Supporting Information Cascade reaction based rapid and ratiometric

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1. Calculation of the detection limit: Figure S1: Fl. Intensity ratio (I462 nm/ I359 nm) Vs. .... 450. 500. 550. 600. 0.0. 5.0x105. 1.0x106. 1.5x106. 2.0x106. 2.5x106.

Electronic Supplementary Material (ESI) for RSC Advances. This journal is © The Royal Society of Chemistry 2014

Supporting Information Cascade reaction based rapid and ratiometric detection of H2S/S2over bio-thiols with live cell-imaging: demasking of ESIPT approach Shyamaprosad Goswami, Abhishek Manna, Monalisa Mondal and Debasish Sarkar

CONTENTS 1. Calculation of the detection limit……………………………………………….....2 2. Calculation of rate constant………………...……..……………………………….3 3. Fluorescence responses of FBBP + interfering species + S2- …………………….3 4. 1H-NMR, 13C-NMR and Mass spectra…………………………………………..4-7 5. Fluorescence spectra of FBBP with different guest spices ………………….....8-10 6. Fluorescence emission spectra of FBBP with Na2S at λex=405 nm…………….11 7. MTT assay of the probe FBBP…………………………………………………...12 8. References…………………………………………………………………….…...12

1. Calculation of the detection limit: 1.1 1.0

y=85402x+0.1096 2 R =0.99

0.9

I462/I359

0.8 0.7 0.6 0.5 0.4 0.3 -6

4.0x10

-6

6.0x10

-6

8.0x10 2-

-5

1.0x10

-5

1.2x10

[S ]

Figure S1: Fl. Intensity ratio (I462 nm/ I359 nm) Vs. Conc. of S2- plot. The detection limit DL of FBBP for S2- was determined from the following equation. [S1] DL = K* Sb1/S Where K = 2 or 3 (we take 2 in this case); Sb1 is the standard deviation and S is the slope of the calibration curve. From the graph we get slope = 85402, and Sb1 value is 0.021958. Thus using the formula we get the Detection Limit = 0.51 µM i.e. FBBP can detect S2- in this minimum level.

2

2. Calculation of rate constant: From Fl. Intensity ratio (I462/I359) vs. time (sec.) plot using first order rate equation (Figure S2), we get rate constant K = slope × 2.303 = 0.0086 × 2.303 = 1.9 × 10-2 sec-1

+ 0.177 1.4 y=0.0086x 2 R = 0.98 1.6

1.2 1.4 1.2 1.0

I462/I359

I462/I359

1.0 0.8 0.6

0.8 0.6 0.4 0.2

0.4

0.0 0

0.2 0

20

40

60

80

100

120

140

20

40

160

60

80 100 120 140 160 180 200

Time (sec.)

Time (sec.) Figure S2: The time vs. Fl. Intensity ratio (I462 nm/ I359 nm) plot between 20 to 160 secend and the full plot is in inset. 3. Fluorescence responses of FBBP + various species + S2-:

1.4 1.2

I 462nm /I 359nm

1.0 0.8 0.6 0.4

3

P + + S 2 S 2- + +C S 2- y + s G + S S 2- H + + S 2- F + + CN + S 2- S 2- + + N + SC 3 S 2- N + +H S 2- O 2 + +O 2 S 2- C + +O l S 2- H . + S 2 O 2

F

B

B

0.0

-

0.2

Figure S3: Ratiometric fluorescence responses (I462nm/I359nm) of FBBP (c = 1.0 x 10–5 M) to S2- (1 equiv) containing 10 equiv of various metal species.

3

4. 1H NMR, 13C NMR and ESI MS spectra: 1 H NMR spectrum and its expansion of Receptor i.e. FBBP in CDCl3:

Expansion of the NMR:

4

1

H NMR spectrum and its expansion of Receptor i.e. FBBP in d6-DMSO:

13

C NMR spectrum of Receptor i.e. FBBP:

5

ESI MS Mass Spectrum of FBBP:

1

H NMR spectrum and its expansion of FBBP + Na2S in d6-DMSO:

6

ESI MS Spectrum of FBBP + Na2S:

7

5. Fluorescence emission spectra of FBBP with different species such as Cysteine, Glutathione, F-, CN-, N3-, SCN-, SO32-, S2O32-, H2O2, HSO3-, Hydroxyl radical, Hypochlorite, Ascorbic Acid, NH4+, Co2+, Al3+, Hg2+ and Cu2+ in CH3CN : H2O (2:8, v/v) (The solutions of anions and oxidants were prepared from Cysteine, Glutathione, TBAF, KCN, NaN3, KSCN, KSO3, K2S2O3, NaHSO3, H2O2, Femton’s reagent, NaOCl, Ascorbic Acid, NH4Cl, CoCl2, Al(NO3)3, 9H2O, HgCl2 and CuCl2 respectively in H2O). (a)

(b) 6

3.5x10 6

6

Cysteine

6

2.5x10

3.0x10

Fl. Intensity (a.u.)

Fl. Intensity (a.u.)

3.0x10

6

2.0x10

6

1.5x10

6

1.0x10

Glutathione

6

2.5x10

6

2.0x10

6

1.5x10

6

1.0x10

5

5.0x10

5

5.0x10

0.0

0.0 350

400

450

500

550

350

600

(c)

400

450

500

550

600

Wavelength (nm)

Wavelength (nm)

(d) 6

3.5x10

6

3.5x10

-

Fl. Intensity (a.u.)

Fl. Intensity (a.u.)

