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Development and validation of an immunochromatographic assay for rapid multi-residues detection of cephems in milk.
- Source :
-
Analytica chimica acta [Anal Chim Acta] 2009 Feb 16; Vol. 634 (1), pp. 129-33. Date of Electronic Publication: 2008 Dec 06. - Publication Year :
- 2009
-
Abstract
- A one-step immunochromatographic assay (ICA) was developed for the detection of seven kinds of cephems in milk. Polyclonal antibodies (PcAb) with group-specific to cephems were raised in rabbits after immunization with cephalexin-keyhole limpet hemocyanin (KLH) conjugate. The specificity of anti-sera was determined by indirect competitive enzyme-linked immunosorbent assay (icELISA), and the 50% inhibitions (IC(50)) of cephalexin and cefadroxil were obtained at 1.5 ngmL(-1); IC(50) of cefatiofur, cefapirin, cefazolin, cefalothin and cefotaxine were 4, 3.7, 3.2, 4.5 and 5 ngmL(-1), respectively. The PcAb against cephems were conjugated to colloidal gold particles as the detection reagent for ICA strips to test for cephems. This method achieved semi-quantitative detection of cephems in <5 min, with high sensitivity to cephalexin and cefadroxil (both 0.5 ngmL(-1)). At the same time, cefatiofur, cefapirin, cefazolin, cefalothin and cefotaxine were detected at <100 ngmL(-1) in spiked processed-milk samples. This method was compared with an enzyme-linked immunosorbent assay by testing 40 milk samples, and the positive samples were validated by a high-performance liquid chromatographic method, with an agreement rate of 100% for both comparisons. In conclusion, the method was rapid and accurate for the multi-residue detection of cephems in milk.
- Subjects :
- Animals
Antibodies chemistry
Antibodies immunology
Antibodies, Immobilized chemistry
Antibodies, Immobilized immunology
Cephalexin immunology
Cephalosporins immunology
Gold Colloid chemical synthesis
Rabbits
Sensitivity and Specificity
Cephalosporins analysis
Chromatography, High Pressure Liquid methods
Immunoassay methods
Milk chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1873-4324
- Volume :
- 634
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Analytica chimica acta
- Publication Type :
- Academic Journal
- Accession number :
- 19154821
- Full Text :
- https://doi.org/10.1016/j.aca.2008.12.004