Sensors and Actuators B 191 (2014) 246–251

Contents lists available at ScienceDirect

Sensors and Actuators B: Chemical

journal homepage: www.elsevier.com/locate/snb

Dopamine functionalized CuInS2 quantum dots as a fluorescence

probe for urea
Siyu Liu, Fanping Shi, Lu Chen, Xingguang Su ∗
Department of Analytical Chemistry, College of Chemistry, Jilin University, Changchun 130012, China

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

Article history: In this paper, a simple, convenient and selective urea-biosensing system with dopamine-functionalized
Received 27 June 2013 CuInS2 quantum dots (QDs) as the fluorescence probe was developed. Water-soluble CuInS2 QDs capped
Received in revised form by 3-mercaptopropionic acid (MPA) was directly synthesized in aqueous solution, and then it was linked
10 September 2013
to dopamine to form the dopamine-functionalized CuInS2 QDs (DA-CuInS2 QDs) fluorescence probe.
Accepted 11 September 2013
When the pH value of DA-CuInS2 QDs solution was adjusted from neutral to alkalinity, the dopamine
Available online 25 September 2013
on the surface of QDs would change into quinone, which would lead to the fluorescence quenching of
DA-CuInS2 QDs due to the charge transfer interactions between QDs and the proximal quinone. Based on
Keywords:
CuInS2 quantum dots
the fact that the hydrolysis of urea in the presence of urase would release OH− to change the pH value of
Dopamine the solution, so the fluorecence intensity of DA-CuInS2 QDs could be linked to the enzymatic degradation
Fluorescence quenching of urea. The novel urea-biosensing system could effectively probe urea in the dynamic concentration
Urea range from 0.2 to 6 mmol/L.
Urase © 2013 Elsevier B.V. All rights reserved.

1. Introduction urase as a catalyst and CdSe/ZnS QDs capped by mercaptosuccinic

Urea, as a fertilizer, and animal feeding stuff additive, is mass-
produced every year and widely applied in agriculture and chemical
industry [1,2]. And not only that, urea is the major excretory prod-
uct of protein metabolism and non-protein nitrogenous substance
in body [3]. Urea in blood or in urine is an important marker in the
diagnosis of renal and liver diseases [4]. It is necessary to perform
the analysis of urea content in serum or urine before diagnosing
the different dysfunctions of the patients. The urea concentration
in serum of healthy adults may be in the range from 30 to 80 mmol/L
[5].
Due to the important effects of urea on human health, to
develop a sensitive and fast method for the determination of
urea has attracted more attention. So far, urea can be measured
by several techniques, such as, conductance [6,7], potentiome-
try [8,9], amperometry [10,11], and spectrophotometry [1,12].
However, complicated operation, long operation time and high
cost were found in these methods. In recent years, a fluorescence
measurement has been successfully applied in the determina-
tion of some small biological molecules owing to its operational
simplicity, high sensitivity and real-time monitoring [13–24]. So
far, there are only a few reports on the fluorescence probe for the
determination of urea. Huang et al. proposed an assay system with
∗ Corresponding author. Tel.: +86 431 85168352.
E-mail address: suxg@jlu.edu.cn (X. Su).
acid (MSA) as an indicator for quantitative analysis of urea, and
the fluorescence intensity of CdSe/ZnS QDs could be enhanced
in alkaline environment after the enzyme–urea reaction [25].
Ruedas-Rama developed an enzyme-linked nanosphere sensor to
probe urea based on fluorescence of QDs responds to the pH value
changes after the enzyme–urea reaction [4].
As well-known, the urase-catalyzed hydrolysis of urea releases
NH4 + , OH− and HCO3 − ions as products, and the pH value of the sys-
tem gradually increase with the urea hydrolysis. Therefore, a high
sensitive pH sensor is the key to develop the fluorescence measure-
ments for urea. As previous reports, QDs capped with some kinds
of mercaptoalkanoic acids provided an obvious route to obtain pH
sensor [25–29], and some other alternative ligands were introduced
to control pH sensitivity of QDs [28,29]. Liu and coworkers reported
the fluorescence of a mercaptoacetic acid-capped CdSe/ZnSe/ZnS
QDs solution increased by around fivefold when the pH changed
from 4 to 10 [27]. Tomasulo et al. developed a strategy to switch the
fluorescence of CdSe/ZnS core–shell QDs with pH changes of sys-
tems that is based on the photoinduced transfer of either energy
from CdSe/ZnS QDs to [1,3]oxazine ligands or electrons from the
organic to the inorganic components [28].
Dopamine (DA), as one of the most significant catecholamine
neurotransmitters, influences a wide variety of human’s moti-
vated behaviors, attention span, and neuronal plasticity [30–33].
Dopamine and some of its derivatives exhibit complex redox prop-
erties with pH-tunable oxidation and reduction potentials [34–36].
