Embed Size (px)
Citation preview
S1
Pyrazoline-based colorimetric and fluorescent probe for detection of
sulphite
Supplementary data
Tomasz Uchacza*, Gabriela Jajko
a, Andrzej Danel
b*, Paweł Szlachcic
b, Szczepan Zapotoczny
a
a Faculty of Chemistry, Jagiellonian University, ul. Gronostajowa 2,
30-387 Kraków, Poland
b Department of Chemistry and Physics, Agricultural University, ul. Balicka 122,
31-149 Kraków, Poland
* Corresponding authors.
E-mail address: [email protected] (Tomasz Uchacz), [email protected] (Andrzej Danel)
Table of contents Page number
Figure S1. ......................................................................................................................................................................................................... S2
Figure S2. ......................................................................................................................................................................................................... S3
Figure S3. ......................................................................................................................................................................................................... S4
Figure S4. ......................................................................................................................................................................................................... S5
Figure S5. ......................................................................................................................................................................................................... S6
Figure S6. ......................................................................................................................................................................................................... S7
Figure S7. ......................................................................................................................................................................................................... S8
Figure S8. ....................................................................................................................................................................................................... S9
Figure S9. ....................................................................................................................................................................................................... S10
Figure S10. ..................................................................................................................................................................................................... S11
Figure S11. ..................................................................................................................................................................................................... S12
Figure S12. ..................................................................................................................................................................................................... S13
Figure S13. ................................................................................................................................................................................................... S14
Figure S14. ..................................................................................................................................................................................................... S15
Figure S15. ..................................................................................................................................................................................................... S16
Figure S16. ..................................................................................................................................................................................................... S17
Figure S17. ..................................................................................................................................................................................................... S18
Figure S18. ..................................................................................................................................................................................................... S19
Figure S19. ..................................................................................................................................................................................................... S20
Figure S20. ..................................................................................................................................................................................................... S21
Figure S21. ..................................................................................................................................................................................................... S22
Figure S22. ..................................................................................................................................................................................................... S23
Figure S23. ..................................................................................................................................................................................................... S24
Figure S24. ..................................................................................................................................................................................................... S25
Table S1. ......................................................................................................................................................................................................... S26
Electronic Supplementary Material (ESI) for New Journal of Chemistry.This journal is © The Royal Society of Chemistry and the Centre National de la Recherche Scientifique 2018
S2
Figure S1. 1H NMR (top) and
13C NMR (bottom) spectra of 4a in CDCl3.
1.04
1.08
1.05
1.95
2.56
1.08
2.05
3.08
2.00
1.03
2.02
1.98
2.00
1.00
7.11
7.12
7.24
7.25
7.26
7.26
7.28
7.28
7.29
7.29
7.30
7.30
7.31
7.34
7.35
7.37
7.40
7.41
7.41
7.41
7.42
7.46
7.47
7.48
7.55
7.55
7.55
7.56
7.56
7.57
7.57
7.58
7.58
7.75
7.76
7.76
7.77
7.77
7.81
7.81
7.81
7.82
7.82
7.92
7.93
7.96
5.43
5.43
5.45
5.45
3.90
3.92
3.93
3.95
3.23
3.24
3.26
3.27
43.6
63.3
103.0
113.2
122.3
125.5
126.3
128.2
128.4
128.8
129.0
129.5
129.9
131.6
132.5
134.1
137.1
140.9
147.9
151.1
155.5
189.9
122.3
125.5
126.3
128.2
128.4
128.8
129.0
129.5
129.9
131.6
132.5
134.1
137.1
140.9
S3
Figure S2. Probable fragmentation path analysis of [4a+Na]+ by means of mass spectrometry.
S4
Figure S3. 1H NMR (top) and
13C NMR (bottom) spectra of 4b in CDCl3.
