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Citation: WANG Li, YE Hua, SANG Hong-Qing, WANG Dan-Dan. Aptamer-based Fluorescence Assay for Detection of Isocarbophos and Profenofos[J]. Chinese Journal of Analytical Chemistry, ;2016, 44(5): 799-803. doi: 10.11895/j.issn.0253-3820.150601 shu

Aptamer-based Fluorescence Assay for Detection of Isocarbophos and Profenofos

College of Food and Drug, Anhui Science and Technology University, Fengyang 233100, China

Received Date: 28 July 2015
Revised Date: 13 January 2016

Fund Project: This work was supported by the National Natural Science Foundation of China (No.31201369), the Key Project of College Excellent Talents Support Plan in Anhui Province (No. gxyqZD2016219) and the Natural Science Research Project of Anhui Science and Technology University (No. ZRC2014395).

A fluorescence method for detection of isocarbophos and profenofos was established based on the specific recognition of aptamer. The aptamer recognizing isocarbophos and profenofos was labeled with fluorescent label FAM at 5' end (F-ssDNA). When it bound to complementary DNA chain labeled with quenching group DABCYL at 3' end (Q-ssDNA), the double-stranded structure would be formed and the fluorescence signals became very weak due to the effect of fluorescence resonance energy transfer. While the Q-ssDNA would be released from the double-stranded structure when the aptamer specifically recognized and bound the targets, and the fluorescence signals of the system would recover. Based on this, isocarbophos and profenofos could be quantitatively detected. The optimized experimental conditions were as follows:firstly, 25 nmol/L F-ssDNA and 50 nmol/L Q-ssDNA-2 were incubated for 20 min at 25℃, then an equal volume of pesticide was added and incubated for another 60 min, then the fluorescence intensity of the system was determined. Under the optimal conditions, the change of the fluorescence intensity of the system (ΔI) showed a linear relationship with the isocarbophos or profenofos concentration in the range of 50 to 500 μmol/L. The limit of detection (LOD, σ) was 11.4 μmol/L for isocarbophos with the relative standard deviation (RSD) of 5.8% (n=10). The LOD was 14.0 μmol/L for profenofos with RSD of 4.9% (n=10). The recovery ranged from 85.8% to 95.3% when this method was applied to detect isocarbophos and profenofos in real water samples.
- Aptamer,
- Fluorescence,
- Isocarbophos,
- Profenofos
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