Citation: ZHANG Liang, WANG Quan-Zhen, WANG Ning, LIU Shu-Zhao. Determination of Uniconazole Residue in Apple by Direct Competitive Enzyme-linked Immunosorbent Assay and High Performance Liquid Chromatography[J]. Chinese Journal of Analytical Chemistry, ;2012, 40(11): 1730-1734. doi: 10.3724/SP.J.1096.2012.20123 shu

Determination of Uniconazole Residue in Apple by Direct Competitive Enzyme-linked Immunosorbent Assay and High Performance Liquid Chromatography

1.
¹ College of Horiculture and Plant Protection, Yangzhou University, Yangzhou 225009, China;

Corresponding author: LIU Shu-Zhao,
Received Date: 15 February 2012
Available Online: 22 May 2012
Fund Project: 本文系863专题课题(No.2009AA03Z331) (No.2009AA03Z331)国家自然科学基金(No.20777064)资助项目 (No.20777064)

With the high specific affinitive antibody to uniconazole prepared in our lab, the method of antibody-immobilized direct competitive ELISA was employed to determine uniconazole residue in apple. Under the optimized conditions the antibody of 4 mg/L dissolved in 0.02 mol/L pH7.6 PB was immobilized on microplate,the peroxidase-hapten conjugate was 105 times diluted in the reaction medium of 0.02 mol/L pH 6.8 PBS containing 0.15 mol/L NaCl, the liner concentrations ranged from 1 mg/L to 10^-4 mg/L, the half-maximal inhibition (IC₅₀) was 1.31 μg/L with relative standard deviation (RSD) of 8.4%, the IC₂₀ was 0.022 μg/L. The recoveries were 84.6%-101%, 99.6%-117% and 80.8%-92.8% with the RSD of 8.6%, 6.9% and 6.7% at the spiked level of 1.0, 0.1, 0.01 mg/kg in apple respectively. The limit of detection for ELISA was 0.024 μg/kg, and the limit of detection for HPLC-ultraviolet visible detector was 0.25 mg/kg.
- Uniconazole,
- Apple,
- Direct competitive enzyme-link immunosorbent assay,
- High-performance liquid chromatography
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