Mycotoxins In Milk Research Articles

Quantitative risk ranking of mycotoxins in milk under climate change scenarios

Mycotoxins are toxic fungal metabolites that may occur in crops. Mycotoxins may carry-over into bovine milk if bovines ingest mycotoxin-contaminated feed. Due to climate change, there may be a potential increase in the prevalence and concentration of mycotoxins in crops. However, the toxicity to humans and the carry-over rate of mycotoxins from feed to milk from bovines varies considerably. This research aimed to rank emerging and existing mycotoxins under different climate change scenarios based on their occurrence in milk and their toxicity to humans. The quantitative risk ranking took a probabilistic approach, using Monte-Carlo simulation to take account of input uncertainties and variabilities. Mycotoxins were ranked based on their hazard quotient, calculated using estimated daily intake and tolerable daily intake values. Four climate change scenarios were assessed, including an Irish baseline model in addition to best-case, worst-case, and most likely scenarios, corresponding to equivalent Intergovernmental Panel on Climate Change (IPCC) scenarios. This research prioritised aflatoxin B1, zearalenone, and T-2 and HT-2 toxin as potential human health hazards for adults and children compared to other mycotoxins under all scenarios. Relatively lower risks were found to be associated with mycophenolic acid, enniatins, and deoxynivalenol. Overall, the carry-over rate of mycotoxins, the milk consumption, and the concentration of mycotoxins in silage, maize, and wheat were found to be the most sensitive parameters (positively correlated) of this probabilistic model. Though climate change may impact mycotoxin prevalence and concentration in crops, the carry-over rate notably affects the final concentration of mycotoxin in milk to a greater extent. The results obtained in this study facilitate the identification of risk reduction measures to limit mycotoxin contamination of dairy products, considering potential climate change influences.

Determination of 22 mycotoxins in milk by liquid chromatography-quadrupole/orbitrap mass spectrometry

Mycotoxins have carcinogenic, mutagenic, hepatotoxic, nephrotoxic, immunotoxic, neurotoxic, and teratogenic properties. Thus, these substances have attracted significant attention because they pose a threat to human health. As research on mycotoxins deepens, new structural analogues of mycotoxins are constantly being discovered. In this study, a method based on high performance liquid chromatography-quadrupole/orbitrap mass spectrometry was established for the simultaneous determination of 22 mycotoxins in milk. A simple, effective, and rapid pretreatment method was optimized by focusing on the solvent type, extractant volume, and extracting salt based on the characteristics of the mycotoxins and sample matrix. The analytes were extracted using 0.5% formic acid acetonitrile solution and added with sodium chloride to separate fats from water. The samples were centrifuged at 8000 r/min (4 ℃) for 5 min using a centrifuge and then concentrated using nitrogen. The dry residue was dissolved with 50% methanol aqueous solution. Twenty-two mycotoxins were separated on a ZORBAX Eclipse Plus C18 chromatographic column (100 mm×2.1 mm, 1.7 μm), and quantitative analysis was performed using the isotope internal standard method. The analytes were determined by liquid chromatography-quadrupole/orbitrap mass spectrometry in positive electrospray ionization mode. Qualitative analyses of the compounds were performed in full mass spectrometry/data-dependent tandem mass spectrometry (MS/dd-MS2) mode. Good linearities in the range of 0.5-100.0 μg/L were observed for the 22 mycotoxins, and the correlation coefficients (R2) were greater than 0.999. The limits of detection (S/N=3) and quantification (S/N=10) ranged from 0.3 to 0.5 μg/kg and from 1.0 to 1.5 μg/kg, respectively. The average recoveries of the 22 mycotoxins at three spiked levels of 1.5, 5.0, and 15 μg/kg were between 84.7% and 100.8%, with relative standard deviations (RSDs) of 1.2%-9.9%. These findings indicate that the method has high sensitivity and accuracy as well as good precision. Finally, the method was applied to the detection and analysis of mycotoxins in 25 actual commercial milk samples. The results revealed that the selected samples were not contaminated with any of the mycotoxins analyzed. Thus, the proposed method is useful as a quick preprocessing and confirmatory method for the simultaneous determination of mycotoxins in milk.

