Journal Pre-proof

A Smartphone-integrated Ratiometric Fluorescence

Sensor for Visual Detection of Cadmium Ions

Wenting Li, Xinai Zhang, Xuetao Hu, Yongqiang

Shi, Zhihua Li, Xiaowei Huang, Wen Zhang, Di
Zhang, Xiaobo Zou, Jiyong Shi

PII: S0304-3894(20)32863-6
DOI: https://doi.org/10.1016/j.jhazmat.2020.124872
Reference: HAZMAT124872

To appear in: Journal of Hazardous Materials

Received date: 26 August 2020
Revised date: 9 December 2020
Accepted date: 11 December 2020
Please cite this article as: Wenting Li, Xinai Zhang, Xuetao Hu, Yongqiang Shi,
Zhihua Li, Xiaowei Huang, Wen Zhang, Di Zhang, Xiaobo Zou and Jiyong Shi,
A Smartphone-integrated Ratiometric Fluorescence Sensor for Visual Detection
of Cadmium Ions, Journal of Hazardous Materials, (2020)
doi:https://doi.org/10.1016/j.jhazmat.2020.124872
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A Smartphone-integrated Ratiometric Fluorescence Sensor for Visual Detection

of Cadmium Ions

Wenting Li, 1,a Xinai Zhang, 1,a Xuetao Hu, 1 Yongqiang Shi, 2 Zhihua Li, 1 Xiaowei Huang, 1 Wen Zhang, 1 Di

Zhang, 1 Xiaobo Zou, 1 Jiyong Shi, 1, *

1. Agricultural Product Processing and Storage Lab, School of Food and Biological Engineering, Jiangsu

University, Zhenjiang, Jiangsu 212013, China

2. School of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, Zhejiang 310058,

China.

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a. The first two authors contributed equally

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*Corresponding author:

Prof. Jiyong Shi

Tel: +86-511-88780201; E-mail: Shi_jiyong@ujs.edu.cn

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Abstract:
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A novel fluorescence sensing platform was fabricated for visual detection of cadmium ions

(Cd2+) with excellent stability and portability. In this protocol, dual-emission ratiometric
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fluorescence probe were constructed based on silicon oxide-coated copper nanoclusters

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(CuNCs@SiO2) as a signal reference and cadmium telluride quantum dots (CdTe QDs) as signal
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response, thereby greatly improving the accuracy of test results. The level of Cd2+ can be reported
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within a wide linear range from 0.010 mg·L-1 to 2.0 mg·L-1 with a sensitive detection limit of 1.1

μg·L-1 (2.75 μg·kg-1) and a quick sample-to-answer monitoring time of 6 min, which was quite

qualified for regularly monitoring Cd2+. Moreover, aiming to attain portable analysis, the

smartphone as colorimetric reader and analyzer were also utilized for rapidly analyzing Cd2+ by

capturing the change in fluorescence color. Additionally, benefiting from the strong combination of

1, 10-phenanthroline (Phen) and Cd2+, the fluorescence probe showed excellent anti-interference

activities for Cd2+ assay in complex oyster matrix. Overall, the sensing platform had significant
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stability, specificity and sensitivity, offering a promising potential for conveniently evaluating the

quality of marine bivalves polluted with Cd2+.

Keywords:

Cadmium pollution, Oyster samples, Fluorescence sensor, Smartphone-based colorimetry, Visual

detection

1. Introduction

Cadmium ions (Cd2+) are a highly poisonous heavy metals with the carcinogenicity and

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teratogenicity, which severely impacts kidneys, liver, and reproductive organs of human (Lee et al.,

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2019; Mahalik et al., 1995; Waalkes, 2000). Moreover, it’s easily accumulated in human body

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through the food chain due to their non-degradable properties (López_Marzo et al., 2013; Wang et
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al., 2020a; Zhang et al., 2020c). Some studies point to excessive Cd2+ in the marine bivalves, rice,
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and other food samples (Chan and Wang, 2019; Deng et al., 2019). According to Codex

Alimentarius Commission (CAC, CODEX STAN 193-1995), the maximum contaminant level of
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Cd2+ in marine bivalves is 2.0 mg·kg-1. Therefore, accurate and rapid monitoring of Cd2+ pollution
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in food samples is crucial to ensure public health and food safety.

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To date, some kinds of quantitative analysis, such as inductively coupled plasma-mass

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spectrometry (ICP-MS) (Cao et al., 2017), atomic absorption spectroscopy (AAS) (Adam and

Anthemidis, 2009), electrochemical methods (Ghanei-Motlagh and Taher, 2017; Li et al., 2019c),

and fluorescent sensor methods (Noh et al., 2017; Sun et al., 2020), have been used to detect trace

amounts of Cd2+. Among them, fluorescent sensor has received wide attention for the assay of Cd2+

due to its unique properties of convenience, low cost, and high sensitivity. (Hu et al., 2019a; Li et al.,

2019b; Wang and Wu, 2019; Zhou et al., 2020). For example, Noh’s group reported a simple

CdNCs sensor and developed a selective and sensitive method for the homogeneous detection of
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Cd2+ based on the Cd2+ concentration-dependent formation of fluorescent CdNCs (Noh et al., 2017).

Wang’s group discovered a novel peptide-based fluorescent chemosensor (DGC) based on soft-hard

acid-base theory induced “turn on” responses to Cd2+ (Wang et al., 2020b). However, all of them

exhibit monochromatic fluorescence signals, and the fluorescence fluctuation generating from

varied reagent concentrations and intrinsic background fluorescence leads to poor stability and low

repeatability for detection results (Hu et al., 2019a; Li et al., 2019b). In order to compensate for the

above drawback, ratiometric fluorescence probes have been proposed to gain the strong and stable

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fluorescent signal for accuracy enhancement.

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Ratiometric fluorescence probes with double emission fluorescence characteristics have

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attracted growing research interest because of their self-calibration function for improving the
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stability and accuracy of fluorescent sensors (He et al., 2018; Hu et al., 2019b; Li et al., 2019b;
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Zhang et al., 2020b). In order to construct a successful ratiometric sensor, it is crucial to design an

appropriate reference probe for further enhancing the assay accuracy (Wang et al., 2020d). At
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present, copper nanoclusters (CuNCs) with the advantages of wide source of raw materials, high
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quantum yield, and low cost, were gradually applied to establish ratiometric fluorescence probes
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(Bai et al., 2020; Li et al., 2020; Maruthupandi et al., 2020; Yang et al., 2019). Compared with
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single CuNCs, functionalized CuNCs present distinctive nature of characteristics, in which

functionalized material takes on long-term stability and biocompatibility, and CuNCs exhibits

specific optical activity to be detected (Hu et al., 2016). Therefore, functionalized CuNCs can be

more suitable for acting as reference signal of ratiometric fluorescence probes.

