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Development of a sensitivity-improved immunoassay for the determination of carbaryl in food samples
Tingting Dong1, Jingwei Sun, Bing Liu
1Key Laboratory of Food Nutrition and Safety, Ministry of Education, Tianjin University of Science and Technology, Tianjin 300457, China.
Background:
With the aim of developing a highly sensitive immunoassay for carbaryl, a hapten which had high similarity to carbaryl was synthesised using a safer and more practical approach. After it was conjugated to horseradish peroxidase, direct competitive heterologous enzyme-linked immunosorbent assay (CD-ELISA) was optimised and characterised. The assay performance conditions were investigated in details. Enhanced chemiluminescence ELISA (ECL-ELISA) was also used in preliminary studies.
Results:
The assay obtained an IC(50) value (the concentration causing 50% inhibition) of 2 microg kg(-1), which was 12-fold more sensitive than previous results of homologous CD-ELISA. In ECL-ELISA, the IC(50) was further decreased 10-fold to 0.2 microg kg(-1). The CD-ELISA developed was applicable for broad conditions, and could be applied on various food samples with a more convenient pre-treatment. Average recoveries were in a range of 88.3-101.7%. The results correlated well with those obtained using high-performance liquid chromatography (HPLC) analysis (R(2) = 0.989).
Conclusion:
The ELISA developed was a great improvement in the determination of carbaryl, showing that the immunoassay developed was a simple, rapid and efficient method that was reliable for the detection of carbaryl and suitable for rapid quantitative or qualitative determination in food samples.
