Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Application of Competitive ELISA Method for Estimation of Urinary Aflatoxin M1 Level

ELISA 방법을 이용한 요중 아플라톡신 M1 측정

  • Kim, Yong-Dae (Department of Preventive Medicine and Medical Research Institute, College of Medicine, Chungbuk National University) ;
  • Kim, Heon (Department of Preventive Medicine and Medical Research Institute, College of Medicine, Chungbuk National University)
  • 김용대 (충북대학교 의과대학 예방의학교실 및 의학연구소) ;
  • 김헌 (충북대학교 의과대학 예방의학교실 및 의학연구소)
  • Received : 2013.01.02
  • Accepted : 2013.02.05
  • Published : 2013.02.28
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Abstract

We compared the efficacy of the competitive ELISA method for measuring the level of urinary aflatoxin M1 (AFM1) with that of the HPLC-fluorescence detector (HPLC-FLD) method. The recovery rate of AFM1 with the ELISA method was 105% (73-124%), and the coefficient of variation of the analysis was 6.85%. The ELISA method showed a 0.20 pg/ml and 0.62 pg/ml limit of detection and limit of quantitation, respectively. In correlation analysis, the two methods showed a very strong and statistically significant correlation (R=0.96, p<0.01). However, in spite of the strong correlation, the ELISA method tended to overestimate the urinary AFM1 concentration compared to the HPLC-FLD method. These results suggest that the competitive ELISA method may be a useful technique for measuring the AFM1 level in high-throughput urine samples, but it needs to be corrected with a regression equation from regression analysis with the HPLC-FLD method.

본 연구는 요중 아플라톡신 M1 (AFM1)의 농도를 측정할 수 있는 competitive ELISA 방법의 특성을 HPLC-fluorescence detector (HPLC-FLD) 방법과 비교하여 평가하였다. ELISA 방법에서의 AFM1의 회수율은 105% (73-124%)였고 측정의 변이계수는 6.85%로 나타났다. ELISA 방법에서의 검출한계와 정량한계는 각각 0.20 pg/ml과 0.62 pg/ml로 조사되었으며, 두 방법을 이용하여 측정한 요중 AFM1 농도는 상관계수 0.96의 매우 높은 상관성이 있는 것으로 확인되었다(p<0.01). 그러나, 이렇게 높은 상관성에도 불구하고, ELISA 방법을 이용한 요중 AFM1의 농도는 HPLC-FLD 방법으로 측정한 값에 비해 상대적으로 높게 나타나는 경향을 보여 ELISA를 이용한 방법이 단시간에 많은 시료를 분석할 수 있는 장점은 있으나 그 결과는 HPLC-FLD 방법을 이용해서 얻은 회귀식을 이용하여 보정을 한 후 제시할 필요가 있는 것으로 판단된다.

Keywords

References

  1. Applebaum, R. S., Brackett, R. E., Wiseman, D. W. and Marth, E. H. 1982. Aflatoxin: oxicity to dairy cattle and occurrence in milk and milk products. A Review J Food Prot 45, 752-777.
  2. Cheng, Z., Root, M., Ran, W., Chen, J. and Campbell, T. C. 1997. Use of an improved method for analysis of urinary aflatoxin M1 in a survey of Mainland China and Taiwan. Cancer Epidem Biomarker Prev 6, 523-529.
  3. Everley, R. A., Ciner, F. L., Zhan, D., Scholl, P. F., Groopman, J. D. and Croley, T. R. 2007. Measurement of aflatoxin and aflatoxin metabolites in urine by liquid chromatographytandem mass spectrometry. J Anal Toxicol 31, 150-156. https://doi.org/10.1093/jat/31.3.150
  4. Hu, W. G., Tien, C. C., Wang, Y. H., Ting, T. Y., Fong, Y. C., Loung, Y. T. and Huang, S. S. 1983. Urinary excretion of aflatoxin M1 in inhabitants of region with high liver cancer incidence in Guangxi. Hygiene Invest 27, 152-154.
  5. Jonsyn-Ellis, F. E. 2001. Seasonal variation in exposure frequency and concentration levels of aflatoxins and ochratoxins in urine samples of boys and girls. Mycopathologia 152, 35-40. https://doi.org/10.1023/A:1011950512675
  6. Kim, E. K., Shon, D. H., Ryu, D., Park, J. W., Hwang, H. J. and Kim, Y. B. 2000. Occurrenceofaflatoxin M1 in Korean dairy products determined by ELISA and HPLC. Food Addit Contam 17, 59-64. https://doi.org/10.1080/026520300283595
  7. Kim, H. J., Kwak, B. Y. and Shon, D. H. 2009. Detection of Aflatoxin M1 in Human and Porcine Urine and Its Risk Assessment. Korean J Food Sci Technol 41, 215-221.
  8. Leong, Y. H., Latiff, A. A., Ahmad, N. I. and Rosma, A. 2012. Exposure measurement of aflatoxins and aflatoxin metabolites in human body fluids. A short review. Mycotoxin Res 28, 79-87. https://doi.org/10.1007/s12550-012-0129-8
  9. Neal, G. E., Eaton, D. L., Judah, D. J. and Verma, A. 1998. Metabolism and toxicity of aflatoxins M1 and B1 in human- derived in vitro systems. Toxicol Appl Pharmacol 151, 152-158. https://doi.org/10.1006/taap.1998.8440
  10. Park, J. W., Kim, E. K. and Kim, Y. B. 2004. Estimation of the daily exposure of Koreans to flatoxin B1 through food consumption. Food Addit Contam 21, 70-75. https://doi.org/10.1080/02652030310001622782
  11. Wogan, G. N., Kensler, T. W. and Groopman, J. D. 2012. Present and future directions of translational research on aflatoxin and hepatocellular carcinoma. A review Food Addit Contam Part A Chem Anal Control Expo Risk Assess 29, 249-257. https://doi.org/10.1080/19440049.2011.563370
  12. Zarba, A., Wild, C. P., Hall, A. J., Montesano, R., Hudson, G. J. and Groopman, J. D. 1992. Aflatoxin M1 in human breast milk from The Gambia, west Africa, quantified by combined monoclonal antibody immunoaffinity chromatography and HPLC. Carcinogenesis 13, 891-894. https://doi.org/10.1093/carcin/13.5.891

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