Patent Publication Number: US-2023148597-A1

Title: Varnish

Description:
FIELD OF INVENTION 
     The invention relates to a varnish. In particular to a varnish for a banknote. 
     BACKGROUND OF THE INVENTION 
     Money, in particular banknotes, is in continuous circulation, passing between many different people. This means that it can easily become contaminated with microorganisms, such as  Staphylococcus , for example  S. aureus , and  Pseudomonas , for example  P. aeruginosa.    
     Banknotes can be varnished to increase the cleanliness of the banknote. However, while this may improve the cleanliness, it doesn&#39;t reduce the level of the microbes. 
     There is thus a need for improved varnishes for banknotes and resulting varnished banknotes that have improved resistance to microorganisms, for example resulting in reduced levels of microorganisms on the banknotes and/or a more bio-film resistant banknote. 
     SUMMARY OF THE INVENTION 
     We have found that by incorporating a lactam into the varnish for the banknote, the resulting banknote has improved resistance to microorganisms. 
     The invention relates in a first aspect to a varnished banknote comprising from 0.0015 to 2.5 wt. % of a lactam. 
     Preferably the lactam is present at a level of from 0.0015 to 1 wt. %. 
     Preferably the lactam is of formula (I) or (II): 
     
       
         
         
             
             
         
       
     
     wherein: 
     R 1  and R 2  are each independently selected from hydrogen, halogen, alkyl, cycloalkyl, alkoxy, oxoalkyl, alkenyl, heterocyclyl, heteroaryl, aryl and aralalkyl; and 
     R 3  is selected from hydrogen, hydroxyl, alkyl, cycloalkyl, alkoxy, oxoalkyl, alkenyl, heterocyclyl, heteroaryl, cycloalkyl, aryl, aralalkyl, —C(O)CR 6 ═CH 2 , and (CH 2 ) n N + (R a ) 3 , where n is an integer from 1 to 16, preferably 2 to 8, and where each R a  is independently H or C 1-4  alkyl; 
     R 4  and R 5  are independently selected from hydrogen, aryl, heterocyclyl, heteroaryl, and arylalkyl; and 
     R 6  is selected from hydrogen and methyl; and 
     R 7  is selected from hydrogen and —C(O)CR 6 ═CH 2 ; and 
     preferably, at least one of R 4  and R 5  is hydrogen. 
     It is preferred the in the lactam of formula (I) or (II), R 1 , R 4  and R 5  are H; R 3  is H, or (CH 2 ) n N + (CH 3 ) 3 , where n is an integer from 1 to 16, preferably 2 to 8; and R 2  is a phenyl group, or a mono-substituted phenyl group; preferably R 2  is selected from phenyl, 4-fluorophenyl, 2-fluorophenyl, 4-chlorophenyl, 3-chlorophenyl, 4-bromophenyl and 4-methylphenyl. 
     Preferably the lactam is a lactam selected from: 
     
       
         
         
             
             
         
       
     
     Where the lactam is cationic in nature, the cation can be used or with a suitable counterion (e.g. iodide). 
     More preferably the lactam is: 
     
       
         
         
             
             
         
       
     
     Most preferably the lactam is: 
     
       
         
         
             
             
         
       
     
     Preferably the lactam is in encapsulated form. 
     Preferably the varnish is a water-based varnish or an ultraviolet drying varnish. 
     Preferably the varnished banknote comprises a material selected from: cellulosic substrate, preferably cotton, or cotton in a blend with flax, abaca or eucalyptus pulp; polymer substrate, preferably polypropylene; or plastic substrate. 
     In a second aspect, the invention relates to the use of a lactam to either impart anti-biofilm properties to a banknote, or to inhibit biofilm growth on a banknote substrate. 
     Preferably, in these uses, the lactam is of formula (I) or (II), R 1 , R 4  and R 5  are H; R 3  is H, or (CH 2 ) n N + (CH 3 ) 3 , where n is an integer from 1 to 16, preferably 2 to 8; and R 2  is a phenyl group, or a mono-substituted phenyl group; preferably R 2  is selected from phenyl, 4-fluorophenyl, 2-fluorophenyl, 4-chlorophenyl, 3-chlorophenyl, 4-bromophenyl and 4-methylphenyl. 
     Preferably, in these uses, the lactam has the following structure: 
     
       
         
         
             
             
         
       
     
     Where the lactam is cationic in nature, the cation can be used or with a suitable counterion (e.g. iodide). 
     More preferably the lactam is: 
     
       
         
         
             
             
         
       
     
     Most preferably the lactam is: 
     
       
         
         
             
             
         
       
     
     DETAILED DESCRIPTION OF THE INVENTION 
     The indefinite article “a” or “an” and its corresponding definite article “the” as used herein means at least one, or one or more, unless specified otherwise. 
     It will be appreciated that, except where expressly provided otherwise, all preferences are combinable. 
    