6

6

-

3.0x10

F

6

2.5x10

6

2.0x10

6

1.5x10

6

1.0x10

CN

3.0x10

6

2.5x10

6

2.0x10

6

1.5x10

6

1.0x10

5

5

5.0x10

5.0x10

0.0

0.0 350

400

450

500

550

600

350

N3

-

SCN

6

3x10

Fl. Intensity (a.u.)

Fl. Intensity (a.u.)

3.0x10

500

550

600

(f)

6

3.5x10

6

450

Wavelength (nm)

Wavelength (nm)

(e)

400

6

2.5x10

6

2.0x10

6

1.5x10

6

1.0x10

-

6

2x10

6

1x10

5

5.0x10

0.0

0 350

400

450

500

Wavelength (nm)

550

600

350

400

450

500

550

600

650

Wavelength (nm)

8

(g)

(h) 6

3.5x10

6

3.5x10

Fl. Intensity (a.u.)

6

6

2.5x10

6

2.0x10

6

1.5x10

6

1.0x10

S 2 O3

3.0x10

Fl. Intensity (a.u.)

SO3

6

3.0x10

2-

2-

6

2.5x10

6

2.0x10

6

1.5x10

6

1.0x10

5

5

5.0x10

5.0x10

0.0 350

400

450

500

550

600

0.0 350

400

Wavelength (nm)

450

(i)

550

600

(j) 6

6

3.5x10

3.5x10

H2O2

-

3.0x10

6

2.5x10

6

2.0x10

6

1.5x10

6

1.0x10

HSO3

6

3.0x10

Fl. Intensity (a.u.)

Fl. Intensity (a.u.)

6

6

2.5x10

6

2.0x10

6

1.5x10

6

1.0x10

5

5

5.0x10

5.0x10

0.0

0.0 350

400

450

500

550

600

350

400

450

500

550

600

Wavelength (nm)

Wavelength (nm)

(k)

(l)

6

3.5x10

6

3.5x10

.

OH

6

Fl. Intensity (a.u.)

3.0x10

Fl. Intensity (a.u.)

500

Wavelength (nm)

6

2.5x10

6

2.0x10

6

1.5x10

6

1.0x10

OCl

6

3.0x10

-

6

2.5x10

6

2.0x10

6

1.5x10

6

1.0x10

5

5

5.0x10

5.0x10

0.0

0.0 350

400

450

500

Wavelength (nm)

550

600

350

400

450

500

550

600

Wavelength (nm)

9

(m)

(n) 6

6

4.0x10

Ascorbic Acid

6

3.0x10

NH4

6

3.5x10

+

6

3.0x10

Fl. Intensity (a.u.)

Fl. Intensity (a.u.)

3.5x10

6

2.5x10

6

2.0x10

6

1.5x10

6

2.5x10

6

2.0x10

6

1.5x10

6

6

1.0x10

1.0x10

5

5.0x10

5

5.0x10

0.0

0.0

350

350

400

450

500

550

400

600

450

500

550

600

Wavelength (nm)

Wavelength (nm) (o)

(p) 6

4.0x10

6

4.0x10

3.5x10

Co

3.5x10

Al

6

2+

6

3+

6

3.0x10

Fl. Intensity (a.u.)

Fl Intensity (a.u.)

6

3.0x10

6

2.5x10

6

2.0x10

6

1.5x10

6

2.5x10

6

2.0x10

6

1.5x10

6

1.0x10

6

1.0x10

5

5.0x10

5

5.0x10

0.0 0.0

350 350

400

450

500

550

400

600

450

500

550

600

Wavelength (nm)

Wavelength (nm)

(q)

(r) 6

6

4.0x10

4.0x10

Hg

6

2+

6

3.5x10

6

6

3.0x10

Fl. Intensity (a.u.)

Fl. Intensity (a.u.)

3.5x10

Cu

2+

6

2.5x10

6

2.0x10

6

1.5x10

6

3.0x10

6

2.5x10

6

2.0x10

6

1.5x10

6

1.0x10

1.0x10

5

5

5.0x10

5.0x10

0.0

0.0

350

400

450

500

Wavelength (nm)

550

600

350

400

450

500

550

600

Wavelength (nm)

10

Figure S4-S22: Fluorescence emission spectra of FBBP with different species such as Cysteine, Glutathione, F-, CN-, N3-, SCN-, SO32-, S2O32-, H2O2, HSO3-, Hydroxyl radical, Hypochlorite, Ascorbic Acid, NH4+, Co2+, Al3+, Hg2+ and Cu2+ in CH3CN : H2O (2:8, v/v) respectively.

6. Fluorescence emission spectra of FBBP with Na2S (excitation at 405 nm): 6

3.5x10

6

Na2S

3.0x10

6

Fl. Intensity (a.u.)

2.5x10

6

2.0x10

6

1.5x10

6

1.0x10

5

5.0x10

0.0 450

500

550

600

Wavelength (nm)

Figure S23: Fluorescence emission spectra of FBBP (c = 1.0 x 10–5 M) with Na2S (c = 1.0 x 10–5 M) at pH 7.5 in CH3CN: H2O (2:8, v/v) (excitation at 405 nm).

11

7. MTT assay of the probe FBBP: 100

Absorbance (%)

80

60

40

20

0 10

20

30

40

50

60

70

80

90 100

[FBBP] in µM

Figure S24: MTT assay with different concentration of probe (FBBP) after 24h.

8. References: [S1] M. Zhu, M. Yuan, X. Liu, J. Xu, J. Lv, C. Huang, H. Liu, Y. Li, S. Wang, D. Zhu, Org. Lett. 2008, 10, 1481-1484.

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