FiberMed11 28-30 June 2011, Tampere, Finland

FUNCTIONALIZED NANOFIBER MEMBRANES FOR DISINFECTION OF WATER

A. Goethals & K. De Clerck Ghent University, Department of Textiles, Zwijnaarde (Gent), Belgium N. Daels1, I. Scampers1 & S.W.H. Van Hulle1,2 University College West Flanders, Research Group EnBichem, Departement of Industrial Engineering and Technology, Kortrijk, Belgium 2 Ghent University, BIOMATH, Department of Applied Mathematics, Biometrics and Process Control, Gent, Belgium
1

Annelies.Goethals@UGent.be

ABSTRACT
Nowadays, water is often disinfected with biocides which can result in the formation of harmful byproducts. Therefore new developments in water treatment are needed. One of such new developments is presented in this study. Nanofiber membranes produced by the electrospinning technique offer great potential in water filtration. Nanofiber structures have a small pore size, a large specific surface area and interconnected open pore structure which makes them appropriate for water filtration. However, the pathogen removal efficiency is not yet satisfactory. To improve this, disinfectants as a functional agent on nanofiber membranes were used in this study. The pathogen removal of the functionalized membranes was evaluated with wastewater from a general hospital with ca. 107 culturable organisms per 100 ml before filtration. Pathogen removal of about 4 to 5,5 log10 is possible with selected biocides. This is higher than a nanofibers membrane without disinfectant, which gives a removal of 2 log10 and commercial membranes with a removal of 3 to 4 log10. In addition to this, the biocides stay immobilized inside the membrane and only a limited fraction (10%) leaches into the water.

INTRODUCTION

A widely used method to inactivate pathogenic micro-organisms in water and wastewater and for preventing waterborne infectious diseases throughout the world is the use of oxidation biocides such as chlorination [1]. However, some studies have reported that the effectiveness of the process is reduced by turbidity, suspended solids and the presence of nitrogen compounds such as ammonia and nitrite [2]. The use of chlorine in water treatment gives rise to undesirable byproducts suspected to pose a hazard to humans and the environment (e.g. tetrachloroethene and trichloroethene) [3], but also the rise in resistant pathogens is considered as being very problematic [4]. For this reason, new developments in water treatment are necessary. Nanofibrous structures, which are produced via electrospinning, have unique characteristics such as high porosity, high absorption capacity, small pore size and high specific surface area. Thanks to this, they have great potential for use in water filtration. The use of nanofiber membranes in water filtration has been tested in previous studies [5-7]. Nanofiber membranes have high clean water permeability (CWP) values compared to other available microfiltration membranes. However the pathogen removal efficiency is currently not satisfactory.

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In this study, nanofibrous non-wovens of polyamide (PA) were produced via electrospinning to develop antimicrobial filtration membranes. In literature mainly (nano)silver particles are used to obtain this antimicrobial effect [8,9], while in this study different biocides are tested as functionalizing agents.

2 2.1

EXPERIMENTAL Electrospinning of functionalized nanofiber membranes

The standard setup for nozzle electrospinning consists of a syringe with a metallic needle, a syringe pump, a high-voltage power supply, and a grounded collector [10]. An electric field is applied across a polyamide 6 solution and a collector plate. As the solution jet travels, it is bend and/or split by the electric forces while the solvent evaporates. This mechanism leads to the formation of fibers which are attracted to the grounded collecting plate (figure 1).

Figure 1 Scheme of the electrospinning process and SEM image of the resulting nanofiber membrane

Scanning electron microscope (SEM) was used to study the influence of electrospinning parameters on the morphology of the membrane [10]. This production process results in a flat sheet nonwoven nanofiber membrane with a mean pore size of 0.4 m, a fiber diameter between 50 and 100 nm and a thickness of 120 m [11]. The applied biocides (WSCP, DPNBA, nAg and bronopol) act with different mechanisms. According to Shrivastava et al. the antibacterial property of silver nanoparticles result from bacterial cell lysis and the interaction with the phosphotyrosine profile of putative bacterial peptides, which could affect the transduction of the bacterial signal and inhibit growth of the organism [12]. Silver is an electrophyl agent, similar to bronopol. DBNPA is an oxidation agent, and works directly via radical-mediated reactions to oxidize organic compounds [13]. Quaternary ammonium compounds, such as WSCP, are most effective against algae and bacteria in alkaline pH ranges because of their cationic surface activity. Their cationic charge forms an electrostatic bond with the negatively charged sites on a bacterial cell wall. This causes stress, leading to cell lyses and death. The quaternary ammonium salts also cause denaturation of proteins, by deforming the permeability of the cell wall and reducing the normal flow of critical nutrients into the cell which leads to cell death [14]. Quaternary ammonium compounds such as WSCP are the primary non-oxidising biocides

FiberMed11 28-30 June 2011, Tampere, Finland

and are mostly applied directly into the water [1]. Silver nanoparticles are currently implemented in a wide variety of consumer products for antimicrobial control [15]. WSCP has an effect on both Grampositive and Gramnegative bacteria, in contrast to silver nanoparticles that have only effect on Gramnegative bacteria [16,17]. The membranes were pre-functionalized by adding the functional agents to the polymer solution before the electrospinning process starts. In this way the biocide is impregnated into the fibers. The added concentrations are expressed as on mass fiber percent (omf%). As such 5 omf% WSCP equals 5 mg WSCP on 100 mg membrane.