1.07
1.10
1.08
3.12
1.04
1.10
1.09
2.16
3.02
0.99
2.01
2.09
1.03
1.00
7.12
7.14
7.15
7.16
7.17
7.18
7.25
7.27
7.27
7.30
7.32
7.32
7.35
7.36
7.37
7.38
7.40
7.40
7.42
7.43
7.45
7.46
7.68
7.69
7.70
7.70
7.76
7.76
7.77
7.78
7.78
7.95
7.98
8.18
8.24
8.24
8.25
8.25
5.43
5.45
5.46
5.48
3.90
3.93
3.95
3.98
3.23
3.25
3.28
3.29
43.7
63.5
101.0
113.4
119.8
122.7
125.7
126.5
128.4
128.5
128.9
129.7
130.0
131.8
133.8
134.4
134.6
141.0
143.1
148.0
151.3
155.8
178.9
119.8
122.7
125.7
126.5
128.4
128.5
128.9
129.7
130.0
131.8
133.8
134.4
134.6
S5
Figure S4. Probable fragmentation path analysis of [4b+Na]+ by means of mass spectrometry.
S6
Figure S5. Optimized molecular structure of 4a.
S7
Figure S6. Optimized molecular structure of 4b.
S8
Figure S7. Optimized molecular structure of 4a+sulphite adduct
S9
Figure S8. Optimized molecular structure of 4b+sulphite adduct
S10
Figure S9. DPV curve of 4a dye
-2.4 -2.0 -1.6 -1.2 -0.8 -0.4 0.0 0.4 0.8 1.2
-0.4
-0.2
0.0
0.2
0.4
0.6
0.8
1.0
DPV of 4a in DMF
Cu
rren
t [ µ
A]
E [V] vs Fc/Fc+
S11
Figure S10. DPV curve of 4b dye
-2.4 -2.0 -1.6 -1.2 -0.8 -0.4 0.0 0.4 0.8 1.2
-0.4
-0.2
0.0
0.2
0.4
0.6
0.8
1.0
Cu
rre
nt
[ µA
]
E [V] vs Fc/Fc+
DPV of 4b in DMF
S12
Figure S11. The selectivity of 4a towards detection of sulphite anion. The changes of color induced by the
presence of 45 equivalents of the different anions monitored in DMF/water solution.
S13
Figure S12. Excitation spectra of 4a upon addition of 45 equivalents of sulphite. Conditions: emission
monochromator set at 475 nm (blue line) and 587 nm (red line).
320 340 360 380 400 420 440 460 480 500 520 540 560
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
No
rma
lize
d e
xcit
ati
on
[a
.u.]
wavelength [nm]
4a without sulphite
4a + 45 eq. of sulphite
S14
Figure S13. Excitation spectra of 4b upon addition of 45 equivalents of sulphite. Conditions: emission
monochromator set at 478 nm (blue line) and 605 nm (red line).
300 350 400 450 500 550
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
No
rma
lize
d e
xcit
ati
on
[a
.u.]
wavelength [nm]
4b without sulphite
4b + 45 eq. of sulphite
S15
Figure S14. Effect of pH on the fluorescence of sulphite-4b product and sulphite free 4b probe (inset)
monitored at 475 nm.
5 6 7 8 9 10 11 12 13 14
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5 6 7 8 9 10 11
0
50
100
150
200
Flu
ore
sc
en
ce
at
47
5 n
m
pH
4b - free pobe
I su
lph
ite/I
0
pH
4b +sulphite
S16
Figure S15. Effect of pH on the fluorescence of sulphite-4a product and sulphite free 4a probe (inset)
monitored at 475 nm.
5 6 7 8 9 10 11 12 13 14
6
8
10
12
14
16
18
5 6 7 8 9 10 11
0
20
40
60
80
100
120
140
Flu
ore
sc
en
ce
at
47
5 n
m
pH
4a - free probe
4a + sulphite
I su
lph
ite/I
0
pH
S17
Figure S16. Effect of pH on the absorption of 4b and 4b+sulphite monitored at 490 nm.
