Accepted Manuscript

Title: A Versatile Fluorimetric Chemosensor for Mercury (II)

Assay Based on Carbon Nanodots

Authors: Xiao-Chun Liang, Shuqin Chen, Jianmei Gao,

Hongyu Zhang, Yu Wang, Ji-Hui Wang, Liang Feng

PII: S0925-4005(18)30206-5
DOI: https://doi.org/10.1016/j.snb.2018.01.182
Reference: SNB 24031

To appear in: Sensors and Actuators B

Received date: 14-9-2017

Revised date: 16-1-2018
Accepted date: 23-1-2018

Please cite this article as: Xiao-Chun Liang, Shuqin Chen, Jianmei Gao, Hongyu
Zhang, Yu Wang, Ji-Hui Wang, Liang Feng, A Versatile Fluorimetric Chemosensor
for Mercury (II) Assay Based on Carbon Nanodots, Sensors and Actuators B:
Chemical https://doi.org/10.1016/j.snb.2018.01.182

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A Versatile Fluorimetric Chemosensor for Mercury (II) Assay

Based on Carbon Nanodots

Xiao-Chun Liang,a, b Shuqin Chen,b Jianmei Gao,b Hongyu Zhang,a, b Yu Wang,b Ji-

Hui Wang*a and Liang Feng*b

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a) School of Biological Engineering, Dalian Polytechnic University, Dalian,

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Liaoning 116034, P. R. China.
b) Key Lab of Separation Science for Analytical Chemistry, Dalian Institute of
Chemical Physics, Chinese Academy of Sciences, Dalian, Liaoning 116023, P. R.
China.
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Corresponding authors: wangjh@dlpu.edu.cn; fengl@dicp.ac.cn
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Highlights
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 2,6-pyridinedicarboxylic acid-derived carbon nanodots (PECDs)have been

fabricated.
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 PECDs were used for Hg2+sensing based on photoluminescence quenching.

 The structural characterization of as-obtained PECDs was thoroughly

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investigated.
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 ThePECDs werecapable of rapidly detecting Hg2+inreservoir water, tap water,

female and male human urines.
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Abstract

Human urine mercury level functions as a pre-warning signal for mercury

explosure, either in acute or chronic events. Unlike common hydrargyrism due to

organic mercury uptake overproof via aquatic products, the risk of inorganic mercury

toxication would be much higher for residents living in mercury-related industry areas

due to direct or indirect mercury wastewater emission to vicinal water system. In this

work, we reported a novel fluorimetric chemosensor, 2,6-pyridinedicarboxylic acid-

derived carbon nanodots (PECDs), which exhibits excellent versatility in direct

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inorganic Hg2+ detection. The fabrication was proved facile, inexpensive and eco-

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friendly. The morphology, chemical and spectral properties of PECDs were

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thoroughly investigated by using HRTEM, XPS, ATR-FTIR, UV-Vis and fluorescent

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spectrum. PECDs showed a broad range of linearity between fluorescence intensity

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and Hg2+ concentration. The limit of detection was calculated as low as 75 nM. The
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sensing mechanism was investigated and assumed as fluorescence quenching
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originated from the formation of PECDs-Hg2+ complex, which facilitates the non-
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radiative relaxation of surface emission of PECDs. In Hg2+ assay for real samples,
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considerable recoveries (RSD<6%) and good linearity (R2>0.99) were obtained in all

four kinds of samples, involving reservoir water, tap water, female and male human
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urines. Thus, we believe our method would make certain contribution to the
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development of applicable versatile sensing platform for Hg2+.

