Citation: YAO Chan-Yuan, YANG Jin-Yi, XU Zhen-Lin, WANG Hong, LEI Hong-Tao, SUN Yuan-Ming, TIAN Yuan-Xin, SHEN Yu-Dong. Indirect Competitive Enzyme-Linked Immunosorbent Assay for Detection of Tylosin in Milk and Water Samples[J]. Chinese Journal of Analytical Chemistry, ;2018, 46(8): 1275-1281. doi: 10.11895/j.issn.0253-3820.181057 shu

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

1.
Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, China;
2.
School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China

Received Date: 10 April 2018
Revised Date: 6 May 2018

Fund Project: This work was supported by the Science and Technology Planning Project of Guangzhou (No. 201704020082), the National Key Research and Development Program of China (No. 2016YFE0106000), the Science and Technology Planning Project of the Guangxi Zhuang Autonomous Region (No.2017AB47020), the National Natural Science Foundation of China (No. 31371769) and Guangdong Province Universities and Colleges Pearl River Scholar Funded Scheme (2017).

To address the issues of tylosin residues in food and environment, a good specific polyclonal antibody was prepared by animal immunization based on a new hapten achieved by oxidizing the aldehyde group on the side chain of tylosin to carboxyl. Further, an indirect competitive enzyme-linked immunosorbent assay (icELISA) was established. The main experimental conditions were as follows:the coating antigen and the antibody were respectively diluted by 6000 times and 2000 times, the reaction buffer was PBST (pH 7.4), and the Tween concentration and ionic concentration were 0.1% and 0.01 mol/L, respectively. The dilution ratio of the secondary antibody was 4000 times and the secondary reaction time was 30 min. Under the optimized conditions, the IC₅₀ and the linear range of this method were 1.39 ng/mL and 0.17-11.0 ng/mL, respectively. The detection limit was 0.07 ng/mL, and the cross-reactivity with analogues was less than 0.1%. The recoveries of spiked raw milk and water samples were 78.4%-105.6% with relative standard deviation less than 15%. Besides, the detection results had good correlation with HPLC method (R²=0.97). This method is suitable for rapid and specific screening of tylosin residues in milk and water samples.
- Indirect competitive enzyme-linked immunosorbent assay,
- Tylosin,
- Milk,
- Water
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沈阳化工大学材料科学与工程学院沈阳 110142