scholarly journals Detection Methods for Aflatoxin M1 in Dairy Products

2020 ◽  
Vol 8 (2) ◽  
pp. 246 ◽  
Author(s):  
Andreia Vaz ◽  
Ana C. Cabral Silva ◽  
Paula Rodrigues ◽  
Armando Venâncio

Mycotoxins are toxic compounds produced mainly by fungi of the genera Aspergillus, Fusarium and Penicillium. In the food chain, the original mycotoxin may be transformed in other toxic compounds, reaching the consumer. A good example is the occurrence of aflatoxin M1 (AFM1) in dairy products, which is due to the presence of aflatoxin B1 (AFB1) in the animal feed. Thus, milk-based foods, such as cheese and yogurts, may be contaminated with this toxin, which, although less toxic than AFB1, also exhibits hepatotoxic and carcinogenic effects and is relatively stable during pasteurization, storage and processing. For this reason, the establishment of allowed maximum limits in dairy products and the development of methodologies for its detection and quantification are of extreme importance. There are several methods for the detection of AFM1 in dairy products. Usually, the analytical procedures go through the following stages: sampling, extraction, clean-up, determination and quantification. For the extraction stage, the use of organic solvents (as acetonitrile and methanol) is still the most common, but recent advances include the use of the Quick, Easy, Cheap, Effective, Rugged, and Safe method (QuEChERS) and proteolytic enzymes, which have been demonstrated to be good alternatives. For the clean-up stage, the high selectivity of immunoaffinity columns is still a good option, but alternative and cheaper techniques are becoming more competitive. Regarding quantification of the toxin, screening strategies include the use of the enzyme-linked immunosorbent assay (ELISA) to select presumptive positive samples from a wider range of samples, and more reliable methods—high performance liquid chromatography with fluorescence detection or mass spectroscopy—for the separation, identification and quantification of the toxin.

Author(s):  
G.M. Goryainova ◽  
◽  
E.A. Denisova ◽  
L.V. Arsen`eva ◽  
V.S. Babunova ◽  
...  

The article is devoted to the study of control issues and methods for detecting residual amounts of antibacterial substances in milk and dairy products. The presence of a large number of antibacterial drugs on the Russian market and their use in dairy farming makes it an urgent problem to detect their residual amount, both in raw milk and in dairy products. The lack of standardized detection methods, sufficiently equipped and accredited laboratories, and the high cost and duration of research are obstacles to monitoringthe content of the residual amount of antibiotics., including state control. Currently, the state monitoring system for screening antibiotics uses the enzyme-linked immunosorbent assay method (ELISA), which is the official method for monitoring animal products adopted in the European Union. To confirm the quantitative content of antibiotics, a high-performance liquid chromatography (HPLC) method with various types of detectors is used, which is reliable but very time-consuming for screening a large number of samples. Using the patented Randox Biochip technology as a method for detecting the residual amount of antibiotics in milk and dairy products, it would be possible to determine up to 25 antibacterial substances in one sample, including those that are not often used in veterinary practice.


2020 ◽  
Vol 13 (3) ◽  
pp. 433-439 ◽  
Author(s):  
Sarah Mohammedi-Ameur ◽  
Mohammedi Dahmane ◽  
Carlo Brera ◽  
Moustafa Kardjadj ◽  
Meriem Hind Ben-Mahdi

Background and Aim: Aflatoxins are metabolites of molds that exert potentially toxic effect on animals and humans. This study aimed to investigate the occurrence of aflatoxin M1 (AFM1) in raw cow milk collected during 1 year (2016-2017) from different regions of Algeria and risk factors associated with the contamination. Materials and Methods: During the survey period, 84 samples of raw milk were collected in three regions of Algeria (northeast, north center, and northwest) during four seasons. AFM1 levels were analyzed by competitive enzyme-linked immunosorbent assay. Results: AFM1 was detected in 39 (46.43%) samples (total mean concentration, 71.92 ng/L; range, 95.59-557.22 ng/L). However, the AFM1 levels exceeded the maximum tolerance limit set by the Food and Drug Administration in the USA (500 ng/L) in only 1 sample (1.19%). Statistical analysis revealed significant differences (p<0.005) between AFM1 levels in milk samples collected in the spring and autumn. The mean AFM1 levels in samples collected in the spring were significantly higher than those in samples collected in autumn. Conclusion: The survey indicates that farmers involved in milk production should be made aware of the adverse effects of aflatoxin contamination in animal feed. A systematic control program of supplementary feedstuff for lactating cows should be introduced by the public health authorities.