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Keywords: Mercury; fluorimetric; chemosensor; carbon nanodots

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1. Introduction

Over the past few decades, mercury and its inorganic salts have attracted

continuous attentions due to their irreplaceability in various industrial activities with

severely neurological, psychological toxicity [1]. More importantly, they also possess

high bioavailability through which inorganic mercury salts can convert to more toxic

and stable, bio-accumulating organic mercury via the methylation of anaerobic

bacteria [2-5]. Unlike common hydrargyrism due to organic mercury uptake

overproof via aquatic products, the risk of inorganic mercury toxication would be

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much higher for residents living in mercury-related industry areas due to direct or

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indirect mercury wastewater emission to vicinal water system [6].A well quoted

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example of environmental contamination which led to death of nearly 600 persons

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took place in Minamata Japan in 1950s [7]. The monitoring of Hg2+ level in various

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samples that are easily affected by mercury wastewater emission is, therefore, very
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important in aquatic ecosystems [8, 9]. As such, the intracorporal mercury level of
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correlating residents should also be periodically examined to avoid the unconscious

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ingestion either through atmospherical or aqueous pathway. A versatile Hg2+ sensing

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platform for in different media detection is highly desirable.

As it is well known that the ingestion of organic mercury (mainly MeHg) can be
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detected from both human blood and urine (via demethylation of MeHg during
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metabolism), whilst exposure to inorganic mercury is predominately accumulated in

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kidneys and excreted in urine [10], which makes urinary system becomes an

importance metabolic pathway of mercury [11, 12]. As a result, mercury level in urine
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has become an important indicator of mercury uptake in human body either via acute

or chronic way. The World Health Organization (WHO) has set a threshold limit value of 50

µg/g creatinine in urine for the occupational exposure to mercury, which is corresponded to about
50 µg/L (0.25 µmol/L), the levels of urinary mercury of non-occupational people are below 20

µg/L (0.1 µmol/L), and there are no clinical or subclinical effects when urinary mercury are below

20 µg/L [13]. Additionally, upon collaborating with the assay of homologous blood

mercury level, the total mercury urinary level can also provide specific information

for searching mercury contamination sources, which would be very helpful for the

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prevention of further exposure and in clinical therapy.

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Comparing to hidebound and jargon-filled instrumental analyses including cold-

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vapor atomic absorption spectrometry(CV-AAS) [14-16], cold-vapor atomic

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fluorescence spectrometry (CV-AFS) [17, 18], and inductively coupled plasma mass

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spectrometry (ICP-MS) [19, 20], fluorescent chemosensors developed from small
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molecules, polymers, proteins, DNA/RNA and some nanomaterials exhibit more
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brilliant perspectives due to their suitability in the establishment of inexpensive,

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personalized, portable pre-warning apparatus [21-26]. Although legions of

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chemosensors with splendid sensitivity or selectivity to Hg2+ have been reported,

drawbacks from tedious synthesis, low yield, high cost, poor robustness and
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solubility, or repeated contamination impeded their applicability.

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To design a capable and versatile chemosensor for Hg2+, an important step is to

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select an eligible Hg2+-receptor which generally consists of metal chelators. Amid a

variety of potential candidates, 2,6-pyridinedicarboxylic acid (PDCA) is remarkable

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because PDCA-functionalized golden nanoparticles have been demonstrated with

excellent sensitivity and selectivity toward Hg2+ [27-30]. However, this kind of probes

also suffered from forgoing problems. A new protocol which can iron out such
problems but retain the sensitivity and selectivity of PDCA-based chemosenor, will be

highly desirable.

In this work, an intentional manipulation by changing PDCA-based sensor matrix

from golden nanoparticles to PDCA-derived carbon nanodots (denoted as PECDs)

was employed. On the one hand, carbon nanodots are widely acknowledged as flaring

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matrices for fluorimetric chemosensors due to their outstanding characteristics

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involving low-cost, easy-fabrication, prominent solubility, superior chemical and

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photophysical stability [31-38]. On the other hand, the residual at the surface of as-

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synthesized PECDs serves the function of natural Hg2+ receptors, giving intrinsic

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sensitivity and selectivity toward Hg2+ without further surface decoration. For
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fabrication, PDCA and ethylenediamine were chosen as reactants which firstly
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supplied sufficient carbon matrix via polymerization before carbonization process.