Some researchers have reported that the oxidation product

0925-4005/$ – see front matter © 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.snb.2013.09.056
 S. Liu et al. / Sensors and Actuators B 191 (2014) 246–251
benzoquinone of dopamine could effectively quench the fluo-
rescence of QDs, and such fluorescence quenching process could
be attributed to charge transfer between the QDs and proximal
benzoquinone [34,36–39].
However, the traditional cadmium-based QDs are toxic for bio-
logical systems and eventually will cause serious environmental
problems due to the leak of cadmium. Recently, a novel kind of
I-III-VI CuInS2 QDs that does not contain any toxic Class A ele-
ment (Cd, Pb, and Hg) or B element (Se and As) has attracted
considerable interests [40–43]. In previous report, we presented
the one-pot synthesis of water-soluble CuInS2 QDs capped with
3-mercaptopropionic acid (MPA) [44].
In this paper, MPA capped CuInS2 QDs are covalently bonded
to dopamine via 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide
hydrochloride (EDC) and N-hydroxysuccinimide (NHS) as coupling
reagents. The fluorescence of the obtained dopamine functional-
ized CuInS2 QDs (DA-CuInS2 QDs) dramatically reduced with the
increased pH value of the environments. Based on the fact that the
urease-catalyzed hydrolysis of urea could release OH− to increase
the pH value of the environments, we developed a simple, conve-
nient and highly sensitive fluorescence method for the detection
of urea utilizing the pH sensitive dopamine functionalized CuInS2
QDs as the fluorescence probe.
2. Experiment
2.1. Apparatus
The fluorescence spectra were obtained by using a Shimadzu
RF-5301 PC spectrofluorophotometer equipped with a xenon
lamp using right-angle geometry. UV–vis absorption spectra were
obtained by a Varian GBC Cintra 10e UV-vis spectrometer. In both
experiments, a 1 cm path-length quartz cuvette was used.
2.2. Reagents
All reagents were of at least analytical grade. The water used
in all experiments had a resistivity higher than 18 M cm−1 .
Copper (II) chloride dihydrate (CuCl2 ·2H2 O), sodium hydroxide
(NaOH), sulfourea (CS(NH2 )2 ), urea, tetrahydrofuran (THF), sodium
dehydrogenized phosphate (NaH2 PO4 ) and disodium hydrogen
phosphate (Na2 HPO4 ) were purchased from Shanghai Qingxi Tech-
nology Co., Ltd. Dopamine, indium (III) chloride tetrahydrate
(InCl3 ·4H2 O), MPA, EDC, NHS, and urease (1 U/mg) were purchased
from Sigma–Aldrich Corporation.
2.3. Preparation of MPA-capped CuInS2 QDs
MPA-capped CuInS2 QDs were synthesized in aqueous solu-
tion, according to our previous report [44]. 0.15 mmol CuCl2 ·2H2 O
and InCl3 ·4H2 O were dissolved in 10.5 mL distilled water, and then
1.8 mmol MPA was injected into the solution. The pH value of the
mixture solution was adjusted to 11.3 by adding 2 mol/L NaOH
solution with stirring during this process. After stirring for 10 min,
0.30 mmol CS(NH2 )2 was dissolved in above solution. The previous
process was all finished at room temperature, and then the mixture
solution was finally transferred into a 15 mL autoclave. Then it was
heated and maintained at 150 ◦ C for 21 h and then cooled down
to room temperature by a natural process. Enthanol was added to
the stock solution to obtain CuInS2 QDs precipitate, and the process
was repeated three times. The unreacted residues were removed by
the cycled washing. The purified CuInS2 QDs were dissolved in PBS
(2 mmol/L, pH 7.0), and stored in the darkroom. The final CuInS2
QDs solution concentration was 0.15 mmol/L.
247
2.4. Preparation of dopamine functionalized CuInS2 QDs
The conjugation of dopamine to CuInS2 QDs was performed uti-
lizing EDC and NHS as coupling reagents according to previous
reports [39,45]. Briefly, the CuInS2 QDs solution was mixed with
EDC and NHS at room temperature, and then dopamine was added
into the solution, stirring for more than 3 h (the mole ratio of QD to
NHS, EDC, dopamine was 1:1500:1500:1000). The DA-CuInS2 QDs
were purified by precipitation with addition of THF to the reaction
system, and the purified DA-CuInS2 QDs powder were dissolved in
aqueous solution, and stored in the darkroom.
2.5. Detection of urea by DA-CuInS2 QDs
For urea determination, 200 ␮L urase solution (12.5 mg/mL, dis-
solved in 10 mmol/L PBS pH 7.0) and different amount of urea were
added in to a 2 mL calibrated test tube, shaken thoroughly for 1 min.
And then 25 ␮L DA-CuInS2 QDs solution (0.12 mg/mL) was added
into the test tube, diluted to the mark with deionized water, shaken
thoroughly and equilibrated for 30 min. The fluorescence spectra of
the mixture were recorded from 580 nm to 850 nm, and the samp-
ling interval was 1.0 nm. The slit widths of excitation and emission
were both 10 nm. The fluorescence (FL) intensity of the maximum
emission peak was used for the quantitative analysis of the urea.