Regulated and Emerging Mycotoxins in Bulk Raw Milk: What Is the Human Risk?

Mycotoxins are abiotic hazards whose contamination occurs at the pre- and post-harvest stages of the maize value chain, with animal exposure through contaminated feed leading to their excretion into milk. Currently, only aflatoxin M1 is regulated in milk products. Since feed materials and complete feed present a multi-mycotoxin composition and are the main mycotoxin source into milk, it is important to recognize the occurrence of multiple toxins and their co-occurrence in this highly consumed food product. The aim of this study was to determine the content of regulated and emerging mycotoxins in milk samples, which allowed for evaluating the occurrence and co-occurrence patterns of different mycotoxins known to contaminate feed materials and complete animal feed. Human exposure considering the occurrence patterns obtained was also estimated. Aflatoxins, fumonisins, zearalenone, and emerging mycotoxins were among the mycotoxins found to be present in the 100 samples analyzed. Concentrations ranged from 0.006 to 16.3 μg L-1, with no sample exceeding the AFM1 maximum level. Though several mycotoxins were detected, no exceeding values were observed considering the TDI or PMTDI. It can be concluded that the observed exposure does not pose a health risk to milk consumers, though it is important to recognize vulnerable age groups.

Simultaneous Detection and Quantification of Aflatoxin M1, Eight Microcystin Congeners and Nodularin in Dairy Milk by LC-MS/MS

Dairy milk holds a prominent position as a widely consumed food, particularly among infants and children. However, it is crucial to address the presence of multiple natural toxic compounds that may co-occur in dairy milk to ensure its safety prior to consumption. Aflatoxin M1 (AFM1), an emerging mycotoxin of interest, is a potential contaminant in the milk of animals who ingest aflatoxin B1 (AFB1). The toxin is regulated in the European Union under Commission Regulation No 1881/2006. Unfortunately, there is a notable lack of data concerning the transfer of various emerging microbial contaminants into dairy milk and, therefore, their natural occurrences. In this study, a simple and sensitive LC-MS/MS method was developed and validated for the quantification of multiple cyanotoxins (microcystin congeners and nodularin) and AFM1 by the main analytical guidelines. Toxins are extracted with methanol 80%, followed by an SPE clean-up step before LC-MS/MS analysis. The LOQ was fixed at 1 µg/L for the nine cyanotoxins and 0.05 µg/L for AFM1. Recoveries were measured between 82.67% and 102%. To the best of our knowledge, there are no other LC-MS/MS methods available for the simultaneous quantification of cyanotoxins and mycotoxins in milk.

Mycotoxins in milk: Occurrence and evaluation of certain detoxification attempts.

Milk contaminated with mycotoxins is a significant issue affecting human health, especially in infants. The current study aimed to investigate the presence of mycotoxins in milk collected from women farmers' vendors (WFV), and to evaluate certain herbal plant fibers as green mycotoxin binders. Moreover, explore the binding efficiency ratios of mycotoxins using shaking or soaking process incorporated with herbal extracts. Furthermore, compare the taste evaluations of tested milk are enriched with herbal extracts. Results indicated that the fumonisins were not detected in the collected cow milk samples but realized a 25% occurrence ratio in buffalo's milk samples. A high occurrence ratio of aflatoxin M1 (aflaM1) was observed in buffalo and cow milk samples. The soaking process of plant fibers in contaminated milk overnight significantly degrades and adsorbs mycotoxins particles. The shacking process incorporated with plant fibers exhibited more effectiveness in mycotoxins degradation than soaking or shacking processes alone. The speed of shacking process played an important role in the mycotoxin's binding process. All the tested plant fibers effectively reduced all mycotoxin presence in contaminated milk, especially green tea, during the soaking or shacking process. Moreover, the shacking process incorporated with plant fibers promoted and supported the mycotoxins degradation process.

