scholarly journals Determination of Aflatoxin M1 in Raw Milk Using an HPLC-FL Method in Comparison with Commercial ELISA Kits—Application in Raw Milk Samples from Various Regions of Greece

2021 ◽  
Vol 8 (3) ◽  
pp. 46
Author(s):  
Martha Maggira ◽  
Maria Ioannidou ◽  
Ioannis Sakaridis ◽  
Georgios Samouris
Keyword(s):  
Limit Of Detection ◽  
Reference Method ◽  
Raw Milk ◽  
Aflatoxin M1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fluorescence Detector ◽  
Elisa Kit ◽  
Milk Samples ◽  
Of Greece

The highly toxic Aflatoxin M1 (AFM1) is most often detected in milk using an Enzyme-Linked-Immunosorbent Assay (ELISA) for screening purposes, while High-Performance Liquid Chromatography with Fluorescence Detector (HPLC-FL) is the reference method used for confirmation. The aim of the present study was the comparison between three commercially available ELISA kits and a newly developed HPLC-FL method for the determination of the AFM1 in milk samples. The developed HPLC-FL method was validated for the AFM1 and Aflatoxin M2 (AFM2), determining the accuracy, precision, linearity, decision limit, and detection capability with fairly good results. All three ELISA kits were also validated and showed equally good performance with high recovery rates. Moreover, the Limit Of Detection (LOD) and Limit Of Quantification (LOQ) values were found to be significantly lower than the Maximum Residue Limit (MRL) (50 ng kg−1). After the evaluation of all three commercial kits, the ELISA kit with the optimum performance along with the HPLC method was used for the determination of AFM1 in raw cow’s, goat’s, and sheep’s milk samples (396) obtained from producers in different regions of Greece. The evaluation of both methods showed that this ELISA kit could be considered as a faster and equally reliable alternative method to HPLC in routine analysis for the determination of AFM1 in milk.

scholarly journals Ultra-high performance liquid chromatographic determination of aflatoxin M1 in urine

2015 ◽  
Vol 8 (4) ◽  
pp. 405-413 ◽  
Author(s):  
S. Mohd Redzwan ◽  
R. Jamaluddin ◽  
A.M. Mohd Sokhini ◽  
A.R. Nurul Aqilah ◽  
A. Zuraini ◽  
...  
Keyword(s):  
Biological Samples ◽  
Analytical Methods ◽  
High Performance ◽  
Limit Of Detection ◽  
Extraction Procedure ◽  
Aflatoxin M1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fluorescence Detector ◽  
The Mean

The development of analytical methods to detect aflatoxin B1 (AFB1) in foodstuffs and its metabolites in human biological samples is useful for risk assessment. The latter methodology, i.e. the measurement of AFB1 biomarkers, has become important to assess human aflatoxin exposure. AFB1-lysine adduct, AFB1-DNA adduct and urinary aflatoxin M1 (AFM1) are some of the AFB1 biomarkers that can be measured by several analytical methods, such as enzyme-linked immunosorbent assay, radioimmunoassay, and high performance liquid chromatography (HPLC). HPLC coupled to a fluorescence detector is useful and preferable due to its high degree of sensitivity, but the analysis may take time and consume large amount of solvents. Therefore, the present study extrapolated the HPLC method to ultra-HPLC for the determination of urinary AFM1. After the extraction procedure with an immunoaffinity column, chromatographic separation was done using a high performance 1.8 μm microparticulate C18 column. The mean recovery from urine samples spiked with 0.5, 1.0 and 2.0 ng/ml AFM1 was 84.4±4.0%, with acceptable recovery values, interday (6.0±5.3%) and intraday (2.6±0.6%) coefficients of variation. The retention time was 5.7 min. This method was used to measure urinary AFM1 in 71 subjects, of which 13 had AFM1 levels above the limit of detection (0.018 ng/ml). The mean urinary AFM1 level of the positive samples was 18.8±28.6 pg/ml, ranging from 2.4 to 100.4 pg/ml. As this is one of the few studies investigating the occurrence of aflatoxin biomarkers in human biological samples in Malaysia, a study with a larger sample size is necessary to investigate the magnitude of aflatoxin exposure among the population.