2.6. Preparation of serum samples
Drug-free human blood samples were collected from healthy
volunteers and then centrifuged at 10 000 rpm for 10 min after
standing for 2 h at room temperature. 1 mL obtained serum sam-
ples and 2 mg urase were mixed and incubated for 24 h at 35 ◦ C
to hydrolyze the inherent urea in the serum. And then 3 mL 15%
trichloroacetic acid was added into the serum to destroy the activ-
ity of urase, which would irreparably lose its catalytic activity under
strong acid condition, and precipitate proteins of the serum [46,47].
After vigorously shaking for 15 min, the mixture was centrifuged
at 10 000 rpm for 10 min at 4 ◦ C. The obtained supernatant was
adjusted to pH 7.0 using NaOH solution and then diluted by 50
times with deionized water. Different concentrations of urea were
added to the diluted serum samples to prepare the spiked samples.
3. Results and discussions
3.1. DA-CuInS2 QDs fluorescence probe
As well-known that, the fluorescence properties of QDs had a
close connection to their surface capped layers [13,48]. As shown
in Scheme 1, in this study, we used dopamine to modify MPA capped
CuInS2 QDs to form the dopamine-functionalized CuInS2 QDs fluo-
rescence probe. The urea would release OH− with urase as catalysts
that could induce changes from dopamine to its corresponding
quinone. And the energy transfer between the QDs and proximal
quinone would lead to the fluorescence quenching of DA-CuInS2
QDs, so the DA-CuInS2 QDs fluorescence probe could be utilized
for the determination of urea. Fig. 1 shows the UV–vis absorption
and fluorescence emission spectra of MPA-capped CuInS2 QDs and
DA-CuInS2 QDs respectively. It could be seen that after conjuga-
tion with dopamine, the fluorescence emission peak exhibited a
blue-shift from 660 nm to 630 nm, which was attributed to that
the surface capping layer of the CuInS2 QDs was changed from the
negatively charged acetate to the electron donor dopamine [39,45].
Fig. S1A shows the FT-IR spectra of the MPA-capped CuInS2 QDs
(curve a) and DA-CuInS2 QDs (curve b). As curve a in Fig. S1A
shown, the majority of MPA functional groups could be clearly
found through the C O stretching peak (1730 cm−1 ), and C H
248 S. Liu et al. / Sensors and Actuators B 191 (2014) 246–251
Scheme 1. The schematic illustration of the synthetic process of DA-CuInS2 QDs and the detection of urea.
stretching mode (2850 cm−1 , 2920 cm−1 , 2960 cm−1 ). The charac-
teristic peak of S H did not appear within 2550–2680 cm−1 , which
might be caused by the covalent bonds between thiols and metal.
As curve b in Fig. S1A shown, the amide I band (1660 cm−1 ), amide
II (1540 cm−1 ) and amide III (1400 cm−1 ) band for CONH group
appeared, which was due to the conjugation of DA to the MPA
capped CuInS2 QDs. UV–vis absorption spectra (230–400 nm) of
the DA-CuInS2 QDs were performed. As shown in Fig. S1B, the typ-
ical UV–vis absorption peak around 280 nm belonging to DA was
observed in the DA-CuInS2 QDs, and not observed in the original
MPA-capped CuInS2 QDs. It was further confirmed the presence of
dopamine after the conjugation,
3.2. The influence of pH on the fluorescence of DA-CuInS2 QDs
The charge transfer interactions between fluorescent QDs and
redox active dopamine have been extensively investigated [34,36].
In this work, we studied the effect of pH on the fluorescence prop-
erties of DA-CuInS2 QDs. As shown in Fig. 2, it could be seen that
the fluorescence intensity of DA-CuInS2 QDs decreased with the
increasing of the pH from 7.0 to 9.93, and when the pH increased to
be 9.93, the fluorescence was almost entirely quenched. The signif-
icant fluorescence quenching was due to that the oxygen-catalytic
oxidation of dopamine potentials was gradually reduced as the
increased pH value from 7.0 to 9.93, and the equilibrium favored
the transformation of dopamine to its oxidation product quinone at
high pH. And the transfer of electrons from a photoexcited QDs to
the lowest unoccupied molecular orbital of the proximal quinone
acceptor would result in the fluorescence quenching of DA-CuInS2
QDs [34,36]. As shown in Fig. S2, under the acid conditions, the flu-
orescence intensity of DA-CuInS2 QDs did not exhibit significant
variation with the decreasing pH value of the system.