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The adoption of one-step solvent-free thermal treatment ensures the simplicity and
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time-saving characteristics for this chemosensor. After thoroughly determination of

spectral properties and sensing ability of as-obtained PECDs, the limit of detection
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(LOD) was calculated as 75 nM, indicating the considerable sensitivity of PECDs

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towards Hg2+. The sensing mechanism was assumed as fluorescence quenching

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originated from the formation of PECDs-Hg2+ complex, which substantially enhanced

the non-radiative relaxation rate of surface emission of PECDs [39], as shown in

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Scheme 1. In application, we successfully demonstrated the versatility of PECDs in

the detection of Hg2+ level in natural water, tap water and human urine samples,

where all results exhibited considerable recoveries (RSD<6%) and excellent linearity
with (R2>0.99). We believe our method will make certain contribution in the

establishment of applicable versatile Hg2+ sensing platform.

2. Experimental

2.1 Materials

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Pyridine-2,6-dicarboxylic acid (PDCA) and ethylenediamine anhydrous (C2H8N2)

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were used to synthesize PECDs. PDCA and CuCl2·2H2O, MgCl2·6H2O, ZnCl2,

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AlCl3·6H2O, CdCl2, AgNO3, NiCl2·6H2O, CoCl2·6H2O, FeCl3·6H2O were purchased

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from Alfa Aesar (Shanghai, China). Ethylenediamine anhydrous, NaCl, KCl, CaCl2

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and acetate buffer solution (ABS, 0.2 M, pH=5.6) were obtained from Tianjin Kemiou
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Chemical Reagent Co., Ltd (Tianjin, China). HgCl2 were bought from Beijing
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Chemical Works. Pure-A-Lyzer Dialysis kits (3500 kDa), was purchased from Sigma
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Aldrich. All reagents were of analytical grade and used as received without further
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purification. All metal cations stock solutions were prepared by using ABS. Deionized

water was used throughout the standard experiments. The natural water samples were
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collected from Xishan reservoir which is drinking-water resources of Dalian city. The
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tap water samples were collected from our laboratory (Shahekou district, Dalian). The
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urine samples were collected from healthy volunteers.

2.2 Instruments
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The UV-Vis absorption spectra were performed by Persee TU-1901 UV-Vis

spectrophotometer. Fluorescent spectra were recorded by F-4600 fluorescence

spectrophotometer. Transmission electron microscopy (TEM) images were

photographed by JEM-2100 microscope in the case of a voltage of 200 kV. The

Thermo Nicolet IS50 attenuated total reflectance Fourier transform infrared (ATR-

FTIR) spectrometer was used to record infrared spectra of as-prepared PECDs and

PDCA. The X-ray photoelectron spectroscopy (XPS) of the PECDs was measured by

Thermo Scientific K-Alpha instrument.

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2.3 Synthesis of PECDs

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The PECDs were synthesized through a one-step solvent-free thermal treatment of

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pyridine-2,6-dicarboxylic acid and ethylenediamine anhydrous. In general, 500 mg

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pyridine-2,6-dicarboxylic acid and 2ml ethylenediamine were added into a 20 mL

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vial ,then the mixture was sonicated for 20 min to form a homogeneous solution. After
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that, the solution was fully transferred to a 25 mL Teflon-lined stainless autoclave, and
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was heated to 250 ℃ for 4 hours. After the reaction, the autoclave was cooled down
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to room temperature. Then 10 mL distilled water was added to dissolve the products,
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and then stirred for a while. The product solution was purified by dialysis against

distilled water within a dialysis kit (3500kDa) for 48 hours. The centrifugal treatment
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was performed to remove unreacted PDCA precipitates and over-carbonized by-

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products. Supernate was collected and dried to give PECDs. The as-obtained PECDs
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were re-dispersed in ABS at a concentration of 1 mg/mL for further application.

2.4 Quantum yield measurement

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The quantum yield (QY) of the PECDs was determined and calculated by using

quinine sulfate (QY=0.54 at 360 nm) as reference[40]. The quinine sulfate was

dissolved in 0.1 M H2SO4 (refractive index is 1.33). The PECDs were dispersed in
ABS (refractive index is 1.33). The absorbance at 360 nm was kept below 0.1 to

minimize the re-absorption effect. The quantum yield was then calculated by the

following equation:

𝐹𝑐 × 𝐴𝑆 × 𝑛𝑐2
𝑄𝑐 = 𝑄𝑠 ×
𝐹𝑠 × 𝐴𝑐 × 𝑛𝑠2

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Wherein “Q” is the quantum yield, “F” is the integrated emission intensity, “n” is the

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refractive index of the solvent, and “A” is the absorbance at 360 nm. The subscript

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“c” and “s” refer to PECDs and quinine sulfate, respectively. The QY of the PECDs

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was calculated as 4.5%.