scholarly journals Reduction of Aflatoxin M1 Levels during Ethiopian Traditional Fermented Milk (Ergo) Production

2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
Tsige Shigute ◽  
Alemayehu P. Washe
Keyword(s):  
Incubation Time ◽  
Raw Milk ◽  
Fermented Milk ◽  
Aflatoxin M1 ◽  
Chemical Components ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bacterial Cells ◽  
Milk Samples ◽  
Significant Difference

In this study, the reduction of aflatoxin M1 (AFM1) levels during lab-scale ergo production was investigated through determination of the residual levels of AFM1 using Enzyme Linked Immunosorbent Assay. The results showed gradual and incubation time dependent reduction of AFM1 level in the raw milk samples being fermented to ergo. The maximum reductions of 57.33 and 54.04% were recorded in AFM1 in natural and LAB inoculums initiated fermentations, respectively, in 5 days of incubation. Although a significant difference (P=0.05) in the AFM1 decrease in the two types of fermentations was recorded, such findings could vary with milk samples depending on initial load of the microorganisms as determined by hygienic conditions. However, the level of AFM1 in control (sterilized) samples showed only a 5.5% decrease during the entire period of incubation. Microbiological investigation showed increasing LAB counts with incubation time. A gradual decrease in pH of the milk samples was observed during fermentation. Considering the fact that both viable and dead bacterial cells could remove AFM1 during ergo production, the mechanism is proposed as predominantly involving noncovalent binding of the toxin with the chemical components of the bacterial cell wall.

Detection of proteolysis in raw milk stored at low temperature by an inhibition ELISA

1994 ◽  
Vol 61 (3) ◽  
pp. 395-404 ◽  
Author(s):  
Catherine Picard ◽  
Isabelle Plard ◽  
Dominique Rongdaux-Gaida ◽  
Jean-Claude Collin
Keyword(s):  
Proteolytic Activity ◽  
Limit Of Detection ◽  
Bacterial Count ◽  
Raw Milk ◽  
Enzyme Linked Immunosorbent Assay ◽  
Psychrotrophic Bacteria ◽  
Milk Samples ◽  
Inhibition Elisa ◽  
Different Strains

SummaryAn inhibition ELISA (enzyme-linked immunosorbent assay) was developed for the determination of caseinomacropeptide (CMP) in order to estimate the proteolysis of κ-casein due to the enzymes of psychrotrophic bacteria in bulk raw milk. The CMP present in milk was quantified specifically by an antibody. The limit of detection was ∽ 0·1 μg/ml and the CV was < 10%. This method was used to study the proteolytic activity of three strains of psychrotrophic Pseudomonas fluorescens in raw milk and to analyse different raw milk samples supplied by four dairy plants. The proteolytic activity for different strains of psychrotrophs and for different milk samples varied considerably, but no correlation was established between the level of microbial flora and κ-casein proteolysis. It is thus not possible to determine the extent of proteolysis from the bacterial count alone. However, by CMP determination in bulk raw milk samples after 6 d storage at 4°C, the mean κ-casein proteolysis was ∽ 4%. Among the milk samples analysed that contained < 107 cfu psychrotrophs/ml, 30% exhibited a proteolysis of κ-casein < 0·5%, i.e. < 5μg CMP/ml.

A survey on the occurrence of aflatoxin M1 in raw milk samples in Bafq and Bahabad, Iran

2021 ◽  
Author(s):  
Vahid Safavizadeh ◽  
Mozhgan Mojkar
Keyword(s):  
Aspergillus Parasiticus ◽  
Raw Milk ◽  
National Standard ◽  
Aflatoxin M1 ◽  
Cow’S Milk ◽  
Elisa Method ◽  
Cow's Milk ◽  
Milk Samples ◽  
The City

Aflatoxins are a group of mycotoxins mostly produced by the fungi called Aspergillus flavus, Aspergillus parasiticus, and Aspergillus nomium. Aflatoxin M1 (AFM1) is the major metabolite of aflatoxin B1 and is a hepatotoxic and carcinogenic toxin. The aim of this study was to determine the level of contamination of cow's milk with aflatoxin M1 in Bafq and Bahabad. For this study, samples of raw cow's milk were collected randomly from milk collection centers around the city of Bafq and Bahabad from March to April. The determination of aflatoxin M1 levels was based on the ELISA method. Contamination was observed in 100% of milk samples. According to the results of the study, the rate of contamination with aflatoxin M1 in 43.3% of milk samples was above the acceptable level (50 ng/L) in Iranian national standard. It is concluded that further monitoring of milk production should be carried out in the spring and winter seasons.