Detection of Antibiotic Residues and Mycotoxins in Milk Using Competitive Immunochromatographic Tests

Abstract As milk should be free from harmful substances before leaving the farm, this study provides the results of the analysis of the presence of β-lactam and tetracycline residues and Aflatoxin M1 (AFM1) in milk samples obtained within one year from Ireland and Slovakia. To evaluate the presence of β-lactam and tetracycline residues, Duplex BT Scan assay, produced by Zeulab S. L. was used. For the detection of AFM1, AflaM1 Scan (Zeulab S. L.) tests were used. Of a total of 69 raw cow’s milk samples analysed, 40 samples were obtained from the farms in Slovakia and 29 samples from the farms in Ireland. Among the 69 analysed samples, 17 (24.6 %) samples were tested positive for the presence of β-lactam residues, 12 (17.4 %) samples for tetracycline residues and 59 (88.4 %) samples for AFM1. Milk samples positive for antibiotic residues and mycotoxins were not acceptable and it is important that the reputation of milk as a healthy and safe food is protected worldwide. Dairy farmers and consumers want to be confident that milk and milk products are of high quality and free of all pharmacologically active substances and toxins.

Incidence and Levels of Aflatoxin M1 in Artisanal and Manufactured Cheese in Pernambuco State, Brazil.

Cheese is one of the most susceptible dairy foods to accumulating aflatoxins due to their high affinity to caseins. The consumption of cheese contaminated with high levels of aflatoxin M1 (AFM1) can be highly harmful to humans. The present work, based on high-performance liquid chromatography (HPLC), highlights the frequency and levels of AFM1 in coalho and mozzarella cheese samples (n = 28) from the main cheese-processing plants in Araripe Sertão and Agreste in the state of Pernambuco, Brazil. Of the evaluated cheeses, 14 samples were artisanal cheeses and the remaining 14 were industrial (manufactured) cheeses. All samples (100%) had detectable levels of AFM1, with concentrations ranging from 0.026 to 0.132 µg/kg. Higher levels (p < 0.05) of AFM1 were observed in artisanal mozzarella cheeses, but none of the cheese samples exceed the maximum permissible limits (MPLs) of 2.5 µg/kg established for AFM1 in cheese in Brazil and 0.25 µg/kg in the European countries by the European Union (EU). The high incidence of low levels of AFM1 found in the evaluated cheeses underscores the need for stringent control measures to prevent this mycotoxin in milk used for cheese production in the study area, with the aim of protecting public health and reducing significant economic losses for producers.

Effects of Dietary Natural Mycotoxins Exposure on Performance, Biochemical Parameters and Milk Small Molecule Metabolic Pathways of Lactating Cows

The presence of mycotoxins in feed has the potential to cause significant detriment to animal and human health, and even severe economic implications. Previous studies on the effects of mycotoxins mainly focused on the addition of commercially available mycotoxins into feeds in animals. In the present study, corn meal and cottonseed were kept in warm and humid conditions to allow for mycotoxins produced and then used to substitute 50% and 100% of normal corn meal and cottonseed in diets for lactating cows for 14 days. The results showed that aflatoxin M1, deoxynivalenol, aflatoxin B1, and zearalenone were primary mycotoxins in milk from cows fed the diets. Compared with the control group, feeding the diets containing mildewy corn meal and cottonseed reduced feed intake, milk yield, and milk fat, protein and lactose productions (p &gt; 0.05). No significant difference was observed in the acetate and valerate concentrations, acetate to propionate ratio, and the calculated CH4 production in rumen fluid (p &gt; 0.05), whereas, the propionate, butyrate, isovalerate concentrations were affected (p &lt; 0.05) depending on the content and type of natural mycotoxins. Serum creatinine and total glyceride concentrations were influenced with corn meal and cottonseed fully replaced with the mildewy feeds. Metabolic pathways for small molecule metabolites in milk were altered by dietary mycotoxin exposures, and the changes were mainly associated with amino acid metabolism, glucose metabolism, and energy metabolism. However, cows exposed to natural mycotoxins in the diets were still in healthy conditions and had low somatic cell count in milk.

