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Determination of Aflatoxins in Yogurt by Dispersive Liquid–Liquid Microextraction and HPLC with Photo-Induced Fluorescence Detection

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Abstract

A high-performance liquid chromatography (HPLC) method with fluorescence detection for the determination of aflatoxins B1, B2, G1, G2, and M1 in yogurt using dispersive liquid–liquid microextraction as alternative sample treatment has been developed. To enhance the fluorescence of aflatoxins B1 and G1, a post-column photochemical derivatization has been proposed, avoiding the use of derivatization reagents. The method was validated using natural yogurt as representative matrix, showing a good linearity in the studied range (25–500 ng kg−1) and limits of quantification below the maximum level established by European Union in milk for the manufacture of milk-based products. Satisfactory recoveries ranging from 69.4 to 99.7 %, with relative standard deviations lower than 11.2 %, were obtained for all the compounds. The proposed method is simple, rapid, with low solvent consumption, inexpensive, and environmentally friendly.

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References

  • Afzali D, Ghanbarian M, Mostafavi A, Shamspur T, Ghaseminezhad S (2012) A novel method for high preconcentration of ultra-trace amounts of B1, B2, G1 and G2 aflatoxins in edible oils by dispersive liquid-liquid microextraction after immunoaffinity column clean-up. J Chromatogr A 1247:35–41

    Article  CAS  Google Scholar 

  • Amini A, Afzali-Grooh D, Chamsaz M (2014) Determination of aflatoxins B1 and B2 in powdered milk using modified liquid chromatography method. J Kerman Univ Med Sci 21:321–331

    Google Scholar 

  • Amoli-Diva M, Taherimaslak Z, Allahyari M, Pourghazi K, Manafi MH (2015) Application of dispersive liquid–liquid microextraction coupled with vortex-assisted hydrophobic magnetic nanoparticles based solid-phase extraction for determination of aflatoxin M1 in milk samples by sensitive micelle enhanced spectrofluorimetry. Talanta 134:98–104

    Article  CAS  Google Scholar 

  • Andrade PD, da Silva JLG, Caldas ED (2013) Simultaneous analysis of aflatoxins B1, B2, G1, G2, M1 and ochratoxin A in breast milk by high-performance liquid chromatography/fluorescence after liquid-liquid extraction with low temperature purification (LLE-LTP). J Chromatogr A 1304:61–68

    Article  CAS  Google Scholar 

  • Anfossi L, Calderara M, Baggiani C, Giovannoli C, Arletti E, Giraudi G (2008) Development and application of solvent-free extraction for the detection of aflatoxin M1 in dairy products by enzyme immunoassay. J Agric Food Chem 56:1852–1857

    Article  CAS  Google Scholar 

  • Bakirdere S, Bora S, Bakirdere EG, Aydin F, Arslan Y, Komesli OT, Aydin I, Yildirim E (2012) Aflatoxin species: their health effects and determination methods in different foodstuffs. Cent Eur J Chem 10:675–685

    CAS  Google Scholar 

  • Beltrán E, Ibáñez M, Sancho JV, Cortés MÁ, Yusà V, Hernández F (2011) UHPLC–MS/MS highly sensitive determination of aflatoxins, the aflatoxin metabolite M1 and ochratoxin A in baby food and milk. Food Chem 126:737–744

    Article  Google Scholar 

  • Bognanno M, La Fauci L, Ritieni A, Tafuri A, De Lorenzo A, Micari P et al (2006) Survey of the occurrence of aflatoxin M1 in ovine milk by HPLC and its confirmation by MS. Mol Nutr Food Res 50:300–305

    Article  CAS  Google Scholar 

  • Campone L, Piccinelli AL, Celano R, Rastrelli L (2011) Application of dispersive liquid–liquid microextraction for the determination of aflatoxins B1, B2, G1 and G2 in cereal products. J Chromatogr A 1218:7648–7654

    Article  CAS  Google Scholar 

  • Campone L, Piccinelli AL, Celano R, Russo M, Rastrelli L (2013) Rapid analysis of aflatoxin M1 in milk using dispersive liquid-liquid microextraction coupled with ultrahigh pressure liquid chromatography tandem mass spectrometry. Anal Bioanal Chem 405:8645–8652