As well-known, the urea could release NH4 + , OH− and HCO3 −
ions as products with urase as the catalyst. In order to exclude the
Fig. 1. The UV–vis absorption and fluorescence emission spectra of MPA-capped
CuInS2 QDs (dash line) and DA-CuInS2 QDs (solid line).
other possible interference on the fluorescence of DA-CuInS2 QDs,
we respectively investigated the effect of urea, NH4 + , HCO3 − and
urase on the fluorescence of DA-CuInS2 QDs. Fig. S3 shows the flu-
orescence intensity of DA-CuInS2 QDs with different concentration
of urea, NH4 + , HCO3 − (in the range from 0 to 10 mmol/L) or urase
(in the range from 0 to 1.25 mg/mL). It can be seen that urea, only
NH4 + , HCO3 − or urase itself cannot induce obviously fluorescence
quenching of the DA-CuInS2 QDs. So it was further confirmed that
we could detect urea based on the fluorescence quenching of DA-
CuInS2 QDs induced by OH− that was released from the hydrolysis
of urea in the presence of urase.
3.3. Optimization for urea detection
As described in Eq. (1), the urea could release ionic product NH4 + ,
HCO3 − and OH− in the presence of urase.
CO(NH2 )2 + 3H2 O = 2NH4 + + HCO3 − + OH− (1)
As shown in Fig. 2, the fluorescence of DA-CuInS2 QDs would be
quenched when pH of the system changed from neutral to alkalin-
ity. Therefore, DA-CuInS2 QDs could be utilized as a fluorescence
probe for the determination of urea with urase as catalyst. As well-
known that the urase plays a key role in the hydrolysis of urea.
Therefore, choosing the appropriate concentration of urase could
improve the sensitivity for urea detection by DA-CuInS2 QDs probe.
In this work, the fluorescence intensity ratio F/F0 (F and F0 are
the fluorescence intensity of DA-CuInS2 QDs with or without urea)
of DA-CuInS2 QDs around 630 nm quenched by 2 mmol/L urea in the
presence of different urase concentration (0.08–1.25 mg/mL) were
studied, and the results were shown in Fig. 3. It can be seen that
Fig. 2. The fluorescence (FL) spectra of DA-CuInS2 QDs after the addition of differ-
ent amounts of NaOH with the pH value of the system changed from 7.00 to 9.93
(pH = 7.00, 7.16, 7.27, 7.58, 7.96, 8.55, 8.95, 9.31, 9.70, 9.93). Inset: the plot of flu-
orescence quenching ratios of F/F0 at 630 nm versus the pH value of the system at
room temperature.
 S. Liu et al. / Sensors and Actuators B 191 (2014) 246–251
Fig. 3. The relationship between the fluorescence intensity ratio F/F0 of DA-CuInS2
QDs and the reaction time. F and F0 are the fluorescence intensity of DA-CuInS2 QDs
with or without urea. Conditions: 2 mmol/L urea and various urase concentrations
(0.08, 0.21, 0.42, 0.63, 1.25 mg/mL).
the fluorescence intensity ratio F/F0 of DA-CuInS2 QDs decreased
slightly with the increasing of reaction time in the presence of
low concentration of urase (0.08 or 0.21 mg/mL). When the urase
concentration increased in the assay system (0.42–1.25 mg/mL),
more dramatic changes of fluorescence intensity ratios F/F0 were
observed. The gradually decreased fluorescence intensity ratios
F/F0 with the increasing of reaction time clearly suggests that the
urea was gradually decomposed to release OH− by catalysis of
urase, and the increased concentrations of OH− in the assay sys-
tem would result in the more obvious fluorescence quenching of
DA-CuInS2 QDs. As Fig. 3 showed when the urase cencentration
increased from 0.63 mg/mL to 1.25 mg/mL, the fluorescence inten-
sity ratios (F/F0 ) after 30 min of enzyme hydrolysis almost reached
the same. The results indicated that the urase concentration of
1.25 mg/mL was sufficient in this urea molecules hydrolysis system,
and it was corresponding with previous reports [4,25].
In this paper, we investigated systematically the effect of the
reaction temperature on this urea detection system, and the results
were shown in Fig. S4. As curve a in Fig. S2 shown, the fluoresce
intensity of DA-CuInS2 QDs decreased gradually with the increase
of reaction temperature from 15 ◦ C to 40 ◦ C, which was due to that
the increase of reaction temperature would increase the rate of oxi-
dation and reduce the dopamine potential for oxidation and shifted
the equilibrium toward the production of quinones [36,49–51].
Urase is an enzyme in organisms, and its activities increased grad-
ually with the increase of temperature at a given range that was
corresponding with fluorescence intensity changes of DA-CuInS2
QDs–urea–urase system (curve b). Considering the stability of DA-
CuInS2 QDs, the activities of urase and being simple to operate,
room temperature 20 ◦ C was chosen as the reaction temperature
in our urea detection system.
