Abstract

In the present work, we demonstrate the activity of certain synthesised phenolic resins presenting -OH and NH- groups on Escherichia-Coli bacteria, synthesise was carried with phenols and formaldehyde in catalysed medium. Characterisation of these resins by IR and NMR spectroscopy

shows the presence of -C[H.sub.2]- for novolac and substituted resins, and -C[H.sub.2]O- for resol resin. The study of the antibacterial activity by the membrane filtration method (Q = 0.4[micro]) by varying resin mass and time gives a better results with the resins containing -[N.sup.+] [R.sub.2]- group than the others with only -OH group.

Introduction

The micro-organism which is generally considered as an indicator of the fecal pollution is Escherichia coli [1], Isolated first by Escherich in 1885, it is known since long time as host for digestive tube and pathogenic for the urinary tract [2], during last decades, the role of certain categories of E.coli in the diarrhoea syndromes [3] was specified and the mechanisms of this pathogenic capacity were analyzed, in the intestine [4] coli is the aerobic species quantitatively most important, and present at a rate of [10.sup.7] at [10.sup.9] bacterial bodies per gram of salts. The search for E.coli in feed water is made to appreciate its portability and its presence in water is the witness of a recent fecal contamination, rendering it unsuitable for consumption [5] one can also find them on the level of various mucous membranes at the man and the animals. The new-born babies sown during the childbirth by contact with the cutaneous flora perineale which comes from the faecal flora. The oral flora of the new-born child comprises regularly Escherichia coli, the fast colonization of the digestive tract results from this. This sowing is proportional to the duration of the childbirth, in particular with the time between the rupture of the membranes and the birth, it is completely inevitable under the natural conditions. The acquisition of E- coli is also inevitable short-term for the children born by Caesarean. In this case the initial contamination is brought by the contact with the mother or the personnel and comes primarily from other new-born babies carrying the germ [6]. The presence of E- coli in the medium surrounding or food signs a faecal contamination, but not obligatorily a human contamination [7]. In general all the warm-blooded animals shelter it. Escherichia coli is one of the principal bacteria responsible for diarrhoea [2], both in the developed countries and the countries in the process of development. The various clinical syndromes are due to different E.coli bacteria. The majority of the urinary infections of young woman observed in medical practice [3] is due to E.coli. The kind coming from the faecal flora contaminate the urine by ascending way; a third of neo-native Meningitides is due to E.coli all these infections, if they are not sufficiently treated can be at the origin of septicemia [6] and can be also in question of the peritonitis, the cholecystites, the salpingites and the post-operative suppuration, playing the role of bacteria pyogenes and pyelonephritis (infection of the kidney with fever and bacteriemy).

Certain kind of the bacteria producing toxin [8] or having invasive properties are pathogenic for the animals and cause diarrhoeas in calves or piglets. These diarrhoeas, by their frequencies of mortality cause significant economic losses.

