Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 156 (2016) 89–97

Contents lists available at ScienceDirect

Spectrochimica Acta Part A: Molecular and Biomolecular

Spectroscopy
journal homepage: www.elsevier.com/locate/saa

Colorimetric determination of melamine in milk using unmodified

silver nanoparticles
Naveen Kumar ⁎, Harish Kumar, Bimlesh Mann, Raman Seth
Dairy Chemistry Division, National Dairy Research Institute, Karnal 132001, Haryana, India

a r t i c l e i n f o a b s t r a c t

Article history: Melamine is nitrogen rich chemical compound used as an adulterant in dairy products by unscrupulous people to
Received 14 July 2015 increase the apparent protein content. This incident prompted the researchers to develop simple methods for
Received in revised form 18 November 2015 easy detection of melamine in food samples. In the present paper, we report a simple and sensitive colorimetric
Accepted 25 November 2015
method for detection of melamine in milk based on silver nanoparticles. This method relies upon the principle
Available online 26 November 2015
that melamine causes the aggregation of silver nanoparticles, resulting in abrupt color change from yellow to
Keywords:
red under optimized conditions. The concentration of melamine in adulterated sample can be quantitated by
Silver nanoparticles (Ag NPs) monitoring the absorption spectra of silver nanoparticles using ultraviolet–visible (UV–Vis) spectrometer. The
Melamine present colorimetric method which utilizes silver nanoparticles of 35 nm can reliably detect melamine down
Milk to a concentration of 0.04 mg l−1.
Adulteration © 2015 Elsevier B.V. All rights reserved.
Aggregation

1. Introduction methodologies based on gas chromatography/mass spectrometry (MS)

Melamine is a synthetic chemical compound widely used in the
manufacture of melamine-formaldehyde resin in chemical industries.
It is also used in production of adhesives, flame retardant material,
and thermosetting plastics [1]. Melamine is rich in nitrogen (66%), and
hence was used to adulterate protein rich products like pet foods, milk
and infant formula to increase the apparent protein content [2,3]. In
2007 and 2008, cases of melamine adulteration were reported in
pet food and infant formula, respectively. The origin of melamine adul-
teration was traced to mainland China which has caused tremendous
health effects and devastating economic loss. Consumption of melamine
contaminated products caused development of kidney stones and other
renal problems. The adulterated products were exported to other coun-
tries also, making it an international issue. The methods (Kjeldahl and
Dumas) used for protein estimation in food samples, measure nitrogen
content and not directly the protein content, thus found unsuitable to
distinguish between nitrogen of protein source or non-protein source
(melamine) [4,5]. Therefore, melamine adulterated food products easily
passed the quality checks.
At that time, it was challenge to find methods for detection
of melamine in food and feed products as it was never thought that
such incident could happen. Later on, researchers developed various
Abbreviations: Ag NPs, silver nanoparticles; TEM, transmission electron microscopy;
DLS, dynamic light scattering; LOD, limit of detection; LOQ, limit of quantification.
⁎ Corresponding author at: School of Bioengineering and Food Technology, Shoolini
University, Solan, HP, India.
E-mail address: nkft87@gmail.com (N. Kumar).
[6,7] high-performance liquid chromatography/MS [8–11], capillary
zone electrophoresis/MS [12,13], chemiluminescence [14,15] and immu-
noassay [16] methods. These methods are sensitive and reliable, but in-
volve high cost, sophisticated instrumentations and are time consuming.
In recent years, use of nanotechnology is gaining momentum to
develop very fast, accurate and cost effective tests for adulteration de-
tection in food products. Nanoparticles are successfully applied to de-
velop colorimetric probe for detection of microbial contamination and
metal ions [17–20]. Nanoparticle based colorimetric methods have
also been reported for the detection of melamine [21–27]. Nanoparticles
are very suitable entity to develop colorimetric probe because of their
strongly distance-dependent optical properties and extremely high
extinction coefficient.
In the present work, we report a simple and cost effective colorimetric
test for the determination of melamine in milk using silver nanoparticles.
Silver nanoparticles are straw yellow in color when dispersed in solution,
but upon aggregation caused by melamine changes to red/orange in color.
2. Materials and methods
2.1. Chemicals
Melamine (99%) and sodium borohydride (NaBH4) were purchased
from Sigma-Aldrich (St Louis, MO, USA). Sodium citrate was purchased
from Glaxo Laboratories (India) Ltd. (Mumbai, India). Sodium hydrox-
ide (NaOH) was purchased from Thermo-Fisher Scientific India Pvt.
Ltd. (Mumbai, India). Silver nitrate (AgNO3) was purchased from