3.4. Fluorescence detection of urea using the DA-CuInS2 QDs
As shown in Fig. 4, the fluorescence of DA-CuInS2 QDs decreased
as the urea concentration increased from 5 ␮mol/L to 12 mmol/L in
the presence of 1.25 mg/mL urase as catalyst. Fig. 4 inset describes
the relationship between the fluorescence intensity ratio F/F0 (F and
F0 are the fluorescence intensity of DA-CuInS2 QDs with or without
urea) and the logarithm of urea concentration (0.005–12 mmol/L,
logarithm value −2.30 to 1.08). It could be seen that when the
concentration of urea was 12 mmol/L, the fluorescence inten-
sity of DA-CuInS2 QDs would decrease to about 20% of original
249
Fig. 4. The fluorescence (FL) spectra of DA-CuInS2 QDs upon the addition of differ-
ent concentration of urea in the range of 0–12 mmol/L (0, 0.005, 0.020, 0.050, 0.10,
0.20, 0.50, 1.0, 2.0, 4.0, 6.0, 8.0, 10.0, 12.0 mmol/L) in the presence of 1.25 mg/mL
urase. Inset: the plot of fluorescence quenching ratios of F/F0 at 630 nm versus the
concentration of urea at room temperature.
fluorescence intensity. The obviously fluorescence quenching
induced by urea with urase as catalysts was due to the hydrolysis
of urea, which released OH− to change the pH value of the system,
and it favored easier transformation of dopamine to its oxidation
product quinone. And then the transfer of electrons from QDs to
the proximal quinone acceptor would result in the fluorescence
quenching of DA-CuInS2 QDs. The proposed urea-biosensing
system could effectively probe urea in the dynamic range from 0.2
to 6 mmol/L.
As shown in Table 1, we compared the detecting limits and
dynamic range of urea with various methodologies including
potentiometry [10,52], amperometry [8], fluorescence measure-
ments [4,25] and our proposed detection system. It can be seen
that the proposed method offered a similar detecting limit and
wide dynamic range for the detection of urea. Compared with previ-
ous reports about urea sensors, the DA-CuInS2 QD-based biosensor
offered many advantages such as simple operations, low cost, with-
out enzyme immobilization, good selectivity and sensitivity.
3.5. Effect of foreign substance
In this paper, we studied the effect of some physiological ions
and molecules on our proposed urea fluorescence detection system.
It could be found that from Fig. S5A, the fluorescence intensity of
DA-CuInS2 QDs with 1.25 mg/mL urase added by 2 mmol/L foreign
substance K+ , Na+ , Ca2+ , Mg2+ , glucose (Glu), glucine (Gly), threon-
ine (Thr) or lysine (lys) remained nearly constant, and only after
the addition of urea, the fluorescence intensity showed obvious
decreasing. The results indicated that the proposed urea detection
system showed the high selectivity to urea. In Fig. S5B, we fur-
ther investigated the fluorescence response of DA-CuInS2 QDs to
2 mmol/L urea with 1.25 mg/L urase in the presence of 2 mmol/L
foreign substance K+ , Na+ , Ca2+ , Mg2+ , Glu, Gly, Thr or lys. It could
be seen from Fig. S5B that even in the presence of 2 mmol/L inter-
ference, the DA-CuInS2 QDs sensor still work the same. The results
showed that the proposed urea detection system could provide
well ability of resisting interference from physiological ions and
molecules.
3.6. Detection of urea in real samples
In order to test the applicability of the proposed method, it
was applied to determinate urea in human serum samples which
spiked with different concentration of urea. The results obtained by
250 S. Liu et al. / Sensors and Actuators B 191 (2014) 246–251
Table 1
Comparison of performance of different urea sensors.
Type Sensing system
Potentiometry Poly(N-vinylcarbazole)/stearic acid Langmuir–Blodgett films
Mercaptohydroquinone-modified gold electrode
Amperometry Urease + polyurethane–acrylate polytoluidine blue film
Fluorescence Urea/MSA-CdSe/ZnS QDs
Mercaptopropionic capped QDs–PAH-CaR
Dopamine functionalized CuInS2 QDs
standard addition method were shown in Table S1. The accuracy of
the proposed method was evaluated by determining the recoveries
of urea in real samples. It could be found that the average recover-
ies of urea in the real samples was in the range of 98–102% and the
RSD was lower than 3.2%. The above results demonstrated that the
potential applicability of the DA-CuInS2 QD-based fluorescence
probe for the detection of urea in human serum samples.
4. Conclusion
We have demonstrated the dopamine functionalized CuInS2
QDs that was sensitive to the pH values changes can be utilized
as an effective fluorescence probe for the determination of urea.
The fluorescence of the DA-CuInS2 QDs was quenched by urea in
the range from 0.2 to 6 mmol/L with urase as the catalyst. The pro-
posed method was successfully applied to the detection of urea in
human serum sample with satisfactory results.
Acknowledgements
This work was financially supported by the National Natural
Science Foundation of China (Nos. 20875036 and 21075050) and
the Science and Technology Development Project of Jilin Province,
China (No. 20110334).
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.snb.2013.09.056.
References
[1] B. Kovács, G. Nagy, R. Dombi, K. Tóth, Optical biosensor for urea with improved
response time, Biosensors and Bioelectronics 18 (2003) 111–118.