The antibacterial polymers act and show an antibacterial activity only on the surfaces of cell bacteria, without penetration in the cells because the external membrane of the negative Gram bacteria consists of phospholipids and shows certain single characteristics where the molecules of LPS are found in external layers of external membrane LPS molecules bring several characters to the Gram negative bacteria (create a clear negative charge on the surface of the cell). Resins with phenol derivatives containing one, two or three hydroxyl groups were reported to exhibit antibacterial activity, and insoluble phenolic resins in water which contain a quaternary ammonium have a high antibacterial activity [9] Whereas it is well known that polymers such as polyesters and polyurethanes are susceptible to microbial attack [10], for example polyurethanes can be degraded by many types of fungi [11] and bacteria [12], mainly due to the hydrolysis of the -NHCOO groups when they are exposed to atmosphere. To overcome these problems is to develop polymer materials with biocide activities [13]. A number of phenolic monomers and aromatic amines have been polymerized in different reaction media, including organic solvent and microemulsions [14]. Polymer structure and properties have been studied and near complete monomer conversions, quantitative polymer yields and high molecular weights have been reported [15]. Polyphenols are commercially produced as novolacs and resoles by condensing phenol with formaldehyde at different molar ratios depending on the type of polymer desired [16]. Random pre-polymers of phenol/formaldehyde are prepared by reacting phenol on the ortho and para positions in the boxing ring with bifunctional formaldehyde, where the bases catalyzed reaction produce a mixture of methylol phenols where the composition of the mixture can be varied by altering the phenol to formaldehyde ratio Inorganic catalysts such as copper halides are known to catalyze phenol polymerization in the presence of molecular oxygen [17]. Another method use enzymes to produce polyphenols and is a potential alternative to chemical process that is currently used in industry, as in the case of enzymatic catalysis, the inorganic catalysis does not require the use of formaldehyde, and the resultant poly (phenylene oxides) exhibit good thermal and stability characteristics, there is a significant fundamental difference in the structure of polyphenols synthesized by the two previous methods. In the case of enzymatic methods the polymer has a continuous conjugated backbone, whereas in the case of inorganic catalysts, the conjugation is broken by methylene bridges and ether links. It is now possible to prepare macromolecules with predominant structures, discrete molecular weights and with specific functional groups. Compounds with hydroxyls and amines handed groups are used widely for anti-bacterial

Experimental

Synthesis of the Phenolic Resins

Resol

A phenol/formaldehyde molar ratio of 0.9 was used with sodium hydroxyl as a catalyst. 1 mol of phenol, 1.1 mol of formaldehyde and 1.5 g of sodium hydroxide were put in a kettle flask then stirred and refluxed at 100[degrees]C for 3 h. The resulting yellow mixture was left until formation of two layers. The aqueous layer was taken off, and the organic layer was distilled at 100[degrees]C in order to eliminate the remaining water. To avoid self-hardening, it is essential to keep distillation temperature lower than 110[degrees]C, since resol can be cured by heating to about 160-200[degrees]C. The crosslinked resin is a hard yellow solid, which is insoluble in all solvents.

Novolac

Phenol/formaldehyde molar ratio of 1.1 was used with oxalic acid as a catalyst. In a kettle flask, 1.2 g of oxalic acid was dissolved in 0.9 mol of formaldehyde, then 1mol of phenol was added to the mixture under stirring and reflux at 100[degrees]C for 3 h. The obtained colourless mixture was left until formation of two layers, the aqueous one was taken off, and the organic one was distilled under vacuum between 100-110[degrees]C in order to remove the remaining water. A colourless viscous resin is obtained and is not self-curing, thus a curing agent such as hexamethylenetetramine is needed along with heating up to 110[degrees]C for 10 min.

p-cresol- formaldehyde (PCF)

The procedure is similar to that of novolac. 1 mol of p-cresol, 0.9 mol of formaldehyde and 1.62 g of oxalic acid were put into a kettle, the mixture was left at room temperature for 30 min under stirring, and then refluxed at 100[degrees]C for 3 h. The obtained mixture forms 2 layers, the aqueous one was taken off and the organic one was distilled at T = 100[degrees]C for the time required to eliminate all water present, then the p-cresol formaldehyde resin was poured into a beaker while still hot.

p-aminophenol- formaldehyde (PAPF)

1 mole of p-amino phenol solution in 200 [cm.sup.3] hot water, 0.9 mole of formaldehyde, and a catalytic amount of oxalic acid were put in a kettle and stirred vigorously at T = 80[degrees]C for 1h20min. The mixture obtained forms 2 layers where the aqueous one is taken off, and the organic one is concentrated by vacuum distillation until the formation of a brown solid (p-amino phenol formaldehyde) soluble in DMSO solvent.

