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UPA Perpustakaan Universitas Jember

Development of an Enzyme-Linked Immunosorbent Assay for the Detection of Difenoconazole Residues in Fruits and Vegetables

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An indirect, competitive enzyme-linked immunosorbent
assay (ic-ELISA) for the detection of difenoconazole was
developed. Two haptens were designed and successfully synthesized.
Hapten 1 had a particular moiety of difenoconazole, while
hapten 2 had its full structure. The polyclonal antibodies against
hapten-protein conjugates were prepared by immunizing rabbits.
After optimization of the conditions, the detection limit (IC15)
and sensitivity (IC50) were 4.58 and 29.10 μg L−1, respectively.
The cross-reactivities of the antibody with 11 triazole fungicides
were all less than 0.1%, which showed that the antibody had
excellent specificity. The recoveries of difenoconazole from the
spiked samples ranged from 89.70 to 102.31% with good accuracy.
The matrix effect was easily removed using a simple, rapid,
and efficient extraction method on fruits and vegetables. The
detection limit was all 229 μg kg−1 in fruits and vegetables. To
validate the ic-ELISA, samples were spiked with difenoconazole
at three different concentrations and simultaneously analyzed
using high-performance liquid chromatography (HPLC). The
results showed a good correlation between the ic-ELISA and
HPLC data (R2 = 0.9970). As a result, the developed immunosorbent
assay is suitable for the quantitative determination of
difenoconazole in fruits and vegetables.

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