    Article  CAS  Google Scholar 

  • Creppy EE (2002) Update of survey, regulation and toxic effects of mycotoxins in Europe. Toxicol Lett 127:19–28

    Article  CAS  Google Scholar 

  • European Commission (2006a) Commission Regulation (EC) No 401/2006 of 23 February 2006 laying down the methods of sampling and analysis for the official control of the levels of mycotoxins in foodstuffs. Off J Eur Union L 70/12

  • European Commission (2006b) Commission Regulation 1881/2006 of 19 December 2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs. Off J Eur Union L 364/15

  • European Commission (2010) Commission regulation 165/2010 of 26 February 2010 amending regulation (EC) No 1881/2006 setting the maximum levels for certain contaminants in foodstuffs as regards aflatoxins. Off J Eur Union L 50/12

  • Gürbay A, Sabuncuoğlu SA, Girgin G, Şahin G, Yiğit Ş, Yurdakök M, Tekinalp G (2010) Exposure of newborns to aflatoxin M1 and B1 from mothers’ breast milk in Ankara, Turkey. Food Chem Toxicol 48:314–319

    Article  Google Scholar 

  • Hoyos Ossa DE, Hincapié DA, Peñuela GA (2015) Determination of aflatoxin M1 in ice cream samples using immunoaffinity columns and ultra-high performance liquid chromatography coupled to tandem mass spectrometry. Food Control 56:34–40

    Article  CAS  Google Scholar 

  • Iha MH, Barbosa CB, Duarte Favaro RM (2011) Chromatographic method for the determination of aflatoxin M1 in cheese, yogurt, and dairy beverages. J AOAC Int 94:1513–1518

    Article  CAS  Google Scholar 

  • International Agency for Research on Cancer (IARC) (2002) IARC monographs on the evaluation of carcinogenic risks to humans. In traditional herbal medicines, some mycotoxins, napthalene and styrene (Vol. 82). Lyon

  • International Agency for Research on Cancer (IARC) (2012) IARC monographs on the evaluation of carcinogenic risks to humans. Chemical agents and related occupations (Vol. 100F). Lyon, France.

  • Iqbal SZ, Asi MR (2013) Assessment of aflatoxin M1 in milk and milk products from Punjab, Pakistan. Food Control 30:235–239

    Article  CAS  Google Scholar 

  • Iqbal SZ, Asi MR, Jinap S (2013) Variation of aflatoxin M1 contamination in milk and milk products collected during winter and summer seasons. Food Control 34:714–718

    Article  CAS  Google Scholar 

  • Iqbal SZ, Jinap S, Pirouz AA, Ahmad Faizal AR (2015) Aflatoxin M1 in milk and dairy products, occurrence and recent challenges: a review. Trends Food Sci Tech 46:110–119

    Article  CAS  Google Scholar 

  • Jan D, Samson RA (2007) A multifaceted approach to fungi and food. In: Dijksterhuis J, Samson RA (eds) Food mycology, 1st edn. CRC Press, New York, pp 121–134

    Google Scholar 

  • Jiang W, Wang Z, Nölke G, Zhang J, Niu L, Shen J (2013) Simultaneous determination of aflatoxin B1 and aflatoxin M1 in food matrices by enzyme-linked immunosorbent assay. Food Anal Methods 6:767–774

    Article  Google Scholar 

  • Kok WT (1994) Derivatization reactions for the determination of aflatoxins by liquid chromatography with fluorescence detection. J Chromatogr B 659:127–137

    Article  CAS  Google Scholar 

  • Kos J, Lević J, Đuragić O, Kokić B, Miladinović I (2014) Occurrence and estimation of aflatoxin M1 exposure in milk in Serbia. Food Control 38:41–46

    Article  CAS  Google Scholar 

  • Lee D, Lee KG (2015) Analysis of aflatoxin M1 and M2 in commercial dairy products using high-performance liquid chromatography with a fluorescence detector. Food Control 50:467–471