Charged Resin

p-chloromethylphenol monomer

A solution of phenol 10N is added to warm solution of NaOH 25N under strong agitation (in order to avoid formation of phenoxide crystals), then medium temperature is adjusted to 60-65 [degrees]C while 0.5 mole of dichloromethane is added directly into the solution in three parts each one after 15 min. After 1h of reaction, the mixture is cooled and neutralized with sulphuric acid.

p-chloromethylphenol-phenol- formaldehyde

in a kettle equipped with stirrer and refrigerator we put p-chloromethylphenol, phenol and formaldehyde in a molar ratio of 1:1:0.9, in the presence of catalytic quantity of oxalic acid as catalyst and for 4h at temperature between 70-75 [degrees]C, we obtain a yellow solid insoluble in water.

Amination of the resin

To 1.2 g of the resin prepared previously, we add 0.55 mol of [Na.sub.2]C[O.sub.3] and 1 mol of diethylamine NH[([C.sub.2][H.sub.5]).sub.2] the mixture is left for almost 5 min under a strong agitation at room temperature, then the obtained ammoniated resin is treated and quaternised.

Quaternisation of the Ammoniated Resin

To 0.01 mol of the amino resin prepared previously diluted by 10 ml of ethanol, 0.01 mol of methyl iodide was added drop wise under stirring and room temperature, then the mixture is left for almost 5 min under these conditions. The obtained resin is then purified and characterized.

Characterization of the Synthesized Phenolic Compounds

Chemical and Spectroscopic Analysis p-chloromethylphenol monomer

The p-chloromethylphenol monomer is checked by chemical reaction, 1 mol of obtained monomer is added to 1 mol of sodium cyanide, then the mixture is heated by addition of hot water, we note a release of C[O.sub.2], when it passes through a solution of NaOH becomes turbid. The thin layer chromatography technique with (50% water, 50% ethanol) eluant shows the presence of only one product and characteristic tests of odor and melting point.

Synthesized Resins

The resins prepared previously were characterized by IR spectroscopy, NMR[sup.1]H, NMR [sup.13]C and viscosimetry. It will be noted that spectroscopic measurements of IR were done on a spectrometer models Perkin apparatus, NMR on a "Brucker 300MHz" and viscosimetry on a capillary viscometer standard Ubelohde with one tenth poise flow at set temperature of 25 [+ or -] 0.1 [degrees]C.

Bacteriological Analysis

The technique used is filtration on membrane, which was adopted by the bacteriological analysis of water in 1951 after it showed that it could give results comparable to those of FTM technique. This method consists by filtering the water sample using a filter which retains the bacteria, incubate the filter in an adapted selective/differential medium during 24h, and then count the fecal colonies of coliformes or coliforme.

Materials and Methods

After putting the resin in the raw water, start the filtration pump, flamb the support surface of the filtration as well as the porous plate (by opening the tap to suck the flame), let cool, then take a membrane with asterile grip and put it in Petri's box at T=37[degrees]C for 24 h of incubation in ENDO and TSI medium to confirm the presence of Coliformes

Search of Fecal Coliformes

By using a BCPL and medium Schubert tubes at 44[degrees]C for 24 h of incubation, when adding the Kovak's reagent it appears a red ring which confirm the presence of fecal coliformes E-Coli bacteria.

Results and Discussion

Chemistry of the Reactions phenol-formol

The reaction phenol-formol corresponds to an electrophile substitution in both acid and alkaline medium where the attack on the para position is supported by polar solvent and the acid conditions whereas the attack on the position ortho is supported by non-polar solvent in the alkaline medium [18]. The phenolic resins are usually made starting from the polymerisation of condensation of phenol and formaldehyde.

Mechanism of Hydroxymethylation Reaction in Alkaline Medium

In alkaline catalysis of phenol-formol the most used catalysts are NaOH, [Na.sub.2] C[O.sub.3] N[H.sub.4] OH, In this case the reaction of the hydroxymethylation in the formaldehyde solution is presented in the methylene glycol shape, the phenol reacts quickly with the alkaline medium by forming the ion phenoxide (figire 1) which is stabilised by resonance (figure.2) [19]

[FIGURE 1 OMITTED]

The alkylation of carbon in the ortho and para substitution occurs, however substitution in meta is exclusively not obvious, the phenol reacts with the methanal in the reaction of hydroxymethylation (figure 3) where the condensation aldol takes place but here the balance of tautomerism is in favour of the form phenol which is most stable due to its aromatic character.