http://dx.doi.org/10.1016/j.saa.2015.11.028
1386-1425/© 2015 Elsevier B.V. All rights reserved.
90 N. Kumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 156 (2016) 89–97

Fig. 1. Absorption spectra of Ag NPs in the absence of melamine (green line) and in the presence of melamine (red line). Insert is the photograph showing visual color change of silver
nanoparticles. Experimental conditions: 600 μl Ag NPs + 400 μl H2O, 600 μl Ag NPs + 400 μl melamine; level of addition of melamine was 1 mg/l and incubation time was 20 min.
(For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Fig. 2. TEM images of Ag NPs A. 600 μl Ag NPs + 400 μl H2O (dispersed); B. 600 μl Ag NPs + 400 μl melamine (0.5 mg/l); C. 600 μl Ag NPs + 400 μl melamine (1 mg/l); and D. 600 μl Ag
NPs + 400 μl melamine (10 mg/l) (aggregated).
 N. Kumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 156 (2016) 89–97 91

Fig. 3. DLS characterization of Ag NPs. (A) Size distributions of Ag NPs (average size 35 nm); (B) size distribution of Ag NPs upon addition of 1 ppm melamine (average size 160 nm);
(C) zeta potential of Ag NPs (−23 mV); (D) zeta potential of Ag NPs upon addition of 1 ppm melamine (−14 mV).