Occurrence of Aflatoxin in Dairy Cow Feed Ingredients and Total Mixed Ration

2019 ◽  
Vol 35 (5) ◽  
pp. 679-686 ◽  
Author(s):  
Zeinab Mohammadi Shad ◽  
Mehrdad Ghavami ◽  
Griffiths G. Atungulu
Keyword(s):  
Dairy Cows ◽  
Dairy Cow ◽  
Rainy Season ◽  
Raw Milk ◽  
Farm Management ◽  
Average Level ◽  
Aflatoxin M1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Feed Ingredients ◽  
Milk Samples

Abstract. The level of aflatoxin B1 (AFB1) in dairy cow feed ingredients and Total Mixed Rations (TMRs) procured at two farms for low- and high-yielding dairy cows were surveyed. Raw milk from the two groups of cows at each farm was sampled 24 h after feeding the cows with examined feedstuffs during both the rainy and the non-rainy season. The aflatoxin M1 (AFM1) level in the raw milk samples was measured 12-24 h later. The levels of AFB1 in feed and AFM1 in milk were determined by validated enzyme linked immunosorbent assay (ELISA). The influence of farm management and type of feeding system on aflatoxin occurrence were considered. AFB1 and AFM1 were detected in 100% of feed and milk samples, respectively. The average level of AFB1 in the feed ingredients and TMRs were in the range of 1.6-104.7 µg/kg and 11.0-56.0 µg/kg, respectively. The average level of AFM1 in milk samples was 77.0 ng/L. The average concentrations of AFB1 in feeds and AFM1 in milk procured in the rainy season were significantly greater than those procured in the non-rainy season (p&lt;0.05). Of the studied feed, maize silage was determined as the most contaminated feed ingredient in terms of AFB1 content. Furthermore, the AFM1 in 75% of milk samples obtained from high-yielding dairy cows and 25% of milk samples obtained from low-yielding dairy cows indicated AFM1 level higher than the maximum allowable Europe Commission limit of 50 ng/L. The results also showed that the occurrence of AFB1 in feed varied with farm feed management. The extent of translocation to AFMI in milk samples was dependent on type of cow, whether low- or high- milk yielding. This study suggests regular risk analysis and using good farm management practices are important to control aflatoxin contamination in feed and milk. Keywords: Aflatoxin B1, Aflatoxin M1, Dairy cows, Feed, Milk yield.

scholarly journals Presence of moulds and aflatoxin M1 in milk

2009 ◽  
pp. 63-68 ◽  
Author(s):  
Vesna Jankovic ◽  
Jelena Vukojevic ◽  
Brankica Lakicevic ◽  
Radmila Mitrovic ◽  
Dejan Vukovic
Keyword(s):  
Dairy Products ◽  
Raw Milk ◽  
Direct Result ◽  
Aflatoxin M1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Contamination Level ◽  
Infant Food ◽  
Fungal Contamination ◽  
Milk Samples ◽  
Feed Samples

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.

scholarly journals Survey of Deoxynivalenol Contamination in Agricultural Products in the Chinese Market Using An ELISA Kit

Toxins ◽  
2018 ◽  
Vol 11 (1) ◽  
pp. 6 ◽  
Author(s):  
Ming Li ◽  
Mingna Sun ◽  
Xia Hong ◽  
Jinsheng Duan ◽  
Daolin Du
Keyword(s):  
Self Assembly ◽  
Preventive Measures ◽  
Limit Of Detection ◽  
Agricultural Products ◽  
Enzyme Linked Immunosorbent Assay ◽  
Agricultural Product ◽  
Chinese Market ◽  
Elisa Kit ◽  
Distillers Dried Grains