[FIGURE 2 OMITTED]

[FIGURE 3 OMITTED]

The compound obtained in the reaction of hydroxymethylation can lose water by heating. One obtains an intermediary product called quinomethane. The reaction which implies the positions ortho and para are catalysed by ions [H.sup.+] or O[H.sup.-], the preceding compound reacts easily with a phenol excess according to a reaction of the Michael type (addition combined on an unsaturated [alpha]- [beta] carbonyl compound) (Figure 4) [20]. At temperature between 60[degrees]C to 100[degrees]C in a strong alkaline medium, two reactions of condensation are possible. (figure 5)[21]

[FIGURE 4 OMITTED]

Mechanism of Hydroxymethylation Reaction in Acid Medium

In acidic medium the most acids used are catalysts HCl, [H.sub.2]S[O.sub.4], [H.sub.3]P[O.sub.4], the reaction of phenol-formol comes from the condensation of methanol (formol) on a phenolic cycle, the reaction is a substitution electrophile in ortho/para on aromatic activated, one can summarised the mechanism of the formation of the prepolymere [21] as follow: the protonation of formol gives an electrophile (figure 6), the condensation of the electrophile on phenol giving a benzene alcohol (figure.7) and the methylol group is unstable under the acid conditions, it condenses on another phenol molecule (figure.8). As there are three positions of substitution one obtains a rigid building two-dimensional. this compound is thermohardening. It was marketed under the name of Bakelite which has many and varied uses (laminated, plastic moulded, coatings of electrical heads of rockets and like insulator replacing the porcelain) (figure9).

[FIGURE 5 OMITTED]

[FIGURE 6 OMITTED]

[FIGURE 7 OMITTED]

Mechanism of Dichloromethane with Phenol Reaction in Alkaline Medium

In this case the halogenation reaction is followed in the same manner as when treated with bromine water where an aqueous solution of phenol gives an immediate precipitate of 2,4,6-tribromophenol, owing to the powerfully activating influence of the negatively charged oxygen in the phenoxide ion [22] (figure 10).

[FIGURE 8 OMITTED]

[FIGURE 9 OMITTED]

[FIGURE 10 OMITTED]

