Aflatoxins and ochratoxin A in export quality raisins collected from different areas of Pakistan

During 2012–2014, 170 samples of export quality raisins were collected from different vendors in Pakistan. The collected samples were analysed for the presence of aflatoxins (AFs) and Ochratoxin A (OTA) contamination using high-performance liquid chromatography technique. The limit of detection and limit of quantification of AFs/OTA were 0.12/0.10 and 0.36/0.30 µg kg^?1, respectively. Only 5% of the samples were contaminated with AFs, ranging 0.15–2.58 µg kg^?1 with a mean of 0.05 ± 0.26 µg kg^?1. None of the raisin samples exhibited AFs contamination above the maximum limit (ML = 4 µg kg^?1) as set by the European Union (EU). About 72% of the samples were contaminated with OTA, ranging 0.14–12.75 µg kg^?1 with a mean of 2.10 ± 1.9 µg kg^?1. However, in 95.3% of the tested samples, OTA level was lower than the ML of 10 µg kg^?1 as regulated by the EU. Apparently, a strict and continuous monitoring plan, including regulatory limits, improves food safety and quality for all types of commodities.     

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).     

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.     

Aflatoxins in dairy cow feed,raw milk and milk products from Turkey

This study aims to detect aflatoxins (AFs) in dairy cow feed, milk and milk products using a high-performance liquid chromatography coupled with fluorescence detection (HPLC-FLD) method. All the validation parameters met the method performance criteria of the European Union. The samples comprised 76 dairy cow feeds and 205 milk and milk products (including yoghurt and yoghurt-based beverage, ayran). AFs were present in 26.3% of the feed samples. Two feed samples exceeded the maximum limit (ML) of 5 µg kg^?1 for AFB₁ as established by the EU. Nineteen milk samples (21.1%) contained aflatoxin M₁ (AFM₁) of which three exceeded the EU ML of 0.05 µg l^?1. In addition, only two yoghurt samples and one ayran sample contained AFM₁, but the levels were lower than the EU ML.     

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.     

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.     

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.     

Aflatoxins and heavy metals in animal feed in Iran

The occurrence of aflatoxin (aflatoxin B₁, aflatoxin B₂, aflatoxin G₁ (AFG₁) and aflatoxin G₂ (AFG₂)) and heavy metal (Pb, Cd, As and Hg) contamination was determined in 40 industrially produced animal feed samples which were collected from the southwest of Iran. The results indicated that 75% of samples were contaminated by four aflatoxins and the level of AFB₁ and sum of aflatoxins were higher than the permissible maximum levels in Iran (5 and 20 µg kg^?1, respectively) in all feed samples. A positive correlation was found between four types of aflatoxins in all the tested samples (p < 0.01) and the positive correlation between AFG₁ and AFG₂ was significant (r² = 0.708). All feed samples had lead concentrations lower than the maximum EU limit, while 5%, 17% and 42.5% of feed samples had As, Cd and Hg concentrations higher than the maximum limits, respectively.     

Mycotoxin contamination of home-grown maize in rural northern South Africa (Limpopo and Mpumalanga Provinces)

The aim of this study was to assess mycotoxin contamination of crops grown by rural subsistence farmers over two seasons (2011 and 2012) in two districts, Vhembe District Municipality (VDM, Limpopo Province) and Gert Sibande District Municiality (GSDM, Mpumalanga Province), in northern South Africa and to evaluate its impact on farmers’ productivity and human and animal health. A total of 114 maize samples were collected from 39 households over the two seasons and were analysed using a validated liquid chromatography-tandem mass spectrometry mycotoxins method. Aflatoxin B₁ (AFB₁) occurrence ranged from 1 to 133 µg kg^?1 in VDM while AFB₁ levels in GSDM were less than 1.0 µg kg^?1 in all maize samples. Fumonisin B₁ levels ranged from 12 to 8514 µg kg^?1 (VDM) and 11–18924 µg kg^?1 (GSDM) in 92% and 47% positive samples, respectively, over both seasons. Natural occurrence and contamination with both fumonisins and aflatoxins in stored home-grown maize from VDM was significantly (p < 0.0001) higher than from GSDM over both seasons.     

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.     

Survey of aflatoxins in chillies from Pakistan produced in rural,semi-rural and urban environments

Chilli peppers from Pakistan are consumed locally and also exported. Their quality is compromised by aflatoxins (AF) contamination. AF in chillies from rural, semi-rural and urban areas of the Punjab region of Pakistan were determined. Twenty-three (52.3%), 22 (50%) and 29 (65.9%) samples from rural, semi-rural and urban areas respectively, contained levels of aflatoxins which exceeded the European Union limits of >5?µg?kg^?1 for AFB₁ and >10?µg?kg^?1 for total AF that apply to spices. Mean values for AFB₁ in ground samples were 23.8, 14.8 and 14.0?µg?kg^?1 for rural, semi-rural and urban areas, respectively. Mean total AF in ground samples were 27.7, 17.7 and 16.2?µg?kg^?1 from equivalent locations. Eleven (50%), 12 (54.5%) and 14 (63.6%) whole samples from rural, semi-rural and urban areas, respectively, contained total levels of AF that exceeded European Union limits. The data indicate that individual localities have particular problems. In conclusion, the concentrations were often greater than the statutory limits set by the European Union.     

