Indirect Competitive Enzyme-Linked Immunosorbent Assay for Detection of Tylosin in Milk and Water Samples

Publication date: August 2018Source: Chinese Journal of Analytical Chemistry, Volume 46, Issue 8Author(s): Chan-Yuan YAO, Jin-Yi YANG, Zhen-Lin XU, Hong WANG, Hong-Tao LEI, Yuan-Ming SUN, Yuan-Xin TIAN, Yu-Dong SHENAbstractA specific polyclonal antibody was prepared based on a new hapten with stable structure. Based on this, an indirect competitive enzyme-linked immunosorbent assay (icELISA) was established for determination of tylosin residues in food and environmental samples. The experimental conditions were optimized as follows: the coating antigen and antibody were respectively diluted by 6000 times and 2000 times, and the competitive reaction time was 40 min. Under the optimized conditions, the method showed a detection limit of 0.07 ng mL−1, an IC50 of 1.39 ng mL−1 and a linear range of 0.17–11.0 ng mL−1. The recoveries of spiked raw milk and water samples were ranged from 78.4% to 105.6%, with the RSDs of less than 15%. Good correlation between icELISA and HPLC method was obtained for spiked samples (R2 = 0.97). This method was suitable for the determination of tylosin residues in milk and water samples.Graphical AbstractA hapten with simple and stable structure was synthesized based on tylosin, and specific polyclonal antibodies were obtained through animal immunity experiments. Based on this antibody, an indirect competitive enzyme-linked immunosorbent assay (icELISA) for the detection of tylosin residues in pure milk and water samples was established. This ...
Source: Chinese Journal of Analytical Chemistry - Category: Chemistry Source Type: research