Spectroscopy Results

Phenolic resins prepared previously and characterized by IR spectroscopy show a difference between the spectra of the resol (figure 11) and the other polymers (Figures 14;17;20;23) which is the absorption band of -C[H.sub.2]O- group at 1001[cm.sup.-1], on the other hand, the band of phenolic -OH varies according to the molecular structures of the resin, for resol (3360 [cm.sup.-1]), which is lower than that of novolac (3368 [cm.sup.-1]). The absorption bands in the area lower than 900 [cm.sup.-1] indicate that the benzene ring is bounded by ortho-ortho, para-para and ortho-para methylene bridge. By comparison between the spectra of novolac and the other substituted resins, it is noticed that the band of phenolic -OH in PCF (3265.3 [cm.sup.-1]), PAPF (3240.2 [cm.sup.-1]) and PPF (AQ) (3328.9 [cm.sup.-1]) is lower than that of novolac which is due to the intermolecular hydrogen bond. In the area 1300 [cm.sup.-1] which represents the deformation in the plan of the -OH, the absorption bands of free normal -OH are near 1250 [cm.sup.-1], and that of -OH present in polymer is in the field of 1300 [cm.sup.-1] -1400 [cm.sup.-1]. The band of associated -OH increases with the increase of the hydrogen bond force, however, the bands of the synthesized resins Novolac (figure 14), Resol (figure 11), PCF(figure 17), PAPF(figure 20) and PPF (AQ) (figure 23) are, respectively, 1363.6, 1365.5, 1369.4, 1380.9 and 1369.4[cm.sup.-1] as it is recapitulated in table 1. The NMR [H.sup.+] spectra of the Resol and Novolac indicate that these resins contain various types of links (o-o, o-p and p-p), the resol (figure 12) contains the -C[H.sub.2]O- (4.8 ppm) and the -C[H.sub.2]- bridges (3.4-4.0 ppm), whereas, the others contain only orho-ortho -C[H.sub.2]- bridge, Novolac (3.3-3.9 ppm) (figure 15), PCF (3.8 ppm) (figure 18), PAPF(3.5 ppm) (figure 21), PPF(AQ) (3.8 ppm) (figure 24), since the para position is occupied by -C[H.sub.3] (2.2 ppm) in the case of PCF, -N[H.sub.2] (4.2 ppm) in the case of PAPF, -C[H.sub.2] N- (4.2 ppm), N[([C.sub.2][H.sub.5]).sup.+] (1.1-2.35 ppm), N-C[H.sub.3] (1.3 ppm) in the case of PPF(AQ) as it is recapitulated in table 2. The same thing with the NMR [sup.13]C spectra. Resol and Novolac indicate that these resins contain various types of links (o-o, o-p and p-p), the resol (figure 13) contains the -C[H.sub.2]O- (65 ppm) and the -C[H.sub.2]- (35-40 ppm) bridges, whereas the others contain only orho-ortho -C[H.sub.2]- bridge, Novolac(30-40 ppm) (figure 16), PCF(30 ppm) (figure 19), PAPF(41-50 ppm) (figure 22), PF(AQ) (30 ppm) (figure 25) since the para position is occupied by -C[H.sub.3] (21.5 ppm) in the case of PCF, by -N[H.sub.2] (110-120 ppm) in the case of PAPF, by -C[H.sub.2] [N.sup.-] (60 ppm), N[([C.sub.2][H.sub.5]).sup.+] (52 ppm) and N-C[H.sub.3]. (49 ppm) in the case of PF(AQ) as it is recapitulated in table 3.

[FIGURE 11 OMITTED]

[FIGURE 12 OMITTED]

[FIGURE 13 OMITTED]

[FIGURE 14 OMITTED]

[FIGURE 15 OMITTED]

[FIGURE 16 OMITTED]

[FIGURE 17 OMITTED]

[FIGURE 18 OMITTED]

[FIGURE 19 OMITTED]

[FIGURE 20 OMITTED]

[FIGURE 21 OMITTED]

[FIGURE 22 OMITTED]

[FIGURE 23 OMITTED]

[FIGURE 24 OMITTED]

[FIGURE 25 OMITTED]

Viscosimetric Measurements

Viscosity of the Quaternary Ammonium Resin

To determine the intrinsic viscosity, many authors proposed empirical relationships by extrapolating the results of the reduced viscosity to the null concentration. For that it is convenient to add a simple electrolyte (salt) to screen the intermolecular electrostatic repulsion to obtain a linear relationship between the reduced viscosity and the polymer concentration, thus, this allows comparing with the neutral polymers case. According to the results of the reduced viscosity versus concentration of charged polymer ([C.sub.0]) in aqueous ethanol solution at T=25[degrees]C show that the polymer behaves as a charged one, where the relationship of FUOSS and STRAUSS is applied. The inverse of reduced viscosity versus concentration fit a straight line graph, by extrapolation to C = 0 we determine the intrinsic viscosity [q] of the charged resin and which is [[eta]] = [10.sup.5] g/ml, this viscosity is too high relatively to that of novolac which is [[eta]] = 40 g/ml, and that of Resol which is [[eta]] = 90 g/ml. this difference is due to the pendant substituted group on the para position of the charged resin. The following graph Figure 26 shows clearly a straight line of inverse reduced viscosity as function of square root of concentration.

[FIGURE 26 OMITTED]

Antibacterial Activity of the Synthesized Phenolic Resins.

