Supporting Information

Transcription

1 Supporting Information Biocatalyst and Colorimetric/Fluorescent Dual Biosensors of H 2 O 2 Constructed via Hemoglobin-Cu 3 (PO 4 ) 2 Organic/Inorganic Hybrid Nanoflowers Jiaojiao Gao, Hui Liu *, Lingyan Pang *, Kai Guo, Junqi Li School of Materials Science and Engineering, Shaanxi University of Science and Technology, Xi an , P. R. China * Corresponding author. Tel: ; Fax: address: liuhui@sust.edu.cn (Hui Liu) * Corresponding author. Tel: ; Fax: address: panglingyan@sust.edu.cn (Lingyan Pang) S-1

2 Figure S1. XRD patterns of Hb-Cu 3 (PO 4 ) 2 HNFs and pure Cu 3 (PO 4 ) 2 3H 2 O nanosheets. Inset: photograph of Hb-Cu 3 (PO 4 ) 2 HNFs (top) and pure Cu 3 (PO 4 ) 2 3H 2 O nanosheets (down) S-2

3 Figure S2. XPS spectra of Cu 2p of pure Cu 3 (PO 4 ) 2 3H 2 O nanosheets. S-3

4 Figure S3. XPS spectra of Fe 2p of Hb powder. S-4

5 Figure S4. The Brunauer-Emmett-teller surface areas images of Hb-Cu 3 (PO 4 ) 2 HNFs. S-5

6 Figure S5. EDX pattern of Hb-Cu 3 (PO 4 ) 2 HNFs. S-6

7 Figure S6. FT-IR spectra of Cu 3 (PO 4 ) 2 3H 2 O, free Hb and the Hb-Cu 3 (PO 4 ) 2 HNFs formed with different time. Experimental conditions: 0.1 mg/ml Hb, 120 mm Cu 2+, 0.1 M PBS at ph=7.4, and incubation different time(1 h, 24 h, 48 h, and 72 h) at 25. S-7

8 Figure S7. UV-vis spectra of the supernatant from the Hb-Cu 3 (PO 4 ) 2 HNFs suspension. Experimental conditions: 0.1 mg/ml Hb, 120 mm Cu 2+, 0.1 M PBS at ph=7.4, and incubation different time(1 h, 24 h, 48 h, and 72 h) at 25. S-8

9 Figure S8. SEM images of different Hb concentrations on the morphologies of the Hb-Cu 3 (PO 4 ) 2 HNFs. (a 1-2 ) 0.0 mg/ml; (b 1-2 ) HNFs-1: 0.05 mg/ml; (c 1-2 ) HNFs-2: 0.1 mg/ml;(d 1-2 ) HNFs-3: 0.5 mg/ml; other conditions: 120 mm Cu 2+, 0.1 M PBS (Na 2 HPO 4, NaH 2 PO 4 ) at ph 7.4, reaction at 25 for 72 h. S-9

10 Figure S9. TG spectrum of the Hb-Cu 3 (PO 4 ) 2 HNFs. The weight loss between RT and 150 was 11.25%, corresponding to the loss of crystallization water of Cu 3 (PO 4 ) 2 3H 2 O. As temperature increased from 150 to 600, the weight loss was 20.1%, resulting from pyrolytic decomposition of Hb. S-10

11 Figure S10. The UV-vis absorption spectra (red curve) and fluorescence emission spectra (black curve) of Rh6G. S-11

12 Figure S11. Normalized time-dependent absorbance changes (λ ab = 527 nm) recorded for Hb-Cu 3 (PO 4 ) 2 HNF biocatalytic reaction systems upon addition of varying concentrations of H 2 O 2. (Hb-Cu 3 (PO 4 ) 2 HNF =3.21mg/mL, room temperature). S-12

13 Figure S12. UV-vis absorption spectra of the (a) tap water and (c) waste water with different concentrations of H 2 O 2 ; Photo inset: visual color change of the tap water and waste water; Inset: plot of absorbance versus H 2 O 2 concentrations and visual color change. Fluorescence emission spectra of the (b) tap water and (d) waste water with different concentrations of H 2 O 2 ; Photo inset: visual fluorescence change of the tap water and waste water; Inset: plot of emission intensities versus H 2 O 2 concentration and fluorescence change. S-13

14 Figure S13. Relative activity of Hb-Cu 3 (PO 4 ) 2 HNF over the course of eight rounds of successive reaction (a) fluorescence method, (b) UV method. Storage stabilities of Hb-Cu 3 (PO 4 ) 2 HNFs and free Hb in deionized water at room temperature (c) fluorescence method, (d) UV method. S-14

15 Figure S14. SEM images of the Hb-Cu 3 (PO 4 ) 2 HNFs (a) after eight rounds of successive catalytic reaction and (b) stored in deionized water for 35 days. S-15

16 Table S1. Weight percentage of Hb in the HNFs were determined by gravimetric methods Sample number Hb-Cu 3 (PO 4 ) 2 hybrid nanoflowers (mg) After calcination treatment (mg) Weight percentage (%) HNFs % HNFs % HNFs % S-16

17 Sample number Table S2. The actual encapsulation yield of Hb in the HNF was further determined by UV-Vis spectrophotometer. Theoretical concentration (mg ml -1 ) Actual concentrat ion (mg ml -1 ) Absorbance (405 nm) The supernatant concentration (mg ml -1 ) The supernatant absorbance (405 nm) Encapsulato n yield (%) HNFs % HNFs % HNFs % S-17

18 Table S3. Comparison of the reported sensor for H 2 O 2 detection Method Fluorogenic Multianalyte Biosensors Linear range of Detection Reference detection limit 0 50 μm 0.35 μm (Wang et al. 2014) Visual detection 0-20, μm (Lin et al. 2014) μm Mediator-free μm 10 nm (Liu et al. 2017) biosensor Ion chromatography mg/l mg/l (Song et al. 2017) with UV detector NF- UV detection 2-10, ppb This work ppb NF-fluorescence detection 2-10, ppb 0.01 ppb This work S-18

19 Table S4. Influence of coexisting substance (fluorescence detection) Coexisting substances Tolerance [CS/H 2 O 2 ] Relative error(%) Coexisting substances Tolerance[ CS/H 2 O 2 ] Relative error (%) Ascorbic 100: Ca : acid Glucose 100: Mg : Uric acid 100: Co : Dopamine 100: Mn : Citric acid 100: NO 4 100: Fe : NH 4 100: Na + 100: SO 4 100: Ni : Cl - 100: Fe : S-19

20 Table S5. Influence of coexisting substance (UV detection) Coexisting substances Tolerance [CS/H 2 O 2 ] Relative error(%) Coexisting substances Tolerance [CS/H 2 O 2 ] Relative error(%) Ascorbic 100: Ca : acid Glucose 100: Mg : Uric acid 100: Co : Dopamine 100: Mn : Citric acid 100: NO 4 100: Fe : NH 4 100: Na + 100: SO 4 100: Ni : Cl - 100: Fe : S-20

21 Table S6. Recovery of H 2 O 2 in different water with this proposed method. (UV detection) Sample Added (ppb) Found (ppb) Recovery (%) RSD (%) (n=3) Rainwater ± ± Tap water ± ± waste water ± ± S-21

22 Table S7. Recovery of H 2 O 2 in different water with this proposed method. (Fluorescence detection) Sample Added (ppb) Found (ppb) Recovery (%) RSD (%) (n=3) Rainwater ± ± Tap water ± ± waste water ± ± S-22