5 6 7 8 9 10 11
0.00
0.02
0.04
0.06
0.08
0.10
0.12
0.14
0.16
0.18 4b probe
4b probe + sulphite
Ab
so
rban
ce a
t 4
90 n
m
pH
S18
Figure S17. Effect of pH on the absorption of 4a and 4a+sulphite monitored at 482 nm.
5 6 7 8 9 10 11
0.00
0.02
0.04
0.06
0.08
0.10
0.12
0.14
0.16
0.18
0.20
0.22
4a probe
4a probe + sulphite
Ab
so
rba
nce
at
48
2 n
m
pH
Figure S18. 1H NMR spectra of 4a in CDCl
S19
in CDCl3 (top) and 4a-SO32-
adduct in D2O/DMSO
/DMSO-d6 (bottom).
S20
Figure S19. The synthesis procedure for model molecule.
(E)-2-benzoyl-3-[4-(N,N-dimethylamino)phenyl]prop-2-enenitrile (model):
4-(N,N-dimethylamino)benzaldehyde (328 mg, 2.2 mmol) and 3-oxo-3-phenylpropanenitrile (320 mg, 2.2
mmol) were dissolved in 96% ethanol (5 mL) in a 25 mL round-bottomed flask. To the mixture 3 drops of
DBU were added and the reaction mixture was heated under reflux for six hours. After cooling, the
precipitate was filtered off and crystallized from ethanol.
Light-orange needles, 503 mg, 83%, mp. 162-163 °C (ethanol). 1H NMR (600 Hz, CDCl3, δ/ppm, Figure
S20, top): 8.01 (s, 1Hvinyl); 8.01-7.98 (m, 2H), 7.85-7.82 (m, 2H), 7.59-7.55 (m, 1H), .50-7.47 (m, 2H), 7.71
(broad d, J = 9.1 Hz, 2H), 3.13 (s, 6H, NMe2). 13
C NMR (150 Hz, CDCl3, δ/ppm, Figure S21): 190.3, 155.9,
154.0, 137.5, 134.7, 132.5, 129.1, 128.5, 119.8, 119.3, 111.7, 101.7, 40.2. Anal. Cald. for C18H16N2O: C,
78.24; H, 5.84; N, 10.14. Found: C, 78.30; H, 5.75; N, 10.09.
Figure S20. 1H NMR spectra of model
S21
model in CDCl3 (top) and model-SO32-
adduct in D
adduct in D2O/DMSO-d6 (bottom).
Figure S21. 13
C NMR spectra of model
S22
model in CDCl3.
S23
Figure S22. Molecular mass analysis of [4b+Na]+ (top) and [4b-SO3]
– adduct (bottom) by means of HR
mass spectrometry .
S24
Figure S23. Molecular mass analysis of [4a+Na]+ (top) and [4a-SO3+2Na]
+ adduct (bottom) by means of
HR mass spectrometry .
S25
Figure S24. Sulphite concentration dependent fluorescence observed for 4a and 4b dissolved in HEPES (pH
= 8)/DMF (blue line - 0 equivalents, green 50 equivalents of sulphite).
420 450 480 510 540 570 600 630 660 690 720 750
04080
120160200240280320
420 450 480 510 540 570 600 630 660 690 720 750
0
100200
300
400
500600
700
800
Flu
ore
sc
en
ce
in
ten
sit
y [
a.u
.]
wavelength [nm]
4b
Flu
ore
sc
en
ce
in
ten
sit
y [
a.u
.]
wavelength [nm]
4a
S26
Table S1. A comparison table about the detection limits of sulphite/bisulphite anions.
Solvent Time of response Detection limit references
Water/DMSO 30 s 0.106 µM 35
PBS/CTAB 7.7 min (50 eq.) 0.23 µM 36
PBS/EtOH 20 min (500 eq.) 2.76 µM 20
HEPES/DMF 9 min 4.87 µM This work
citric acid buffer - 6.30 µM 37
Buffer/DMSO a few min 28.2 µM 38
PBS - 30 µM 39