2.5 Fluorescence detection of Hg2+

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Fluorometric detection of Hg2+ was performed in distilled water at room
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temperature. 50 μL of 1 mg/ml PECDs was added in 850 μL distilled water, and then
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100 μL of HgCl2 with different concentrations was added. The mixtures were shaken

up and incubated for 10 minutes to ensure complete reaction. The fluorescent spectra
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of all samples were recorded at the excitation wavelength of 340 nm.

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2.6 Interference Test

Thirteen common metal cations were used for interference experiments under the
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same condition, including K+, Cu2+, Na+, Mg2+, Zn2+, Ca2+, Al3+, Cd2+, Ag+, Ni2+,

Co2+, Fe3+, and Hg2+. The concentration of all metal cations was adjusted to 100 μM
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in the interference experiment. After adding different metal cations, the mixtures were

shaken up and incubated for 10 minutes to ensure complete reaction. In the

fluorometric measurement, the excitation wavelength for all samples was 340 nm.

2.7 Detection of Hg2+ in real samples

 The natural water samples were collected from Xishan reservoir. The samples

were centrifuged at 12500 rpm for 20 min and then filtered through a cellulose acetate

membrane with 0.22 µm pore diameter. The resultant water samples were spiked with

Hg2+ at different concentrations as stock solutions for further use.

The tap water samples were firstly centrifuged at 12500 rpm for 20 min to remove

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large impurities. A 0.22 µm cellulose acetate membrane was used for further

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purification of tap water sample. The PECDs were dispersed in tap water and spiked

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with Hg2+, and then incubated for 10 minutes at the room temperature. The change of

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fluorescence intensity of mixtures at 420 nm was measured to evaluate practice usage

of PECDs for Hg2+ detection in tap water samples.

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The urine samples were collected from healthy volunteers. 2.50 mL of each urine
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sample and 4 mL nitric acid were added in Teflon vessels. The vessels were sealed
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and put into the microwave digestion system. The parameters set in microwave
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digestion process were shown in table S1 (see the Supporting Information, SI). The

digested solution was poured in round flask, and the excessive nitric acid was
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removed by distill method. The exhaust NO2 was bubbling within condensation
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NaOH solution. Clear solutions were finally obtained after acid removal process. The
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urine samples were diluted 10-fold. The ABS solution with Hg2+ at different

concentrations were spiked into each the urine sample for further experiments.
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3. Results and discussion

3.1 Characterization of the PECDs

The morphology and size distribution of PECDs were investigated by

Transmission electron microscopy (TEM). As shown in Fig. 1, the as-prepared

PECDs were monodispersed nanoparticles with near spherical morphology (down

inset of Fig. 1), and were well separated from each other. The average diameter was

determined 5.49 nm, which is similar with CDs previously reported otherwhere[9,

41].

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In order to obtain specific chemical composition and elemental analysis of PECDs

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X-ray photoelectron spectroscopy (XPS) were performed..As shown in Fig 2.A,

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PECDs are typical N-doped carbon dots. The C/N/O atom ratio of PECDs was

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calculated as C16.61:N5.01:O3.38. Since in pure PDCA, the N:O atom ratio would be

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much lower (N8:O38), ethylenediamine-terminated polymerization
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(>N20.28:O15.44) was assumed to take place before the carbonization step. The C1s
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spectrum showed three peaks at 284.3、285.4、287.5 eV which were attributed to C-

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C/C=C、C-N、C=O bonds, respectively (Fig 2 B). The peaks at 399.0、400.7、

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401.0 eV in N1s spectrum were ascribed to pyridyl N, -(C=O)-(N-H)- and N-H

groups, respectively(Fig 2 C). Only one peak at 531.0 eV was found in O1s spectrum
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(Fig 2 D), and was assigned to the C=O bands. These results strongly demonstrated
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that amide groups were formed in PECDs.