Merck Ltd. (Mumbai, India). Melamine stock solution was prepared
by dissolving 10 mg of melamine in 1 l of water. All the solvents
and reagents were of analytical grade and were used without further
purification. Millipore Milli-Q ( MΩ cm) water was used in all the exper-
iments. The raw milk was obtained from the cattle yard in the university
campus.
2.2. Preparation of silver nanoparticles
All the glassware used in preparation of silver nanoparticles were
dipped in freshly prepared aqua regia (1:3, HNO3:HCl) and rinsed thor-
oughly with water. Silver nanoparticles (Ag NPs) were prepared by
chemical reduction method as described by Mehrgardi and Ahangar
92 N. Kumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 156 (2016) 89–97
[28] with some modifications. Briefly, 12 ml of silver nitrate solution
(1 mM) was added drop by drop to 28 ml of 2 mM sodium borohydride
(NaBH4). The reaction mixture was stirred vigorously in an ice bath
using a magnetic stirrer. The solution turned light yellow after the addi-
tion of 4 ml of silver nitrate which showed the initiation of silver nano-
particle synthesis and a straw yellow when all of the silver nitrate
(12 ml) had been added. Then 5 mg of sodium citrate tri-hydrate was
added to the above solution as a stabilizing reagent to prevent the
aggregation of Ag NPs. The bright yellow colored Ag NPs were then
stored in dark bottle at 4 °C for further use.
2.3. Sample extraction and detection of melamine
Milk constituents (such as casein and fats) may interfere in mela-
mine detection using silver nanoparticles; therefore, samples were
treated before analysis to precipitate the protein and remove fat. Sample
extraction procedure is described here. 10 ml of raw milk sample was
taken in a centrifuge tube. 1.5 ml potassium hexacyanoferrate (II)
trihydrate (3.6% aq.) solution was added in the sample and vortexed
for 1 min. Then 1.5 ml zinc sulfate (7.2% aq.) solution was added and
vortexed for 1 min. The mixture was then centrifuged at 10,000 rpm
for 5 min. The clear supernatant was taken in another centrifuge tube
and adjusted to pH 8 with 1 M NaOH. It was filtered through 0.44 μm fil-
ter and filtrate was used for colorimetric detection of melamine using
silver nanoparticles. 400 μl of sample filtrate was mixed with 600 μl of
silver nanoparticles and absorption spectra were recorded to carry out
recovery study. Spiked milk samples were made by the addition of
stock melamine solution at different concentrations (0, 0.2, 0.4, 0.8,
1.6 mg/l) and extracted following the above described procedure.
2.4. Instrumentation
Absorption spectra were recorded on a SPECORD 200 UV–Vis
Spectrophotometer (Analytikjena Germany) at room temperature.
Fig. 4. Schematic illustration of melamine detection and visual color change of Ag NPs after addition of melamine. Insert is the photograph of visual color change of Ag NPs upon addition of
1 mg/l of melamine.
Transmission electron microscopy (TEM) measurements were
made on a TECNAI F20 (FEI Co., Holland) high resolution transmis-
sion electron microscope operated at an accelerating voltage of
200 kV. The samples for TEM characterization were prepared by
placing a drop of colloidal solution on carbon-coated copper grid
and dried at room temperature. Size of nanoparticles were deter-
mined on Malvern Zetasizer ver. 7.03 (Malvern instruments ltd.,
United Kingdom) equipped with a 633 nm He–Ne laser. Dynamic
Light Scattering is used to measure particle size. The pH measurements
were carried out on digital pH meter (Electronic Corporation of India
Ltd.). The centrifugation was performed on a KUBOTA 6800 centrifuge
(KUBOTA Corporation, Japan). The photographs were taken with
Samsung SL-502 digital camera.
3. Results and discussion
3.1. Characterization of silver nanoparticles
The silver nanoparticles synthesized in the present study were
bright yellow in color. UV–Vis spectra showed that the dispersed silver
nanoparticles exhibited strong absorption maxima around 397 nm.
After interaction with melamine molecules, silver nanoparticles aggre-
gated and the yellow color changed to orange/reddish with appearance
of new absorption maxima around 540 nm (Fig. 1). The aggregation of
silver nanoparticles was confirmed by TEM images and dynamic light
scattering (DLS) study. Fig. 2 shows the dispersed silver nanoparticles
in the absence of melamine and aggregation of silver nanoparticles in
the presence of melamine. With the increase in concentration of
melamine, aggregation of silver nanoparticles also increased (Fig. 2).
DLS results determined the average size of silver nanoparticles as
35 nm which upon aggregation changed to 160 nm and zeta potential
changed from −23 mV in the absence of melamine to −14 mV in the
presence of melamine (Fig. 3).
 N. Kumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 156 (2016) 89–97 93

Fig. 5. (A) Time-dependent change in absorption spectra of Ag NPs in the presence of melamine (0.5 mg/l). (B) Effects of reaction time on the absorption ratio (A540/A397) (n = 3).