Meanwhile, considering the importance of Cd2+ assays in practical applications, process

simplification is typically required to achieve a rapid-detection of Cd2+. As the most promising

portable and point-of-care sensing platform for onsite monitoring, a smartphone-based colorimetric
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or fluorescence strategy has currently been used for the rapid detection of some hazardous

substances (Hou et al., 2019; Ye et al., 2020; Zhang et al., 2020a). Smartphone as sample-to-answer

analyzers is portable and easy to be operated and can act as signal reader with installing programs

provided by the third-party service providers (Li et al., 2019a; Wang et al., 2019; Wang et al.,

2020c). The key to successful application of automatic colorimetry is to develop sensitive

recognition cell as an attachment to be integrated into a smartphone which employs the built-in

camera as optical sensor to read out the analyte concentration with image processing technique (Sun

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et al., 2019). There have been studies for the rapid colorimetric detection of tetracycline (Wang et

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al., 2020c) and glucose (Wu et al., 2020) based on smartphone-integrated ratiometric fluorescence

sensor.
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Herein, a novel ratiometric and colorimetric fluorescent probe based on silicon oxide-coated
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CuNCs (CuNCs@SiO2) as reference probe was proposed for detecting Cd2+ in oyster samples.

Cadmium telluride quantum dots (CdTe QDs) as response signal in this probe, upon exposure to
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Cd2+, the fluorescence intensity of CdTe QDs recovered because the coordination between 1,
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10-phenanthroline (Phen) and Cd2+ terminated the photo-induced hole transfer (PHT) between CdTe
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QDs and Phen. A smartphone-integrated fluorescence sensing platform was developed by

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combining the proposed fluorescence probe. Overall, this strategy offers a powerful solution for

monitoring Cd2+ pollution in marine bivalves with excellent stability, portability, and sensitivity.

2. Experimental

2.1. Reagents

Thioglycolic acid (TGA), ascorbic acid, sodium hydroxide (NaOH), tetraethyl orthosilicate

(TEOS), Tris (hydroxymethyl) aminomethane (Tris), hydrochloric acid (HCl), Phen, tellurium

powder (99%), 1-hexyl alcohol, cyclohexane, cadmium chloride (CdCl2), cupric sulfate (CuSO4),
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and sodium borohydride (NaBH4) were purchased from Aladdin Industrial Corporation (Beijing,

China). Cd2+ standard solution was purchased from Sinopharm Chemical Reagent Co., Ltd.

(Shanghai, China). All the reagents used were of analytical grade. All solutions were prepared using

double-distilled water from Millipore (Milli-Q, 18.2 MΩ·cm).

2.2. Apparatus

All fluorescence spectra were obtained using a fluorescence spectrophotometer (F-98,

Lengguang, China). Fourier transform infrared (FT-IR) spectroscopy was recorded using an FT-IR

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spectrometer (Nicolet FT-IR spectrometer 5700, USA). Transmission electron microscopy (TEM)

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was conducted using a transmission electron microscope (JEM-1400, JEOL, Japan). The signal of

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the fluorescence color was read and analyzed using Color Picker software. Fluorescence colors
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were captured by the camera of an iPhone in a sealed dark box configured with an ultraviolet lamp
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(λex=365 nm). Some other instruments such as ICP-MS (XSeries II, ThermoFisher, USA),

Microwave Digestion System (TOP wave, Analytik Jena, Germany), and electric hot plate (GS,
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Zhenghong, China) were used for sample pretreatment and actual sample detection.
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2.3. Synthesis of CdTe QDs

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CdTe QDs were prepared following previously reported procedures with slight modifications
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(Hu et al., 2014; Wang et al., 2018). In brief, tellurium powder (0.5 mmol) was mixed with NaBH4

(1.5 mmol) in 4 mL of deionized water under violent stirring, and mixture was deaerated with N2 in

an ice-cold for 6 h to obtain NaHTe. Then, 1 mmol·L-1 CdCl22.5H2O and 2.4 mmol·L-1 TGA were

dissolved in 50 mL of deionized water. After the solution became clear and transparent, 0.1 mol·L-1

NaOH was used to adjust pH of the solution to 10.5, and the solution was deoxygenated with N2 for

25 min. Then, 2 mL of NaHTe was rapidly injected into this solution under vigorous stirring and

refluxing for 10 h to obtain the orange CdTe QDs solution. CdTe QDs were centrifuged at 8000
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 rpm for 25  min and washed two times with ethanol and double-distilled water, respectively. Next,

the mixture was purified with a dialysis membrane (2000 Da) for 48 h to remove unreacted

precursors. Finally, the solution was diluted to 50 mL with double-distilled water and preserved at

4 °C until further use.

2.4. Preparation of CuNCs@SiO2

CuNCs were synthesized in the aqueous phase following a previously described method with

some modification (Momeni et al., 2017). Typically, CuSO4 (19 mL, 6.5 mmol·L-1) and NaOH (1

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mL, 0.5 mol·L-1) were mixed at room temperature. Next, ascorbic acid (20 mL, 0.1 mol·L-1) was

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added to the mixture. The mixture was stirred and mixed for 10 min at 50 °C, and the color of the

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mixture changed from yellow to dark pink. Subsequently, NaOH solution (0.5 mol·L-1) was added
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drop-wise until the color of the mixture gradually changed from dark pink to pale yellow, and the
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corresponding pH was in the range of 8-9. The mixture was heated at 50 °C for 15 h to obtain

CuNCs. Then, CuNCs were centrifugated at 8000 rpm for 10 min and dialyzed using a dialysis
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membrane (1000 Da) for approximately 24 h. Finally, the solution was diluted to 50 mL with
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double-distilled water and preserved at 4 °C until further use.

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CuNCs@SiO2 were synthesized using the reverse microemulsion method (Gai et al., 2012; Lv
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et al., 2018). Purified CuNCs (5 mL) were mixed with TritonX-100 (0.9  mL), 1-hexyl alcohol (1.0

mL), and cyclohexane (3.5 mL), and the suspension was stirred for 20 min in a flask. After the

addition of TEOS (5 mL), the reaction was allowed to proceed under vigorous stirring for 48 h for

silica growth at room temperature. As-synthesized CuNCs@SiO2 were precipitated by the addition

of 10 mL of ethanol and were centrifuged at 8000  rpm for 25  min. CuNCs@SiO2 were further

purified through repeated centrifugation cycles twice in double-distilled water and ethanol,

respectively. Finally, aqueous dispersions of CuNCs@SiO2 were obtained through sonication for 10
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min and diluted into 100 mL with double-distilled water and preserved at 4 °C until further use.

2.5 Preparation of the ratiometric probe and the application for Cd2+ detection

In this study, the ratiometric probe consisting of CdTe QDs, CuNCs@SiO2, and Phen was

developed for Cd2+ detection. The probe was synthesized through two steps: preparation of CdTe

QDs-Phen (the CdTe QDs probe) and fabrication of the CuNCs@SiO2- CdTe QDs probe (the

ratiometric probe). As shown in Scheme 1, CdTe QDs (10 mL, 50 mmol·L-1) and Phen (10 mL, 550

mmol·L-1) were mixed to prepare the CdTe QDs probe, and the ratiometric probe was prepared by

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mixing the CdTe QDs probe (10 mL, 50 mmol·L-1) and CuNCs@SiO2 (15 mL). For Cd2+ detection,

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0.5 mL of the ratiometric probe (27 mmol·L-1) or the CdTe QDs probe (27 mmol·L-1) and 1 mL of

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the Cd2+ standard solution at different concentrations were mixed at 25 °C for 6 min. Fluorescence
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signals were measured and recorded using a fluorescence spectrophotometer (λex=365 nm). The
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fluorescence intensity ratio (I580/I436) between CdTe QDs (I580) and CdTe QDs (I436) was calculated

for quantitative analysis of the Cd2+ content. Moreover, for colorimetric aaslysis, the mixture was
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transferred to a 1.5 mL centrifuge tube and placed in a dark box, and under UV illumination
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(λex=365 nm), the fluorescence color image was captured by the camera of the smartphone.
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2.6 Cd2+ detection in oyster samples

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Oyster samples were bought from Changshen supermarket (Zhenjiang, Jiangsu Province,

China). Oyster samples removed viscus and cleaned with physiological saline, and dried to a

constant weight at 55 °C. The samples were ground into a fine powder by using an ultra-micro

pulverizer and were stored at 4 °C. Then, the oyster samples underwent pretreatment with the

microwave digestion method. The specific procedure was shown in Supplementary Material. For

Cd2+ detection in the oyster samples, 0.5 mL of the ratiometric probe and 1 mL of digestion solution

samples were mixed at 25 °C for 6 min. Fluorescence signals were measured and recorded using the
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fluorescence spectrophotometer. The ICP-MS method was employed as the standard method, and

the optimal instrumental parameters of ICP-MS are presented in Table S2 (Li et al., 2020).