2009 ◽  
Vol 72 (4) ◽  
pp. 885-889 ◽  
Author(s):  
YAŞAR KESKIN ◽  
RUHTAN BAŞKAYA ◽  
SEHER KARSLI ◽  
TÜRKAN YURDUN ◽  
OĞUZ ÖZYARAL

This survey was undertaken to determine the extent of aflatoxin M1 (AFM1) contamination in human breast milk and raw cow's milk in Istanbul, Turkey. Samples of human and raw cow's milk were collected randomly and analyzed for AFM1 using an enzyme-linked immunosorbent assay and high-performance liquid chromatography with fluorescence detection in which the samples were cleaned up with immunoaffinity columns. In this study, AFM1 was detected in 8 (13.1%) of 61 human breast milk samples examined (mean ± SD level, 5.68 ± 0.62 ng/liter; range, 5.10 to 6.90 ng/liter) and 20 (33.3%) of 60 raw cow's milk samples examined (range, 5.40 to 300.20 ng/liter). Five (8.3%) of the positive raw cow's milk samples had AFM1 levels (153.52 ± 100.60 ng/liter; range, 61.20 to 300.20 ng/liter) that were higher than the maximum tolerance limit (0.05 ppb) stipulated by regulations in Turkey and some other countries.


2016 ◽  
Vol 56 (10) ◽  
pp. 1637 ◽  
Author(s):  
N. Aslam ◽  
I. Rodrigues ◽  
D. M. McGill ◽  
H. M. Warriach ◽  
A. Cowling ◽  
...  

The objectives of this study were to observe the extent of transfer of aflatoxin B1 in feed to the aflatoxin M1 metabolite in milk in Nili-Ravi buffaloes and to evaluate the efficacy of a commercial mycotoxin binder (Mycofix, Biomin Singapore) incorporated into feed to minimise this transfer. Multiparous animals (n = 28) were randomly distributed to four groups corresponding to two treatments each with two levels of aflatoxin B1. Individual animals were exposed to naturally contaminated feed providing a total of 1475 µg/day (Groups A and B) or 2950 µg/day (Groups C and D) of aflatoxin B1. Groups B and D were given 50 g of mycotoxin binder daily mixed with feed whereas Groups A and C were kept as controls. Feed samples were analysed by reverse phase high performance liquid chromatography for aflatoxin B1 and milk samples were evaluated by enzyme-linked immunosorbent assay for the liver metabolite aflatoxin M1. The mean value of total daily aflatoxin M1 excretion for animals fed 2950 µg/day of aflatoxin B1 (112.6 µg/day) was almost double (P < 0.001) than the excretion in buffaloes fed 1475 µg/day (62.2 µg/day). The mean daily concentration of aflatoxin M1 in milk of animals from both treatment groups supplemented with 50 g/day of mycotoxin binder was 76.5 µg/day, nearly 22 µg lower than those without binder at 98.3 µg/day (s.e.d. = 5.99: P < 0.01). The interaction of binder and treatment was not significant i.e. the 50 g/day of binder was able to sequester aflatoxin B1 with the same efficiency in groups fed with high and low concentrations of aflatoxin B1. Carry over was (3.44%) lower (P = 0.001) in animals supplemented with 50 g/day of mycotoxin binder than those fed no binder (4.60%). Thus buffaloes are highly efficient at transferring aflatoxins in feed to the aflatoxin M1 metabolite in milk, whereas mycotoxin binder is capable of alleviating without preventing this contamination risk.


2016 ◽  
Vol 9 (2) ◽  
pp. 305-315 ◽  
Author(s):  
E.D. Womack ◽  
D.L. Sparks ◽  
A.E. Brown

Aflatoxin M1 (AFM1) is associated with carcinogenicity, genotoxicity, mutagenicity, and teratogenicity and as a result, represents a human health problem worldwide. This review will detail the toxicity, analytical methodology, occurrence, and prevention and control of AFM1 in milk and milk products. The probable daily intakes (PDI) per bodyweight (bw) worldwide ranged from 0.002 to 0.26 ng/kg bw/day for AFM1. Nevertheless, the high occurrence of AFM1 demonstrated in this review establishes the need for monitoring to reduce the risk of toxicity to humans. The recommended extraction method of AFM1 from milk is liquid-liquid with acetonitrile because of the acceptable recoveries (85-97%), compatibility with the environment, and cleanest extracts. The recommended analytical technique for the determination of AFM1 in milk is the high performance-liquid chromatography-fluorescence detector (HPLC-FLD), achieving a 0.001 µg/kg detection limit. The HPLC-FLD is the most common internationally recognised official method for the analysis of AFM1 in milk. The suggested extraction and analytical method for cheese is dichloromethane (81-108% recoveries) and ELISA, respectively. This review reports the projected worldwide occurrence of AFM1 in milk of 2010-2015. Of the 7,841 samples, 5,873 (75%) were positive for AFM1, 26% (2,042) exceeded the maximum residue levels (MRL) of 0.05 µg/kg defined by the European Union and 1.53% (120) exceeded the MRL of 0.5 µg/kg defined by the US Food and Drug Administration. The most effective way of preventing AFM1 occurrences is to reduce contamination of AFB1 in animal feed using biological control with atoxigenic strains of Aspergillus flavus, proper storage of crops, and the addition of binders to AFB1-contaminated feed. Controllable measures include the addition of binders and use of biological transforming agents such as lactic acid bacteria applied directly to milk. Though the one accepted method for the control of AFM1 in milk and milk products is the enforcement of governmental MRL.


