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. 2024 Dec 16;13(24):4057.
doi: 10.3390/foods13244057.

Development of a Dual-Readout Multicolor Immunoassay for the Rapid Analysis of Isocarbophos in Vegetable and Fruit Samples

Zijian Chen  1   2   3 , Wei-Xuan Huang  4 , Hongwu Wang  1   2   3 , Meiling Zhang  1 , Kai Chen  1 , Hao Deng  5
Affiliations

Development of a Dual-Readout Multicolor Immunoassay for the Rapid Analysis of Isocarbophos in Vegetable and Fruit Samples

Zijian Chen et al. Foods. .

Abstract

Multicolor immunoassay is a powerful tool for rapid analysis without the use of bulky instruments owing to various color conversions, which is suitable for on-site visual analysis for pesticides. Herein, this study developed a multicolor immunoassay for the rapid detection of isocarbophos. After competitive immunoassay, the secondary antibody (GAM-ALP) catalyzed ascorbyl-2-phosphate (AAP) into ascorbic acid (AA). The AA can reduce K3[Fe(CN)6] into K4[Fe(CN)6]. The latter can react with Fe3+ to form Prussian blue; otherwise, the orange AAP-Fe3+ complex was generated. Therefore, the multicolor immunoassay achieved a color conversion of orange-green-blue in response to isocarbophos, allowing for rapid semiquantitative analysis by the naked eye. After parameter optimization, the multicolor immunoassay was developed depending on the ratiometric absorbance between the Prussian blue and AAP-Fe3+ complex. Moreover, a smartphone was used to measure the RGB value of the color conversion for the development of portable visual, quantitative analysis. Both the absorbance-based and RGB-based multicolor immunoassays showed good accuracy and practicability in the recovery test. This study provided a common approach for the development of dual-readout multicolor immunoassay, which can be used for on-site rapid screening by quantitative or visual semiquantitative analysis.

Keywords: RGB analysis; alkaline phosphase; isocarbophos; multicolor immunoassay; pesticide.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
(A) The schematic diagram of the multicolor immunoassay; (B,C) the color reaction of the reagents for the development of multicolor immunoassay; (D) the absorption spectrum and (E) A715/A425 ratio for the color conversion of multicolor immunoassay.
Figure 2
Figure 2
The (A) SEM and (B) TEM graph of Prussian blue; (C) the DLS analysis for Prussian blue; (D) the high-resolution image for Prussian blue and the insert graph indicated the lattice fringe of Prussian blue nanocube; the (E) FTIR and (F) XRD analysis of Prussian blue.
Figure 3
Figure 3
The XPS analysis of Prussian blue; (A) survey XPS spectra; (B) C 1s; (C) N 1s; (D) Fe 2p.
Figure 4
Figure 4
(A) The GAM-ALP dilution optimization; (B) the AAP concentration optimization; (C) the titer for multicolor immunoassay with different concentrations of coating antigen; (D) the optimization of the concentration of coating antigen. The red asterisk indicates the optimized parameter.
Figure 5
Figure 5
The (A) calibration curve and (B) linear range of multicolor immunoassay using the microplate reader determination.
Figure 6
Figure 6
(A) The schematic diagram of the RGB mode for the analysis of multicolor immunoassay; (B) the B/G mode for the RGB analysis; (C) the B/R mode for the RGB analysis.
Figure 7
Figure 7
The (A) calibration curve and (B) linear range of multicolor immunoassay using the RGB value determination.
Figure 8
Figure 8
The study of matrix effect with different dilution ratios.
See this image and copyright information in PMC

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References

    1. Hassani S., Momtaz S., Vakhshiteh F., Maghsoudi A.S., Ganjali M.R., Norouzi P., Abdollahi M. Biosensors and their applications in detection of organophosphorus pesticides in the environment. Arch. Toxicol. 2016;91:109–130. doi: 10.1007/s00204-016-1875-8. - DOI - PubMed
    1. Aragay G., Pino F., Merkoçi A. Nanomaterials for sensing and destroying pesticides. Chem. Rev. 2012;112:5317–5338. doi: 10.1021/cr300020c. - DOI - PubMed
    1. Balali-Mood M., Balali-Mood K., Moodi M., Balali-Mood B. Health aspects of organophosphorous pesticides in asian countries. Iran. J. Public Health. 2012;41:1–14. - PMC - PubMed
    1. Canli A.G., Surucu B., Ulusoy H.I., Yilmaz E., Kabir A., Locatelli M. Analytical methodology for trace determination of propoxur and fenitrothion pesticide residues by decanoic acid modified magnetic nanoparticles. Molecules. 2019;24:4621. doi: 10.3390/molecules24244621. - DOI - PMC - PubMed
    1. Zhao F.N., Wang L., Li M.Y., Wang M., Liu G.Y., Ping J.F. Nanozyme-based biosensor for organophosphorus pesticide monitoring: Functional design, biosensing strategy, and detection application. TrAC Trends Anal. Chem. 2023;165:117152. doi: 10.1016/j.trac.2023.117152. - DOI

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