Comparative Performance of Fluorometry and High Performance Liquid Chromatography in the Detection of Alfatoxin M1 in Two Commercial Cheeses
Aflatoxin M1 (AFM1) is frequently found in milk and dairy products. It is a metabolite formed in cows from aflatoxin B1 (AFB1), contained in animal feeds. In cheese production AFM1 distributes between curds and whey. In this study, cows were fed 64 µg/AFB1/d for the high treatment, and 5 µg/AFB1/d f...
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ndltd-UTAHS-oai-digitalcommons.usu.edu-etd-17312019-10-13T05:56:05Z Comparative Performance of Fluorometry and High Performance Liquid Chromatography in the Detection of Alfatoxin M1 in Two Commercial Cheeses Pena, Gustavo Aflatoxin M1 (AFM1) is frequently found in milk and dairy products. It is a metabolite formed in cows from aflatoxin B1 (AFB1), contained in animal feeds. In cheese production AFM1 distributes between curds and whey. In this study, cows were fed 64 µg/AFB1/d for the high treatment, and 5 µg/AFB1/d for the low treatment, to obtain milk contaminated with AFM1 over the 0.5 µg/L and under 0.05 µg/L restrictions, respectively. Cheese was manufactured with milk contaminated with AFM1 at 0.8 and 0.03 ìg/kg by the higher and lower treatment, respectively. Two commercial cheeses were elaborated: a hard-aged cheese (cheddar cheese) and soft high moisture cheese (fresco cheese) to evaluate whether the cheese type had any impact on AFM1 analysis. AFM1 was extracted from cheese using immunoaffinity columns. Analyses were carried out by using high pressure liquid chromatography (HPLC) as the reference method and fluorometry as a method of validation. Analysis was by 2-way fixed factor analyses. AFM1 was detected in all samples by both methods of analysis. There were no detectable statistical differences between cheese types (P>0.05). AFM1 content was significantly different between the high and low concentration of AFB1 used to make the cheese type (P<0.01). Our regression model shows a linear relationship between fluorometry and HPLC methods; R2 = 0.9141 from cheddar cheese and R2 = 0.9141 from fresco cheese. There were no statistical differences between methods of analysis (P>0.05). Carryover of AFM1 in cheese detected by fluorometry in cheddar cheese was 163% and 80% for high and low treatments, respectively, and in fresco cheese was 119 and 133 for high and low treatments, respectively. These carryovers are below that reported in the literature. Results suggest that fluorometry is a simple and reliable AFM1 detection method for screening samples of complex matrices such as cheese. 2010-05-01T07:00:00Z text application/pdf https://digitalcommons.usu.edu/etd/735 https://digitalcommons.usu.edu/cgi/viewcontent.cgi?article=1731&context=etd Copyright for this work is held by the author. Transmission or reproduction of materials protected by copyright beyond that allowed by fair use requires the written permission of the copyright owners. Works not in the public domain cannot be commercially exploited without permission of the copyright owner. Responsibility for any use rests exclusively with the user. For more information contact Andrew Wesolek (andrew.wesolek@usu.edu). All Graduate Theses and Dissertations DigitalCommons@USU aflatoxin cheese fluorometry animal culture and nutrition animal pathology agricultural chemistry Agronomy and Crop Sciences Animal Sciences Food Science |
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aflatoxin cheese fluorometry animal culture and nutrition animal pathology agricultural chemistry Agronomy and Crop Sciences Animal Sciences Food Science Pena, Gustavo Comparative Performance of Fluorometry and High Performance Liquid Chromatography in the Detection of Alfatoxin M1 in Two Commercial Cheeses |
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Aflatoxin M1 (AFM1) is frequently found in milk and dairy products. It is a metabolite formed in cows from aflatoxin B1 (AFB1), contained in animal feeds. In cheese production AFM1 distributes between curds and whey. In this study, cows were fed 64 µg/AFB1/d for the high treatment, and 5 µg/AFB1/d for the low treatment, to obtain milk contaminated with AFM1 over the 0.5 µg/L and under 0.05 µg/L restrictions, respectively. Cheese was manufactured with milk contaminated with AFM1 at 0.8 and 0.03 ìg/kg by the higher and lower treatment, respectively. Two commercial cheeses were elaborated: a hard-aged cheese (cheddar cheese) and soft high moisture cheese (fresco cheese) to evaluate whether the cheese type had any impact on AFM1 analysis. AFM1 was extracted from cheese using immunoaffinity columns. Analyses were carried out by using high pressure liquid chromatography (HPLC) as the reference method and fluorometry as a method of validation. Analysis was by 2-way fixed factor analyses. AFM1 was detected in all samples by both methods of analysis. There were no detectable statistical differences between cheese types (P>0.05). AFM1 content was significantly different between the high and low concentration of AFB1 used to make the cheese type (P<0.01). Our regression model shows a linear relationship between fluorometry and HPLC methods; R2 = 0.9141 from cheddar cheese and R2 = 0.9141 from fresco cheese. There were no statistical differences between methods of analysis (P>0.05). Carryover of AFM1 in cheese detected by fluorometry in cheddar cheese was 163% and 80% for high and low treatments, respectively, and in fresco cheese was 119 and 133 for high and low treatments, respectively. These carryovers are below that reported in the literature. Results suggest that fluorometry is a simple and reliable AFM1 detection method for screening samples of complex matrices such as cheese. |
author |
Pena, Gustavo |
author_facet |
Pena, Gustavo |
author_sort |
Pena, Gustavo |
title |
Comparative Performance of Fluorometry and High Performance Liquid Chromatography in the Detection of Alfatoxin M1 in Two Commercial Cheeses |
title_short |
Comparative Performance of Fluorometry and High Performance Liquid Chromatography in the Detection of Alfatoxin M1 in Two Commercial Cheeses |
title_full |
Comparative Performance of Fluorometry and High Performance Liquid Chromatography in the Detection of Alfatoxin M1 in Two Commercial Cheeses |
title_fullStr |
Comparative Performance of Fluorometry and High Performance Liquid Chromatography in the Detection of Alfatoxin M1 in Two Commercial Cheeses |
title_full_unstemmed |
Comparative Performance of Fluorometry and High Performance Liquid Chromatography in the Detection of Alfatoxin M1 in Two Commercial Cheeses |
title_sort |
comparative performance of fluorometry and high performance liquid chromatography in the detection of alfatoxin m1 in two commercial cheeses |
publisher |
DigitalCommons@USU |
publishDate |
2010 |
url |
https://digitalcommons.usu.edu/etd/735 https://digitalcommons.usu.edu/cgi/viewcontent.cgi?article=1731&context=etd |
work_keys_str_mv |
AT penagustavo comparativeperformanceoffluorometryandhighperformanceliquidchromatographyinthedetectionofalfatoxinm1intwocommercialcheeses |
_version_ |
1719267125432942592 |