FOOD SCIENCE ›› 2020, Vol. 41 ›› Issue (10): 304-310.doi: 10.7506/spkx1002-6630-20190123-293

• Safety Detection • Previous Articles     Next Articles

Development of an Indirect Competitive Enzyme Linked Immunoassay for the Analysis of Amantadine in Chicken Muscle

TAN Shu, YANG Jinyi, XU Jihua, SHEN Yudong, ZENG Daoping, SU Xiaona, ZHONG Cuili, XU Xiaoxuan, SUN Yuanming   

  1. (1. Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University, Guangzhou 510642, China; 2. Forensic Science Institute of Ji’an Municipal Bureau of Public Security, Ji’an 343000, China; 3. Wens Food Group Co. Ltd., Yunfu 527499, China)
  • Online:2020-05-25 Published:2020-05-15

Abstract: An indirect competitive enzyme linked immunoassay (ic-ELISA) for the detection of amantadine residue in chicken muscle was developed in this paper. The hapten was synthesized via the reaction of N-(1-adamantylamino) carbamide with glyoxylic acid, and then conjugated to the carrier proteins keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA) by the active ester method to generate an immunogen and a coating antigen, respectively. BALB/c mice were immunized with the immunoantigen for antibody generation. Subsequently, a specific monoclonal antibody against amantadine was obtained. The optimal dilution of coating antigen and antibody concentration were 80 000 and 122.50 μg/L, respectively. Under the optimized experimental conditions, the ic-ELISA showed an IC50 of 0.69 μg/L with a limit of detection (LOD) of 0.21 μg/L. The linear range was 0.07–6.15 μg/L, and the recoveries for spiked chicken muscle were in the range from 101.69% to 108.71% with coefficients of variation less than 15%. Furthermore, the results of ic-ELISA were highly correlated with those of high performance liquid chromatography-mass spectrometry (HPLC-MS) (R2 = 0.97), which demonstrated that the established ic-ELISA is of high accuracy and good reliability. Therefore, the ic-ELISA proved to be a suitable method for the rapid detection of amantadine in real chicken samples.

Key words: chicken muscle, amantadine, indirect competitive enzyme linked immunosorbent assay, monoclonal antibody

CLC Number: