Study On The Enrofloxacin Colloidal Gold Test Strip In Aquatic Products

Enrofloxacin is the derivatives of modified quinolone, due to its high antibacterial ability often used as a highly efficient, broad-spectrum antimicrobial agents. With the widespread use of fluoroquinolones, in clinical and aquaculture, the residual drug of fluoroquinolones will slowly accumulate in animal body which directly threaten people’s lives and safety. At present, the common methods used for detecting Enrofloxacin is HPLC and so on, although this method with high sensitivity, but it needs the complex operation and long time of analysis, unfavorable testing on the scene that unable to keep up with demand of the market. The GICA strip has “simple, rapid, sensitive”, is the main means of detection the future trend of development.By studied the factors on Enrofloxacin colloidal gold test strip, determined the process of preparing strip and optimal condition. The results are as follows: In 100 m L 0.01% chloroauric acid was added 2.0 m L 1% sodium citrate. The boiling reaction time was 10 min, stirring speed was 200 r/min, using magnetic stirrer with heating that can be prepared by colloidal gold solution with better quality. According to the empirical formula of colloidal gold, can be calculated the particle of colloidal gold was about 14.60 nm. In the colloidal gold labeled, the optimal amount of protein marker was 9.6 μg/m L, the optimum p H was 9.0 markers. By optimizing the preparation of enrofloxacin colloidal gold test strips conditions, determination of coating antigen and anti-mouse antibody concentration respectively was 2.0 mg/m L and 0.5 mg/m L, the gold labeled antibody optimal was 4 times. The best centrifugal speed of gold labeled antibody was 11000 r/min; The heavy resuspension was 0.01 mol/L p H=9.0 borate buffer solution contained 1% sucrose、0.1%BSA; The labeled antibody dilute solution was 0.01mol/L p H=7.4 PB contained 1%BSA、5% sucrose、0.02%Na N3. Comparative and study of different materials, the final choice of the water absorption pad type was H5072、the cellulose nitrate film was Millipore180、the conjugate pad type was Ahlstrom8964 and the sample pad type was GL-b02. Through the performance testing of strip: The sensitivity of test strip was 60 ng/m L, strip of ENR has good specificity and no cross reaction to other structural analogues; The test strip can be sealed and stored at 4 degrees Celsius for 4 months, greatly improves the stability of test strip. Through the optimization of sample pretreatment and the actual samples, it was able to detect the positive samples of 16 μg/kg, on site rapid detection of enrofloxacin residue is of great practical significance.