Development of a Specifically Enhanced Enzyme-Linked Immunosorbent Assay for the Detection of Melamine in Milk

Molecules

Hongtao Lei¹  Rui Su¹  Simon A. Haughey¹  Qiang Wang¹  Zhenlin Xu¹  Jinyi Yang¹  Yudong Shen¹  Hong Wang¹  Yueming Jiang¹ 

[1]¹Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, Guangdong Province, China

关键词: melamine;    ELISA;    specificity;    milk;   

DOI  :  10.3390/molecules16075591

来源: mdpi

【 摘 要 】

An indirect competitive enzyme-linked immunosorbent assay (icELISA) with enhanced specificity for melamine in milk was developed. Three haptens of melamine with different spacer-arms were used to prepare different plate coating antigens. It was found that the icELISA show best sensitivity and specificity to melamine when using the coating antigen prepared by coupling 3-(4,6-diamino-1,6-dihydro-1,3,5-triazin-2-ylthio)propanoic acid (Hapten C) with ovalbumin (OVA). The 50% inhibitory concentration (IC₅₀) value was 35.4 ng·mL⁻¹, the limit of detection (LOD) was 8.9 ng·mL⁻¹ and the detectable working range (20–80% inhibitory concentration) was from 14.9 to 108.5 ng·mL⁻¹, respectively. Compared to the ELISA results previously reported, the developed icELISA in the present study showed a much lower cross-reactivity to cyromazine, a fly-killing insecticide widely used in vegetables and stables. Recoveries obtained from milk samples in this study were in agreement with those obtained using the HPLC-MS method, indicating the detection performance of the icELISA could meet the requirement of the residue limit set by the Codex Alimentarius Commission. Therefore, the developed immunoassay can be applied for the analysis of melamine presented in milk.

【 授权许可】

CC BY   
This is an open access article distributed under the Creative Commons Attribution License (CC BY) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.