3. Results and discussion

3.1. Working principle of the ratiometric probe

The preparation of the ratiometric probe involved two steps (Scheme 1). The CdTe QDs probe

was prepared by mixing purified CdTe QDs and Phen. Then, obtained CuNCs@SiO2 were mixed

with the CdTe QDs probe to obtain the ratiometric probe. In this system, CuNCs@SiO2 provided

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the reference signal, and the CdTe QDs probe provided the response signal. Scheme 1 describe the

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principle of the developed fluorescence sensing platform for Cd2+ detection. First, the strong affinity

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between the two nitrogen atoms of Phen and the cadmium atoms of CdTe QDs lead to the formation
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of low-fluorescent CdTe QDs-Phen complexes (the CdTe QDs probe) (Hu et al., 2014). Previous
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research has shown the number of Phen attached on the surface of CdTe QDs depends on the size of

CdTe QDs (Zhang et al., 2013). Firstly, the size of CdTe QDs with the diameter of 3.2 nm were
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calculated according to the method of earlier study (Yu et al., 2003). The specific calculation
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process was shown in Supplementary Material. And the calculation results demonstrated that, one
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CdTe QDs could at most absorb 34 molecules of Phen for the developed probe. This value was in
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agreement with an estimated required space of 1 nm2 for one upright ligand on CdTe QDs surface

(Volker et al., 2010). As shown in Fig. S1, during the attachment of Phen, the shape and peak

position of the quenched fluorescence of CdTe QDs remains unaltered. Besides, the UV/Vis

absorption of CdTe QD+Phen did not shift compared to CdTe QDs (Fig. S3). These observations

indicated that the quenching mechanism of Phen toward the fluorescence of CdTe QDs was not by

“surface chemistry” interaction (Chen et al., 2013; Gui et al., 2013). To investigate the charge

transfer process for fluorescence quenching of CdTe QDs, the energetic positions of valence band
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(VB) and conduction band (CB) of CdTe QDs with respect to Phen were determined based on

electrochemical redox of the isolated component (Zhang et al., 2013; Ye et al., 2020; Zhang et al.,

2011). Typical CV measurements were conducted for estimating the energy levels of CdTe QDs and

Phen, and the specific procedure is shown in section 2.8 (Supplementary Material) (Hu et al., 2014;

Zhang et al., 2013). The onset potentials of oxidation for CdTe QDs and Phen are 0.62 and 0.45 eV,

respectively, indicating that the valence band of CdTe QDs is ∼0.2 eV lower than the highest

occupied molecular orbital of Phen (Fig. S4). Therefore, the electronic reaction occurred between

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CdTe QDs and Phen is a photo-induced hole transfer (PHT) process: upon excitation, the

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photogenerated holes on the CdTe QDs preferentially transfered to the Phen ligands and become

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trapped, preventing the effective hole/electron recombination (the inset of Fig. S4) (Benvidi et al.,
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2010; Qian et al., 2017). Furthermore, the fluorescence lifetimes of CdTe QDs, CdTe QDs+Phen,
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and CdTe QDs+Phen+Cd2+ were studied. As shown in Fig. S5, the negligible changes in the average

lifetimes of CdTe QDs in the absence and presence of Phen were observed by fitting with a
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double-exponential component, indicative of a static fluorescence quenching process (Chen et al.,

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2013). When Cd2+ was presented, Phen had higher affinity to Cd2+ than to Cd atoms of CdTe QDs;
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Phen detached from the CdTe QDs-Phen complexes and formed [Cd(Phen)2(H2O)2]2+ and CdTe
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QDs (Benvidi et al., 2010; Hu et al., 2014; Qian et al., 2017). Moreover, SiO2 prevented CuNCs

from reacting with other substances; therefore, CuNCs@SiO2 provided steady and creditable

reference signals. Moreover, the ratiometric probe showed well-separated characteristic

fluorescence emission peaks at 440 and 580 nm, which were assigned to CuNCs@SiO2 and the

CdTe QDs probe, respectively. At different concentrations of Cd2+, the ratiometric probe showed

various fluorescent colors due to the fluorescence intensity of dual-emission fluorophore changes.

Based on the aforementioned principles, the ratiometric and colorimetric sensing platform could
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realize the detection the content of Cd2+.

Scheme 1. Schematic of the ratiometric probe for Cd2+ detection.

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3.2. Characterization of CdTe QDs, CuNCs, CuNCs@SiO2, and the ratiometric

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probe

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The morphology and size of CdTe QDs, CuNCs, and CuNCs@SiO2 were characterized

through TEM. As shown in Fig. 1A, the average diameter of CdTe QDs was approximately 3 nm,
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with near-spherical morphology. The high-resolution transmission electron microscopy (HRTEM)
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image revealed individual islands in the crystal lattice with an interplanar spacing of 0.316 nm (Fig.
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1B), the finding of the (101) lattice plane of CdTe QDs conformed to the result of a previous study
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(Pan et al., 2019). As shown in Fig. 1C, CuNCs exhibited favorable dispersion, with an average
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diameter of approximately 2 nm. After functionalization of CuNCs with SiO2, CuNCs@SiO2 were
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found to be coated with silica (the mean diameter was approximately 10 nm), as indicated in Fig.

1D. This observation confirmed the existence of SiO2 on the outer surface of CuNCs. Then,

compared the ratiometric probe before and after reaction with Cd2+, the morphology and crystal

lattice of CdTe QDs were almost unchanged (Fig. S6 & S7). As shown in Fig. S7 & S8, CdTe QDs

still remained monodisperse in the CdTe QDs probe and the ratiometric probe. These results were

consistent with previous studies (Hu et al., 2014), indicating that the ratiometric probes were

successfully prepared.