Co-occurrence and distribution of deoxynivalenol,nivalenol and zearalenone in wheat from Brazil

Fusarium mycotoxins deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEN) were investigated in wheat from the 2009 and 2010 crop years. Samples (n = 745) from commercial fields were collected in four wheat producing regions (WPR) which differed in weather conditions. Analyses were performed using HPLC-DAD. Contamination with ZEN, DON and NIV occurred in 56, 86 and 50%, respectively. Also, mean concentrations were different: DON = 1046 µg kg^?1, NIV < 100 µg kg^?1 and ZEN = 82 µg kg^?1. Co-occurrence of ZEN, DON and NIV was observed in 74% of the samples from 2009 and in 12% from 2010. Wet/cold region WPR I had the highest mycotoxin concentration. Wet/moderately hot region WPR II had the lowest mycotoxin levels. Furthermore, the mean concentration of each mycotoxin was higher in samples from 2009 as compared with those from 2010. Precipitation during flowering or harvest periods may explain these results.     

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.     

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.     

Raman Spectroscopic and Mass Spectrometric Determination of Aflatoxins

This paper reported on selective Raman spectroscopic and mass spectrometric determination of functionalized aflatoxins. Performance parameter methods encompassed six component mixtures of AFs, B₁, G₁, M₁ and B_2A, G_2A, and M_2A at concentration range of ∈ 0.09₆–21.58₁?ng g^?1. Limits of detection (LODs) and limits of quantitation (LOQ) are 0.13₅ and 0.45 ng g^?1, respectively. The matrix effect of AFB_2A, AFG_2A, and AFM_2A in multicomponent mixtures is evaluated. The LODs of 0.11–0.09 ng g^?1 is achieved. Raman spectroscopic method performance parameters at concentration range of ∈ 0.10–5,000 μg kg^?1 for six component mixtures (r ²?=?0.9765₁) are LOD?=?10.13 μg kg^?1 and LOQ?=?33.7₆?μg kg^?1. The designed experimental schedule and sampling model is tested on contaminated grain samples. As far as the biochemistry and genetic engineering of aflatoxins and their major metabolites involved the high regioselectivity and binding affinity to N7 guanine residue at DNA, causing carcinogenic and mutagenic effects, the data reported are of emergency on the frontier fields of analytical chemistry, environmental bioengineering, monitoring, and assessment of human health risk from aflatoxins contamination. Aflatoxins are considered as biochemical weapons. Therefore, the essence of the study is on the instrumental methodology, which would be of interest and encompassed public security research and related scientific elaborations of more effective methods for (1) prevention, (2) reducing of the damages, and/or (3) acquisition of the outgrowths from biological (chemical) terrorist attacks.     

Survey on acrylamide in roasted coffee and barley and in potato crisps sold in Italy by a LC–MS/MS method

ABSTRACT

A survey on the occurrence of acrylamide (AA) in roasted coffee, barley, and potato crisps was carried out using an intra-lab validated liquid chromatography (LC)–MS (mass spectrometry)/MS method. Over the years 2015–2016, 66 samples of coffee, 22 of roasted barley, and 22 of potato crisps were collected from retail outlets in Italy. AA was detected in almost all samples. In roasted coffee, the level exceeded 450 µg kg^?1, the limit recommended by the European Commission (EC), in 36.4% of the samples. In roasted barley, mean contamination was slightly lower than in coffee and no sample exceeded the EC limit of 2000 µg kg^?1. The AA contamination in potato crisps was remarkable. A percentage of 36.4 (n = 8) showed a value higher than the EC limit of 1000 µg kg^?1. Considering the average consumption of coffee and potato crisps by Italian people, AA exposure is significant and should be decreased.     

Determination and surveillance of hydrocortisone and progesterone in livestock products by liquid chromatography-tandem mass spectrometry

A simple analytical method for the determination of hydrocortisone and progesterone in bovine, swine, and chicken muscle and eggs was developed. Hydrocortisone and progesterone were extracted with acetonitrile and subsequently cleaned-up using an Oasis^® HLB mini-cartridge. The method was validated in accordance with Japanese guidelines and exhibited trueness from 86.6% to 104.3% and precision (relative standard deviations (RSDs) of repeatability and within reproducibility were under 8.7% and 11.7%, respectively). The method was applied to 103 bovine muscle, 137 swine muscle, 69 chicken muscle and 52 egg samples that were commercially available in Tokyo, Japan. The hydrocortisone concentration was 0.9–41.2 µg kg^?1 in all bovine muscle samples, with an average of 7.7 µg kg^?1 and a median of 6.2 µg kg^?1. The progesterone concentration in 50 samples exceeded the limit of quantification (LOQ) and reached a maximum of 95.4 µg kg^?1. Hydrocortisone was also detected in all swine muscle samples at concentrations of 2.0–56.0 µg kg^?1. Its average and median concentrations amounted to 13.1 and 11.3 µg kg^?1, respectively. Twenty-three samples contained progesterone levels surpassing the LOQ, with a maximum concentration of 107.0 µg kg^?1. No chicken muscle samples contained any of the analytes. The progesterone concentration was 15.5–200.0 µg kg^?1 in all egg samples, with an average of 95.4 µg kg^?1 and a median of 90.5 µg kg^?1.     