[2] J.M. Garrido, R. Mendez, J.M. Lema, Treatment of wastewaters from a
formaldehyde-urea adhesives factory, Water Science and Technology 42 (2000)
293–300.
[3] R. Doong, H.M. Shih, Array-based titanium dioxide biosensors for ratiometric
determination of glucose, glutamate and urea, Biosensors and Bioelectronics
25 (2010) 1439–1446.
[4] M.J. Ruedas-Rama, E.A.H. Hall, Analytical nanosphere sensors using quantum
dot–enzyme conjugates for urea and creatinine, Analytical Chemistry 82 (2010)
9043–9049.
[5] J.C. Chen, J.C. Chou, T.P. Sun, S.K. Hsiung, Portable urea biosensor based on
the extended-gate field effect transistor, Sensors and Actuators: B Chemical
91 (2003) 180–186.
[6] P.S. Chaudhari, A. Gokarna, M. Kulkarni, M.S. Karve, S.V. Bhoraskar, Porous sil-
icon as an entrapping matrix for the immobilization of urease, Sensors and
Actuators: B Chemical 107 (2005) 258–263.
[7] W.Y. Lee, S.R. Kim, T.H. Kim, K.S. Lee, M.C. Shin, J.K. Park, Sol–gel-derived thick-
film conductometric biosensor for urea determination in serum, Analytica
Chimica Acta 404 (2000) 195–203.
[8] I. Vostiar, J. Tkac, E. Sturdik, P. Gemeiner, Amperometric urea biosensor based
on urease and electropolymerized toluidine blue dye as a pH-sensitive redox
probe, Bioelectrochemistry 56 (2002) 113–115.
[9] A.P. Soldatkin, J. Montoriol, W. Sant, C. Martelet, N. Jaffrezic-Renault, A novel
urea sensitive biosensor with extended dynamic range based on recombinant
urease and ISFETs, Biosensors and Bioelectronics 19 (2003) 131–135.
[10] F. Mizutani, S. Yabuki, Y. Sato, Voltammetric enzyme sensor for urea using
mercaptohydroquinone-modified gold electrode as the base transducer,
Biosensors and Bioelectronics 12 (1997) 321–328.
Dynamic range (mmol/L) Detection limit (mmol/L) Reference
10–68 5 [52]
0.2–5 0.2 [10]
0.2–0.8 0.02 [8]
0.01–120 0.01 [25]
0.4–25 0.4 [4]
0.2–6 0.1 This work
[11] Y.C. Luo, J.S. Do, Urea biosensor based on PANi (urease)-Nafion® /Au composite
electrode, Biosensors and Bioelectronics 20 (2004) 15–23.
[12] H.C. Tsai, R.A. Doong, Preparation and characterization of urease-encapsulated
biosensors in poly(vinyl alcohol)-modified silica sol–gel materials, Biosensors
and Bioelectronics 23 (2007) 66–73.
[13] C.C. Carrión, S. Cárdenas, B.M. Simonet, M. Valcárcel, Selective quantification of
carnitine enantiomers using chiral cysteine-capped CdSe(ZnS) quantum dots,
Analytical Chemistry 81 (2009) 4730–4733.
[14] Y.J. Chen, X.P. Yan, Chemical redox modulation of the surface chemistry of
CdTe quantum dots for probing ascorbic acid in biological fluids, Small 5 (2009)
2012–2018.
[15] S. Huang, Q. Xiao, R. Li, H.L. Guan, J. Liu, X.R. Liu, Z.K. He, Y. Liu, A simple and
sensitive method for l-cysteine detection based on the fluorescence intensity
increment of quantum dots, Analytica Chimica Acta 645 (2009) 73–78.
[16] H. Huang, Y. Gao, F.P. Shi, G.N. Wang, Shah S. Mazhar, X.G. Su, Determination
of catecholamine in human serum by a fluorescent quenching method based
on a water-soluble fluorescent conjugated polymer–enzyme hybrid system,
Analyst 137 (2012) 1481–1486.
[17] W.T. Wu, T. Zhou, A. Berliner, P. Banerjee, S.Q. Zhou, Glucose-mediated
assembly of phenylboronic acid modified CdTe/ZnTe/ZnS quantum
dots for intracellular glucose probing, Angewandte Chemie 122 (2010)
6704–6708.
[18] J.P. Yuan, W.W. Guo, E.K. Wang, Quantum dots–bienzyme hybrid system for
the sensitive determination of glucose, Biosensors and Bioelectronics 23 (2008)
1567–1571.
[19] N. Malashikhina, Valeri Pavlov, DNA-decorated nanoparticles as nanosensors
for rapid detection of ascorbic acid, Biosensors and Bioelectronics 33 (2012)
241–246.
[20] R. Gill, L. Bahshi, R. Freeman, I. Willner, Optical detection of glucose and
acetylcholine esterase inhibitors by H2 O2 -sensitive CdSe/ZnS quantum dots,
Angewandte Chemie International Edition 47 (2008) 1676–1679.
