Colorimetric detection of melamine based on p-chlorobenzenesulfonic acid-modified AuNPs

  • Jianfang Li
  • Pengcheng Huang
  • Fangying WuEmail author
Research Paper


A highly selective and sensitive method is developed for colorimetric detection of melamine using gold nanoparticles (AuNPs) functionalized with p-chlorobenzenesulfonic acid. The addition of melamine induced the aggregation of AuNPs, as evidenced from the morphological characterizations and the color changed from red wine to blue, which could also be monitored by the UV–visible spectrometer and even naked eyes. This process caused a significant increase in the absorbance ratio (A650nm/A520nm) of p-chlorobenzenesulfonic acid–AuNPs. Under optimized conditions, the system exhibited a linear response to melamine in the range of 6.0 × 10−7–1.5 × 10−6 mol L−1 with a correlation coefficient of 0.997, and the limit of detection can even be 2.3 nM, which was much lower than some other methods and the safe limits (20 μM in both the USA and EU, 8.0 μM for infant formula in China, 1.2 μM in the CAC (Codex Alimentarius Commission) review for melamine in liquid infant formula). More importantly, the developed method presented excellent tolerance to coexisting common metal ions such as Ca2+, Zn2+, whose concentration is 1000 times of melamine, so that it had been applied to the analysis of melamine in liquid milk and milk powder with the recovery of 97.0–101 % and 100–103 %, respectively, indicating that the proposed method is quite a highly effective means to determine melamine in milk products.


Melamine Gold nanoparticles p-Chlorobenzenesulfonic acid Colorimetric Health effects 



This work was financially supported by Natural Science Foundation of China (No. 21365014, 21505067).

Supplementary material

11051_2016_3465_MOESM1_ESM.doc (254 kb)
Supplementary material 1 (DOC 254 kb)


