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1 Supporting Information Applying arbon Dots-Metal Ions Ensembles as a Multichannel Fluorescent Sensor Array: Detection and Discrimination of Phosphate Anions Shan Sun,, Kai Jiang, Sihua Qian, Yuhui Wang, and Hengwei Lin*, Key Laboratory of Graphene Technologies and Applications of Zhejiang Province, Ningbo Institute of Materials Technology & Engineering (NIMTE), hinese Academy of Sciences (AS), Ningbo 31521, P. R. hina University of hinese Academy of Sciences, eijing, 149, P. R. hina * linhengwei@nimte.ac.cn; Tel: ; Fax: Table of contents Figure S1. S-2 Figure S2..S-2 Figure S3..S-3 Table S1....S-3 Figure S4..S-4 Figure S5..S-4 Figure S6..S-5 Figure S7..S-5 Figure S8..S-6 Figure S9..S-7 Figure S1....S-8 Figure S11....S-9 Figure S S-9 Table S S-1 S 1
2 A.21nm Frequency% Diameter (nm) Absorbance Excitation Emission Wavelength (nm) Normalized FL intensity PL Intensity (a.u.) D 48 nm 5 nm 52 nm 54 nm 56 nm 58 nm 6 nm Wavelength (nm) Figure S1. (A) TEM and () AFM images of the Ds; () UV-Vis absorption, FL excitation and emission spectra of the Ds aqueous dispersion [Inset: photographs of the Ds aqueous dispersion under daylight (left) and the excitation of 54 nm (right)]; (D) emission spectra of the Ds aqueous solution under different excitation wavelengths. Intensity (a.u.) A O 1s N 1s 1s Si 2s Si 2p Intensity (a.u.) 1 4 Measured Fitting = 3 -N -O =O 2 O=-OH inding energy (ev) inding energy (ev) Figure S2. (A) Wide scan XPS spectrum of the Ds, and () high resolution XPS 1s of the Ds and fitting results. S 2
3 1 Transmittance (%) Figure S3. FT-IR spectrum of the Ds Wavenumbers (cm -1 ) Table S1. Fluorescence quenching efficiencies of the as-prepared carbon dots (3 μg/ml) to different metal ions (5 μm) under specific conditions. e 3+ Fe 3+ r 3+ Zn 2+ Fe 2+ u 2+ Hg 2+ Ag + Pb 2+ ph=7. Tris 48% 58% 42% 7% 1% 44% 49% 17% 16% ph=5. HAc-NaAc 22% 64% 3% 5% 24% 19% 89% 16% 22% ph=7.4 HEPES 8% 45% 4% % % 96% 9% 8% 29% S 3
4 [e3+] [ e 3+ ]/μm.8.4 [Fe3+ ] 4 8 [ Fe3+ ]/μm 2 1 μm N-FL FL intensity (a.u.) A N-FL FL intensity (a.u.) 6 1 μm N-FL FL intensity (a.u.) [u2+ ] 2 6 Wavelength (nm) Wavelength (nm) 1 2 [ u2+ ] /μm 2 μm Wavelength (nm) A Normalized FL intensity 1. e3+ (2μM) Fe3+ (5μM) u2+ (1μM) Normalized FL intensity Time (min) Time (min) Time (min) 15 D Normalized FL intensity Normalized FL intensity Figure S4. Fluorescence (FL) titrations of the Ds (3 μg/ml) with e3+ (A), Fe3+ () and u2+ (), respectively. Inset: normalized fluorescence intensities (N-FL) of the Ds at 64 nm with the changes of the three metal ions (all the excitation wavelength at 54 nm) Time (min) 2 Figure S5. (A) Time-dependence of normalized fluorescence intensities of the Ds with the addition of e3+, Fe3+ and u2+ (2, 5 and 1 μm, respectively). (-D) Incubation time effects of the Ds-e3+ (), Ds-Fe3+ () and Ds-u2+ (D) ensembles with the addition of phosphate anions (5 μm), respectively. S 4
5 PPI PI.4.2. N-FL intensity 1. N-FL intensity 1. N-FL intensity A1. PPI PI PPI PI Figure S6. Normalized fluorescence intensities of the Ds in the absence (gray bar) and presence (black bar) of five phosphate anions (2 μm) in ph 7 Tris (A), ph 5 HAc-NaAc (), and ph 7.4 HEPES () buffers, respectively. A E D F Figure S7. TEM images of the Ds with the addition of e3+ (A), Fe3+ (), and u2+ (E), respectively; TEM images of the Ds-e3+ ensemble in the presence of (), Ds-Fe3+ensemble in the presence of (D), and Ds-u2+ ensemble in the presence of (F), respectively. S 5