Influence of grains postharvest conditions on mycotoxins occurrence in milk and dairy products

Abstract Mycotoxins are secondary metabolites produced by some filamentous fungi, which can cause toxic effects in humans and animals. The purpose of the study was to report the influence of postharvest conditions of grains and animal feed on the occurrence of mycotoxins in milk and dairy products which are widely consumed worldwide and have several health benefits for consumers. Among the most toxic mycotoxins are aflatoxins (AFs), with AFB1 being the most toxic and present in grains and cereals used in the feeding of dairy cows. After ingestion, AFB1 is converted to AFM1, which is excreted in milk, being a source of direct contamination to consumers of this product and dairy products. Thus, knowledge about this substance and the care that can be taken to reduce the consumption of contaminated food is a matter of great importance concerning public health and food safety. This review reports that the temperature and humidity at which the grains are stored, are the main conditions that can be controlled during storage, aiming to reduce the growth of fungi and the production of mycotoxins. This work reinforces that the continuous control of mycotoxins in milk and dairy products is of vital importance to obtain a safe product. Besides, strategies to mitigate the development of fungal contamination have been carefully revised to prevent the formation of these toxic substances.

Qualitative-quantitative analysis of multi-mycotoxin in milk using the high-performance liquid chromatography-tandem mass spectrometry coupled with the quick, easy, cheap, effective, rugged and safe method.

High-performance liquid chromatography-tandem mass spectrometry coupled with the quick, easy, cheap, effective, rugged and safe method was established for the qualitative and quantitative detections of 20 mycotoxins in milk. The linear range of this method was 0.01-10 μg/L and the correlation coefficients were all greater than or equal to 0.9933. At three levels of addition, the spiked recoveries ranged from 80.00 to 112.50%, relative standard deviations were 2.67-14.97%, limits of quantitation were 0.02-4.00 μg/kg, and limits of detection were 0.007-1.300 μg/kg. This developed procedure for the identification and quantitation of mycotoxins provided prospective support for quality regulation.

Multianalyte method for the determination of regulated, emerging and modified mycotoxins in milk: QuEChERS extraction followed by UHPLC–MS/MS analysis

A simple method for the quantification of 40 mycotoxins in milk was developed. This method is based on a QuEChERS extraction followed by the ultra-high liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) detection, and allows the simultaneous analysis of regulated, emerging, and modified mycotoxins. A sample treatment procedure was optimized to include a concentration step for the analysis of some compounds such as aflatoxin M1. The method was in-house validated in terms of limits of detection (LODs), limits of quantification (LOQs), linearity, recoveries, and precision. LOQs lower than 10 ng/mL were obtained, and recoveries ranged from 61% to 120% with a precision, expressed as the relative standard deviation, lower than 15%. Therefore, acceptable performance characteristics were obtained fulfilling European regulations. The method was successfully applied for the quantification of mycotoxins in raw milk. It can be highlighted high occurrence of beauvericin and enniatins were found in low amounts.

Polyethyleneimine-modified magnetic carbon nanotubes as solid-phase extraction adsorbent for the analysis of multi-class mycotoxins in milk via liquid chromatography-tandem mass spectrometry.

Polyethyleneimine-modified magnetic multi-walled carbon nanotubes were developed to extract 10 mycotoxins. Simple modification of polyethyleneimine was achieved on the magnetic substrate by using an epoxy-containing silane agent as a linker. The resultant magnetic adsorbent was integrated with reverse phase and anion exchange interaction sites. Under optimal extraction conditions, only 20.0mg adsorbent was used to extract the mycotoxins from 50.0mL loading solution. The maximum adsorption capacities of the adsorbent toward the mycotoxins ranged from 4.9 to 10.2mg/g. Adsorption and desorption were completed within 3.0 and 2.0min, respectively. The adsorbent could be used for six repeated runs without evident change in extraction performance. The adsorbent combined with liquid chromatography-tandem mass spectrometry was applied further to analyze the mycotoxins in milk. The absolute recoveries of the 10 mycotoxins ranged from 88.3 to 103.5% with relative standard deviations that ranged from 2.4 to 6.5%, and their limits of detection were 0.003 to 0.442μg/kg. The proposed adsorbent has great potential in the routine analysis of mycotoxins in ordinary analytical chemistry laboratory.