  1. Cai HH, Yu X, Dong H, Cai J, Yang PH (2014) Visual and absorption spectroscopic detections of melamine with 3-mercaptopriopionic acid-functionalized gold nanoparticles: a synergistic strategy induced nanoparticle aggregates. J Food Eng 142:163–169CrossRefGoogle Scholar
  2. Cao BY, Yang H, Song J, Chang HF, Li SQ, Deng AP (2013) Sensitivity and specificity enhanced enzyme-linked immunosorbent assay by rational hapten modification and heterogeneous antibody/coating antigen combinations for the detection of melamine in milk, milk powder and feed samples. Talanta 116:173–180CrossRefGoogle Scholar
  3. Chen CR, Men GW, Bu WH, Liang CS, Sun HC, Jiang SM (2015) A colorimetric and fluorescent probe for multiple transition metal ions (Cu2+, Zn2+ and Ni2+): fast response and high selectivity. Sens Actuators B 220:463–471CrossRefGoogle Scholar
  4. Chudzinski MG, McClary CA, Taylor MS (2011) Anion receptors composed of hydrogen- and halogen-bond donor groups: modulating selectivity with combinations of distinct noncovalent interaction. J Am Chem Soc 133:10559–10567CrossRefGoogle Scholar
  5. Early RJ, Yu H, Mu XYP, Xu HY, Guo L, Kong QX, Zhou JM, He B, Yang XY, Huang HL, Hu E, Jiang Y (2013) Repeat oral dose toxicity studies of melamine in rats and monkeys. Arch Toxicol 87:517–527CrossRefGoogle Scholar
  6. Ghosh A, Talukdar S, Ghosh K, Das T, Das D (2014) Colorimetric and fluorescence recognition of tryptophan and histidine using phthalaldehyde based probe: experimental, computational, cell imaging and fish tissue analysis. RSC Adv 4:55286–55289CrossRefGoogle Scholar
  7. Jin RC, Wu GS, Li Z, Mirkin CA, Schatz GC (2003) What controls the melting properties of DNA-linked gold nanoparticle assemblies? J Am Chem Soc 125:1643–1654CrossRefGoogle Scholar
  8. Lachenmeier DW, Humpfer E, Fang F, Schu B, Dvortsak P, Spraul M (2009) NMR-spectroscopy for nontargeted screening and simultaneous quantification of health-relevant compounds in foods: the example of melamine. J Agric Food Chem 57:7194–7199CrossRefGoogle Scholar
  9. Lam CW, Lan L, Che XY, Tam S, Wong SSY, Chen Y, Jin J, Tao SH, Tang XM, Yuen KY, Tam PKH (2009) Diagnosis and spectrum of melamine-related renal disease: plausible mechanism of stone formation in humans. Clin Chim Acta 402:150–155CrossRefGoogle Scholar
  10. Li H, Rothberg L (2004) Colorimetric detection of DNA sequences based on electrostatic interactions with unmodified gold nanoparticles. Proc Natl Acad Sci USA 101:14036–14039CrossRefGoogle Scholar
  11. Li JP, Chen ZQ, Li YP (2011) A strategy for constructing sensitive and renewable molecularly imprinted electrochemical sensors for melamine detection. Anal Chim Acta 706:255–260CrossRefGoogle Scholar
  12. Li W, Li H, Zhang J, Tian XW (2015a) Effect of melamine toxicity on tetrahymena thermophila proliferation and metallothionein expression. Food Chem Toxicol 80:1–6CrossRefGoogle Scholar
  13. Li Y, Xu JY, Sun CY (2015b) Chemical sensors and biosensors for the detection of melamine. RSC Adv 5:1125–1147CrossRefGoogle Scholar
  14. Liang R, Zhang RM, Qin W (2009) Potentiometric sensor based on molecularly imprinted polymer for determination of melamine in milk. Sens Actuators B 141:544–550CrossRefGoogle Scholar
  15. Liang XS, Wei HP, Cui ZQ, Deng JY, Zhang ZP, You XY, Zhang XE (2011) Colorimetric detection of melamine in complex matrices based on cysteamine-modified gold nanoparticles. Analyst 136:179–183CrossRefGoogle Scholar
  16. Liu JX, Zhong YB, Liu J, Zhang HC, Xi JZ, Wang JP (2010) An enzyme linked immunosorbent assay for the determination of cyromazine and melamine residues in animal muscle tissues. Food Control 21:1482–1487CrossRefGoogle Scholar
  17. Lu QJ, Zhao JN, Xue SY, Yin P, Zhang YY, Yao SZ (2015) A “turn-on” fluorescent sensor for ultrasensitive detection of melamine based on a new fluorescence probe and AuNPs. Analyst 140:1155–1160CrossRefGoogle Scholar
  18. Ma Y, Niu HY, Zhang XL, Cai YQ (2011) One-step synthesis of silver/dopamine nanoparticles and visual detection of melamine in raw milk. Analyst 136:4192–4196CrossRefGoogle Scholar
  19. Miao P, Han K, Sun HX, Yin J, Zhao J, Wang BD, Tang YG (2014) Melamine functionalized silver nanoparticles as the probe for electrochemical sensing of clenbuterol. ACS Appl Mater Interfaces 6:8667–8672CrossRefGoogle Scholar