6 A.3 e 3+ Fe 3+ u 2+ Normalized(F-F ) PPI PI.2.1 Amino acid P2 (22.%). -.1 Metal ions, anions and control P1 (73.5%) Thr Try Gly ys ontrol Na 2 SO 4 Kl a 2+ NaAc Squared Euclidean Distance.96 Figure S8. (A) FL responses of the triple-channel sensor array to five phosphate anions (all at 5 μm), in which F and F represent the array s FL intensities of the Ds-metal ions ensembles in the presence and absence of phosphate anions, respectively; () principal component analysis (PA) plot for the discrimination of the phosphate anions and other interfering substances (anions and metal ions are 5 μm; amino acids are 5 μm) based on the FL signal changes of the sensor array; () hierarchical cluster analysis (HA) plot for the discrimination of the phosphate anions and other interfering substances based on the FL signal changes of the sensor array. S 6
7 A Normalized(F-F ) e 3+ Fe 3+ u PPI PI.2 P 2 (18.9%) Metal ions, anions and control Gly Thr Amino acid Try ys ontrol Kl P 1 (75.%) Na 2 SO 4 NaAc a 2+ ys Thr Try Gly Squared Euclidean 4.8 Figure S9. (A) FL responses of the triple-channel sensor array to five phosphate anions (all at 5 μm), in which F and F represent the array s FL intensities of the Ds-metal ions ensembles in the presence and absence of phosphate anions, respectively; () principal component analysis (PA) plot for the discrimination of the phosphate anions (5 μm) and other interfering substances (5 μm) based on the FL signal changes of the sensor array; () hierarchical cluster analysis (HA) plot for the discrimination of the phosphate anions (5 μm) and other interfering substances (5 μm) based on the FL signal changes of the sensor array. S 7
8 A Normalized(F-F ) Respond to 5 μm add Respond to 1 μm 2 μm e 3+ 5 μm e 3+ e 3+ concentration (μm) Ds + 2 μm e 3+ Ds + 5 μm e 3+ Figure S1. (A) FL responses of the Ds-e 3+ ensembles (prepared by 2 and 5 μm e 3+, respectively) to different concentrations of ; () the photographs of the Ds with the addition of different concentrations of e 3+ and then different concentrations of (a further aggregation or disaggregation process can be easily distinguished from their colors). S 8
9 A).2 control P 2 (21.3%) Serum + Serum Serum + Serum + Serum + Serum P 1 (78.2%) ) ontrol Serum.6.12 Squared Euclidean Distance.36 Figure S11. PA (A) and HA () plots for the discrimination of five phosphate anions (all at 5 μm) in the presence of fetal bovine serum based on the FL signal changes of the as-developed triple-channel sensor array. Normalized (F-F ) e 3+ Fe 3+ u Sample # Figure S12. FL responses of the triple-channel sensor array to 2 blind samples, in which F and F represent the array s FL intensities of the Ds-metal ions ensembles in the presence and absence of phosphate anions, respectively. S 9
10 Table S2. Identification of 2 blind samples. Sample # Identification Verification Sample # Identification Verification lank lank lank lank lank lank 9 lank lank 19 1 lank lank 2 1% identified, % false positives S 1
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