Rapid Detection Methods Based On Surface Enhanced Raman Scattering(SERS) For Accurately Detecting Chloramphenicol Antibiotics

Chloramphenicol(CAP)is a highly effective broad-spectrum antibiotic which has several analogues including Thiamphenicol(TAP)?Florfenicol(FFC)and other derivatives.The Chloramphenicol antibiotics(CAPs)are widely used to prevent and treat infectious diseases of animals,humans and poultry,because of their good effects of sterilization and cheapness.CAPs have great side effects on human health when they are absorbed in human body and then they have been banned for good.So riching CAPs Raman spectrums and establishing a high specific,sensitive and rapid immunology detection has very important theoretical and realistic meanings.The study included three parts which were the quick identification of the raman spectrums of chloramphenicol antibiotics,the assembly and the assessment of colloidal gold immune chromatography test and the highly specific,quick and quantitative Surface Enhanced Raman Scattering(SERS)detection technology which combined the raman label with immune chromatography.The research is as follows:1.Rapid identification of chloramphenicol antibiotics based on SERSThrough the calculation of the structure and the calculation of enhancement factor(EF)of gold(Au),silver(Ag)and silver @ gold nanoparticles(Ag@Au),the nanoparticles with good dispersibility and enhanced effects were selected.The CAP was chosen as the research object and the gold colloid was selected as the raman enhance substrate.The Raman signal of chloramphenicol was well detected when the ethanol was selected as the solvent and tin foil was selected as the solid phase carrier.With chloramphenicol antibiotics concentration as the abscissa,the height of the raman peak of chloramphenicol 1348 cm–1,thiamphenicol1593 cm-1,florfenicol 1142 cm-1 as the ordinate,the calibration curve had a good linear relationship within the range of the concentration of 1?M-0.1M.2.Assembly,optimization and evaluation of immunochromatography colloidal gold testThe titer of the CAPs monoclonal antibody were measured by indirect ELISA and indirect competitive ELISA which is more than 64000.The IC50 of chloramphenicol,thiamphenicol,florfenicol were 0.87?g/m L,0.23?g/m L and 2.5 ?g/m L,and the ELISA limits of detection(LOD)were 0.21,0.035,0.48?g/m L.The assembly of the colloidal gold immunochromatography test was made up of sample pad GL-b02 and combination pad Ahlstrom8964,and then its treating fluid was optimized.The concentration of the testing sample was the limits of detection of the immunochromatography test when the test had a significant color.The LODs of the chloramphenicol,thiamphenicol,florfenicol were 50ng/m L,25 ng/m L,50ng/m L.which had lower limits of detection,And the repeatability,stability,specificity of the test were good.3.Assembly,optimization and evaluation of raman label immunochromatography test4-Mercaptobenzoic acid(4 – MBA)was chosen as the Raman label molecular.On the basis of stability and Raman enhancement effect,the Ag@Au was selected as the enhanced substrates between Au,Ag and Ag@Au.The raman label immunochromatography test were assembled immune chromatography test with the highly specific,quick and quantitative Surface Enhanced Raman Scattering,and were evaluated.The results showed that there was an excellent linear tendency between logarithm of CAPs concentration and the inhibition rate in the measured range of 0 ng/m L ~256 ng/m L,the standard curve of the CAP was y==-0.3069x+1.7431,R2=0.994,IC50 was 18.4ng/m L and LOD was 2.6 ng/ m L;the standard curve of the TAP was y==-0.3069x+1.7431,R2=0.994,IC50 was 13.9 ng/m L and LOD was 2.0 ng/ m L;the standard curve of the FFC was y==-0.3069x+1.7431,R2=0.994,IC50 was 18.9ng/m L and LOD was 3.1ng/ m L.In addition,the detection system was evaluated,including precision,specificity and stability.The results showed that there was a good precision with intra-assay coefficient of variation of 3.7% ~ 9.1%,interassay coefficient of variation of 5.8% ~13.8% in detecting CAPs.The cross-reactivity rates of the system with other major chemicals in research was less than 1%,which had a good specificity.The detection system was finally applied in the detection of CAP in aquatic products.The recovery rate result of CAP was 91.5% ~105%.The raman label immunochromatography test can improve the sensitivity and specificity,and it has an important practical application value in food safety detection.