Next Article in Journal
Unsymmetrical Mesoporphyrinic Complexes of Copper (II) and Zinc (II). Microwave-Assisted Synthesis, Spectral Characterization and Cytotoxicity Evaluation
Previous Article in Journal
Intramolecular Conversions of (Aminoferrocenylpenta-1,4-dienyl)-ferrocenylcarbenes: Synthesis of Diferrocenylmono-, bi-, tricycles and Amino(diferrocenyl)hexa-1,3,5-trienes
Article Menu

Export Article

Open AccessArticle
Molecules 2011, 16(7), 5591-5603;

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

Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, Guangdong Province, China
Institute of Agri-Food and Land Use, Queen’s University Belfast, Belfast BT9 5AG, Northern Ireland, UK
Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, Guangdong Province, China
Authors to whom correspondence should be addressed.
Received: 17 May 2011 / Revised: 13 June 2011 / Accepted: 15 June 2011 / Published: 30 June 2011
(This article belongs to the Section Organic Chemistry)
Full-Text   |   PDF [328 KB, uploaded 18 June 2014]   |  


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. View Full-Text
Keywords: melamine; ELISA; specificity; milk melamine; ELISA; specificity; milk

Figure 1

This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

Share & Cite This Article

MDPI and ACS Style

Lei, H.; Su, R.; Haughey, S.A.; Wang, Q.; Xu, Z.; Yang, J.; Shen, Y.; Wang, H.; Jiang, Y.; Sun, Y. Development of a Specifically Enhanced Enzyme-Linked Immunosorbent Assay for the Detection of Melamine in Milk. Molecules 2011, 16, 5591-5603.

Show more citation formats Show less citations formats

Related Articles

Article Metrics

Article Access Statistics



[Return to top]
Molecules EISSN 1420-3049 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top