[21] Y. Gao, Yan. Li, X. Zou, H. Huang, X.G. Su, Highly sensitive and selective detec-
tion of biothiols using graphene oxide-based molecular beacon-like fluorescent
probe, Analytica Chimica Acta 731 (2012) 68–74.
[22] P. Wu, X.P. Yan, Ni2+ -modulated homocysteine-capped CdTe quantum dots as
a turn-on photoluminescent sensor for detecting histidine in biological fluids,
Biosensors and Bioelectronics 26 (2010) 485–490.
[23] L. Zhou, Y.H. Lin, Z.Z. Huang, J.S. Ren, X.G. Qu, Carbon nanodots as fluorescence
probes for rapid, sensitive, and label-free detection of Hg2+ and biothiols in
complex matrices, Chemical Communication 48 (2012) 1147–1149.
[24] F. Pu, Z.Z. Huang, J.S. Ren, X.G. Qu, Ligand/lon-based ensemble for fluores-
cence turn on detection of cysteine and histidine with tunable dynamic range,
Analytical Chemistry 82 (2010) 8211–8216.
[25] C.P. Huang, Y.K. Li, T.M. Chen, A highly sensitive system for urea detection
by using CdSe/ZnS core-shell quantum dots, Biosensors and Bioelectronics 22
(2007) 1835–1838.
[26] H.D. Duong, J.I. Rhee, Use of CdSe/ZnS luminescent quantum dots incorporated
within sol–gel matrix for urea detection, Analytica Chimica Acta 626 (2008)
53–61.
[27] Y.S. Liu, Y. Sun, P.T. Vernier, C.H. Liang, S.Y.C. Chong, M.A. Gundersen,
pH-sensitive photoluminescence of CdSe/ZnSe/ZnS quantum dots in human
ovarian cancer cells, Journal of Physical and Chemistry C 111 (2007) 2872–2878.
[28] M. Tomasulo, I. Yildiz, S.L. Kaanumalle, F.M. Raymo, pH-sensitive ligand for
luminescent quantum dots, Langmuir 22 (2006) 10284–10290.
[29] A.S. Susha, A.M. Javier, W.J. Parak, A.L. Rogach, Luminescent CdTe nanocrystals
as ion probes and pH sensors in aqueous solutions, Colloids Surface A 281 (2006)
40–43.
[30] M. Darvas, R.D. Palmiter, Restricting dopaminergic signaling to either dorsolat-
eral or medial striatum facilitates cognition, Journal of Neuroscience 30 (2010)
1158–1165.
[31] D.L. Robinson, B.J. Venton, M.L.A.V. Heien, R.M. Wightman, Detecting subsecond
dopamine release with fast-scan cyclic voltammetry in vivo, Clinical Chemistry
49 (2003) 1763–1773.
[32] V. Hefco, K. Yamada, A. Hefco, L. Hritcu, A. Tiron, T. Nabeshima, Role of the
mesotelencephalic dopamine system in learning and memory processes in the
rat, European Journal of Pharmacology 475 (2003) 55–60.
[33] P. Damier, E.C. Hirsch, Y. Agid, A.M. Graybiel, The substantia nigra of the human
brain II. Patterns of loss of dopamine-containing neurons in Parkinson’s disease,
Brain 122 (1999) 1437–1448.
[34] X. Ji, G. Palui, T. Avellini, H. Bin, C.Y. Na, K.K.L. Yi Jr., H. Mattoussi, On the
pH-dependent quenching of quantum dot photoluminescence by redox active
dopamine, Journal of the American Chemical Society 134 (2012) 6006–6017.
 S. Liu et al. / Sensors and Actuators B 191 (2014) 246–251
[35] A. Klegeris, L.G. Korkina, S.A. Greenfield, Autoxidation of dopamine: a compar-
ison of luminescent and spectrophotometric detection in basic solutions, Free
Radical Biology and Medicine 18 (1995) 215.
[36] I.L. Medintz, M.H. Stewart, S.A. Trammell, K. Susumu, J.B. Delehanty, B.C.
Mei, J.S. Melinger, J.B. Blanco-Canosa, P.E. Dawson, H. Mattoussi, Quantum-
dot/dopamine bioconjugates function as redox coupled assemblies for in vitro
and intracellular pH sensing, Nature Materials 9 (2010) 676–684.
[37] S.J. Clarke, C.A. Hollmann, Z.J. Zhang, D. Suffern, S.E. Bradforth, N.M. Dim-
itrijevic, W.G. Minarik, J.L. Nadeau, Photophysics of dopamine-modified
quantum dots and effects on biological systems, Nature Materials 5 (2006)
409–417.
[38] R. Gill, R. Freeman, J.P. Xu, I. Willner, S. Winograd, I. Shweky, U. Banin, Pro-
bing biocatalytic transformations with CdSe-ZnS QDs, Journal of the American
Chemical Society 128 (2006) 15376.