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 To further confirm the observation, FT-IR spectra were used to characterize the functional

groups of CuNCs, CuNCs@SiO2, CdTe QDs, and CdTe QDs-Phen (Fig. 1E). Compared with the

FT-IR spectrum of CuNCs, the spectrum of CuNCs@SiO2 revealed absorption peaks of stretching

vibrations of Si-O at 1100  cm-1, suggesting that CuNCs were encapsulated by SiO2 (Huang et al.,

2018). For CdTe QDs-Phen and CdTe QDs, the FTIR spectrum of CdTe QDs-Phen showed a band

at 729 cm-1 and 1517 cm-1, which was assigned to aromatic rings, indicating that Phen was coupled

to CdTe QDs (Hu et al., 2014). The above results showed that, CuNCs@SiO2 and the CdTe QD

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probe were successfully synthesized

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The optical properties of CuNCs, CuNCs@SiO2, and CdTe QDs are shown in Fig. 1F. The

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maximum fluorescence emission of CuNCs was detected at 436 nm (λex=365 nm), and CuNCs
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emitted intense blue fluorescence under a 365 nm UV lamp. The fluorescence peak position and
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fluorescence color of CuNCs@SiO2 were almost invariant compared with CuNCs; this finding

indicated that the silica coating did not change the fluorescence properties of CuNCs. CdTe QDs
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showed the maximum fluorescence emission at 580 nm (λex=365 nm), and the corresponding
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solution displayed yellow fluorescence.

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Fig. 1. (A) TEM image of CdTe QDs; (B) HRTEM image of CdTe QDs, (C) TEM image of CuNCs, (D)
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TEM image of CuNCs@SiO2; (E) FT-IR spectra of CuNCs, CuNCs@SiO2, CdTe QDs-Phen, and CdTe QDs; (F)
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the fluorescence emission spectra of CuNCs, CdTe QDs, and CuNCs@SiO2 (ex=365nm).

3.3. Optimization of experimental conditions

The fluorescence properties of the ratiometric probe were studied. To identify the optimum

ratio of CuNCs@SiO2 and the CdTe QDs probe in the ratiometric probe, the fluorescence intensity

ratio (I580/I436) and the fluorescence color were studied by determining the fluorescence signals of

the system before and after Cd2+ addition. As shown in Fig. 2A and 2B, without Cd2+ addition, the

I580/I436 value decreased as the proportion of CuNCs@SiO2 increased. The corresponding

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fluorescence color changed from yellow to blue, indicating that the fluorescence color of the

composite system could be tuned by changing the relative intensity of fluorescent peaks. After Cd2+

addition, the I580/I436 value increased as the Cd2+ concentration increased; this is because the

fluorescence intensity of the CdTe QDs probe (response signal) increased gradually, and the

fluorescence intensity of CuNCs@SiO2 (built-in reference signal) remained almost constant. Fig.

2B presents the fluorescence color of the ratiometric probe under different ratio of CuNCs@SiO2

and the CdTe QDs probe and different Cd2+ concentrations. The fluorescent color of a composite

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system mainly depends on the fluorescence properties of fluorescent chromophores (He et al., 2018).

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Hence, in this study, the ratio of CuNCs@SiO2 and the CdTe QDs probe played an important role in

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the fluorescence color of the composite system. When the volume ratio of the CdTe QDs probe to
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CuNCs@SiO2 was 2:3, the chromatic aberration of fluorescent colors could easily be identified by
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the eyes at different Cd2+ concentrations. Therefore, the volume ratio of the CdTe QDs probe to

CuNCs@SiO2 was 2:3 in follow-up experiments. Then, to obtain optimal experimental results, the
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pH and reaction time were optimized. The specific procedure is shown in Supplementary Material.
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As shown in Fig. 2C and 2D, the optimal pH and reaction time were 7.5 and 6 min, respectively.
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Fig. 2. (A) Fluorescence intensity ratio (I580/I436) of the CdTe QDs probe and CuNCs@SiO2 under different ratio

(2:1, 2:2, 2:3, 2:4, 2:5, and 2:6) and different Cd2+ concentrations (0, 0.5, 1.0, 1.5, and 2.0 mg·L-1); (B) the

corresponding photographic fluorescence images; the effects of pH (C) and reaction time (D) on the performance

of the ratiometric probe.

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3.4. Ratiometric detection of Cd2+

To evaluate the analytical performance of the ratiometric probe, its sensitivity was studied and

compared with that of the the CdTe QDs probe. Under optimal conditions, the CdTe QDs probe was

incubated with different concentrations of Cd2+ at room temperature for 6 min. As shown in Fig. 3A,

with an increase in the Cd2+ concentration, the fluorescence intensity at 580 nm gradually increased,

and the intensity of monochromatic yellow fluorescence became stronger. Upon exposure to Cd2+,

coordination between Phen and Cd2+ terminated PHT between CdTe and Phen, resulting in the

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fluorescence recovery of CdTe QDs (Hu et al., 2014; Qian et al., 2017). As shown in Fig. 3B, the

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fluorescence restoration was proportional to the Cd2+ concentration, exhibiting two consecutive

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linear relationships: one is in the range of 0.020 mg·L-1˗0.40 mg·L-1 with a good correlation
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coefficient of 0.9901, and the other linear regression is in the range of 0.40 mg·L-1˗1.8 mg·L-1
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(R2=0.9892). The limit of detection (LOD) was calculated to be 1.8 μg·L-1 by using the following

formula: 3σ/k (where σ denotes the standard deviation obtained by conducting 11 repeated
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detections of a blank solution and k is the slope of the calibration curve) (He et al., 2018).
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For the ratiometric probe, as the concentration of Cd2+ increased, the fluorescence intensity
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gradually recovered at 580 nm and almost unchanged at 436 nm. Moreover, the fluorescence color
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gradually changed from blue to red as the Cd2+ concentration increased. As stated earlier, because of

the competitive binding of Cd2+ to Phen, Phen detached from CdTe QDs, leading to the

fluorescence recovery of CdTe QDs. Simultaneously, due to the protection provided by SiO2, the

fluorescence intensity of CuNCs@SiO2 remained unchanged. As shown in Fig. 3D, the I580/I436

value linearly increased as the Cd2+ concentration increased, which could be expressed by two

consecutive linear relationships: Y=1.2601X+0.3694 (R2=0.9764) and Y=0.6715X+0.6277

(R2=0.9962), in the range of 0.010 mg·L-1˗0.40 mg·L-1 and 0.40 mg·L-1˗2.0 mg·L-1, respectively.
14
The limit of detection (LOD) was calculated to be 1.1 μg·L-1 by using the following formula: 3σ/k.

More importantly, as shown in the insert of Fig. 3C, as the Cd2+ concentration increased, the

fluorescence color change could be distinguished by the naked eyes; thus, this method provides a

convenient visual analysis of Cd2+. This was mainly because the fluorescence intensity of the dual

fluorescent chromophore transformed (Li et al., 2019b). Compared with the CdTe QDs probe, the

ratiometric probe has a lower detection limit, a wider detection range, and could easily identify

fluorescence color changes. As shown in Table S3, the performance parameters of the ratiometric

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probe proposed in this study are comparable or more favorable than those of reported probes.

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Moreover, using CuNCs@SiO2 as reference signal greatly improves the detection stability and

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reduces the detection cost of the ratiometric fluorescence probe. Besides, the fluorescence color
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gradually changed from blue to yellow with the increase of Cd2+ concentration, and the change of
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fluorescence color could be distinguished by the naked eye, verifying the feasibility of colorimetric

analysis. The above results revealed the promising potential of the ratiometric probe for Cd2+
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detection.
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Fig. 3. (A) Fluorescence spectrum of the CdTe QDs probe at different concentrations of Cd2+ (0.020˗1.8 mg·L-1);

the inset: the fluorescence images of the system at different concentrations of Cd2+ (ex=365nm). (B) Linear

relationship between the fluorescence intensity of the CdTe QDs probe and the Cd2+ concentration. (C)

Fluorescence spectrum of the ratiometric probe at different concentrations of Cd2+ (0.010˗2.0 mg·L-1); the inset:

the fluorescence images of the system at different concentrations of Cd2+ (ex=365nm). (D) Linear relationship

between I580/I436 of the ratiometric probe and the Cd2+concentration.