3.2. Principle of melamine detection using silver nanoparticles
Fig. 4 describes the basic principle of melamine detection using
silver nanoparticles. The silver nanoparticles are stabilized by nega-
tively charged citrate ions which counteract the Van der Walls'
attraction and keep the nanoparticles dispersed in aqueous solution.
When melamine is present, it interacts with negatively charged
silver nanoparticles through hydrogen bonding. Hydrogen bonding
between silver nanoparticles and melamine molecules cause the ag-
gregation of silver nanoparticles. Upon aggregation, the yellow color
of silver nanoparticles changed to red color and absorption spectra
shows a red shift toward 540 nm as shown in Fig. 1. A single molecule
of melamine contains six active sites to form double hydrogen bonds
with a similar site of another melamine molecule [29]. Thus with
increase in the concentration of melamine, more and more silver
nanoparticles aggregate and absorption spectra around 540 nm
increased simultaneously.
3.3. Optimization of assay conditions
The colorimetric method reported here is based upon the melamine
induced aggregation and color change of silver nanoparticles with cor-
responding change in absorption spectra. The aggregation of silver
nanoparticles can be influenced by factors such as media pH and reac-
tion time. Therefore, influence of the media pH (4.0–11.0) and reaction
time on the assay were investigated for making this method highly sta-
ble and reproducible. Media pH were adjusted using 1 M NaOH. It was
found that pH 8.0 was the optimum pH for reproducible results as
under extreme pH conditions, silver nanoparticles may undergo self-
aggregation and false positive results may be obtained. To determine
94 N. Kumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 156 (2016) 89–97

Fig. 6. (A) UV–Vis absorption spectra of Ag NPs in the presence of different concentrations of melamine (0, 0.1, 0.5, 1.0, 1.5 and 2.0 ppm) (n = 3). (B) Calibration curve of melamine de-
tection in the presence of melamine.

the interaction time required for aggregation of nanoparticles, change in
absorption spectra of silver nanoparticles after addition of melamine
(0.5 mg/l) was observed and absorption ratio (A540/A397) was plotted
against time (Fig. 5). Absorption peak around 540 nm showed a gradual
increase with corresponding increase in absorption ratio which became
constant after 18 min. It is assumed that no further aggregation of silver
nanoparticles occurred as all the melamine molecules have been
exhausted. Therefore, the results were taken after 20 min. However,
the color change at higher concentration of melamine was very fast as
compared to lower concentration.
3.4. Calibration curve and sensitivity
Standard melamine solutions were prepared by dissolving the
known amount of melamine in water. Silver nanoparticles were used
to detect the melamine in standard melamine solution under above op-
timized conditions and calibration curve were drawn by plotting absor-
bance ratio (A540/A397) against the melamine concentration (Fig. 6).
Calibration curve was linear over a concentration range of 0.1–2 mg/l
with regression equation y = 0.1615x + 0.1436 and R2 of 0.99. There
was gradual increase in the absorption value of A540/A397 with increase
 N. Kumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 156 (2016) 89–97
Fig. 7. Selectivity of the Ag NPs toward melamine versus other tested adulterants and preservatives. The concentration of melamine is 1 ppm, whereas that of all the other substances is
100 ppm (n = 3).
in concentration of melamine. This calibration curve was used for
quantitative estimation of melamine in spiked milk samples.
3.5. Interference study
To determine the specificity of present colorimetric method for
detection of melamine, we investigated the possible interference of
common adulterants and preservatives. Change in absorption ratio