Dietary exposure to mycotoxins of the Hong Kong adult population from a Total Diet Study

Dietary exposure of Hong Kong adults to mycotoxins and their metabolites including aflatoxins (AFs), ochratoxin A (OTA), fumonisins (FNs), deoxynivalenol (DON), acetyldeoxynivalenols (AcDONs) and zearalenone (ZEA) was estimated using the Total Diet Study (TDS) approach to assess the associated health risk to the local people. Sixty commonly consumed food items, collected in four seasons, were sampled and prepared as consumed. These mycotoxins were primarily found at low levels. The highest mean levels (upper bound) were: AFs, 1.50 µg kg^–1 in legumes, nuts and seed; OTA, 0.22 µg kg^–1 in sugars and confectionery; FNs, 9.76 µg kg^–1 in cereals and their products; DON and AcDONs, 33.1 µg kg^–1 in cereals and their products; and ZEA, 53.8 µg kg^–1 in fats and oils. The estimated dietary exposures of Hong Kong adults to the mycotoxins analysed were well below the respective health-based guidance values, where available. For AFs, the upper-bound exposure for high consumers is 0.0049 µg kg bw^–1 day^–1, which was estimated to contribute to about 7.7 (< 1%) of liver cancer cases when compared with 1222 liver cancer cases per year in Hong Kong. The percentage contributions of the estimated 95th percentile dietary exposures (lower and upper bound) to the health-based guidance values of individual mycotoxins were: ochratoxin A, 3.6–9.2%; fumonisins, 0.04–8.5%; deoxynivalenol and acetyldeoxynivalenols, 21.7–28.2%; and zearalenone 3.3–34.5%. The findings indicate that dietary exposures to all the mycotoxins analysed in this study were unlikely to pose an unacceptable health risk to the Hong Kong population.     

Determination of mycotoxins in biscuits,dried fruits and fruit jams: an assessment of human exposure

ABSTRACT

A reliable, fast and simple method using UHPLC-MS/MS was developed for the determination of aflatoxins B₁ (AFB1), G₁ (AFG1), B₂ (AFB2) and G₂ (AFG2), ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZEA), HT-2 toxin and T-2 toxin in crude extracts of biscuits with fruit filling, cookies, dried fruits and fruit jams. The method was successfully demonstrated on 39 samples of biscuits with fruit filling, 34 cookies, 14 dried fruits and 10 fruit jams. The mycotoxins detected in biscuits samples were ZEA, OTA, T-2 and AFB1 with an average concentrations of positive samples of 2.64, 4.10, 8.13 and 1.32 µg kg^?1, respectively; while the mycotoxins detected in jam samples were AFB1, OTA, T-2 and AFB2 with an average concentrations of positive samples of 2.00, 17.7, 4.37 and 1.15 µg kg^?1, respectively. The results showed that the majority of samples were in compliance with relevant regulations. However in eight samples of biscuits and three samples of fig jam the contents of OTA were higher than the existing OTA limits. The combined dietary exposure of selected mycotoxins was estimated for the first time for children, adolescents and adults. The estimated combined dietary exposures were all lower than the proposed value assumed to predict a possible risk scenario.     

Occurrence of fumonisins and aflatoxins in cereals from markets of Hebei province of China

Fumonisins B₁, B₂ and B₃ (FB₁, FB₂ and FB₃) and aflatoxins B₁ (AFB₁), B₂ (AFB₂), G₁ (AFG₁) and G₂ (AFG₂) are both major mycotoxins of food concern, because of their wide range of concentration and possible co-occurrence. Therefore, a contamination survey in corn and wheat flour by liquid chromatography–tandem mass spectrometry was carried out. Quantification of fumonisins and aflatoxins was based on internal calibration (by the use of ¹³C₃₄-fumonisin) and external calibration, respectively. Fumonisins were detected in 95% of corn samples and in 7% of wheat flour samples, with the mean level (FB₁?+?FB₂?+?FB₃) of 441?µg?kg^?1 and 0.09?µg?kg^?1, respectively. Low levels of aflatoxins were detected in 37% of the samples with a mean level (B₁?+?B₂?+?G₁?+?G₂) of 0.12?µg?kg^?1. Fumonisins and aflatoxins were not detected in 29% of the samples analysed. Simultaneous occurrence of fumonisins and aflatoxins was observed in 12% of samples.