A total of 328 agricultural product samples highly suspected to be contaminated, from flour companies, feed companies, and livestock farms throughout China, were surveyed for deoxynivalenol (DON) contamination using a self-assembly enzyme-linked immunosorbent assay (ELISA) kit. An ELISA kit for DON was developed with a 4.9 ng mL−1 limit of detection (LOD) in working buffer and a 200 ng g−1 LOD in authentic samples. The DON contamination detection rate was 88.7%, concentrations ranged from 200.9 to 6480.6 ng g−1, and the highest DON contamination was found in distillers’ dried grains with solubles with an average of 3204.5 ng g−1. Wheat bran and wheat were found to be the most commonly contaminated samples, and the corn meal samples had the lowest average DON level. This ELISA kit is a powerful alternative method for the rapid, sensitive, specific, accurate, and high-throughput determination of DON and can meet the maximum requirement levels. This survey suggests that DON contamination in the Chinese market is serious, and the contamination risk deserves attention. Essential preventive measures should be implemented to ensure food safety and human health.

scholarly journals AFLATOXIN EXCRETION THROUGH MILK

2008 ◽  
Vol 1 (1) ◽  
pp. 66-70
Author(s):  
Milica Živkov Baloš ◽  
Željko Mihaljev ◽  
Mira Kovačević ◽  
Dejan Bugarski
Keyword(s):  
Thin Layer ◽  
Limit Of Detection ◽  
Dairy Farms ◽  
Raw Milk ◽  
Aflatoxin M1 ◽  
Elisa Method ◽  
Obligatory Control ◽  
Milk Samples ◽  
Two Samples ◽  
Layer Chromatography

In the period January to June 2006 the samples of feed were collected from feed factories in Southern Baåka and Srem district. The samples of raw milk and full mix were taken from 5 dairy farms. A total of 50 raw milk samples was examined. The samples were examined on the presence of aflatoxin B1 using the method of thin layer chromatography (TLC) and simultaneously, using ELISA tests. Milk samples were examined using immunoenzyme tests for the presence of aflatoxin M1. Aflatoxin content in all the examined feed and mix samples was below LOD (limit of detection) of TLC method, also this content was below MRL according to ELISA method. In total of 50 samples of raw milk, aflatoxin M1 was detected in two samples originating from different farms. Aflatoxin was detected in 7.5 ng/l, i.e. 10 ng/l respectively, what is considerably lower than MRL. Based on the obtained results it is considered that obligatory control of raw milk for the presence of aflatoxin is necessary.

A Novel Rapid Enzyme Immunoassay (Fluorophos BetaScreen) for Detection of β-Lactam Residues in Ex-Farm Raw Milk

1998 ◽  
Vol 61 (7) ◽  
pp. 808-811 ◽  
Author(s):  
ÅSE STERNESJÖ ◽  
GÖRAN JOHNSSON
Keyword(s):  
Enzyme Immunoassay ◽  
Limit Of Detection ◽  
Raw Milk ◽  
Penicillin G ◽  
Milk Samples ◽  
Quality Testing ◽  
Response Profile ◽  
Very High ◽  
Higher Sensitivity

A novel, qualitative enzyme immunoassay based on fluorescence detection for determination of (β-lactam antibiotics in raw, commingled milk (Fluorophos BetaScreen E. U. test) was evaluated. A dose-response profile for penicillin G was constructed by analysis of spiked milk samples. The limit of detection, defined as the concentration of penicillin G that resulted in 95% of the samples being evaluated as positive, was 1.8 μg/kg. The repeatability of the assay was very high both within and between the three participating milk quality testing laboratories. In total 5,061 randomly selected tanker milk samples were analyzed with the BetaScreen test and compared with the Delvotest SP. The agreement between the two tests was 99.7%. Probably due to a higher sensitivity to penicillin G, the BetaScreen test detected almost twice as many suspect positive tanker milk samples (0.45%) as the Delvotest SP (0.26%).

Determination of aflatoxin M1 levels in raw milk samples in Gilan, Iran

2013 ◽  
Vol 5 ◽  
pp. 151-156 ◽  
Author(s):  
M. Laleh rokhi ◽  
R. Kazemi Darsanaki ◽  
M. Mohammadi ◽  
M. Hassani Kolavani ◽  
Kh. Issazadeh ◽  
...  
Keyword(s):  
Raw Milk ◽  
Aflatoxin M1 ◽  
Milk Samples
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