Supplementary Data for
A Fluorescent and Colorimetric Chemosensor for Nitric Oxide Based
on 1, 8-Naphthalimide
Min Wang, a Zhaochao Xu*b, Xu Wang, Jingnan Cui*a
a
* State Key Laboratory of Fine Chemicals, Dalian University of Technology, 2 Linggong Road, Dalian 116024,
P.R. China E-mail: [email protected];
b*
Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023, P.R.
China E-mail:[email protected].
800
700
Intensity
600
500
400
300
NOC13
200
100
0
0
200
400
600
800
1000
1200
1400
Time/s
Fig. S1. Kinetic profile of DAN (10 μM) with NOC13 (10 μM) solution. Conditions: Ethanol–Tris-HCl buffer (50
mM, pH = 7.4) solution (1:1, v/v, rt). Excitation at 370 nm.
1000
none
1 µM
2 µM
4 µM
6 µM
8 µM
10 µM
12 µM
14 µM
16 µM
18 µM
20µM
Intensity
800
600
400
200
0
400
450
500
550
Wavelength/nm
Fig. S2. Fluorescence changes of DAN (10 μM) upon addition of NOC13 solution (0, 1, 2, 4, 6, 8, 10, 12, 14, 16,
18, 20 M). Then emission intensity of DAN reached the maximum when 16 M NOC13 solution was added
which did not continue to increase when 18M or 20M NOC13 was added.
a.
32
28
Intensity
24
20
16
12
8
4
0
425
400
475
450
500
575
550
525
Wavelength/nm
20
b.
Y=1.93118+0.04262 X
R=0.98614
16
F454/F0
12
8
4
0
0
50
100
150
200
250
300
350
400
Wavelength/nm
c.
1.0
Y=5.49965+0.71422 X
2
R =0.98342
(Imin-I)/(Imin-Imax)
0.8
0.6
0.4
0.2
0.0
-7.6
-7.4
-7.2
-7.0
-6.8
-6.6
-6.4
Log[NOC13]
Fig. S3. (a). Fluorescence changes of DAN (10 μM) upon addition of NOC13 solution (0, 10, 25, 50, 100, 150,
200, 300, 400 nM). (b). Fluorescence ratio (F454/F0) changes of DAN (10 μM) upon addition of NOC13 (0–400
nM). (c). A linear regression curve was then fitted to these normalized fluorescence intensity data, and the point at
which this line crossed the ordinate axis was considered as the detection limit 1.9 × 10-8 M for NO. Conditions:
Ethanol–Tris-HCl buffer (50 mM, pH = 7.4) solution (1:1, v/v, rt). Excitation at 370 nm.
0.20
none
1 µM
2 µM
4 µM
6 µM
8 µM
10 µM
12 µM
14 µM
16 µM
18 µM
20µM
Absorbance
0.15
0.10
0.05
0.00
500
450
400
350
600
550
Wavelength/nm
25
20
A371/A476
15
10
5
0
-2
0
2
4
6
8
10
12
14
16
[NO]/uM
Fig. S4. (a). Absorbance changes of DAN (10 μM) upon addition of NOC13 solution (0-20 µM). (b). Absorbance
ratio (A371/A476) changes of DAN (10 μM) upon addition of NOC13 (0-14 µM). Conditions: Ethanol–Tris-HCl
buffer (50 mM, pH = 7.4) solution (1 : 1, v/v, rt).
Cell incubation and fluorescence imaging:
HT29 cells (Human colon adenocarcinoma grade II cell line) were cultured in minimum
essential medium (DMEM) supplemented with 10% fetal bovine serum. The cells were incubated
at 37℃ in 5% CO2 and 20% O2 for 24 h. The cells were incubated with 10 μM DAN for 30 min
and then washed with PBS (phosphate buffered saline, pH=7.4) three times. Then the fluorescence
imaging studies were observed under a Leica TCS-SP2 confocal fluorescence microscopy with a
10 × objective lens. In the second group, after the incubation of 10 μM DAN for 30 min, 40 μM
NOC13 solution was inserted into PBS with HT29 cells, then washed with PBS for three times
before the fluorescence imaging were observed.
Reference
1.
F. Li, J. Cui, L. Guo, X. Qian, W. Ren, K. Wang and F. Liu, Bioorg. Med. Chem., 2007, 15, 5114-5121.