15
3.5 Selectivity and stability of the ratiometric probe toward Cd2+

Selectivity is another crucial index to estimate the detection capability of the ratiometric probe.

The I580/I436 value in response to Cd2+ at a concentration of 1.24 mg·L-1 and to possible interfering

analytes (Ca2+, Mg2+, Al3+, Hg2+, Ni2+, Fe3+, Mn2+, K+, Na+, Zn2+ Cu2+, and Fe2+) were determined

under the same conditions. As shown in Fig. 4A, the I580/I436 value of the ratiometric probe

significantly changed in the presence of Cd2+. However, a relatively slight change of in the I580/I436

value was observed in the presence of interfering analytes (310 mg·L-1 for Na+ and K+, 31 mg·L-1

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for Mg2+ and Ca2+, 6.2 mg·L-1 for Zn2+, and 3.1 mg·L-1 for other metal ions), even the

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concentrations of the interference metal ions are 300 times or 10 times higher than that of Cd2+. In

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order to discuss the specificity of the ratiometric probe to Cd2+ in more detail, another interference
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experiment was conducted when other interference coexisted with Cd2+. The specific results are
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shown in Fig. 4A and Fig. 4B. Clearly, the fluorescence response of the ratiometric probe toward

Cd2+ was scarcely interfered by the presence of the coexisting interference ions. The changed in the
a

I580/I436 value was mainly due to the coordination between Phen and Cd2+ for terminating PHT
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between CdTe and Phen, resulting in the fluorescence recovery of CdTe QDs (Benvidi et al., 2010).
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In summary, the ratiometric probe has favorable selectivity and specificity for Cd2+.
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The detection stability of the dual-emission ratiometric probes and the single-emission CdTe

QDs probe was investigated at different measurement temperatures. This investigation was

conducted following the specific operation in our previous research(Li et al., 2020), which is

provided in Supplementary Material. As shown in Fig. 4C, the deviation rate of the ratiometric

probes and the CdTe QDs probe was 0.25 %˗0.79 % and 0.35 %˗1.9 % at different temperatures,

respectively. Compared with the single-emission CdTe QDs probe, the deviation rate of the

ratiometric probe was reduced by 0.10 %˗1.1 %. The results showed that the dual-emission
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ratiometric probe can improve the detection stability of the fluorescence probe.

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Fig. 4. (A) Selectivity of the ratiometric probes for detecting Cd2+ (1.24 mg·L-1) and interfering substances (N = 6)
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Inset is the corresponding fluorescence photograph of the sensor solutions under 365 nm UV lamp. (B)
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Fluorescence response of the ratiometric probes to different metal ions without Cd2+ and with Cd2+ (1.24 mg·L-1)

(N = 6). Inset is the corresponding fluorescence photograph of the sensor solutions under 365 nm UV lamp. (C)
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Deviation rate of the detection results of the ratiometric probes and the CdTe QDs probe at different measurement
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temperatures (N=6). (D) The comparison of fluorescence sensing of Cd2+ in standard solution and Cd2+ in the
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matrix of oyster samples. The content of Cd2+ in oyster samples were measured by ICP-MS and the Cd2+ of
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standard solution were artificially added to the corresponding concentration.

3.6. Applicability in oyster samples

In this study, the performance of the fluorescence probe was estimated by comparing with the

ICP-MS method. Through sensitivity experiment, the LOD of the fluorescence method was 2.75

μg·kg-1 for oyster samples (1 mg·kg-1=0.4 mg·L-1). As shown in Table 1, the contents of Cd2+ in

oyster samples were in the range of 0.545˗3.27 mg·kg-1 and 0.551˗3.21 mg·kg-1, as revealed by the

proposed fluorescence method and the ICP-MS method, respectively. The relative standard
17
deviations (RSD, N=6) of the fluorescence method was less than 4 %. The paired t-test result

revealed no significant difference between the fluorescence and ICP-MS methods. These results

indicated that the ratiometric probe based on CuNCs@SiO2 and the CdTe QDs probe have high

stability and accuracy for detecting Cd2+ in oyster samples. Moreover, compared with the

fluorescence response in standard solution, there is no remarkable change of that for evaluating

Cd2+ in the matrix of oyster samples (Fig. 4D). The results demonstrate that the matrix of oyster

samples has no obvious influence on Cd2+ assay, verifying the high anti-interference ability and

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good selectivity of the sensor for Cd2+ determination in marine product.

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Table 1. Concentration of Cd2+ in oyster samples detected by the fluorescence and ICP-MS methods
Oyster
Samples
Fluorescence method
(mg·kg-1)a
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RSD (%)
(N=6)
ICP-MS method
(mg·kg-1)a
Paired t-test
Pb
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1 0.545 1.87 0.551 0.33
2 0.842 2.24 0.861 0.15
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3 1.36 2.66 1.38 0.25

4 2.13 3.89 2.16 0.28
a

5 3.27 3.62 3.21 0.55

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a
Average of six measurements
b
Paired t-test: P < 0.05, significant difference; P > 0.05, no significant difference
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3.7. Colorimetric analyzer for Cd2+ detection in oyster samples

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To obtain a low cost and rapid determination tool for practical applications, a colorimetric

analyzer was proposed, in which the ratiometric probe was combined with a common smartphone

as a fluorescent color signal reader and substance content analyzer. In this study, oyster samples

were used to evaluate the practicability of the colorimetric analyzer. The specific steps were as

follows (Fig. 5A): first, oyster samples were processed; subsequently, the ratiometric probe was

added to the sample in a centrifuge tube. Then, the treated samples were placed under the UV lamp

with 365 nm, and the smartphone camera captured the fluorescent color. Meanwhile, as shown in
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Fig. 5B, the fluorescence signals were measured and recorded using a fluorescence

spectrophotometer. The content of Cd2+ was calculated by bringing in the previous linear

relationship (I580/I436 < 0.9: Y=1.2601X+0.3694; I580/I436 > 0.9: Y=0.6715X+0.6277). Next, the

Color Picker app converted fluorescent color signals into digital values representing the red, green,

and blue (RGB) color mode. Finally, the ratio of green (G) and red (R) channels (G/R) was

calculated for quantitative analysis of Cd2+. As shown in Fig. 5C, a favorable linear relationship was

found between the value of G/R and the content of Cd2+ (0.014˗1.3 mg·L-1)/(0.035˗3.25 mg·kg-1);

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the relationship could be expressed by Y=4.0134X+1.3501 (R2=0.9972). These results indicated

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that the colorimetric analyzer could provide a quantitative determination of Cd2+. Although the

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sensitive detection of Cd2+ has been realized in this work, there also has a problem in the utilization
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of CdTe QDs after the assays. In view of this problem, the research of desorption of Cd2+ and reuse
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of CdTe QDs will be further explored to realize the environmentally friendly detection of Cd2+ in

the future work.