  20. Panuwet P, Nguyen JV, Wade EL, Souza PED, Ryan PB, Barr DB (2012) Quantification of melamine in human urine using cation-exchange based high performance liquid chromatography tandem mass spectrometry. J Chromatogr B 887–888:48–54CrossRefGoogle Scholar
  21. Paul IE, Rajeshwari A, Prathna TC, Raichur AM, Chandrasekarana N, Mukherjee A (2015) Colorimetric detection of melamine based on the size effect of AuNPs. Anal Methods 7:1453–1462CrossRefGoogle Scholar
  22. Rajapandiyan P, Tang WL, Yang J (2015) Rapid detection of melamine in milk liquid and powder by surface enhanced Raman scattering substrate array. Food Control 56:155–160CrossRefGoogle Scholar
  23. Rovina K, Siddiquee S (2015) A review of recent advances in melamine detection techniques. Food Compos Anal 43:25–38CrossRefGoogle Scholar
  24. Shen JC, Zhang Y, Yang H, Yang Y, Zhou ZG, Yang SP (2014) Detection of melamine by a magnetic relaxation switch assay with functionalized Fe/Fe3O4 nanoparticles. Sens Actuators B 203:477–482CrossRefGoogle Scholar
  25. Shrivastav AM, Mishra SK, Gupta BD (2015) Fiber optic SPR sensor for the detection of melamine using molecular imprinting. Sens Actuators B 212:404–410CrossRefGoogle Scholar
  26. Song J, Wu FY, Wan YQ, Ma LH (2015) Colorimetric detection of melamine in pretreated milk using silver nanoparticles functionalized with sulfanilic acid. Food Control 50:356–361CrossRefGoogle Scholar
  27. Squadrone S, Ferro GL, Marchis D, Mauro C, Palmegiano P, Amato G, Genin EP, Abete MC (2010) Determination of melamine in feed: validation of a gas chromatography-mass spectrometry method according to 2004/882/CE regulation according to 2004/882/CE regulation. Food Control 21:714–718CrossRefGoogle Scholar
  28. Venkatasami G Jr, JRS H (2010) A rapid, acetonitrile-free, HPLC method for determination of melamine in infant formula. Anal Chim Acta 665:227–230CrossRefGoogle Scholar
  29. Wisner JA, Beer PD, Drew MGB (2001) A Demonstration of anion templation and selectivity in pseudorotaxane formation. Angew Chem Int Ed 40:3606–3609CrossRefGoogle Scholar
  30. Wu QQ, Long Q, Li HT, Zhang YY, Yao SZ (2015) An upconversion fluorescence resonance energy transfer nanosensor for one step detection of melamine in raw milk. Talanta 136:47–53CrossRefGoogle Scholar
  31. Xavier SSJ, Karthikeyan C, Kumar GG, Kim AR, Yoo DJ (2014) Colorimetric detection of melamine using β-cyclodextrin-functionalized silver nanoparticles. Anal Methods 6:8165–8172CrossRefGoogle Scholar
  32. Xing HB, Zhan SS, Wu YG, He L, Zhou P (2013) Sensitive colorimetric detection of melamine in milk with an aptamer-modified nanogold probe. RSC Adv 3:17424–17430CrossRefGoogle Scholar
  33. Yazgan NN, Boyac IH, Topcu A, Tamer U (2012) Detection of melamine in milk by surface-enhanced Raman spectroscopy coupled with magnetic and Raman-labeled nanoparticles. Anal Bioanal Chem 403:2009–2017CrossRefGoogle Scholar
  34. Yin RH, Liu J, Li HS, Bai WL, Yin RL, Wang X, Wang WC, Liu BS, Han XH, Han J, He JB, Han XR (2014) The toxic effects of melamine on spleen lymphocytes with or without cyanuric acid in mice. Res Vet Sci 97:505–513CrossRefGoogle Scholar
  35. Yokley RA, Mayer LC, Rezaaiyan R, Manuli ME, Cheung MW (2000) Analytical method for the determination of cyromazine and melamine residues in soil using LC-UV and GC-MSD. J Agric Food Chem 48:3352–3358CrossRefGoogle Scholar
  36. Zeng HJ, Yang R, Wang QW, Li JJ, Qu LB (2011) Determination of melamine by flow injection analysis based on chemiluminescence system. Food Chem 127:842–846CrossRefGoogle Scholar
  37. Zhang XF, Zhao H, Xue Y, Wu ZJ, Zhang Y, He YJ, Li XJ, Yuan ZB (2012) Colorimetric sensing of clenbuterol using gold nanoparticles in the presence of melamine. Biosens Bioelectron 34:112–117CrossRefGoogle Scholar
  38. Zhou CH, Zhao JY, Pang DW, Zhang ZL (2014) Enzyme-induced metallization as a signal amplification strategy for highly sensitive colorimetric detection of avian influenza virus particles. Anal Chem 86:2752–2759CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  1. 1.College of ChemistryNanchang UniversityNanchangChina

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