[39] D.R. Cooper, D. Suffern, L. Carlini, S.J. Clarke, R. Parbhoo, S.E. Bradforth,
J.L. Nadeau, Photoenhancement of lifetimes in CdSe/ZnS and CdTe quantum
dot–dopamine conjugates, Physical Chemistry Chemical Physics 11 (2009)
4298–4310.
[40] H. Nakamura, W. Kato, M. Uehara, K. Nose, T. Mata, S. Otsuka-Yao-Matsuo, M.
Miyazaki, H. Maeda, Tunable photoluminescence wavelength of chalcopyrite
CuInS2 -based semiconductor nanocrystals synthesized in a colloidal system,
Chemistry of Materials 18 (2006) 3330–3335.
[41] R.G. Xie, M. Rutherford, X.G. Peng, Formation of high-quality I-III-VI semicon-
ductor nanocrystals by tuning relative reactivity of cationic precursors, Journal
of the American Chemical Society 131 (2009) 5691–5697.
[42] S.L. Castro, S.G. Bailey, R.P. Raffaelle, K.K. Banger, A.F. Hepp, Nanocrystalline
chalcopyrite materials (CuInS2 and CuInSe2 ) via low-temperature pyroly-
sis of molecular single-source precursors, Chemistry of Materials 15 (2003)
3142–3147.
[43] W.W. Xiong, G.H. Yang, X.C. Wu, J.J. Zhu, Aqueous synthesis of color-tunable
CuInS2 /ZnS nanocrystals for the detection of human interleukin 6, ACS Applied
Materials Interfaces 5 (2013) 8210–8216.
[44] S.Y. Liu, H. Zhang, Y. Qiao, X.G. Su, One-pot synthesis of ternary CuInS2 quan-
tum dots with near-infrared fluorescence in aqueous solution, RSC Advances 2
(2012) 819–825.
[45] S.J. Clarke, C.A. Hollmann, F.A. Aldaye, J.L. Nadeau, Effect of ligand density on
the spectral, physical, and biological characteristics of CdSe/ZnS quantum dots,
Bioconjugate Chemisty 19 (2008) 562–568.
[46] J.H. Peng, Y.H. Wang, J.L. Wang, X. Zhou, Z.H. Liu, A new biosensor for glucose
determination in serum based on up-converting fluorescence resonance energy
transfer, Biosensors and Bioelectronics 28 (2011) 414–420.
251
[47] A.M. Alam, M. Kamruzzaman, S.H. Lee, Y.H. Kim, S.Y. Kim, G.M. Kim, H.J. Jo, S.H.
Kim, Determination of catecholamines based on the measurement of the metal
nanoparticle-enhanced fluorescence of their terbium complexes, Microchimica
Acta 176 (2012) 153–161.
[48] T. Jin, F. Fujii, E. Yamada, Y. Nodasaka, M. Kinjo, Control of the optical properties
of quantum dots by surface coating with calix[n]arene carboxylic acids, Journal
of the American Chemical Society 128 (2006) 9288–9289.
[49] H.A. Laitinen, W.E. Harris, Chemical Analysis an Advanced Text and Reference,
2nd edn., McGraw-Hill, New York, 1975.
[50] F.P. Fehlner, N.F. Mott, Low temperature oxidation, Oxidation of Metals 1 (1970)
59–99.
[51] C. Wang, R. Yuan, Y.Q. Chai, F.X. Hu, Simultaneous determination of hydro-
quinone, catechol, resorcinol and nitrite using gold nanoparticles loaded
on poly-3-amino-5-mercapto-1,2,4-triazole-MWNTs film modified electrode,
Analytical Methods 4 (2012) 1626–1628.
[52] R. Singhal, A. Gambhir, M.K. Pandey, S. Annapoorni, B.D. Malhotra, Immobi-
lization of urease on poly(N-vinyl carbazole)/stearic acid Langmuir–Blodgett
films for application to urea biosensor, Biosensors and Bioelectronics 17 (2002)
697–703.
Biographies
Siyu Liu is currently carrying out his graduate work for his doctor degree under the
guidance of Professor Su in Jilin University. His research focuses on the synthesis
and functionalization of quantum dots and their application on sensors for metal
ion and small molecules.
Fanping Shi is currently carrying out her graduate work for her masters degree
under the guidance of Professor Su in Jilin University. Her research focuses on the
synthesis and functionalization of quantum dots and their application on sensors
for metal ion and small molecules.
Lu Chen received her bachelor degree in Institute of Chemistry Jilin University. Her
research focuses on the application of quantum dots on sensors for small molecules.
Xingguang Su is a professor at the Department of Analytical Chemistry at the College
of Chemistry, Jilin University. She received her master degree from Jilin University
(China) in 1992 and her doctor degree from Jilin University (China) in 1999. Her
research focuses on the synthesis, characterization, functionalization and applica-
tion of quantum dots and quantum dots-tagged microspheres in biomedicine.