Some phenolic compounds (resins with phenol derivatives containing one, two, or three hydroxyls groups) are bactericides, were also brought back to the antibacteriennes activities due to the groups of hydroxyls [23]. The polymers are supposed being adsorbed on surfaces negatively charged with cells by the electrostatic interaction,

In our case, synthesized resins are insoluble in the water containing salts of AQ starting from polymers phenolic (PF) and studied the antimicrobic activity such phenolic resins and their derivatives. Sight that the chemicals based on phenol are among disinfecting most common for environmental surfaces. Instead of phenol or cresol, today it is more common to employ a mixture of the phenolic compounds strongly substituted to carry out their antibacterienne activity.

The results of disaffection are often specified by the time and the necessary amount of disinfecting agent. First, we count the number of colony before and after putting biocide.

The percentage of bacterial reduction of the cells is calculated according to the following formula: % reduction = [ ([N.sub.Rb]-[N.sub.Rt]) / [N.sub.Rb]].100.

Where [N.sub.[R.sub.t]] is the number of the bacterial cells counted after pouting the biocide (resin) and [N.sub.Rb] is the number of the bacterial cells before pouting biocide (resin). From the graph Figure 27 we deduce that all bacteria are exterminated after 45min by all synthesized resins except the PCF one, which can not retain bacteria more than 37.5%, whereas all bacteria are exterminated only after 10min with the quaternary ammonium resin (PPF(AQ)and 25 min with PAPF. From that, the antibacterial activity of the resins decreases as follow: PPF(AQ> PAPF> Novolac> Resol> PCF.

On other hand, rate of surviving bacteria versus concentration graph Figure 28 with fixed time t =10 min, show that with c = 3.0 g/l all bacteria are exterminated by all synthesized resins except PCF one, which eliminate only 12.5% of the holly bacteria, whereas all bacteria are exterminated with only c = 0.05 g/l with quaternary ammonium resin and c = 0.1 g/l with PAPF.

[FIGURE 27 OMITTED]

[FIGURE 28 OMITTED]

From these results we conclude that the antibacterial activity of the resin decreases as follow:

PPF(AQ) > PAPF > Novolac > Resol > PCF

On summarizing the antibacterial activity of PPF(AQ) resin as function of time and concentration simultaneously Table 4, we note that for a concentration of 0.05 g/l and a time lower than 5min, there is a total retention of E-coli bacteria. These results confirm exactly the recent search of Jeong, J and Al which synthesized the poly (styrene alt-anhydride maleic) combined with 4-aminophenol (AP) [24], acids 4-hydroxyphenol and 4-aminobenzoic [25]. The polymers of AP-dependent showed a bactericidal activity towards S dore and E. coli but it was less effective than free AP, probably due to its higher molecular weight compared to polymers of AP containing the quaternary ammonium salt with at least a long alkylic chain are usually very effective opposite a great number of micro-organism such as the bacteria, the algae, the mycetes [26].

Conclusion

This study show clearly that the reduction of E.Coli bacteria is better with the quaternary ammonium resin which contains an -[N.sup.+]R- and -OH groups on its chain compared to the synthesised others containing only -OH groups, with very court time and small concentration resin, which quaternary ammonium resin present also the qualities required as an antibacterial activity materials. The whole of this work enabled us to show that the resins and their derivatives presenting a positive charge or hydroxide ion in their structure have a significant and encouraging antibacterial activity especially toward E coli bacteria.

References

[1] James, P.N., James., B.K., 1998, "Diarrheagenic Eschrichia- Coli.," Clinical. Microbiology. Reviews., 11, pp. 142-201.

