12. - 14. 10. 2010, Olomouc, Czech Republic, EU

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NANOTEXTILES MODIFIED WITH PHTHALOCYANINE DERIVATIVES, PHOTODYNAMIC ACTIVITY AGAINST GRAM-POSITIVE BACTERIA

Marie KARASKOVAa Jan RAKUSANa Oldrich JIRSAKb, Petr JILEKc

a Center of Organic Chemistry Ltd. Rybitvi 296, 533 54 Rybitvi Czech Republic b Technical University of Liberec Halkova 6, 461 17 Liberec 1 Czech Republic

c Charles University Prague, Faculty of Pharmacy, Heyrovskeho 1203, 500 05 Hradec

Kralove Czech Republic Abstract

The following derivatives of phthalocyanine (Pc) were synthesized: HOAlPc (SO3H)2,

ZnPc (OC2H4N+CH3)4 I

-4, ZnPc(SO3H)0-1 (SO2NHCH2 CH2N(C2H5)2)1-3,

HOAlPc(SO3H)0-1(SO2NHCH2CH2NH CH2CH2OH)1-2, and Ag2Pc These phthalocyanine derivatives were

incorporated into polyurethane solutions and subsequently electrospun into nanofibrous layers. Thus,

nanotextiles containing 0,1 and 1 weight per cent of phthalocyanine derivatives were prepared.

Photodynamic bactericidal action of nanotextile samples modified with the particular phthalocyanines were

tested by exposing to the daylight and in the dark, respectively, by using of Gram-positive bacteria

Staphylococcus aureus. All the samples of nanotextile performed antimicrobial effect. The best results were

obtained with the nanotextile sample modified with ZnPc (OC2H4N+CH3)4 I

-4

1. INTRODUCTION

Nanotextiles have attracted considerable interest and have been found to be highly promising candidates for a variety

uses, among others for removing bacteria1 by filtration from the stream of air. In the process, the stream of air is passing

through the filter, made from nanotextile, while bacteria are kept on the surface of filter. On the other hand,

phthalocyanines and their metal complexes also have attracted considerable interest, owing their capability, by so called

photodynamic effect, after the suitable illumination change diatomic oxygen into various reactive oxygen species. As the

reactive oxygen species are highly bactericidal, they could be used for combating bacteria2 on the surface of nanotextile.

Both the effects mentioned above, has become a base for the lower described study, aimed towards the new filtration

material, capable by the photodynamic process kill the bacteria kept

on its surface.

2. EXPERIMENTAL

2.1 The following phthalocyanine derivatives were

prepared by Center of Organic Chemistry Ltd.

M = Ag2 , Zn, HOAl

R1, R2, R3, R4 = indepedantly H, -SO3H, -OC2H4N+CH3)4I

-4,

SO2NHCH2CH2CH2N(C2H5)2,

- SO2NHCH2CH2NH CH2CH2OH

12. - 14. 10. 2010, Olomouc, Czech Republic, EU

1. HOAlPc (SO3H)2

2. ZnPc (OC2H4N+CH3)4 I

-4

3. ZnPc (SO2NHCH2CH2CH2N(C2H5)2)1-2

(SO3H)0-1

4. HOAlPc (SO2NHCH2CH2NH CH2CH2OH)1-2

(SO3H)0-1

5. Ag2Pc

1. Was prepared by sulfonation of hydroxyaluminium phthalocyanine with fuming sulfuric acid.

2. Was prepared by methylation of zinc tetrakis-(3-pyridyloxy)phthalocyanine3 with methyl iodide

3. Was prepared by chlorosulfonation of zinc phthalocyanine with chlorosulfonic acid and subsequent

condensation of zinc phthalocyanine sulfochloride with 3 -(diethylamino)-propylamine

4. Was prepared by chlorosulfonation of hydroxyaluminium phthalocyanine and subsequent

condensation of hydroxyaluminium phthalocyanine with N-2- oxyethyl- etylendiamine

5. Was prepared by reaction of lithium phthalocyanine with AgNO3 in ethanol.4

2.2 Preperation of nanotextiles containing the phthalocyanine derivatives shown above

Phthalocyanine derivatives were incorporated into polyurethane solutions, which were subsequently

electrospun into nanofibrous layers at the Technical university of Liberec. Nanotextiles, made this way on

polyurethane basis (PUB), contained 1% of phthalocyanine derivatives. The control sample of nanotextile,

containing no phthalocyanine, was prepared on polyurethane basis by the same way.

2.3 Photodynamic antimicrobial effect of nanotextiles containing derivatives of phthalocyanine in

nanofibres

The nanotextiles modified with phthalocyanine derivatives were tested in photodynamic antimicrobial activity

at the Faculty of Pharmacy of Charles University.

The strain of Staphylococcus aureus (CCM 4516) was used for tests of photodynamic antimicrobial effect

of nanotextiles modified with phthalocyanine derivatives. Two methods of tests were used.