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The infrared spectra were recorded to elucidate the structural difference between

precursor, PECDs and PECDs/Hg complex, as shown in Fig.3. The formation of -

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(C=O)-(N-H)- was reflected from the shift of C=O stretching vibration peak from

1695 to 1654 cm-1, the presence of strong amide bending peak at 1523 cm-1, the

weakening of C-O stretching at 1247 cm-1 and hydrogen-bonded O-H out of plane
bending peak at 921 cm-1, and the replacement of –OH stretching at 2539 cm-1 by -N-

H stretching at 3288 cm-1. This matched well with the absent peak represented

carboxylic groups and neogenic peak of amide in XPS results. By the comparison of

IR spectra of PECDs and PECDs+Hg2+, we found that peaks relating to amines were

much altered. For instance, pyridyl N at 742 cm-1 was much weakened with the

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presence of the motion of –N-H towards low wave number field from 3288 to 3264

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cm-1. Resultantly, peaks correlating to coordinating N-H vibration in the range from

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2400 to 3200 cm-1 were strengthened (see the SI, Fig. S1). These results imparted that

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the chelate positions of PECDs with Hg2+ would predominately consist of pyridine-

related structures.
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3.2 Spectral properties of PECDs
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The absorption spectrum was recorded, as shown in Fig. 4 A (red line). The 50

µg/mL PECDs in ABS barely showed absorption bands in visible light region, thus
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leading to transparent appearance (inset of Fig. 4 A). Two absorbance peaks centered
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at around 224 and 275 nm, respectively, could be clearly observed on the absorption

spectrum. According to previous literatures about CDs, the peak at 224 nm was
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ascribed to the π→π* transition at conjugated component of carbon backbone. The

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other peak at 275 nm could be attributed to electron transition from non-bonding to π*

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(n→π*) orbital at heteroatoms. Comparing to the absorption peaks of PDCA at 196

nm and 272 nm (in black line in Fig. 4 A), broadened absorption band of PECDs

imparted two things: 1) the broadened and red-shifted peak at 224 implied that new

species containing-π-bond formed during the formation of skeleton of PECDs

involving graphite-like components (orderly hexagon arrangement in Fig.1 inset); 2)

the weakened but broadened peak at 275 nm suggested that the chemical environment

of heteroatoms is significantly changed after synthesis. This assumption was

consistent with the XPS and IR results about the formation of carboxamides and other

aromatic heterocycles which resultantly lowered the possibility of n→π* via the

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decrease of electron long pairs number in skeleton.

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On fluorescent spectra, the precursor PDCA exhibits a characterized emission

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centered at approximate 390 nm under 340 nm excitation. For PECDs, an intense

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emission band centered at 420 nm with a shoulder around 390 nm could be observed

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by the same excitation wavelength. This shoulder peak was by no means originated
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from residue of unreacted precursor, but was proposed stemming from PDCA-
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resembled component in PECDs matrix due to two reasons: First, PDCA has slight
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solubility (about 5 g/L, 5‰) in water, which means after two times dialysis (1:100)
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and following centrifuge treatment, the residual concentration of PDCA would not be

higher than 1 µg/mL in as-prepared 50 µg/mL PECDs stock solution. And this
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theoretical value was evaluated based on the premise that no PDCA was consumed
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during the reaction. In the experiment, the ultimate PDCA residue can be ignored,
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since ethlylenediamine was exceeding in raw ratio. The second reason was concluded

from fluorescence quenching experiment and will be elaborated in the later paragraph.
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When exposing PECDs to a series of excitation wavelength ranging from 310 to

400 nm, we found fluorescence intensity maximum was firstly increased, and then

decreased gradually accompanied with a considerable red-shift from 410 to 465 nm

(Fig. 4 B). The excitation maximum of PECDs is observed at 340nm. According to a

recent report which excitation single-dot spectroscopy was performed for analyzing

excitation-dependent properties of CDs, it stated that the emission sites are related to

the sp2 subregions in the carbon core, as well as the functional groups on the surface.