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Moreover, the performance of the colorimetric analyzer was estimated by comparing with the
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ICP-MS method. As shown in Table 2, the contents of Cd2+ in oyster samples measured by the
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proposed colorimetric method and the ICP-MS method were in the range of 0.337˗3.19 mg·kg-1 and
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0.338˗3.21 mg·kg-1, respectively. The relative standard deviations (RSD, N=6) of the colorimetric

method was less than 5 %. The paired t-test result revealed no significant difference between the

colorimetric and ICP-MS methods. These results indicated that the colorimetric analyzer have high

repeatability and reliability for detecting Cd2+ in oyster samples.

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 Table 2. Concentration of Cd2+ in oyster samples detected by the colorimetric and ICP-MS methods
Oyster Colorimetric method RSD (%) ICP-MS method Paired t-test
Samples (mg·kg-1)a (N=6) (mg·kg-1)a Pb
1 0.337 1.62 0.338 0.211
2 0.731 2.89 0.834 0.354
3 1.461 3.52 1.459 0.198
4 2.379 1.94 2.381 0.308
5 3.19 4.58 3.21 0.437
a
Average of six measurements
b
Paired t-test: P < 0.05, significant difference; P > 0.05, no significant difference

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Fig. 5. (A) Schematic of smartphone-based colorimetry for Cd2+ detection in oyster samples. (B) The fluorescence

spectrum of the ratiometric probe and the treated samples with different contents of Cd2+. (C) Linear relationship
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between the ratio of G/R (blue line)/ the ratio of I580/I436 (red line) and the Cd2+ concentrations.

4. Conclusions

A facile and highly sensitive ratiometric probe and colorimetric analyzer based on

CuNCs@SiO2 and the CdTe QDs probe were developed for detecting trace Cd2+ in oyster samples.

The ratiometric probe showed excellent sensitivity to Cd2+ in the range of 0.010˗2.0 mg·L-1, with

LOD of 1.1 μg·L-1. No significant difference was observed between the fluorescence method and

the ICP-MS method, and RSD of the fluorescence method was < 4 %. Moreover, the colorimetric

20
reader and analyzer were proposed combined with the change in fluorescence color to rapid test the

Cd2+. The respond value displayed favorable linearity to the Cd2+ concentration in the range of

0.014˗1.3 mg·L-1 (R2=0.9972). The fluorescence method was successfully applied to detect Cd2+ in

oyster samples. This study proved the excellent selectivity and stability of the ratiometric and

colorimetric fluorescent sensing platform, which has promising potential for rapid monitoring in

practical applications.

Acknowledgements: This manuscript was edited by Wallace Academic Editing.

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Funding: This work was supported by the National Key Research and Development Program of

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China [grant numbers 2018YFD0400800]; the National Natural Science Foundation of China [grant

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numbers 31772073, 31671844]; the Natural Science Foundation of Jiangsu Province [grant
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numbers BE2019359]; Project of Faculty of Agricultural Equipment of Jiangsu University [grant
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numbers NZXB20200212]; and Priority Academic Program Development of Jiangsu Higher

Education Institutions (PAPD).

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Reference
rn

Adam, I. S. I., Anthemidis, A. N., 2009. Flow injection wetting-film extraction system for flame atomic absorption
spectrometric determination of cadmium in environmental waters, Talanta, 77 1160-1164.
u

https://doi.org/10.1016/j.talanta.2008.08.015
Jo

Bai, H., Tu, Z., Liu, Y., Tai, Q., Guo, Z., Liu, S., 2020. Dual-emission carbon dots-stabilized copper nanoclusters for
ratiometric and visual detection of Cr2O72- ions and Cd2+ ions, Journal of Hazardous Materials, 386 121654.
https://doi.org/10.1016/j.jhazmat.2019.121654
Benvidi, A., Lanjwani, S. N., Ding, Z., 2010. Cd2+ transfer across water/1,2-dichloroethane microinterfaces facilitated
by complex formation with 1,10-Phenanthroline, Electrochimica Acta, 55 2196-2200.
https://doi.org/10.1016/j.electacta.2009.11.056
Cao, Y., Deng, B., Yan, L., Huang, H., 2017. An environmentally-friendly, highly efficient, gas pressure-assisted sample
introduction system for ICP-MS and its application to detection of cadmium and lead in human plasma, Talanta,
167 520-525.
https://doi.org/10.1016/j.talanta.2017.02.057
Chan, C. Y., Wang, W.-X., 2019. Biomarker responses in oysters Crassostrea hongkongensis in relation to metal
contamination patterns in the Pearl River Estuary, southern China, Environmental Pollution, 251 264-276.
https://doi.org/10.1016/j.envpol.2019.04.140
Chen, S., Tian, J., Jiang, Y., Zhao, Y., Zhang, J., Zhao, S., 2013. A one-step selective fluorescence turn-on detection of

21
 cysteine and homocysteine based on a facile CdTe/CdS quantum dots–phenanthroline system, Analytica Chimica
Acta, 787 181-188.
https://doi.org/10.1016/j.aca.2013.05.048
Deng, S., Yu, J., Wang, Y., Xie, S., Ran, Z., Wei, W., 2019. Distribution, transfer, and time-dependent variation of Cd in
soil-rice system: A case study in the Chengdu plain, Southwest China, Soil and Tillage Research, 195 104367.
https://doi.org/10.1016/j.still.2019.104367
Gai, F., Zhou, T., Zhang, L., Li, X., Hou, W., Yang, X., Li, Y., Zhao, X., Xu, D., Liu, Y., Huo, Q., 2012. Silica
cross-linked nanoparticles encapsulating fluorescent conjugated dyes for energy transfer-based white light
emission and porphyrin sensing, Nanoscale, 4 6041.
https://doi.org/10.1039/c2nr31194a
Ghanei-Motlagh, M., Taher, M. A., 2017. Novel imprinted polymeric nanoparticles prepared by sol–gel technique for
electrochemical detection of toxic cadmium(II) ions, Chemical Engineering Journal, 327 135-141.
https://doi.org/10.1016/j.cej.2017.06.091

of
Gui, R., An, X., Huang, W., 2013. An improved method for ratiometric fluorescence detection of pH and Cd2+ using
fluorescein isothiocyanate–quantum dots conjugates, Analytica Chimica Acta, 767 134-140.

ro
https://doi.org/10.1016/j.aca.2013.01.006
He, W., Gui, R., Jin, H., Wang, B., Bu, X., Fu, Y., 2018. Ratiometric fluorescence and visual imaging detection of
dopamine based on carbon dots/copper nanoclusters dual-emitting nanohybrids, Talanta, 178 109-115.
https://doi.org/10.1016/j.talanta.2017.09.019 -p
Hou, L., Qin, Y., Li, J., Qin, S., Huang, Y., Lin, T., Guo, L., Ye, F., Zhao, S., 2019. A ratiometric multicolor fluorescence
re
biosensor for visual detection of alkaline phosphatase activity via a smartphone, Biosensors and Bioelectronics,
143 111605.
https://doi.org/10.1016/j.bios.2019.111605
lP

