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Liposomes loaded with quantum dots for ultrasensitive on-site determination of aflatoxin M1 in milk products

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Abstract

A quantitative fluorescence-labeled immunosorbent assay and qualitative on-site column tests were developed for the determination of aflatoxin M1 in milk products. The use of liposomes loaded with quantum dots as a label significantly increased the assay sensitivity by encapsulating multiple quantum dots in a single liposome and, therefore, amplifying the analytical signal. Two different techniques were compared to obtain aflatoxin–protein conjugates, used for further coupling with the liposomes. The influence of nonspecific interactions of the liposome-labeled conjugates obtained with the surface of microtiter plates and column cartridges was evaluated and discussed. The limit of detection for fluorescence-labeled immunosorbent assay was 0.014 μg kg-1. For qualitative on-site tests, the cutoff was set at 0.05μg kg-1, taking into account the EU maximum level for aflatoxin M1 in raw milk, heat-treated milk, and milk for the manufacture of milk-based products. The direct addition of labeled conjugate to the milk samples resulted in an additional decrease of analysis time. An intralaboratory validation was performed with sterilized milk and cream samples artificially spiked with aflatoxin M1 at concentrations less than, equal to and greater than the cutoff level. It is shown that milk products can be analyzed without any sample preparation, just diluted with the buffer. The rates for false-positive and false-negative results were below 5 % (2.6 % and 3.3 %, respectively).

Scheme of liposome and liposome loaded with water-insoluble quantum dots

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Acknowledgments

This research was financially supported by the Belgian Federal Science Policy Office (BELSPO) in order to promote the S&T cooperation with China (contract BL/02/C58), the Special Research Fund (BOF), Ghent University (01SB2510), and the Russian Foundation of Basic Research (RFBR, project 12-03-91167). We gratefully acknowledge Tom Coenye (Ghent University) for making available the EnVision 2104 multilabel plate reader, S. De Smedt for giving us the opportunity to perform the experiments in his laboratory, and Bart Lucas for his appreciable advice.

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Authors and Affiliations

  1. Laboratory of Food Analysis, Faculty of Pharmaceutical Sciences, Ghent University, Harelbekestraat 72, 9000, Ghent, Belgium

    N. V. Beloglazova & S. De Saeger

  2. “Neuronet” Department, OJSC CSRIT Technomash, I. Franko str. 4, 121108, Moscow, Russia

    P. S. Shmelin

  3. Department of General and Inorganic Chemistry, Chemistry Institute, Saratov State University, Astrakhanskaya 83, 410012, Saratov, Russia

    I. Yu Goryacheva

Authors
  1. N. V. Beloglazova
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  2. P. S. Shmelin
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  3. I. Yu Goryacheva
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  4. S. De Saeger
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Corresponding author

Correspondence to N. V. Beloglazova.

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Published in the topical collection Rapid Detection in Food and Feed with guest editors Rudolf Krska and Michel Nielen.

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Beloglazova, N.V., Shmelin, P.S., Goryacheva, I.Y. et al. Liposomes loaded with quantum dots for ultrasensitive on-site determination of aflatoxin M1 in milk products. Anal Bioanal Chem 405, 7795–7802 (2013). https://doi.org/10.1007/s00216-013-7096-6

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  • DOI: https://doi.org/10.1007/s00216-013-7096-6

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