By the first method the pairs of nanotextile samples modified with phthalocyanines were inserted into Petri

bowls and overlapped with Baird Parker agar base containing Staphylococcus aureus (SA) After agar base

had got stiffen, always one sample of the pair was kept in the dark 3 hours, the other sample of the pair was

kept on the day light 3 hours. All the Petri bowls were subsequently incubated 24 hour at 350 C and

evaluated.

By the second method the pairs of nanotextile samples modified with phthalocyanines were inserted into

Petri bowls and inoculated on surfaces with physiological saline suspension of Staphylococcus aureus

(SA). Afterwards always one sample of the pair was kept wet in the dark 3 hours, the other sample of the

pair was kept wet on the day light 3 hours. All the samples were subsequently overlapped with Baird Parker

agar base, incubated 24 hour at 350 C and evaluated.

12. - 14. 10. 2010, Olomouc, Czech Republic, EU

The pictures below illustrate the results of tests realized with the control sample 0 of nanotextile (PUB)

prepared without phthalocyanine and 5 samples of nanotextiles (PUB), modified with 1% of phthalocyanine

derivatives. The evident black flecks are collonies of proliferated Staphylococcus Aureus.

T

he

sam

ple

Samples overlapped with SA suspension in Parker agar base

Samples inoculated with SA suspension in physiological saline and subsequently

overlapped with Parker agar base

Kept on day light Kept in dark Kept on day light Kept in dark

1

2

3

4

5

contr

ol sa

mp

le

0

12. - 14. 10. 2010, Olomouc, Czech Republic, EU

2.4 Evaluation of photodynamic antimicrobial test of nanotextiles modified with phthalocyanine

derivatives

Samples:

1. PUB nanotextile modified with 1,0 % HOAlPc (SO3H)2

2. PUB nanotextile modified with 1,0 % ZnPc (OC2H4N+CH3)4 I

-4

3. PUB nanotextile modified with 1,0% ZnPc (SO2NHCH2CH2CH2N(C2H5)2)1-2

(SO3H)0-1

4. PUB nanotextile modified with 1,0 % HOAlPc( SO2NHCH2CH2NH CH2CH2OH)1-2

(SO3H)0-1

5. PUB nanotextile modified with 1,0 % Ag2Pc PUB nanotextile without Pc the control sample

3. CONCLUSIONS

The following conclusion can be done from evaluation of photodynamic antimicrobial tests of nanotextiles

modified with phthalocyanine derivatives.

The samples of nanotextiles overlapped with Baird Parker agar base, containing Staphylococcus aureus,

kept in the dark and on the light 3 hours respectively, performed after incubation of 24 hours at 350 C

practically none bactericidal effect both on the light and in the dark. The similar result was obtained with the

control sample 0.

The samples of nanotextiles inoculated on its surfaces with physiological saline suspension of

Staphylococcus aureus, and kept wet in the dark and on the day light 3 hours respectively, and afterwards

overlapped with Baird Parker agar base performed, after incubation of 24 hours at 350 C, bactericidal effect

on the light for the samples 1 – 5. The best one was the sample 2, zinc tetrakis- (3- methylpyridyloxy)

phthalocyanine, the worst one was the sample 5, silver phthalocyanine. The samples 1 - 5 kept in the dark

performed, when compared with the samples kept on the light, worse bactericidal effect except of the

samples 1 and 2. The control sample 0 kept both on day light and in the dark performed none bactericidal

effect.

For the good photodynamic bactericidal effect is therefore necessary to get bacteria in the close contact with

nanotextile, modified with suitable phthalocyanine derivatives. Special filtration membranes, based on

nanotextile, modified with phthalocyanine derivatives, may be developed on the base the results mentioned

above. Such filtration membranes might be useful in various industrial installations and also in health care.

Acknowledgement

The work was supported by the Grant Agency of Ministry of Education, Youth and Physical Training,

no. 2B06104 and by the Grant Agency of the Academy of Sciences of the Czech Republic (grant No.

KAN400720701).

12. - 14. 10. 2010, Olomouc, Czech Republic, EU

REFERENCES

1. JIRSAK O. at all: Textiles containing a least one layer of polymeric nanofibers and method of production of the layer of

polymeric nanofibers from the polymer solution through electrostatic spinning EP 1 869 232

2. POLONY R. at all: Process for combating micro-organisms, and novel phthalocyanine compounds US Patent 4,318,883 (1982)

3. GASPARDT S. A new method for the synthesis of zinc tetrakis(3-methylpyridyloxy)phthalocyanine J.Chem.Soc. , Perkin Trans

2, 383-389 (1989)

4. BARETT P.A., FRYE D.A., LINSTEAD R.P. Phthalocyanines and associated compounds. Part XIV. Further investigations of

metallic derivatives J. Chem. Soc., 1157-1163 (1938).