Author(s):  
S. Senin ◽  
V. Danchuk ◽  
S. Midyk ◽  
V. Ushkalov ◽  
O. Iakubchak

The dairy industry of Ukraine is developing dynamically, its needs for the quality of raw materials are growing significantly. Detection of mycotoxins in raw milk is one of the main indicators of its safety. The high degree of toxicity of mycotoxins is a threat to the health of the lactating animal, so a large number of them are excreted in milk. If we talk about ruminants, the vast majority of mycotoxins are utilized by microorganisms of the pancreas, which does not occur in monogastric animals, so the list of mycotoxins in their milk can be much wider than the secretion of mammalian mammals. To date, the maximum permissible levels (MRLs) of mycotoxins in raw milk and dairy products have been established. Thus, a comprehensive determination of the content of mycotoxins in the secretion of the breast has not only technological but also important diagnostic value. Milk sample preparation is the most important step in the determination of mycotoxins and consists of sampling, extraction and purification from impurities. For the extraction of aflatoxins, the method of liquid extraction with acetonitrile or chloroform is used. Purification of extracts is carried out on immunoaffinity columns, cartridges with special sorbents or using certain manufacturers (MycoSep®).Enzyme-linked immunosorbent assay and high-performance liquid chromatography with fluorescence detection are used to determine aflatoxin B1 and M1 in raw milk of cows. However, all these methods have a number of disadvantages, namely: long and expensive sample preparation and insufficiently high selectivity. Currently, the complex determination of mycotoxins in various matrices by high-performance liquid chromatography with mass spectrometric detection (LC-MS/MS) and the use of modified QuEChERS sample preparation is gaining popularity. The advantage of this technique is the combination of faster and cheaper sample preparation of QuEChERS samples with highly selective LC-MS/MS chromatography. Key words: mycotoxins, raw milk, analytical methods, QuEChERS.


Author(s):  
Vesna Jankovic ◽  
Jelena Vukojevic ◽  
Brankica Lakicevic ◽  
Radmila Mitrovic ◽  
Dejan Vukovic

Aflatoxin M1 (AFM1) appears in milk or dairy products as a direct result of the cattle's ingestion of feed contaminated with aflatoxin B1 (AFB1). This study comprises mycological and mycotoxicological investigations of 23 milk samples (raw, infant food, pasteurized, whey and yoghurt). The mycological testing showed dominant presence of genus Geotrichum. G. candidum was found in 9 samples, with the highest contamination in the raw milk samples. The contamination level of AM1 is defined by using direct competitive enzyme- -linked immunosorbent assay (ELISA). AFM1 was found in 9 samples. AFM1 levels were lower than the recommended limits. However, as AFM1 is considered a probable human carcinogen (2B type), it is necessary to achieve a low level of AFM1 in milk. Therefore, cows' feed samples from various cowsheds are supposed to be evaluated routinely for aflatoxin, and kept away from fungal contamination as much as possible.


Foods ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 2109
Author(s):  
Zifei Wang ◽  
Pengjie Luo ◽  
Baodong Zheng

Aflatoxin B1 (AFB1) is a toxic compound naturally produced by the genera Aspergillus. Distillers’ grains can be used as animal feed since they have high content of crude protein and other nutrients. However, they are easily contaminated by mycotoxins, and currently there are no rapid detection methods for AFB1 in distillers’ grains. In this study, a lateral flow immunoassay (LFIA) based on red fluorescent microsphere (FM), is developed for quantitative detection of AFB1 in distillers’ grains. The whole test can be completed within 15 min, with the cut-off value being 25.0 μg/kg, and the quantitative limit of detection (qLOD) being 3.4 μg/kg. This method represents satisfactory recoveries of 95.2–113.0%, and the coefficients of variation (CVs) are less than 7.0%. Furthermore, this technique is successfully used to analyze AFB1 in real samples, and the results indicates good consistency with that of high-performance liquid chromatography (HPLC). The correlation coefficient is found to be greater than 0.99. The proposed test strip facilitates on-site, cost-effective, and sensitive monitoring of AFB1 in distillers’ grains.


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