Detection Technology Research Of Chloramphenicol In Lateral Flow Immune Chromatography

Chloramphenicol (Chloramphenicol, CAP) is a broad-spectrum antibiotics and it hasbeen widely used in aquaculture industry because of its high efficiency and low price. Butchloramphenicol has serious side effects and the residue in animal food has a serious threatto human health. Therefore, the developed countries have already banned or strictly limitedchloramphenicol (CAP) used in food animals, and made strict standards of CAP’s highestresidues in food. The traditional chloramphenicol residue analysis detection method needsexpensive equipments, professional technical personnel, long analysis cycle and high costs,so it can’t achieve rapid detection. Therefore, it is very necessary to establish a kind ofeconomic, sensitive and efficient CAP residue analysis method. Lateral flow immunechromatography detection method can use the specificity conjugation reaction of antigenand antibody to determine few antigen and antibody. This article was based on the methodof anti-chloramphenicol polyclonal antibody’s separation and purification to develop thethe immune colloidal gold test strip of chloramphenicol, thus this study has great practicalsignificance and business development value.Diazo synthesis method was used in this experiment to compound CAP-OVA andCAP-BSA and the two antigen had couplinged successfully by uv spectrophotometry,infrared spectrometry, polyacrylamide electrophoresis, etc. CAP-OVA was used for theimmune original to immune Balb/C mice， collect antiserum and use affinitychromatography purification to prepare the anti-chloramphenicol polyclonal antibody. Itwas confirmed by ELISA that the anti-serum’s titers were up to1:32000and there was nocross-reaction among streptomycin, tetracycline, oxytetracycline and penicillin analogues,so it has high specificity affinity.In the lateral flow chromatography immunochromatography: rabbit anti-mouse IgGantibody and coated-antigen were immobilised on the lateral flow membrane of thestrip(AE99) and used respectively as the quality control line and detection line. Then,immunochromatography assay papers were assembled. According to the color depth, wegot conclusions that the test strip detection limit of the assay paper was1μg/ml and nosignificant cross-reactivity with chloramphenicol structural analogues happened.