[2] Johson. J.R., 1991, "Virulence factors in Escherichia Coli urinary tract infection", Clin. Microbiol. Rev., 4, pp. 80-128

[3] Nataro, J.P., Kaper, J.B., 1998, "Diarrheagenic Escherichia Coli" Clin. Microbiol. Rev., 11, pp. 142-211

[4] Orskove, I., Orskove, F., 1985 "Escherichia Coli extra-intestinal infections", J. Hyg. Camb., 95, pp. 551-575

[5] Garcia, M. I., Le Bougnenec, C., 1998, "Enteric infection due to Escherichia Coli", Clin. Microbiol. Infect., 4, pp. 38-43

[6] Krueger, K.M., Barbieri, J.T., 1995, "The family of bacterial ADP-ribosilating exotoxins", Clin. Microbiol.Rev., 8, pp. 34-47

[7] Tarr, P., 1995, "Escherichia Coli 0157 H 7: clinical diagnostic, and epidemmiological aspects of human infection", Clin. Infect. dis., 20, pp. 1-10

[8] Jallat, C., Aubel, D., Darfeuille-Michaud, A., Joly, B., 1991, " Toxines et adherence du colibacille dans les diarrhees, "Med. Mal. Infect., 21, pp, 556-561

[9] Denyer, S. P., 1995, "Mechanisms of action of antibacterial biocides," Intl. Biodeterior & Biodegrad., 36, pp. 227-245

[10] Gu., J-D., 2003, "Microbiological deterioration and degradation of synthetic polymeric materials," Intl. Biodeterior & Biodegrad., 52, pp. 69-91

[11] Crabbe., J. R., Campbell., J. R., Thompson., L., Walz, S. L., Shultz., W. W., 1994,. "Biodegradation of colloidal ester based polyurethane by soil Fungi", Intl Biodeterior & Biodegrad., 33, pp. 103-113.

[12] El-Sayed, AHMM., Mahmoud, W. M., Davis, E. M., Coughlin, R. W., 1996, "Biodegradation of polyurethane coatings by hydrocarbon degrading bacteria," Intl. Biodeterior & Biodegrad., 37, pp. 69-79.

[13] Jayukumar, R., Rajkumar, M., Nagendram, R., Nanjundan, S., 2002, "Synthesis and characterisation of metal containing polyurethane with antibacterial activity," Journal of Applied Polymer Science, 85, pp. 1194-1206.

[14] Dordick. J. S., 1987, "Polymerisation of phenol catalysed byperoxidase in non aqueous media," Biotechnology & Bioengineering, 30, pp. 31-36.

[15] Rao, A.M., John., V.T., Gonzalez, R. D., Akkara, J. A., and Kaplan, D.L., 1993, "Catalytic and interfacial aspects of enzymatic polymer synthesis in reversed micellar systems," Biotechnology. Bioengineering., 41, pp, 531-540.

[16] Kopf, P.W., 1985, Encyclopedia of Polymer Science and Engineering., Vol. 11, Wiley, New York, pp. 45.

[17] White, D. M., and Cooper, G.D., 1982, Encyclodedia of Chemical Technology, 3rd edn, vol; 18, Wiley, New York, pp. 594.

[18] COWIE, J.M.G., 1994, "Polymers: chemistry & physics of modern materials" 2nd edition p 45

[19] Bruckner, R., 1999, "Mecanismes reactionnels en chimie organique", De Boeck Universite, pp. 45

[20] Bakales, M., Menges, O., 1988, "Phenolic Resins. Encylcopedia of Polymer Science and Engineering". John Wiley & Sons: New York, Vol. 11, pp. 152.

[21] Knop, A., Pilato, L.A., 1985, "Phenolic Resins: Chemistry, Applications and Performance". Springer-Verlag Berlin Heidelberg:, pp. 254

[22] Sfurniss, B.A., Hannaford, J., SMITH, P.W.G., TATCHELL, A.R., 1991, "Textbook of practical organic chemistry", VOGEL'S , Fifth edition, pp. 976.

[23] Nonaka, T., Uemura, Y., Ohse, K., Jyono, K., Kurihara, S., (1997), J Appl. Polym. Sci, 66, pp, 1621.