(A540/A397) of silver nanoparticles was monitored in the presence of
melamine and other substances. Results are shown in Fig. 7. No major
interference was observed by any of the adulterant or preservative on
detection of melamine using silver nanoparticles.
Interference of selected amino acids, lactose, galactose and amine
bearing compounds was also investigated on melamine detection
using silver nanoparticles. Change in absorption ratio (A540/A397) was
monitored and results are shown in Fig. S1. It was found that the
presence of amino acids, lactose, galactose and diphenylamine
didn't cause any interference in melamine detection, but the pres-
ence of amine containing compounds namely o-phenylenediamine
dihydrocholride and p-phenylenediamine caused the change in absorp-
tion ratio (A540/A397) of silver nanoparticles. Similar results were
obtained in our previous work where gold nanoparticles were used as
sensor for melamine detection in milk [23].
3.6. Analysis of melamine in pure and spiked milk samples
In order to validate the reliability of the proposed method to
specifically detect melamine in real milk samples, known quantities of
melamine were doped into the samples of raw milk before sample pre-
treatment, then the samples were processed and analyzed according to
the prescribed procedures. No distinguishable color change of Ag NPs
was observed upon addition of filtrate from control samples while red
to orange/red color change was observed upon addition of filtrate
from melamine spiked samples and confirmed by the change in absorp-
tion spectra of silver nanoparticles (Fig. 8).
The absorbance ratios (A540/A397) were calculated based on the data
shown in Fig. 8(A), and a good linear relationship (y = 0.1527 × +
0.0136, R2 = 0.99) was obtained between absorption ratio (A540/A397)
and concentration of melamine in the range of 0.2–1.6 mg/l (Fig. 8).
95
Recoveries and precision data are shown in Table 1. The recoveries of
melamine from spiked milk samples varied from 92.5 to 99.4% with an
average recovery ± SD (n = 3) of 96.4 ± 3.24. The relative standard
deviations (RSDs) ranged from 5.26–8.18% (n = 3). Recoveries and pre-
cision data are presented in Table 1.
In general, method accuracy and precision were very good, with
recoveries greater than 92% and relative standard deviations (RSDs)
less than 9%, indicating the acceptable reproducibility of the proposed
colorimetric method.
3.7. Limit of detection (LOD) and limit of quantification (LOQ)
Limits of detection (LOD) and limits of quantification (LOQ) were
calculated as three and ten times, respectively, the standard deviations
of the intercepts of the calibration curves divided by the slopes of
the calibration curves shown in Fig. 6B [30]. Under the optimized
conditions, the proposed method exhibits 0.04 mg/l and 0.12 mg/l
of limit of detection (LOD) and limit of quantification (LOD) respec-
tively. This limit is much lower than the maximum level of melamine
permitted by USFDA for milk and milk products [31]. The recom-
mended limit of melamine concentrations by the US Food and Drug
Administration is 1 mg/l in infant formula and 2.5 mg/l in other
food products.
4. Conclusion
In this paper, we have presented a simple and sensitive colorimetric
method for the detection of melamine in milk samples. Yellow colored
silver nanoparticles of 35 nm size were used to develop the colorimetric
method and melamine, if present in sample, cause the aggregation of
silver nanoparticles resulting visual color change from yellow to red.
The simple color change of silver nanoparticles can be used as an indica-
tion for melamine adulteration omitting the use of sophisticated instru-
ments. The reported method is highly sensitive and reliable to detect
melamine down to a concentration of 0.04 mg l−1.
Supplementary data to this article can be found online at http://dx.
doi.org/10.1016/j.saa.2015.11.028.
96 N. Kumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 156 (2016) 89–97

