Supporting information
S-nitrosothiol tethered polymer hexagons - synthesis, characterisation and antibacterial effect
S. Priya, R. Nithya and Sheela Berchmans*
Electrodics and Electrocatalysis division,
Central Electrochemical Research Institute, Karaikudi, Tamilnadu-630006-INDIA
ESI fig-1 SEM image of raw polymer PVMMA (powder).
ESI fig – 2 SEM image of RSNO conjugated polymer prepared by varying concentration a) 1mg b) 0.1 mg c) 0.08 mg under
the solvent ratio 10:1 (ethanol:water)
0.4
Nitrosated cysteine(RSNO)
PVMMA conjugated RSNO
PVMMA in acetone
0.16
0.14
RSNO
PVMMA-RSNO
0.10
absorbance
Absorbance
0.12
0.2
0.08
0.06
0.04
0.02
0.00
-0.02
400
500
600
nm
0.0
300
400
500
nm
ESI Fig.3 UV-Vis spectra of RSNO in H2O, RSNO conjugated PVMMA and PVMMA in
acetone. Inset shows visible spectra of RSNO and RSNO conjugated PVMMA
Saville assay
Reagents prepared include solution A (1mM ammonium sulfamate in 0.5M HCl), solution
B (1% sulfanilamide in 0.5M HCl), solution C (1% sulphanilamide and 0.2% HgCl2 in 0.5M HCl),
and solution D (0.02% N-(1-naphthyl)-ethylenediamine-dihydrochloride). For each reaction
mixture, 1ml of two samples was mixed with 1ml of solution A and incubated for 10min, followed by
addition of 1ml of solution B to one sample and 1ml of solution C to the other. After incubation for
5min, 1ml of solution D was mixed with both samples, and incubated for another 5min until
formation of the azo dye was complete. The absorbance of each sample was read at 540 nm.
The concentration of RSNO was determined by subtracting the absorbance of samples treated with
solution B from the absorbance of samples treated with solution C.
6
Concentration (M)
5
4
3
2
t1/2 = 16 minutes
1
0
50
100
150
200
250
time (minutes)
ESI Fig. 4 Plot of concentration of nitric oxide vs time calculated from ferrocenyl
cation formation using spectrophotometric method
Weight of the RSNO
Solvent ratio used for the
Amount of NO released
tethered polymer
synthesis of polymer (water:
(M)
(mg)
ethanol)
0.7
1:1
3.4
1.2
1:1
5.7
0.7
1:4
2.1
1.3
1:4
3.5
0.7
10:1
2.5
0.7
2:3
3
SI-Table 1 Amount of NO released from RSNO conjugated PVMMA under different solvent ratio.
Degree of functionalization of RSNO on PVMMA surface
Amount of RSNO cconjugated PVMMA = 1.9 mg (0.0019g)
Weight percentage loss due RSNO = 40 %
Molecular mass of RSNO = 150
Number of moles of RSNO bound to 1.9 mg of PVMMA = 0.00198 X 40/100 = 5.3 x 10 -6 moles
150
ESI -Fig. 5 Time and concentration based bactericidal efficacy of RSNO conjugated PVMMA with E.Coli ccontrol, d1-0.4mg . Inset shows the enlarged plot of bactericidal activity for d2-1mg, d3- 1.9mg and d4-4 mg respectively.
ESI -Fig. 6 Time and concentration based bactericidal efficacy of RSNO conjugated PVMMA with Pseudomonas
aeruginosa c-control, d1- 0.4mg. Inset shows enlarged plot of bactericidal activity for d2-1 mg,d3-1.9 mg and d4-4 mg
respectively.
ESI Fig. 7 Time and concentration based bactericidal efficacy of RSNO conjugated PVMMA with S.aureus c-control, d10.4mg. Inset shows the enlarged plot of bactericidal activity for d2-1,d3-1.9 and d4- 4 mg respectively.
absorbance
0.6
0.4
0.2
0.0
300
400
500
600
Wavelength (nm)
ESI Fig.8 UV-Vis spectra of PVMMA-RSNO before (red) and after NO release (black)
Fluorescence
(a)
(b)
Bright feild
(c)
ESI Fig. 9 Fluorescence and bright field confocal images of (a)E.coli (b) S.aureus (c) Pseudomonas after 5 hour incubation
with RSNO conjugated PVMMA under nutrient condition. Scale bars are 2 m