MYCOTOXINS IN MILK AND DAIRY PRODUCTS

The article gives an overview of modern opinions about the ways of possible contamination of milk and dairy products by mycotoxyn. The key indicator of the presence of mycotoxins in milk and dairy products is the level to which micromycetes affect productive livestock’s feed. Yet, mycromycetes and mycotoxines do not always occur together: some test samples can contain certain micromycetes, but no mycotoxines. Mycotoxines are synthesised by micromycetes only under certain favourable conditions. The multi-chambered stomach ecosystem of lactating cows utilises most mycotoxins occurring in food. Only a small amount of those is excreted in milk. However, some mycotoxins can bind to milk caseins. In this case, cheese and other dairy products can contain far higher amounts of mycotoxins than at the initial stage of milk production. The paper compares the maximum permissible levels of mycotoxins in milk and in dairy products according to the regulations of Ukraine and the EU. It presents the mycotoxins isolated from secretions of mammary glands of humans and productive animals, and describes their effect on the body’s physiology. It also provides a structural diagram of how mycotoxins contaminate milk and dairy products following the path “Animal feed – dairy products.” We suggest four-stage assessment of the risks of mycotoxin contamination of milk and dairy products: Stage I – identifying the producer of mycotoxin (molecule, metabolites in feed, milk, and dairy products). Anamnesis; Stage II – constructing a sequence diagram. Inspecting all production stages to identify the main ways and periods of contamination, determining and describing the symptoms of contamination; Stage III – assessing how the intensity and duration of exposure to a mycotoxin and its metabolites are likely to effect on the body. Modelling the influence of mycotoxins on the body; Stage IV – assessing the risk and determining measures to eliminate or minimise it. Risk scenario forecast

Rapid, on-site, and sensitive detection of aflatoxin M1 in milk products by using time-resolved fluorescence microsphere test strip

Aflatoxin M1 (AFM1), a frequently occurring mycotoxin in milk and dairy products, causes acute and chronic poisoning. In this study, we developed a time-resolved fluorescence microsphere immunochromatographic test strip (TRFM–ICTS) combined with ultraviolet light for the rapid and quantitative detection of AFM1 in milk and its products. Polystyrene microspheres enclosing time-resolved fluorescent europium (III) [Eu(III)-TRFM] were prepared as a label and then conjugated to a monoclonal antibody. Pre-incubation was introduced for competitive recognition. Under optimal conditions, TRFM–ICTS had a linear range of 0.05–2.0 ng/mL with a 50% inhibitory concentration (IC50) of 0.204 ng/mL, which was nearly five–fold lower than those of ICTS in a traditional structure with traditional pretreatment (IC50 = 1.10 ng/mL). This test strip enhanced the sensitivity from 0.3 ng/mL (by using gold nanoparticles-strip method previously reported) to 0.019 ng/mL (by using this TRFM–ICTS method). The improved sensitivity may be due to the detection signals of immunoassay probes (with each fluorescence microsphere coupled with numbers of antibodies). TRFM–ICTS exhibited good recoveries ranging from 84.6% to 119.0% with a coefficient of variation (CV) lower than 8.31% in spiked samples, including milk powder, ultrahigh-temperature (UHT) and pasteurized milk. The results obtained using TRFM–ICTS were consistent with those obtained by ultrahigh–performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) during testing of real milk samples. In addition, TRFM–ICTS has a longer storage period, which basically meets the needs of the commercial market. Owing to its high sensitivity and practicability, the proposed TRFM–ICTS can be used as a sensitive, simple, rapid, and versatile method for the quantitative detection of AFM1 in milk.

Determination of Aflatoxin M1 in Raw Milk from Different Provinces of Ecuador.