Fig. 8. A. Concentration-dependent absorption spectra of the Ag NPs; B. corresponding plot of absorption ratio (A540/A397) versus melamine concentration spiked in raw milk. Inserts are
the photographs of 600 μl Ag NP solution mixed with 400 μl melamine solutions. The concentrations of spiked melamine (from left to right) are 0, 0.2, 0.4, 0.8, 1.6 mg/l respectively.

Acknowledgement

The financial support provided by National Dairy Research Institute,

Table 1 Karnal, India is gratefully acknowledged. Instrumentation facility (TEM
Recovery and precision data for melamine detection in spiked raw cow milk samples using and Zetasizer) availed at NIPER, S.A.S. Nagar, Mohali, India is also
silver nanoparticles.
acknowledged.
Sample Melamine added (mg/l) Recovered (mg/l) Recovery (%) RSD (%)

0.2 0.19 ± 0.01 95.0 5.26

0.4 0.37 ± 0.02 92.5 5.41 References
Raw milk
0.8 0.79 ± 0.06 98.8 7.59
1.6 1.59 ± 0.13 99.4 8.18 [1] Y.N. Wu, Y.F. Zhao, J.G. Li, A survey on occurrence of melamine and its analogues in
tainted infant formula in China, Biomed. Environ. Sci. 22 (2009) 95–99.
Data are presented as mean ± standard deviation (SD) (n = 3), RSD: relative standard [2] M. Lin, A review of traditional and novel detection techniques for melamine and its
deviation. analogues in foods and animal feed, Front. Chem. Sci. Eng. 3 (2009) 427–435.
 N. Kumar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 156 (2016) 89–97
[3] S.A. Tittlemier, Methods for the analysis of melamine and related compounds in
foods: a review, Food Addit. Contam. Part A: Chem. Anal. Control Expo. Risk Assess.
27 (2010) 129–145.
[4] F. Sun, W. Ma, L. Xu, Y. Zhu, L. Liu, C. Peng, L. Wang, H. Kuang, C. Xu, Analytical
methods and recent developments in the detection of melamine, Trends Anal.
Chem. 29 (2010) 1239–1249.
[5] T. Saint-Denis, J. Goupy, Optimization of a nitrogen analyser based on the Dumas
method, Anal. Chim. Acta 515 (2004) 191–198.
[6] X.M. Xu, Y.P. Ren, Y. Zhu, Z.X. Cai, J.L. Han, B.F. Huang, Y. Zhu, Direct determination of
melamine in dairy products by gas chromatography/mass spectrometry with
coupled column separation, Anal. Chim. Acta 650 (2009) 39–43.
[7] H. Miao, S. Fan, P.P. Zhou, L. Zhang, Y.F. Zhao, Y.N. Wu, Determination of melamine
and its analogues in egg by gas chromatography–tandem mass spectrometry using
an isotope dilution technique, Food Addit. Contam. A 27 (2010) 1497–1506.
[8] A. Filazi, U.T. Sireli, H. Ekici, H.Y. Can, A. Karagoz, Determination of melamine in milk
and dairy products by high performance liquid chromatography, J. Dairy Sci. 95
(2012) 602–608.
[9] X.L. Zheng, B.S. Yu, K.X. Li, Y.N. Dai, Determination of melamine in dairy products by
HILIC-UV with NH2 column, Food Control 23 (2012) 245–250.
[10] Y.Y. Chao, C.T. Lee, Y.T. Wei, H.S. Kou, Y.L. Huang, Using an on-line microdialysis/
HPLC system for the simultaneous determination of melamine and cyanuric acid
in non-dairy creamer, Anal. Chim. Acta 702 (2011) 56–61.
[11] A. Khedr, Optimized extraction method for LC–MS determination of bisphenol A,
melamine and di (2-ethylhexyl) phthalate in selected soft drinks, syringes, and
milk powder, J. Chromatogr. B 930 (2013) 98–103.
[12] Y. Wen, H. Liu, P. Han, Y. Gao, F. Luan, X. Li, Determination of melamine in milk
powder, milk and fish feed by capillary electrophoresis: a good alternative to
HPLC, J. Sci. Food Agric. 90 (2010) 2178–2182.
[13] C.Y. Guo, H.Y. Wang, X.P. Liu, L.Y. Fan, L. Zhang, C.X. Cao, Fast and selective determi-
nation of total protein in milk powder via titration of moving reaction boundary
electrophoresis, Electrophoresis 34 (2013) 1343–1351.
[14] H.J. Zeng, R. Yang, Q.W. Wang, J.J. Li, L.B. Qu, Determination of melamine by flow