2.2

Removal of Pathogens

The tests were performed in a flow through system in which the samples (100 ml) were filtered over a functionalized nanofiber membrane (11 cm diameter) with a pressure filter (1-1.5 bar) in a dead-end filtration cell, placed on a filter support (see) [11]. The filtration cell was previously autoclaved at 121C for 15 min. Water samples were collected from a general hospital (107-108 CFU/ 100 ml) and diluted as needed. Further the culturable microorganisms were enumerated by inoculation in a nutrient agar culture medium (CM0003, Oxoid, UK) at 37C (EN ISO 6222, 1999 with the exception that nutrient agar was used) for 48 hours. Each experiment was started with a novel functionalized membrane.

Figure 2 Flow through filtration set-up for determining pathogen removal with electrospun nanofibers First the test was performed on different kinds of functionalized membranes with hospital wastewater. As such, the best performing biocide could be chosen for further tests with inoculated water. Staphylococcus aureus strain LMG 8224 (as Grampositive strain) and Escherichia coli strain LMG 2093 (as Gramnegative strain) were used for the inoculation of this influent water. The cultures were grown in nutrient broth (CM0001, Oxoid, UK) at 37C to stationary phase [18].

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3 3.1

RESULTS AND DISCUSSION Disinfection of hospital waste water

To evaluate the removal of pathogens water samples were taken from waste water from a general hospital (107-108 colony forming units / 100 ml). When the water samples were filtered onto a non functionalized membrane (NF) with a dead-end filtration cell, a 2 log10 reduction in culturable microorganisms was observed in the filtrate (figure 2). The removal with a non-functionalized membrane is not as good as other microfiltration studies. With other commercial membranes a 2 log 10 4 log10 removal is possible [8,19,20]. This is probably due to the deformation of the nanofibers during pressurized filtration. Higher reduction was reached when silver nanoparticles were added to the membrane. A 3 log10 4 log10 was reached by adding respectively 1 3 omf% silver nanoparticles to the spinning solution. Nanofiber membranes with biocides as functionalization agents achieve high reduction rates, as WSCP and bronopol achieve removal of 4 log10 5,5 log10. The results show that due to the nAg and the functionalization with biocides in the functionalized membrane a higher efficiency (3,9 log 10 5,5 log10) could be achieved.
6 Removal of pathogens (log) 5 4 3 NF 2 1 0 NF nAg WSCP Membrane Bronopol DPNBA 1% 3% 5%

Figure 3 Removal rate of pathogens with functionalized and non functionalized membranes. NF = non functionalized, nAg = functionalized with silver nanoparticles, WSCP= functionalized with WSCP (1 or 3 omf%), bronopol= functionalized with bronopol (1 or 3 omf%) and DBNPA= functionalized with DBNPA (1 or 3 omf).

3.2

Removal of specific pathogens

The leaching of WSCP from the membrane has been previously studied, showing that more than 90% is retained on the membrane [21]. Further tests with inoculated water were done using a 5 omf % WSCP functionalised nanofiber membrane, since this gave the best results. S. aureus shows a higher log10 CFU/100ml removal (5.8 log10 CFU/100 ml) then E. coli (4.0 log10 CFU/ 100 ml) (figure 4). In the study of Lebleu et al., S. aureus had a 3 to 2 log10 removal higher than E. coli. In our study a similar difference in both bacterial removal is seen. Lebleu et al. stated that the role of the cell-wall structure is important in the retention of bacteria in microfiltration membranes [22]. Grampositive bacteria have a thicker peptidoglycan layer and therefore less deformable and thus better rejected than Gramnegative ones. It was proven that this wall deformability is more determining in the transfer mechanisms than other properties such as surface zeta potential or hydrophobicity/ hydrophilicity balance. This passing of bacterial through a

FiberMed11 28-30 June 2011, Tampere, Finland

microfiltration membrane is illustrated and calculated by Suchecka et al. [22]. Microbial cells can be carried across the membrane through significantly smaller pores, but this mechanism needs more research. Important factors are the osmotic pressure, cell membrane/polymeric membrane interaction and cell membrane characteristics.

Log10-removal (CFU/100ml)

7,00 6,00 5,00 4,00 3,00 2,00 1,00 0,00 0,5 Volume (l) 25 E. coli S. aureus

Figure 4 Results of long term tests after rinsing the membrane with 0.5 l and 25 l before the inoculated volume (0.5 l) was added. Darkly colored bars represent E. coli, light gray bars represent S. aureus

CONCLUSION

The functionalized nanofiber membranes with 5 omf% WSCP can be used as antibacterial membrane. Short term experiments demonstrated that a 5.2 log10 CFU/100ml removal is possible with hospital waste water. These experiments demonstrate the potential of electrospun nanofiber membranes for water filtration and disinfection with WSCP as case study. Further research should now be conducted towards the reproducibility and controllability of electrospinning of functional nanofibers. These criteria remain a challenge in the engineering of a cost-effective large scale production system. Also other biocides or other nanofiber polymer materials should be tested in further research towards disinfection for obtaining the best combination of membrane material with antibacterial functionalization agent.

ACKNOWLEDGMENTS
The authors would like to thank the Institute for the Promotion of Innovation by Science and Technology in Flanders (IWT) for their financial support (IWT-Tetra project contract numbers 060113 and 080148).

REFERENCES
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