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Detection of Aflatoxin M1 in milk with a Mach–Zehnder Interferometric immunosensor
1, 2 , 3 , 4 , 5 , 2 , 3 , * 1
1  Immunoassays–Immunosensors Lab, INRASTES, NCSR “Demokritos”, Aghia Paraskevi, 15341, Greece
2  Analytical Chemistry Lab, Department of Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis Zografou,15771, Greece
3  Immunoassays-Immunosensors Lab, INRASTES, NCSR “Demokritos”, Athens, Greece
4  Optical sensors Lab, Institute of Nanoscience & Nanotechnology, NCSR “Demokritos”, Athens, Greece
5  Optical Sensors Lab, Institute of Nanoscience & Nanotechnology, NCSR “Demokritos”, Aghia Paraskevi, 15341, Greece
Academic Editor: Jun-Jie Zhu

Abstract:

Aflatoxin M1 (AFM1) is the hydroxylated form of Aflatoxin B1 (AFB1) and is expelled in the milk of both humans and animals following the consumption of AFB1-contaminated food. AFM1 has been categorized as a Group 1 carcinogen by the International Agency for Research on Cancer. Consequently, the European Commission has established a maximum allowable concentration of 50 pg/mL for AFM1 in dairy products and milk. Here, a rapid and sensitive approach for detecting AFM1 in bovine milk is presented. The analytical setup comprises a broad-band white LED, a spectrophotometer, and a silicon photonic probe, all interconnected by a bifurcated optical fiber [1]. Additionally, a laptop powers the system and facilitates signal monitoring through specialized software. The silicon photonic probe is equipped with two Mach–Zehnder interferometers: one functionalized with AFM1-bovine serum albumin conjugate, and the other with bovine serum albumin to serve as a blank. The analysis involves immersing the probe directly into a mixture of anti-AFM1 antibodies and the sample, followed by sequential immersion into biotinylated anti-rabbit IgG antibody and streptavidin solutions. The entire assay process takes 12 minutes, and the limit of detection in undiluted milk is 20 pg/mL, below the EU maximum allowable limit of 50 pg/mL. The assay demonstrates accuracy, with %recovery values ranging from 87.5 to 112%, and repeatability, with intra/inter-assay coefficients of variation below 7.6%. Given its analytical performance and compact instrumentation, the proposed immunosensor proves to be an ideal solution for precise on-site determination of AFM1 in milk samples.

Acknowledgements: This work was financially supported by the European Regional Development Fund of the European Union and Greek national funds through the Operational Program Competitiveness, Entrepreneurship and Innovation, under the call RESEARCH–CREATE–INNOVATE (project code: Τ2ΕΔΚ-01934 FOODSENS).

[1] M. Angelopoulou, et al., Biosens. Bioelectron. 215 (2022) 114570

Keywords: photonic sensor; aflatoxin M1; immunosensor; milk
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