Such excitation-dependent emission property had been widespread reported as an

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important criteria for CDs species[42-44].

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3.3 Sensing of Hg2+

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To illustrate the sensing ability of PECDs towards Hg2+, fluorometric titration

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experiments were conducted. As shown in Fig. 5 A, upon the addition of different

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amounts of Hg2+ into 10 μL PECDs ABS, the fluorescent intensity gradually
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decreased along with increasing the concentration of Hg2+ from 0.1 to 100 µM. The
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calibration curve for determining Hg2+ was obtained by plotting the values of F/F0
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versus the concentrations of Hg2+ (Fig. 5B). The correlation coefficients (R2) were
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calculated as 0.997 and 0.9926 within the two linear concentration ranges of 0.1−10

and 10−100 µM, respectively (see the SI, Fig. S2 and S3). The limit of detection
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(LOD) was estimated to be 75 nM based on a signal-to-noise ratio of 3 at three times

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the standard deviation of 3 measurements of blank samples. This result explicitly

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demonstrated the excellent sensitivity of PECDs to Hg2+ by comparing to those

obtained from other CDs or nanomaterials based Hg2+ chemosensors reported

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previously (Table 1).

For the purpose of understanding quenching mechanism, we also recorded

absorption spectra of PECDs in the presence and absence of Hg2+, as shown in Fig. 5
C. Upon the addition of 100 μM Hg2+, the absorption band centered at 224 nm was

much strengthened, yet the absorption band centered at 275 nm was nearly

unattenuated. This result implied that Hg2+ would be attached to the conjugated

domain of PECDs, rather than to the oxygen-related surface groups. Although the

photoluminescence mechanism of CDs is still an open controversy, the emission

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originated from oxygen-related surface groups are thought making major contribution

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to fluorescence of CDs. In the case of PECDs, the complex formation between

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PECDs and Hg2+ was assumed taking place at conjugated-amine (primarily pyridyl N)

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chelating position of PECDs rather than surface groups according to results from

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absorption and IR spectra. The formed PECDs@Hg2+ complex substantially boosted
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electrons exciting transition at neogenic conjugated component, and to some extents,
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suppressed emission at oxygen-related surface groups via the formation of more

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favorable non-radiative deactivation pathway. This assumption could also explain

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why Hg2+ could not completely quench the fluorescence of PECDs. Another thing

should be noted here is that the decrease of fluorescence intensity of PECDs in the
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presence of Hg2+ was integrated (shoulder peak with main peak), which could exclude
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coexisting the residue of PDCA with PECDs in solution as we mentioned in foregoing

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paragraphs.

The selectivity experiment was performed by using thirteen metal cations as

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interference, including Hg2+, K+, Cu2+, Na+, Mg2+, Zn2+, Ca2+, Al3+, Cd2+, Ag+, Ni2+,

Co2+ and Fe3+. It was seen that the fluorescence intensity changed distinctively upon

the addition of different metal ions at the same concentration (100 µM) at excitation
wavelength of 340 nm, as shown in Fig. 5 D (black). Only the addition of Hg2+ led to

a dramatic quenching of fluorescent intensity of PECDs. Although the addition of

Cu2+ and Fe3+ also gave rise to certain decrease of fluorescent intensity, the

interference experiment results clearly elucidated capability of PECDs for selectively

detecting Hg2+, as shown in Fig. 5 D (red). In the present other metal cations, the

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sensitivity of PECDs towards Hg2+ was merely affected, even coexisting with Cu2+

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and Fe3+. According to the literature, such outstanding selectivity and specificity of

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PECDs to Hg2+ could be explained as the formation of more stable complex by

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PECDs with Hg (II) than with other metal ions due to larger dissociation constants

logK [54].
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3.4 Analytical applications in real samples
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In order to investigate the versatility of PECDs for Hg2+ detection, different real
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samples including reservoir water, tap water and urine samples were tested[6, 9, 49].
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All conditions were kept the same as those in the standard experiment except the

replacement of distilled water by real samples.

