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Establishment of magnetic beads-based enzyme immunoassay for detection of chloramphenicol in milk.

Food chemistry (2013-02-28)
Jing Xu, Weiwei Yin, Yuanyang Zhang, Jian Yi, Meng Meng, Yabin Wang, Huyin Xue, Taichang Zhang, Rimo Xi
ABSTRACT

In this research, magnetic beads-based enzyme immunoassays were investigated for rapid analysis of chloramphenicol (CAP) in milk. To improve sensitivity of CAP determination, two kinds of immunomagnetic separation methods were designed and compared. Magnetic polystyrene microspheres were conjugated with anti-CAP antibody (Method I) or goat-anti-mouse IgG (Method II). The whole determination could be finished in 1.25 h. Both methods showed high sensitivity to CAP in buffer, and obtained an IC(50) value of 0.05 ng mL(-1) for Method I and 0.4 ng mL(-1) for Method II. The methods showed high specificity, only showing a little cross-reaction towards CAP succinate. The two methods were applied to detect CAP in milk. The recovery rates were 80-106% and the coefficients of variation (CVs) were 4.7-15%. The immunomagnetic assay showed promising potential in rapid screening field for CAP analysis. Between the two methods, Method I is more sensitive, and Method II is more suitable for producing a general assay by changing a primary antibody for another analyte.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Chloramphenicol, ≥98% (HPLC)
Sigma-Aldrich
Chloramphenicol, BioReagent, suitable for plant cell culture
Supelco
Chloramphenicol, VETRANAL®, analytical standard
Sigma-Aldrich
Chloramphenicol, meets USP testing specifications
Sigma-Aldrich
Chloramphenicol, γ-irradiated
Chloramphenicol, European Pharmacopoeia (EP) Reference Standard