Development of a Magnetic Nanoparticles-Based Screen-Printed Electrodes (MNPs-SPEs) Biosensor for the Quantification of Ochratoxin A in Cereal and Feed Samples
Abstract
:1. Introduction
2. Results and Discussion
2.1. Optimizations and Comparisons of Three Variants of MNPs-ELISA
2.2. Specificity Study
2.3. Optimization of the Electrochemical Biosensor Immunoassay
2.4. Calibration Curve of the Electrochemical Biosensor Immunoassay
2.5. Recovery Studies
2.6. Commercial Samples Analysis
3. Conclusions
4. Materials and Methods
4.1. Materials
4.2. Equipment
4.3. Synthesis of the OTA and Antibody Conjugates
4.4. Preparation of Magnetic Nanoparticles Conjugates
4.5. Development of MNP-ELISAs
4.6. Optimization of MNPs-ELISAs
4.7. Specificity Study
4.8. Development of the Electrochemical Biosensor Immunoassay
4.9. Recovery Study and Comparison of Detection in Commercial Samples by LC-MS/MS
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Analytical Methods | Final Concentration (μg/mL) | Dilution Ratio | |||
---|---|---|---|---|---|
Anti OTA-HRP | OTA-BSA-HRP | OTA-HRP | MNPs-BSA-OTA | MNPs-Anti OTA | |
Anti OTA-HRP MNPs-ELISA | 0.2 | NU a | NU | 1/50 | NU |
OTA-BSA-HRP MNPs-ELISA | NU | 0.1 | NU | NU | 1/100 |
OTA-HRP MNPs-ELISA | NU | NU | 0.5 | NU | 1/100 |
Analytical Method | LOD (ng/mL) | IC50 (ng/mL) | Detection Range (IC20–IC80, ng/mL) | Regression Equation |
---|---|---|---|---|
Anti OTA-HRP MNPs-ELISA | 0.25 | 1.53 | 0.39–5.94 | y = 0.5083x + 0.4065 (R2 = 0.9908) |
OTA-BSA-HRP MNPs-ELISA | 0.14 | 0.54 | 0.20–1.44 | y = 0.7032x + 0.6884 (R2 = 0.9927) |
OTA-HRP MNPs-ELISA | 0.04 | 0.31 | 0.06–1.13 | y = 0.4836x + 0.7477 (R2 = 0.9906) |
Methods | Matrix | LOD (ng/mL) | Recovery (%) | Detection Range (ng/mL) | Ref. |
---|---|---|---|---|---|
Nanobody-based ELISA | Cereal | 0.16 | 80–105 | 0.27–1.47 | [30] |
dsDNA-base fluorescence method | Corn | 5 | 89.2–94.1 | 0.00–100 | [33] |
Portable Flurescence | Cocoa | 1.25 | 79.05–83.25 | 1.25–10 | [34] |
Silver nanoparticles-based LFIA | Juice and wine | 0.06 | 88.0–110.0 | 0.08–5.0 | [35] |
Fluorescence polarisation aptamer | Wine | 1.1 | 83–113 | - | [36] |
Quantum dots-based aptasensor | Foodstuff | 0.5 | - | 1–30 | [37] |
Fluorescent biosensor | Corn | 2.57 | 96.5–101.4 | 5.0–160 | [38] |
A Label-Free Aptasensor | Corn | 0.012 | 96–106 | 0.04–0.48 | [39] |
A Polyaniline film-based aptasensor | - | 0.1 | - | 0.1–10 | [40] |
Fluorescent aptamer-based sensor | Corn | 0.8 | 83–106 | 1–100 | [41] |
FRET aptasensor | Peanut | 0.0025 | 90–110 | 0.01–20 | [42] |
Impedimetric aptasensor | Wine | 0.002 | 102–107 | 0.002–6 | [43] |
MNPs-SPEs sensor | Cereal and feed | 0.007 | 78.7–113.5 | 0.01–0.82 | This work |
Samples | Concentrations (μg/kg) | Recovery Rate (%) (Mean ± SD a) | CV b (%) | |
---|---|---|---|---|
Spiked | Detected | |||
1 | 1.25 | 1.42 | 113.5 ± 3.6 | 3.2 |
2 | 2.5 | 1.97 | 78.7 ± 7.3 | 9.3 |
3 | 5 | 4.26 | 85.3 ± 9.8 | 11.5 |
4 | 10 | 8.12 | 81.2 ± 5.8 | 7.2 |
5 | 20 | 18.34 | 91.6 ± 7.4 | 8.1 |
Samples | MNPs-SPEs Sensor (μg/kg), Mean ± SD a | LC-MS/MS (μg/kg), Mean ± SD |
---|---|---|
Corn 1 | 12.37 ± 1.86 | 14.87 ± 1.21 |
Corn 2 | 19.75 ± 2.17 | 22.94 ± 1.32 |
Corn 3 | 21.67 ± 1.69 | 25.57 ± 2.01 |
Corn 4 | 2.94 ± 0.67 | 3.98 ± 0.73 |
Corn 5 | 24.28 ± 2.41 | 27.71 ± 2.13 |
Wheat 1 | 27.41 ± 1.61 | 20.99 ± 1.71 |
Wheat 2 | 25.97 ± 2.03 | 20.01 ± 1.61 |
Wheat 3 | 2.35 ± 0.97 | 1.36 ± 0.12 |
Wheat 4 | 5.98 ± 1.03 | 4.18 ± 0.31 |
Feedstuff 1 | 15.86 ± 1.15 | 18.39 ± 1.03 |
Feedstuff 2 | 21.65 ± 1.24 | 15.97 ± 1.41 |
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Zhang, X.; Wang, Z.; Xie, H.; Sun, R.; Cao, T.; Paudyal, N.; Fang, W.; Song, H. Development of a Magnetic Nanoparticles-Based Screen-Printed Electrodes (MNPs-SPEs) Biosensor for the Quantification of Ochratoxin A in Cereal and Feed Samples. Toxins 2018, 10, 317. https://doi.org/10.3390/toxins10080317
Zhang X, Wang Z, Xie H, Sun R, Cao T, Paudyal N, Fang W, Song H. Development of a Magnetic Nanoparticles-Based Screen-Printed Electrodes (MNPs-SPEs) Biosensor for the Quantification of Ochratoxin A in Cereal and Feed Samples. Toxins. 2018; 10(8):317. https://doi.org/10.3390/toxins10080317
Chicago/Turabian StyleZhang, Xian, Zuohuan Wang, Hui Xie, Renjie Sun, Tong Cao, Narayan Paudyal, Weihuan Fang, and Houhui Song. 2018. "Development of a Magnetic Nanoparticles-Based Screen-Printed Electrodes (MNPs-SPEs) Biosensor for the Quantification of Ochratoxin A in Cereal and Feed Samples" Toxins 10, no. 8: 317. https://doi.org/10.3390/toxins10080317