Hu, X., Shi, J., Shi, Y., Li, W., Arslan, M., Zhang, W., Huang, X., Li, Z., Xu, Y., Li, Y., 2019a. A ratiometric
fluorescence sensor for ultra-sensitive detection of trypsin inhibitor in soybean flour using gold
nanocluster@carbon nitride quantum dots, Analytical and Bioanalytical Chemistry, 411 3341-3351.
a

https://doi.org/10.1007/s00216-019-01806-1.
rn

Hu, X., Shi, J., Shi, Y., Zou, X., Tahir, H. E., Holmes, M., Zhang, W., Huang, X., Li, Z., Xu, Y., 2019b. A dual-mode
sensor for colorimetric and fluorescent detection of nitrite in hams based on carbon dots-neutral red system, Meat
Science, 147 127-134.
u

https://doi.org/10.1016/j.meatsci.2018.09.006
Jo

Hu, X., Wang, W., Huang, Y., 2016. Copper nanocluster-based fluorescent probe for sensitive and selective detection of
Hg(2+) in water and food stuff, Talanta, 154 409-415.
https://doi.org/10.1016/j.talanta.2016.03.095
Hu, X., Zhu, K., Guo, Q., Liu, Y., Ye, M., Sun, Q., 2014. Ligand displacement-induced fluorescence switch of quantum
dots for ultrasensitive detection of cadmium ions, Analytica Chimica Acta, 812 191-198.
https://doi.org/10.1016/j.aca.2014.01.006
Huang, L., Mu, Y., Chen, J., Tian, J., Huang, Q., Huang, H., Deng, F., Wen, Y., Zhang, X., Wei, Y., 2018. One-pot
ultrafast preparation of silica quantum dots and their utilization for fabrication of luminescent mesoporous silica
nanoparticles, Materials Science and Engineering: C, 93 679-685.
https://doi.org/10.1016/j.msec.2018.08.035
López_Marzo, A. M., Pons, J., Blake, D. A., Merkoçi, A., 2013. High sensitive gold-nanoparticle based lateral flow
Immunodevice for Cd2+ detection in drinking waters, Biosensors and Bioelectronics, 47 190-198.
https://doi.org/10.1016/j.bios.2013.02.031
Lee, G., Suonan, Z., Kim, S. H., Hwang, D.-W., Lee, K.-S., 2019. Heavy metal accumulation and phytoremediation

22
 potential by transplants of the seagrass Zostera marina in the polluted bay systems, Marine Pollution Bulletin, 149
110509.
https://doi.org/10.1016/j.marpolbul.2019.110509
Li, F., Hu, Y., Li, Z., Liu, J., Guo, L., He, J., 2019a. Three-dimensional microfluidic paper-based device for multiplexed
colorimetric detection of six metal ions combined with use of a smartphone, Analytical and Bioanalytical
Chemistry, 411 6497-6508.
https://doi.org/10.1007/s00216-019-02032-5
Li, W., Hu, X., Li, Q., Shi, Y., Zhai, X., Xu, Y., Li, Z., Huang, X., Wang, X., Shi, J., Zou, X., Kang, S., 2020. Copper
nanoclusters @ nitrogen-doped carbon quantum dots-based ratiometric fluorescence probe for lead (II) ions
detection in porphyra, Food Chemistry, 320 126623.
https://doi.org/10.1016/j.foodchem.2020.126623
Li, W., Shi, Y., Hu, X., Li, Z., Huang, X., Holmes, M., Gong, Y., Shi, J., Zou, X., 2019b. Visual detection of nitrite in
sausage based on a ratiometric fluorescent system, Food Control, 106 106704.

of
https://doi.org/10.1016/j.foodcont.2019.06.030
Li, Y., Xia, T., Zhang, J., Cui, Y., Li, B., Yang, Y., Qian, G., 2019c. A manganese-based metal-organic framework

ro
electrochemical sensor for highly sensitive cadmium ions detection, Journal of Solid State Chemistry, 275 38-42.
https://doi.org/10.1016/j.jssc.2019.03.051
Lv, Y., Li, J., Wu, R., Wang, G., Wu, M., Shen, H., Li, L. S., 2018. Silica-encapsulated quantum dots for highly efficient
-p
and stable fluorescence immunoassay of C-reactive protein, Biochemical Engineering Journal, 137 344-351.
https://doi.org/10.1016/j.bej.2018.06.016
re
Mahalik, M. P., Hitner, H. W., Prozialeck, W. C., 1995. Teratogenic effects and distribution of cadmium (Cd2+)
administered via osmotic minipumps to gravid CF-1 mice, Toxicology Letters, 76 195-202.
https://doi.org/10.1016/0378-4274(95)80003-V
lP

Maruthupandi, M., Thiruppathi, D., Vasimalai, N., 2020. One minute synthesis of green fluorescent copper nanocluster:
The preparation of smartphone aided paper-based kit for on-site monitoring of nanomolar level mercury and
sulfide ions in environmental samples, Journal of Hazardous Materials, 392 122294.
a

https://doi.org/10.1016/j.jhazmat.2020.122294
rn

Momeni, S., Ahmadi, R., Safavi, A., Nabipour, I., 2017. Blue-emitting copper nanoparticles as a fluorescent probe for
detection of cyanide ions, Talanta, 175 514-521.
https://doi.org/10.1016/j.talanta.2017.07.056
u

Noh, Y., Jo, E.-J., Mun, H., Ahn, Y.-d., Kim, M.-G., 2017. Homogeneous and selective detection of cadmium ions by
Jo

forming fluorescent cadmium-protein nanoclusters, Chemosphere, 174 524-530.

https://doi.org/10.1016/j.chemosphere.2017.02.025
Pan, D., Chen, K., Zhou, Q., Zhao, J., Xue, H., Zhang, Y., Shen, Y., 2019. Engineering of CdTe/SiO2 nanocomposites:
Enhanced signal amplification and biocompatibility for electrochemiluminescent immunoassay of
alpha-fetoprotein, Biosensors and Bioelectronics, 131 178-184.
https://doi.org/10.1016/j.bios.2019.02.022
Qian, J., Wang, K., Wang, C., Ren, C., Liu, Q., Hao, N., Wang, K., 2017. Ratiometric fluorescence nanosensor for
selective and visual detection of cadmium ions using quencher displacement-induced fluorescence recovery of
CdTe quantum dots-based hybrid probe, Sensors and Actuators B: Chemical, 241 1153-1160.
https://doi.org/10.1016/j.snb.2016.10.020
Sun, L., Wang, T., Sun, Y., Li, Z., Song, H., Zhang, B., Zhou, G., Zhou, H., Hu, J., 2020. Fluorescence resonance energy
transfer between NH2–NaYF4:Yb,Er/NaYF4@SiO2 upconversion nanoparticles and gold nanoparticles for the
detection of glutathione and cadmium ions, Talanta, 207 120294.
https://doi.org/10.1016/j.talanta.2019.120294

23
Sun, Y., Wei, M., Liu, R., Wang, H., Li, H., Kang, Q., Shen, D., 2019. A smartphone-based ratiometric fluorescent
device for field analysis of soluble copper in river water using carbon quantum dots as luminophore, Talanta, 194
452-460.
https://doi.org/10.1016/j.talanta.2018.10.019
Volker, J., Zhou, X., Ma, X., Flessau, S., Lin, H., Schmittel, M., Mews, A., 2010. Semiconductor nanocrystals with
adjustable hole acceptors: tuning the fluorescence intensity by metal-ion binding, Angewandte Chemie.
International Ed. in English, 49 6865-6868.
https://doi.org/10.1002/anie.201001441
Waalkes, M. P., 2000. Cadmium carcinogenesis in review, Journal of Inorganic Biochemistry, 79 241-244.
https://doi.org/10.1016/S0162-0134(00)00009-X
Wang, H., Da, L., Yang, L., Chu, S., Yang, F., Yu, S., Jiang, C., 2020a. Colorimetric fluorescent paper strip with
smartphone platform for quantitative detection of cadmium ions in real samples, Journal of Hazardous Materials,
392 122506.