injection analysis based on chemiluminescence system, Food Chem. 127 (2011)
842–846.
[15] J. Zhang, M. Wu, D. Chen, Z. Song, Ultrasensitive determination of melamine in milk
products and biological fluids by luminol-hydrogen peroxide chemiluminescence, J.
Food Compos. Anal. 24 (2011) 1038–1042.
[16] T.L. Fodey, C.S. Thompson, I.M. Traynor, S.A. Haughey, D.G. Kennedy, S.R.H. Crooks,
Development of an optical biosensor based immunoassay to screen infant formula
milk samples for adulteration with melamine, Anal. Chem. 83 (2011) 5012–5016.
[17] L. Li, B. Li, Y. Qi, Y. Jin, Label-free aptamer-based colorimetric detection of mercury
ions in aqueous media using unmodified gold nanoparticles as colorimetric probe,
Anal. Bioanal. Chem. 393 (2009) 2051–2057.
97
[18] R.A. Sperling, P.R. Gil, F. Zhang, M. Zanella, W.J. Parak, Biological applications of gold
nanoparticles, Chem. Soc. Rev. 37 (2008) 1896–1908.
[19] D. Wang, W. Dou, G. Zhao, Y. Chen, Immunosensor based on electrodeposition of
gold-nanoparticles and ionic liquid composite for detection of Salmonella pullorum,
J. Microbiol. Methods 106 (2014) 110–118.
[20] Y. Zhang, R. Li, Q. Xue, H. Li, J. Liu, Colorimetric determination of copper (II) using a
polyamine-functionalized gold nanoparticle probe, Microchim. Acta (2015). http://
dx.doi.org/10.1007/s00604-015-1498-4.
[21] H. Ping, M. Zhang, H. Li, S. Li, Q. Chen, C. Sun, T. Zhang, Visual detection of melamine
in raw milk by label-free silver nanoparticles, Food Control 23 (2012) 191–197.
[22] W. Yun, H. Li, S. Chen, D. Tu, W. Xie, Y. Huang, Aptamer-based rapid visual biosens-
ing of melamine in whole milk, Eur. Food Res. Technol. 238 (2014) 989–995.
[23] N. Kumar, R. Seth, H. Kumar, Colorimetric detection of melamine in milk by
citrate-stabilized gold nanoparticles, Anal. Biochem. 456 (2014) 43–49.
[24] J.Y. Xin, L.X. Zhang, D.D. Chen, K. Lin, H.C. Fan, Y. Wang, C.G. Xia, Colorimetric detec-
tion of melamine based on methanobactin-mediated synthesis of gold nanoparti-
cles, Food Chem. 174 (2015) 473–479.
[25] K. Rovina, S. Siddiquee, N.K. Wong, Development of melamine sensor based on ionic
liquid/nanoparticles/chitosan with modified gold electrode for determination of
melamine in milk product, Sens. Bio-Sens. Res. 4 (2015) 16–22.
[26] J. Du, Y. Wang, W. Zhang, Gold nanoparticles-based chemiluminescence resonance
energy transfer for ultrasensitive detection of melamine, Spectrochim. Acta A 149
(2015) 698–702.
[27] H.P. Borase, C.D. Patil, R.B. Salunkhe, R.K. Suryawanshi, B.K. Salunke, S.V. Patil,
Biofunctionalized silver nanoparticles as a novel colorimetric probe for melamine
detection in raw milk, Biotechnol. Appl. Biochem. (2015). http://dx.doi.org/10.
1002/bab.1306.
[28] M.A. Mehrgardi, L.E. Ahangar, Silver nanoparticles as redox reporters for the ampli-
fied electrochemical detection of the single base mismatches, Biosens. Bioelectron.
26 (2011) 4308–4313.
[29] F. Silly, A.Q. Shaw, M.R. Castell, G.A.D. Briggs, M. Mura, N. Martsinovich, L.
Kantorovich, Melamine structures on the Au (111) surface, J. Phys. Chem. C 112
(2008) 11476–11480.
[30] S. Ehling, S. Tefera, I.P. Ho, High-performance liquid chromatographic method for
simultaneous detection of the adulteration of cereal flours with melamine and relat-
ed triazine by-products ammeline, ammelide, and cyanuric acid, Food Addit.
Contam. A 24 (2007) 1319–1325.
[31] US FDA, Interim Safety and Risk Assessment of Melamine and Its Analogues in
Food for Humans, United States Department of Health and Human Services, Food
and Drug Administration, Center for Food Safety and Nutrition, Silver Spring, MD,
2008 (Accessed on 12 November, 2010. http://www.cfsan.fda.gov/~dms/
melamra3.html).