Aflatoxin M1 (AFM1) is a mycotoxin from Aspergillus flavus and A. parasiticus, classified as carcinogenic and hepatotoxic. The objective of the present investigation was to determine its presence in raw milk from north-central Ecuador, constituted by the provinces of Pichincha, Manabí, and Santo Domingo de los Tsáchilas. These areas represent approximately 30% of Ecuadorian milk production. By the end of the investigation, a total of 209 raw milk samples were collected, obtained both during the dry (June and August) and rainy seasons (April and November) of 2019. AFM1 concentrations were measured with lateral flow immunochromatographic assays, and 100% of the samples were positive for this mycotoxin, presenting a mean value of 0.0774 μg/kg with a range of 0.023 to 0.751 μg/kg. These AFM1 levels exceeded the European Union regulatory limit of 0.05 μg/kg in 59.3% (124/209) of samples, while only 1.9% (4/209) exceeded the Ecuadorian legal limit of 0.5 μg/kg. By using non-parametric tests, significant differences were determined (p ≤ 0.05) between the provinces for months of study, climatic season (being higher in the dry season), and climatic region (greater in the coast region). On the other hand, there were no significant differences (p ≥ 0.05) between the types of producers or between production systems. Therefore, AFM1 contamination in raw milk does not present a serious public health problem in Ecuador, but a monitoring and surveillance program for this mycotoxin in milk should be developed to prevent consumer health problems.

Longitudinal assessment of mycotoxin co-exposures in exclusively breastfed infants

Early-life development of infants may be critically affected by man-made or natural contaminants including mycotoxins. However, data on the occurrence of food contaminants in breast milk is scarce and prohibits a comprehensive exposure and risk assessment for mothers and their infants.Here, we present a longitudinal exposure assessment over the first 211 days of a single newborn girl (studyA) by measuring multiple mycotoxins in milk. Eighty-seven consecutive breast milk samples were obtained from the newborn’s mother living in Austria and following a regular mixed diet. Mycotoxins were analyzed by utilizing a highly sensitive LC-MS/MS approach covering 29mycotoxins and key metabolites. In addition to this longitudinal study, three mothers provided breast milk samples each on five consecutive days, for a preliminary comparison of inter-day and inter-individual variation in exposures (studyB). StudyA revealed that mycotoxin occurrence in breast milk was limited to the emerging mycotoxins alternariol monomethyl ether (AME), beauvericin (BEA), enniatins (A, A1, B, B1) and to ochratoxin A (OTA), which is regulated in commercial infant food. These mycotoxins were, if present, mostly detected at very low concentrations (<10 ng/L), except AME which exceeded this concentration on two distinct days by a factor of 3x and 5x. Overall, longitudinal results indicated chronic low-dose exposure to the detected mycotoxins. Other regulated mycotoxins including the carcinogenic aflatoxins or the estrogenic zearalenone and their biotransformation products were absent in all tested samples. StudyB confirmed the results of studyA, with minimal inter-day and inter-individual variation. In addition, a preliminary correlation of OTA levels occurring in breast milk and matched urine samples was found (r = 0.64, p = 0.034) in study B. Based on the data set obtained in studyA, exposure of the infant was estimated. Exposure estimates of individual mycotoxins were on average below 1 ng/kg body weight per day.Our preliminary findings suggest that recommended maximum daily intake levels might not be exceeded in the Austrian population. However, exposure is likely to be higher in populations with lower food safety standards. In the light of co-occurrence of several emerging mycotoxins in breast milk, future studies should address low-dose mixture effects. This also includes other environmental contaminants which may be present in this bio-fluid and should involve an exposome-scale risk assessment. All these efforts must be intended to minimize exposure of mothers and infants in a window of high susceptibility.

Transcriptomic and proteomic profiling reveals the intestinal immunotoxicity induced by aflatoxin M1 and ochratoxin A

Mycotoxins-contaminated milk could threaten human health; therefore, it is necessary to demonstrate the toxicological effect of mycotoxins in milk. Most recently, researchers have paid more attention to the immunotoxic effects of the individual cereal-contaminating mycotoxins, namely, zearalenone and deoxynivalenol. However, there is scant information about the intestinal immunotoxicity of aflatoxin M1 (AFM1), let alone that of a combination of AFM1 and ochratoxin A (OTA), which often co-occur in milk. To reveal the inflammatory response caused by these mycotoxins, expression of inflammation-related genes in differentiated Caco-2 cells was analyzed, demonstrating a synergistic effect of the mixture of AFM1 (4 μg/mL) and OTA (4 μg/mL). Integrative transcriptomic and proteomic analyses were also performed. A cross-omics analysis identified several mechanisms underlying this synergy: (i) compared with stimulation with either compound alone, combined use resulted in stronger induction of proteins involved in immunity-related pathways; (ii) combination of the two agents targeted different points in the same pathways; and (iii) combination of the two agents activated specific inflammation-related pathways. These results suggested that combined use of AFM1 and OTA might exacerbate intestinal inflammation, indicating that regulatory authorities should pay more attention to food contamination by multiple mycotoxins when performing risk assessments.