of
https://doi.org/10.1016/j.jhazmat.2020.122506
Wang, H., Yang, L., Chu, S., Liu, B., Zhang, Q., Zou, L., Yu, S., Jiang, C., 2019. Semiquantitative Visual Detection of

ro
Lead Ions with a Smartphone via a Colorimetric Paper-Based Analytical Device, Analytical Chemistry, 91
9292-9299.
https://doi.org/10.1021/acs.analchem.9b02297
-p
Wang, P., Wu, J., 2019. Highly selective and sensitive detection of Zn(II) and Cu(II) ions using a novel peptide
fluorescent probe by two different mechanisms and its application in live cell imaging, Spectrochimica Acta Part
re
a-Molecular And Biomolecular Spectroscopy, 208 140-149.
https://doi.org/10.1016/j.saa.2018.09.054
Wang, P., Wu, J., Zhao, C., 2020b. A water-soluble peptide fluorescent chemosensor for detection of cadmium (II) and
lP

copper (II) by two different response modes and its application in living LNcap cells, Spectrochimica Acta Part A:
Molecular and Biomolecular Spectroscopy, 226 117600.
https://doi.org/10.1016/j.saa.2019.117600
a

Wang, S.-N., Zhu, J., Li, X., Li, J.-J., Zhao, J.-W., 2018. Fluorescence turn-on sensing of trace cadmium ions based on
rn

EDTA-etched CdTe@CdS quantum dot, Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy,
201 119-127.
https://doi.org/10.1016/j.saa.2018.04.065
u

Wang, T., Mei, Q., Tao, Z., Wu, H., Zhao, M., Wang, S., Liu, Y., 2020c. A smartphone-integrated ratiometric
Jo

fluorescence sensing platform for visual and quantitative point-of-care testing of tetracycline, Biosensors and
Bioelectronics, 148 111791.
https://doi.org/10.1016/j.bios.2019.111791
Wang, X., Yu, S., Wang, J., Yu, J., Arabi, M., Fu, L., Li, B., Li, J., Chen, L., 2020d. Fluorescent nanosensor designing
via hybrid of carbon dots and post-imprinted polymers for the detection of ovalbumin, Talanta, 211 120727.
https://doi.org/10.1016/j.talanta.2020.120727
Wu, X., Wu, P., Gu, M., Xue, J., 2020. Ratiometric fluorescent probe based on AuNCs induced AIE for quantification
and visual sensing of glucose, Analytica Chimica Acta, 1104 140-146.
https://doi.org/10.1016/j.aca.2020.01.004
Yang, J., Li, Z., Jia, Q., 2019. Design of dual-emission fluorescence sensor based on Cu nanoclusters with
solvent-dependent effects: Visual detection of water via a smartphone, Sensors and Actuators B: Chemical, 297
126807.
https://doi.org/10.1016/j.snb.2019.126807
Ye, Y., Wu, T., Jiang, X., Cao, J., Ling, X., Mei, Q., Chen, H., Han, D., Xu, J. J., Shen, Y., 2020. Portable

24
 Smartphone-Based QDs for the Visual Onsite Monitoring of Fluoroquinolone Antibiotics in Actual Food and
Environmental Samples, ACS Applied Materials & Interfaces, 12 14552-14562.
https://doi.org/10.1021/acsami.9b23167
Yu, W. W., Qu, L., Guo, W., Peng, X., 2003. Experimental determination of the extinction coefficient of CdTe, CdSe,
and CdS nanocrystals, Chemistry of Materials, 15 2854-2860.
https://doi.org/10.1021/cm033007z
Zhang, Y, Q, Cao, X, A, 2011. Optical characterization of CdSe quantum dots with metal chalcogenide ligands in
solutions and solids, Applied Physics Letters, 99 023106
https://doi.org/10.1063/1.3610456
Zhang, L., Lin, H., Wang, C., Liu, W.-R., Li, S., Cheng, Y., Xu, J., Gao, H., Li, K., Copner, N., Chen, X., Wang, Y.,
2020a. A Solid-State Colorimetric Fluorescent Pb2+-Sensing Scheme: Mechanically-Driven CsPbBr3
Nanocrystallization in Glass, Nanoscale, 12
https://doi.org/10.1039/D0NR01818J

of
Zhang, L., Zhu, K., Ding, T., Hu, X., Sun, Q., Xu, C., 2013. Quantum dot-phenanthroline dyads: detection of
double-stranded DNA using a photoinduced hole transfer mechanism, Analyst, 138 887-893.

ro
https://doi.org/10.1039/c2an36303h
Zhang, W., Liu, C., Han, K., Wei, X., Xu, Y., Zou, X., Zhang, H., Chen, Z., 2020b. A signal on-off ratiometric
electrochemical sensor coupled with a molecular imprinted polymer for selective and stable determination of
imidacloprid, Biosensors and Bioelectronics, 154 112091.
https://doi.org/10.1016/j.bios.2020.112091
-p
re
Zhang, W., Liu, C., Liu, F. H., Zou, X. B., Xu, Y. W., Xu, X. C., 2020c. A smart-phone-based electrochemical platform
with programmable solid-state-microwave flow digestion for determination of heavy metals in liquid food, Food
Chemistry, 303 7.
lP

https://doi.org/10.1016/j.foodchem.2019.125378
Zhou, J., Li, B., Qi, A., Shi, Y., Qi, J., Xu, H., Chen, L., 2020. ZnSe quantum dot based ion imprinting technology for
fluorescence detecting cadmium and lead ions on a three-dimensional rotary paper-based microfluidic chip,
a

Sensors and Actuators B: Chemical, 305 127462.

rn

https://doi.org/10.1016/j.snb.2019.127462
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Graphical Abstract

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26
CRediT auhor statement

Wenting Li: Conceptualization, Methodology, Writing - original draft.

Xinai Zhang: Formal analysis, Writing - review & editing.

Xuetao Hu: Methodology, Writing - review & editing.

Yongqiang Shi: Investigation, Writing - review & editing.

Zhihua Li: Software, Visualization.

Xiaowei Huang: Formal analysis, Writing - review & editing.

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Di Zhang: Validation, Investigation.

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Wen Zhang: Formal analysis.

Xiaobo Zou: Supervision.

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Jiyong Shi: Supervision, Funding acquisition.
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Declaration of interests

☒ The authors declare that they have no known competing financial interests or personal relationships that
could have appeared to influence the work reported in this paper.

☐The authors declare the following financial interests/personal relationships which may be considered as
potential competing interests:

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Highlights

 The fluorescence sensor with built-in reference probe improved detection

selectivity and accuracy.

 The limit of 1.1 μg·L-1 for Cd2+ detection is qualified for the safety inspection of

Cd2+.

 SiO2-coated CuNCs as reference was designed for further enhancing stability.

 Smartphone-based colorimetry could achieve visual assay of Cd2+ in oyster.

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 The strategy offers a powerful solution for monitoring Cd2+ pollution in marine

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bivalves.

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