An enzyme-linked immunoassay is described for the fluorometric determination of aflatoxin B1 (AFB1). It is based on the use of carbon dots (CDs) synthesized by using Litchi chinensis as the carbon source via a hydrothermal method. The CDs were modified with MnO2 nanosheets upon which their blue fluorescence (with excitation/emission peaks at 340/425 nm) is quenched. In the presence of AFB1, a competitive immunoreaction takes place between (1) AFB1 conjugated to bovine serum albumin and deposited in the wells of a microplate, and (2) antibody against AFB1 and labeled with alkaline phosphatase (ALP). On subsequent addition of ascorbic acid 2-phosphate, it will be hydrolyzed by ALP to form ascorbic acid and phosphate. The ascorbic acid produced reduces MnO2 nanosheets to Mn2+ ions which then are relased from the CDs. This causes the recovery of fluorescence. Under optimum conditions, fluorescence decreases linearly with increasing AFB1 concentration in the range from 1.0 ng·kg−1 to 10 μg·kg−1, and the limit of detection is 0.69 ng·kg−1. The precision of this method (expressed as RSD) is ±10.3%. The accuracy was tested by analyzing both naturally contaminated and spiked food samples, and the results were in good agreement with those obtained by the established ELISA.
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Support by the National Natural Science Foundation of China (Grant nos.: 21475025, 21675029 and 21874022), and the Chongqing Science & Technology Commission of China (Grant no.: CSTC2015JCYJBX0126 and CSTC2016SHMSZX20001) is gratefully acknowledged.
The author(s) declare that they have no competing interests.
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Tang, D., Lin, Y. & Zhou, Q. Carbon dots prepared from Litchi chinensis and modified with manganese dioxide nanosheets for use in a competitive fluorometric immunoassay for aflatoxin B1. Microchim Acta 185, 476 (2018). https://doi.org/10.1007/s00604-018-3012-2
- Fluorescence immunoassay
- Oxidation-reduction reaction
- Contaminated and spiked sample
- Peanut sample
- Litchis fruit