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For natural and tap water samples, all recoveries were calculated based the results
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obtained from the addition of 1 µM, 2µM, 4 µM and 8 µM Hg2+ to reservoir and tap
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water samples, and were listed in Table 2 and Table 3, respectively. The calculated
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results from experiments were closing to those initially spiked within samples, and

showed excellent linearity with R2=0.99 with small RSD values (lower than 6%), as

shown in Fig. 6 A and B. These results indicated that PECDs are capable of detection

Hg2+ level in natural and tap water that are easily affected by mercury wastewater
emission.

We further detected Hg2+ level in male and female urine samples by using PECDs

chemosensor. The preparation of urine samples was demonstrated in experimental

section Fluorescence intensity of PECDs in urine samples was decreased with

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increasing the concentration of Hg2+ from 0.1 to 8 µM. The calibration curves were

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obtained by plotting the values of F/F0 versus the concentrations of Hg2+ (Fig. 6C and

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6D), and exhibited excellent linearity in both samples (R2=0.9977 for male and

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R2=0.9997 for female). The recoveries of Hg2+ in urine samples at different

concentrations were also calculated and shown in Table 4. Although the results

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showed slight fluctuation, the PECDs method for inorganic Hg2+ level detection in
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urine was clearly feasible.
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4. Conclusions
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In summary, we reported a PDCA-based Hg2+-targeted fluorimetric chemosensor

based on a one-step, solvent-free thermal treatment of PDCA with ethylenediamine.

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The morphology and chemical structure of PDCA-based carbon dots (PECDs) were
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thoroughly investigated by using HRTEM, XPS, and ATR-FTIR analysis. The as-
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obtained PECDs showed excitation-dependent emission and blue fluorescence in

aqueous solution. The sensitivity and selectivity of PECDs towards Hg2+ were

thoroughly determined by using fluorescent titration and interference experiments,

and the limit of detection (LOD) was calculated as low as 75 nM. In application,
PECDs showed competitive sensing ability for Hg2+ in different real samples

involving natural water, tap water and human urine samples. We believed that PECDs

strategy characterized as easy-fabrication, good-stability and low-price would offer a

new approach to establish versatile platform for inorganic Hg2+ detection in real use.

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Acknowledgement

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This work was financially supported through the National Natural Science Foundation

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of China (Grant 21677144，31371764). The work was also supported by the Chinese

Academy of Sciences (100 Talents Program).

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Biographies

Xiaochun Liang is currently a master student in Dalian Polytechnic University, and

her research focuses on the preparation of nanomaterials.
Shuqin Chen is currently a PhD student in Dalian Institute of Chemical Physics,
Chinese Academy of Sciences. Her research focuses on the synthesis of metal clusters.

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Jianmei Gao is currently a PhD student in Dalian Institute of Chemical Physics,

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Chinese Academy of Sciences. Her research focuses on the fabrication of sensors.

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Hongyu Zhang received his master degree in 2016 from Dalian Polytechnic

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University. His research focuses on the synthesis of carbon nanodots.
Yu Wang received his PhD degree in 2014 from Kyungpook National University, and

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is currently an assistant professor of Dalian Institute of Chemical Physics in Chinese
Academy of Sciences. His main research focuses on materials preparation and
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characterization and their applications in sensing.
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Jihui Wang received his PhD degree in 2003 from Chungbuk National University.
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Currently he is a professor of Dalian Polytechnic University. His main research

focuses on food safety control.
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Liang Feng received his PhD degree in 2005 from Wuhan University. Currently he is
a professor of Dalian Institute of Chemical Physics. His main research focuses on the
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development of sensors for the determination of environmental pollutants, including

toxic gases, heavy-metal ions, poisonous anions, etc.
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Figure captions