Aflatoxin M1 in UHT and powder milk marketed in the northern area of Morocco

Aflatoxins are secondary metabolites produced by some species belonging to Aspergillus genus. They are considered as natural contaminants in feed and food products. The present study investigates the presence of mycotoxins in milk; it offers an evaluation of aflatoxin M1 (AFM1) occurrence in UHT and powder milk. The method applied for AFM1 analysis in milk was first validated and then used to determine the AFM1 levels in UHT milk (n = 40) and powder milk (n = 7) samples collected from supermarkets and dairy companies in both Tangier and Tetouan cities (northern area of Morocco). AFM1 was determined in milk samples using liquid chromatography coupled to fluorescence detection after immuno-affinity clean up. Validation step showed that the method used was linear at concentrations ranging between 10 and 160 ng/kg; it was reproducible and provided a recovery rate ranging between 99.5% and 114.6%. Analytical results showed that AFM1 was detected in 14 out of 40 UHT milk samples (35%) with an average concentration of 14.76 ± 10.21 ng/kg and detected in all the 7 powder milk samples (100%) with an average concentration of 25.50 ± 12.06 ng/kg. None of the analyzed samples exceeded the maximum limit sets by both Moroccan and European regulations in liquid and powder milk. Based on the results of this study, the consumer seems to be exposed to a lower risk of AFM1. However, a monitoring program should be implemented and enforced.

Aflatoxin M1 in milk in Hisar city, Haryana, India and risk assessment

ABSTRACTAflatoxin M1 (AFM1) contamination in 150 samples of milk, sold in market of Hisar city of Haryana, India, was investigated by using High Pressure Liquid Chromatography (HPLC). Out of these, 40 samples contained AFM1 at a concentration below the limit of detection (LOD) of 0.052 μg/kg. Among the AFM1 contaminated samples, 46 raw milk samples contained a concentration above the LOD but less than the limit of quantitation (LOQ), whereas 64 samples were above the LOQ. Of these samples, 31 exceeded the maximum limit of 0.5 μg/kg prescribed by FSSAI, India. Based on this study, the dietary intake of AFM1 for adults through consumption of milk was estimated. The results indicated that AFM1 contamination can be a food safety issue for raw and pasteurised milk consumed in India. Therefore, there is a need for a national monitoring programme to control the level of mycotoxins in milk.

Target analysis and retrospective screening of mycotoxins and pharmacologically active substances in milk using an ultra-high-performance liquid chromatography/high-resolution mass spectrometry approach

Milk is a nutritious food suitable for infants and adults, and it plays an important role in the human diet. However, it may also be a vehicle for food contaminants. In this report, we developed a method using ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-Q-Exactive Orbitrap HRMS; Thermo Fisher Scientific, Waltham, MA) for simultaneous identification of target pharmacologically active substances and mycotoxins in milk. We also used the Q-Orbitrap operating in full scan mode to identify other possible drugs and microbial metabolites that occurred in samples. Fifty-six commercially available milk samples from the Italian market were analyzed. Investigated analytes were extracted using a QuEChERS (quick, easy, cheap, effective, rugged, and safe) approach. Method detection and quantification limits and performance criteria set by European regulations were fulfilled. Pharmacologically active substances were detected in 49% of samples (range 0.007-4.53 ng/mL), including nontarget mycotoxins. Retrospective analysis allowed us to identify other antibiotics and pharmacologically active substances, as well as nonregulated fungal/bacterial metabolites at a relatively high incidence. From the obtained values, the need for continuous monitoring of contaminants in the milk production chain is clear. This is the first study to assess the presence of pharmacologically active substances, mycotoxins, and other microbial metabolites in Italian milk samples using the UHPLC-Q-Orbitrap HRMS system.