Abstract
In this study, the testing time was reduced to 1 h through optimizing, and an LOD of 0.3±0.15 μg·L-1 and an IC50 of 2.7±0.3 μg·L-1 were achieved by the optimized CD-ELISA. The sample extraction of 1-fold acetonitrile followed by a 20-fold dilution in assay diluted buffer has proven to be sufficient to eliminate the influence of matrix. The CD-ELISA was validated further by comparing with the standard HPLC. The average recovery ratio of 89.7%~112.7% was obtained, and the coefficients of variation were less than 11.0%. In actual samples detection, except for Pleurotus eryngii, carbendazim was not detected, and other mushrooms were all detected, and only carbendazim in Volvariella volvacea exceeded the standard, and the over-standard rate was 16.7 %. In a word, the rapid, sensitive and efficient CD-ELISA for quantifing carbendazim in mushrooms was established in the study.
Highlights
As a broad spectrum benzimidazole fungicide, carbendazim is widely used in agricultural production with the characteristics of high efficiency, low toxicity and the inner attraction [1]
The results show that A0 / IC50 at pH 7.4 is good
In order to demonstrate accuracy and efficiency of the CDELISA, the recovery experiment by adding three-level standard samples, was carried out, as shown in Table 5, the average recovery ratio were 89.7%~112.7%, despite this slight overestimation or underestimation, which is found within the range accepted in an analytical methodology, and all of coefficients of variation (CV) were less than 11.0%, which indicated the developed sample extraction and the CD-enzyme linked immunosorbent assay (ELISA) of detecting carbendazim in food samples could be applied
Summary
As a broad spectrum benzimidazole fungicide, carbendazim is widely used in agricultural production with the characteristics of high efficiency, low toxicity and the inner attraction [1]. In 2015, Ana Ucles et al[12] had established an ELISA for detecting carbendazim, imazalil and thiabendazole residues in vegetable samples, and through 2h’ testing, the 0.5 μg·kg1~13.8 μg·kg-1 of IC 50 value of the calibration curve was obtained, after simple pretreatment, the method has a linearity range of 10 μg·kg-1 ~80 μg·kg. These researches, complex pretreatment method can't meet the needs of the rapid detection. The CD-ELISA could be applied to the rapid quantitative determination of carbendazim in food, especially in mushrooms
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