Time-Resolved Fluorescent Immunochromatography of Aflatoxin B1 in Soybean Sauce: A Rapid and Sensitive Quantitative Analysis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagents and Apparatus
2.2. Conjugation of Eu-Nanospheres to Antibody
2.3. Preparation of the Immunochromatographic Strip
2.4. Evaluation of TRFIA
2.5. Sample Treatment for TRFIA
2.6. Sample Treatment for HPLC Analysis
3. Results and Discussion
3.1. Optimization of Eu-Probe
3.1.1. Optimal pH of Boric Acid Buffer Solution for Conjugation
3.1.2. Optimal Antibody Level Labeled with Eu-Nanospheres
3.2. Optimization of the IC Strip
3.3. Optimization of TRFIA
3.4. Optimization of Sample for TRFIA
3.5. Validation by HPLC
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Sample | Number | Spike µg·kg−1 | Result by TRFIA µg·kg−1 | CV% | Recovery % | |||||
---|---|---|---|---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | Mean | |||||
Soybean sauce | 1 | ND | ND | ND | ND | ND | ND | - | - | - |
2 | 0.80 | 0.99 | 0.97 | 0.90 | 0.79 | 0.92 | 0.91 | 8.6 | 114.3 | |
3 | 4.00 | 4.57 | 3.88 | 3.69 | 4.33 | 4.22 | 4.14 | 8.5 | 103.5 | |
4 | 8.00 | 7.90 | 7.39 | 7.74 | 8.43 | 7.26 | 7.75 | 6.0 | 96.8 | |
5 | 12.00 | 11.45 | 11.98 | 13.01 | 12.89 | 12.87 | 12.44 | 5.7 | 103.7 | |
Soybean sauce | 1 | ND | ND | ND | ND | ND | ND | - | - | - |
2 | 2.00 | 2.10 | 2.07 | 1.93 | 2.20 | 2.29 | 2.12 | 6.4 | 105.9 | |
3 | 5.00 | 5.02 | 5.24 | 5.46 | 5.34 | 5.09 | 5.23 | 3.4 | 104.6 | |
4 | 10.00 | 9.42 | 9.89 | 9.65 | 10.37 | 10.57 | 9.98 | 4.5 | 99.8 | |
5 | 15.00 | 12.98 | 12.07 | 13.58 | 13.12 | 13.68 | 13.09 | 5.0 | 87.2 | |
Average | 6.0 | 110.0 |
Soybean Sauce Sample | HPLC Detected AFB1 µg·kg−1 | Intra-Batch (n = 5) | Inter-Batch (n = 5) | ||
---|---|---|---|---|---|
Average Detected Value µg·kg−1 | Average CV % | Average Detected Value µg·kg−1 | Average CV % | ||
1# | ND | ND | - | ND | - |
2# | 0.8 | 0.69 | 7.9 | 0.68 | 8.7 |
3# | 3.2 | 3.43 | 4.6 | 3.58 | 5.2 |
4# | 8.9 | 9.30 | 5.3 | 9.20 | 5.6 |
5# | ND | ND | - | ND | - |
6# | 0.3 | 0.32 | 8.9 | 0.27 | 9.2 |
7# | 4.7 | 4.43 | 6.6 | 3.98 | 5.9 |
8# | 12.4 | 13.2 | 5.3 | 13.7 | 5.6 |
9# | 6.4 | 6.00 | 6.4 | 5.80 | 7.2 |
10# | 0.8 | 0.69 | 9.1 | 0.68 | 9.4 |
Number | AFB1 Result by HPLC µg·kg−1 | AFB1 Result by TRFIA µg·kg−1 |
---|---|---|
1# | 5.20 | 4.90 |
2# | 6.40 | 6.70 |
3# | 9.80 | 10.7 |
4# | 3.20 | 2.75 |
5# | 2.90 | 2.45 |
6# | 0.80 | 0.70 |
7# | 1.30 | 1.50 |
8# | 3.60 | 4.20 |
9# | 0.31 | 0.44 |
10# | 3.60 | 3.80 |
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Wang, D.; Zhang, Z.; Li, P.; Zhang, Q.; Zhang, W. Time-Resolved Fluorescent Immunochromatography of Aflatoxin B1 in Soybean Sauce: A Rapid and Sensitive Quantitative Analysis. Sensors 2016, 16, 1094. https://doi.org/10.3390/s16071094
Wang D, Zhang Z, Li P, Zhang Q, Zhang W. Time-Resolved Fluorescent Immunochromatography of Aflatoxin B1 in Soybean Sauce: A Rapid and Sensitive Quantitative Analysis. Sensors. 2016; 16(7):1094. https://doi.org/10.3390/s16071094
Chicago/Turabian StyleWang, Du, Zhaowei Zhang, Peiwu Li, Qi Zhang, and Wen Zhang. 2016. "Time-Resolved Fluorescent Immunochromatography of Aflatoxin B1 in Soybean Sauce: A Rapid and Sensitive Quantitative Analysis" Sensors 16, no. 7: 1094. https://doi.org/10.3390/s16071094
APA StyleWang, D., Zhang, Z., Li, P., Zhang, Q., & Zhang, W. (2016). Time-Resolved Fluorescent Immunochromatography of Aflatoxin B1 in Soybean Sauce: A Rapid and Sensitive Quantitative Analysis. Sensors, 16(7), 1094. https://doi.org/10.3390/s16071094