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Fig.1. Transmission electron microscopy (TEM) of the as-synthesized PECDs. The up inset
showed the size distribution of PECDs, and the down inset illustrated the morphology of PECDs.
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Fig.2. (A)XPS spectra of the PECDs. (B-D) High-resolution XPS spectra of C1s (B), N1s (C) and
O1s (D) of the PECDs, respectively.
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Fig. 3. ATR-FTIR spectrum of the PDCA (blue curve), PECDs(red curve) and PECDs+Hg2+
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(black curve).
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Fig.4. (A) UV–vis absorption of the PDCA (black) and PECDs (red), photoluminescence
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excitation (blue) and emission (green) spectra of PECDs in aqueous solutions (50 µg/mL ),
λex=340 nm and λem =420 nm. Insets show the photographs of the obtained PECDs in aqueous
solution under illumination of white (left) and UV (365 nm, right) light; (B) Normalized FL
spectral of the PECDs at different excitation wavelength.
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Fig.5. (A) Fluorescence spectra of PECDs in the prescence of various concentrations of Hg2+ (0,

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0.1, 1, 2, 4, 8, 10, 20, 40, 80, 100 µM), (B) Corresponding relationship between F/F0 and the
concentrations of Hg2+, (C) UV–vis absorption of PECDs in the absence (black) and presence
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(red) of Hg2+ (100 μM), (D) The corresponding fluorescence intensities of PECDs towards
different ions(100 μM) in the absence (black) and presence (red) of Hg2+ (100 μM).
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Fig.6 (A-B) F/F0 of PECDs mixed with different concentrations of Hg2+ (0.1−10 µM) (n = 3) in
the nature (A) and tap water (B). (C-D) F/F0 of PECDs mixed with different concentrations of
Hg2+ (0.1−8µM) (n = 3) in the urine samples of Male(C) and Female (D).
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Scheme 1. Schematic of the preparation of PECDs and their application for Hg2+ sensing.
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 Table 1 Comparison of sensing performance of different fluorescent probes for Hg2+ detection.

Liner Detection
Fluorescent probes Ref.
range(µM) limits(µM)
Polymer Sensor (salen and
1-30 0.728 [45]
perylenyl moieties)
N-Functionalized carbon dots 0.1-2.69 2.69 [46]
N-CQDs(folic acid) 0-2.69 1.3 [47]
CD/Fe3O4 4-16 5.04 [48]

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Thiazole yellow chemical
0.1-32 0.01 [49]

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sensor
N,S -CDs 0-0.3 0.00137 [50]

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PTIQ 0-100 0.098 [6]

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N,S co-doped Carbon dots 0-40 2 [51]
Carbon dots 0-50 0.047 [52]
MPA-PDCA 0.25-0.5 0.1 [53]
PECDs 0-100 0.075 U This work
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Table 2 Determination and recoveries tests of Hg2+ in the nature water by the present methods
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(n=3).

Added Hg2+ (µM) PECDs meana ±SDb(µM) Recoveries (%) RSD (%)
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1 1.00±0.025 100.35 2.49

2 2.10±0.068 105.02 3.24

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4 4.06±0.11 101.51 2.72

8 8.14±0.18 101.77 2.23

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a Mean value of three determinations.

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b standard deviation.
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Table 3 Determination and recoveries tests of Hg2+ in the tap water by the present methods (n=3).
Added Hg2+ (µM) PECDs meana ±SDb(µM) Recoveries (%) RSD (%)
1 1.03±0.053 103.33 5.09

2 2.04±0.10 102.09 4.94

4 4.10±0.17 102.50 4.15

8 7.59±0.37 94.87 4.88

a Mean value of three determinations.

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b standard deviation.

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Table 4 Determination and recoveries tests of Hg2+ in the urine sample by the present methods

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(n=3).

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Samples Added Hg2+ (µM) PECDs meana ±SDb(µM) Recoveries(%) RSD(%)

0.1 0.11±0.0037 108.22 3.46

Female
1 0.93±0.015
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4 3.99±0.15 99.66 3.80
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8 7.96±0.0077 99.44 0.097
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0.1 0.084±0.0010 84.40 1.22

1 0.98±0.048 97.65 4.95

Male
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4 4.25±0.12 106.37 3.02

8 7.65±0.15 95.66 1.9

a Mean value of three determinations.
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b standard deviation.
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