standard deviations of 2.2 to 4.8%. AFB₁ was quantified in 47 samples (22.5%) with a range of 15.4–49.9 μg kg^?1 in 2016 and in 33 samples (15.1%) with 8.8–22.2 μg kg^?1 in 2017, respectively. The AFB₁ contamination in peanut oil was season-depended in western Guangdong with the worst case in spring (24.2–37.9%). Overall, a significant reduction of AFB₁ occurrence was observed in western Guangdong after 2016. It is advisable to control AFB₁ in bulk and self-pressed oil from family workshops and regulate it mandatorily if necessary.     

Aflatoxin occurrence in milk and supplied concentrates of goat farms of north‐eastern Italy

BACKGROUND: There is little information about the occurrence of aflatoxin M₁ in goat milk. A survey involving 17 dairy goat farms of north‐eastern Italy was completed during 2005 and 2006, in order to evaluate the prevalence of milk contamination and its relationship with type and level of concentrate supplied. RESULTS: 132 concentrate and 85 milk samples were collected during five farm visits and analysed for aflatoxins. Aflatoxin B₁ (AFB₁) was > 0.1 µg kg^?1 in two‐thirds of the feeds and > 5 µg kg^?1 in nine. Contamination was higher in maize than in other pure feeds (median: 0.8 versus 0.1 µg kg^?1); complementary feeds showed intermediate values. Aflatoxin M₁ (AFM₁) was > 3 ng kg^?1 in one‐third of milks and > 25 ng kg^?1 in three. All the milk samples were below EU statutory limits. The farm ranks for milk AFM₁ level and the peak of concentrate AFB₁ contamination were significantly correlated (0.642). CONCLUSIONS: Risk to human health was generally found to be absent, with only a few cases involving feed contamination to be monitored. The main aflatoxin risk for goat milk could arise from maize and maize‐based concentrates in the more intensive breeding conditions. Copyright © 2008 Society of Chemical Industry