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Enhanced Competitive Chemiluminescent Enzyme Immunoassay for the Trace Detection of Insecticide Triazophos
Author(s) -
Jin Maojun,
Shao Hua,
Jin Fen,
Gui Wenjun,
Shi Xiaomei,
Wang Jing,
Zhu Guonian
Publication year - 2012
Publication title -
journal of food science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.772
H-Index - 150
eISSN - 1750-3841
pISSN - 0022-1147
DOI - 10.1111/j.1750-3841.2012.02659.x
Subject(s) - chemistry , chemiluminescence , chromatography , immunoassay , detection limit , luminol , high performance liquid chromatography , horseradish peroxidase , chemiluminescent immunoassay , solvent , enzyme , antibody , biochemistry , immunology , biology
A direct competitive chemiluminescent enzyme immunoassay (CLEIA) for triazophos was developed, which was based on the anti‐THHe IgG monoclonal antibody and a heterogeneous enzyme tracer (THHu‐HRP). Several components of chemiluminescent enhanced solution (CES) were optimized. The results showed that 1 mM of p‐iodo‐phenol, 0.625 mM of luminol, and 4 mM of H 2 O 2 had the best performance. Based on the study of CES, the influence of several factors (assay buffer, blocking substance, and solvent) on the immunoassay was investigated. The sensitivity for detection, IC 50 value was 0.87 ng/mL at a practical working concentration range between 0.04 ng/mL and 5 ng/mL and the limit of detection for triazophos was 0.063 ng/mL. The average recovery of triazophos added to lettuce, carrot, apple, water, and soil were 78.71%, 67.52%, 118.3%, 117.2%, and 122.0%, respectively. Finally, comparison between the methods of CLEIA and high‐performance liquid chromatography‐tandem mass spectrum (HPLC‐MS/MS) was performed. The results obtained from CLEIA were in agreement with those of HPLC‐MS/MS. Practical Application: The method developed in the paper could take positive or negative test of vegetable, fruit, and environmental samples for triazophos with high sensitivity and efficiency.