The Development Of The Colloidal Gold Test Strip Applied To Test Two Important Veterinary Drug Residues And The Optimization Of Its Technological Process

Objectives:To optimize and develop commercialized colloidal gold test strip of clenbuterol (CL) and establish stable product research and development platform of colloidal gold test strip. On that basis, to develop colloidal gold test strip for monitoring 1-aminohydantoin (AHD) in meat.Methods:Preparation of colloidal gold,lable,block and so on to optimize the whole technological process for colloidal gold test strip, mass commercialized colloidal gold test strip to detect CL were developed. On the basis of all above optimization, Colloidal gold was prepared by controlled reduction of gold chloride with sodium citrate. Monoclonal antibody against CPAHD purified was conjugated to colloidal gold. AHD's derivates (CPAHD) was synthesized with AHD and 4-Formylbenzoic acid (4-CBA). And then by the method of active ester, CPAHD was conjugated to bovine serum albumin (BSA) as complete antigen named CPAHD-BSA. The sensitivity, specificity, stability and reliability of test strips were determined, according to this feature to develop AHD test strip.Results:The CL-BSA and CPAHD-BSA were synthesized successfully. Colloidal gold were prepared with good size uniformity. The whole colloidal gold technological process was optimized, and mass commercialized colloidal gold test strips were developed. Colloidal gold test strip were developed to detect AHD. The assay could be completed in 3 min and detected 1-aminohydantoin's derivates (CPAHD) at 5 ng mL-1, equivalent to 2.33 ng mL-1 of AHD. The antibody showed no cross reactivity with nitrofuran analogs except for nitrofurantoin at a high concentration. The test strip was stable at room temperature for up to 28 weeks. Parallel analyses of meat samples with test strips and enzyme-linked immunosorbent assay (ELISA) obtained data in good agreement.Conclusions:The whole colloidal gold technological process was optimized, and mass commercialized colloidal gold test strips were developed. A sensitive, specific, stable, convenient and cheap colloidal gold test strip for detection of AHD in meat samples was developed.