    Article  CAS  Google Scholar 

  • Leong MI, Fuh MR, Huang SD (2014) Beyond dispersive liquid-liquid microextraction. J Chromatogr A 1335:2–14

    Article  CAS  Google Scholar 

  • Mulunda M, Mike D (2014) Occurrence of aflatoxin M1 from rural subsistence and commercial farms from selected areas of South Africa. Food Control 39:92–96

    Article  CAS  Google Scholar 

  • Prieto-Simón B, Campàs M (2009) Immunochemical tools for mycotoxin detection in food. Monatsh Chem 140:915–920

    Article  Google Scholar 

  • Reiter E, Zentek J, Razzaz E (2009) Review on sample preparation strategies and methods used for the analysis of aflatoxins in food and feed. Mol Nutr Food Res 53:508–524

    Article  CAS  Google Scholar 

  • Romero-González R, Martínez-Vidal JL, Garrido-Frenich A (2010) Liquid chromatography for the determination of mycotoxins in foods. Food Sci Technol Ser. New York: Nova Science Publisher

  • Saraji M, Boroujeni MK (2014) Recent developments in dispersive liquid-liquid microextraction. Anal Bioanal Chem 406:2027–2066

    Article  CAS  Google Scholar 

  • Sartori AV, Swensson de Mattos J, de Moraes MHP, da Nóbrega AW (2015) Determination of aflatoxins M1, M2, B1, B2, G1, and G2 and ochratoxin A in UHT and powdered milk by modified QuEChERS method and ultra-high-performance liquid chromatography tandem mass spectrometry. Food Anal Methods 8:2321–2330

    Article  Google Scholar 

  • Shephard GS (2009) Aflatoxin analysis at the beginning of the twenty-first century. Anal Bioanal Chem 395:1215–1224

    Article  CAS  Google Scholar 

  • Viñas P, Campillo N, López-García I, Hernández-Córdoba M (2013) Dispersive liquid-liquid microextraction in food analysis. A critical review. Anal Bioanal Chem 406:2067–2099

    Article  Google Scholar 

  • Wang H, Zhou XJ, Liu YQ, Yang HM, Guo QL (2010) Determination of aflatoxin M1 in milk by triple quadrupole liquid chromatography-tandem mass spectrometry. Food Addit Contam 27:1261–1265

    Article  CAS  Google Scholar 

  • Wang Y, Liu X, Xiao C, Wang Z, Wang J, Xiao H et al (2012) HPLC determination of aflatoxin M1 in liquid milk and milk powder using solid phase extraction on OASIS HLB. Food Control 28:131–134

    Article  Google Scholar 

Download references

Acknowledgments

Financial support was from the Project Reference AGL2015-70708-R (MINECO/FEDER, UE) and from the Excellence Project Reference P12-AGR-1647 (Junta de Andalucía). AMH thanks the “Erasmus Mundus–Al Idrisi II program” for a predoctoral grant.

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    Authors and Affiliations

    1. Department of Analytical Chemistry, Faculty of Sciences, University of Granada, Av Fuentenueva s/n, 18071, Granada, Spain

      Ahmed M. Hamed, David Moreno-González, Ana M. García-Campaña & Laura Gámiz-Gracia

    2. Department of Dairy Science, Faculty of Agriculture, Cairo University, 12613, Giza, Egypt

      Ahmed M. Hamed

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    1. Ahmed M. Hamed
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    2. David Moreno-González
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    3. Ana M. García-Campaña
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    4. Laura Gámiz-Gracia
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    Correspondence to Laura Gámiz-Gracia.

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    Ahmed M. Hamed and David Moreno-González contributed equally to this work.

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    Hamed, A.M., Moreno-González, D., García-Campaña, A.M. et al. Determination of Aflatoxins in Yogurt by Dispersive Liquid–Liquid Microextraction and HPLC with Photo-Induced Fluorescence Detection. Food Anal. Methods 10, 516–521 (2017). https://doi.org/10.1007/s12161-016-0611-6

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    • DOI: https://doi.org/10.1007/s12161-016-0611-6

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