[24] Jeong, J.H., Byoun, Y.S., Lee, Y.S., 2002; React. & Funct. Polym, 50, pp, 257

[25] Jeong, J.H., Byoun, Y.S., Lee, Y.S., 2000, J. Ind.. Eng. Chem, 7, pp. 310.

[26] Tashiro, T., 2001, Macromol. Mater. Eng., 286 (63), pp, 7. Baudrion, F., Perichaud, A., Coen, S., 1998, J Appl. Polym. Sci, 70 (8), pp, 2657, Nurdin, N., Helary, G., Sauvet, G. J., 1993, Appl. Polym. Sci, 50, pp, 663.

Messaoud Chaiba (a), * and Safia Miloudi (a)

(a) LCE, BP 78 Universite Ibn-Khaldoun Tiaret 14000 Algeria

* Corresponding author E-mail: lce@mail.univ-tiaret.dz

Table 1: IR Absorption of groups in the different resin.

Displacement\
Absorption
Bandage         [[??].sub.OH]    [[??].sub.CH]    [[??].sub.CH2]

Resol              3360             3016.5           2920
Novolac            3367.9           3020.3           2910.4
PCF                3265.3           3020.0           2920
PAPF               3240.2           3008.7           3020
PF (AQ)            3328.9           2962             2736.8

Displacement\
Absorption
Bandage         [[??].sub.C-O]   [[??].sub.C=C]   [[??].sub.CH3]

Resol              1001             1510.2              -
Novolac               -             1608.5              -
PCF                   -             1608.5           2898.3
PAPF                  -             1618.2              -
PF (AQ)               -             1595                -

Displacement\
Absorption
Bandage         [[??].sub.NH2]   [[??].sub.C-N]   [d.sub.ip](OH)

Resol                 -                -             1365.5
Novolac               -                -             1363.6
PCF                   -                -             1369.4
PAPF               3414.8           1110.9           1380.9
                   3508
PF (AQ)               -             1238.3           1369.4

Table 2: RMN 1H displacement of bonds of the different resins.

Attribution\                                      -CH2O-
[delta] (ppm)    Aromatic        -OH             (bridge)

Resol            6.5-6.3          5.3             4.8
Novolac          6.7-7.3         05               -
PCF              6.6-7.1          4.8             -
PAPF             6.1-7.0          8.5             -
PF (AQ)          6.7-7.5          4.9             -

Attribution\     -CH2-           p-              p-
[delta] (ppm)    (bridge)        CH3             NH2

Resol            3.4-4           -               -
Novolac          3.3-3.9         -               -
PCF              3.8              2.2            -
PAPF             3.5             -               4.2
PF (AQ)          3.8             -               -

Attribution\     p-              N-              N-
[delta] (ppm)    [CH2N.sup.+]    CH2CH3          CH3

Resol            -               -               -
Novolac          -               -               -
PCF              -               -               -
PAPF
PF (AQ)          4.2              2.35/1.1       1.3

Table 3: RMN [sup.13]C displacement of bonds of the different resins.

Attribution                                         -CH2O-
[delta] (ppm)     Aromatic           -OH            (pont)

Resol              120-145            -               65
Novolac            113-120            -               -
PCF                120-135            -               -
PAPF                 130              -               -
PF (AQ)            118-135            -               -

Attribution         -CH2-
[delta] (ppm)      (pont)           p-CH3           p-NH2

Resol               35-40             -               -
Novolac             30-40             -               -
PCF                  30              21.5             -
PAPF                41-50             -
PF (AQ)              30               -               -

Attribution
[delta] (ppm)   p-[CH2N.sup.+]     N-CH2CH3         N-CH3

Resol                 -               -               -
Novolac               -               -               -
PCF                   -               -               -
PAPF                  -               -               -
PF (AQ)              60              51/8             49

Table 4: Rate of surviving bacteria versus time(min) and
concentration(g/l).

t (min)\              5           10          20          25
[C.sub.p] (g/l)

0.0025                50          30          20          20
0.0500                00          00          00          00
0.0780                00          00          00          00