    
     
       DESCRIPTION OF THE FIGURES 
         FIG.  1    is a graph showing the reduction of microorganism  P. aeruginosa  on treatment of cellulosic (paper) and polymer substrates with lactam varnish; 
         FIG.  2    are SEM images showing the reduction of microorganism  P. aeruginosa  on treatment of cellulosic (paper) and polymer substrates with lactam varnish; 
         FIG.  3    is a graph showing the reduction of microorganism  C. albicans  on treatment of cellulosic (paper) and polymer substrates with lactam varnish; 
         FIG.  4    are SEM images showing the reduction of microorganism  C. albicans  on treatment of cellulosic (paper) and polymer substrates with lactam varnish; 
         FIG.  5    is a graph showing the reduction of microorganism  S. aureus  on treatment of cellulosic (paper) and polymer substrates with lactam varnish; 
         FIG.  6    are SEM images showing the reduction of microorganism  S. aureus  on treatment of cellulosic (paper) and polymer substrates with lactam varnish 
     
    
    
     LACTAM 
     A lactam is a cyclic amide. Preferred lactams are γ-lactams which have 5 ring atoms. 
     Preferably the lactam is of formula (I) or (II): 
     
       
         
         
             
             
         
       
     
     wherein: 
     R 1  and R 2  are each independently selected from hydrogen, halogen, alkyl, cycloalkyl, alkoxy, oxoalkyl, alkenyl, heterocyclyl, heteroaryl, aryl and aralalkyl; and 
     R 3  is selected from hydrogen, hydroxyl, alkyl, cycloalkyl, alkoxy, oxoalkyl, alkenyl, heterocyclyl, heteroaryl, cycloalkyl, aryl, aralalkyl, —C(O)CR 6 ═CH 2 , and (CH 2 ) n N + (R a ) 3 , where n is an integer from 1 to 16, preferably 2 to 8, and where each R a  is independently H or C 1-4  alkyl; 
     R 4  and R 5  are independently selected from hydrogen, aryl, heterocyclyl, heteroaryl, and arylalkyl; and 
     R 6  is selected from hydrogen and methyl; and 
     R 7  is selected from hydrogen and —C(O)CR 6 ═CH 2 ; and 
     Preferably, at least one of R 4  and R 5  is hydrogen. 
     It will be appreciated that, where appropriate groups may be optionally substituted. Optional substituents may include halogens, C 1-4 alkyl, C 1-4 haloalkyl (for example, CF 3 ) and C 1-4 alkoxy. 
     Alkyls may, for example, be C 1-12 alkyls, such as C 1-6 alkyls. Aryls may, for example, be C 6-10 aryls, for example, phenyls. 
     Preferably, at least one of R 1  and R 2  is selected from heterocyclyl, heteroaryl, aryl and arylalkyl. 
     Preferably, R 1  is hydrogen. Preferably, R 3  is hydrogen, or (CH 2 ) n N + (R a ) 3 , where n is an integer from 1 to 16, preferably 2 to 8, and where each R a  is independently H or C 1-4  alkyl, more preferably R a  is CH 3 ; Preferably, R 4  is hydrogen. Preferably, R 5  is hydrogen. Preferably, R 6  is hydrogen. Preferably, R 7  is hydrogen. Preferably, R 2  is aryl or aralalkyl. More preferably, R 2  is a phenyl group or a substituted phenyl group, for example, a mono-substituted phenyl group. Substitution may be ortho, meta, or para. Preferred substituents include halogen and methyl. For example, and without limitation, R 2  may be selected from phenyl, 4-fluorophenyl, 2-fluorophenyl, 4-chlorophenyl, 3-chlorophenyl, 4-bromophenyl and 4-methylphenyl. 
     More preferably in the lactam of formula (I) or (II), R 1 , R 4  and R 5  are H; R 3  is H, or (CH 2 ) n N + (CH 3 ) 3 , where n is an integer from 1 to 16, preferably 2 to 8; and R 2  is a phenyl group, or a mono-substituted phenyl group; preferably R 2  is selected from phenyl, 4-fluorophenyl, 2-fluorophenyl, 4-chlorophenyl, 3-chlorophenyl, 4-bromophenyl and 4-methylphenyl. 
     Even more preferably the lactam is of formula (I), R 1 , R 4  and R 5  are H; R 3  is H, or (CH 2 ) n N + (CH 3 ) 3 , where n is an integer from 1 to 16, preferably 2 to 8; and R 2  is a phenyl group, or a mono-substituted phenyl group; preferably R 2  is selected from phenyl, 4-fluorophenyl, 2-fluorophenyl, 4-chlorophenyl, 3-chlorophenyl, 4-bromophenyl and 4-methylphenyl. 
     Where the lactam is cationic in nature, it can be used as such, or suitably with a counterion (e.g. iodide) 
     Most preferably the lactam is a lactam selected from: 
     
       
         
         
             
             
         
       
     
     Where the lactam is cationic in nature, the cation can be used or with a suitable counterion (e.g. iodide). 
     More preferably the lactam is: 
     
       
         
         
             
             
         
       
     
     Most preferably the lactam is: 
     
       
         
         
             
             
         
       
     
     Preferably the lactam is encapsulated. 
     Suitably, the encapsulated lactam is a polymer encapsulated lactam. 
     The encapsulated lactam may be encapsulated in a polymer selected from a poly urea polymer, a melamine-formaldehyde copolymer; a urea formaldehyde copolymer and mixtures thereof. 
     Suitably the polymer is a condensation polymer. For example, the polymer may be a condensation polymer of produced from a diamine and a diisocyanate. 
     For example, the polymer may be or may comprise a polyurea of Formula P1: 
     
       
         
         
             
             
         
       
     
     wherein R P1  comprises a phenylene and R P2  is an alkylene. 
     For example, R P1  may be —CH 2 -phenylene; in other words, the polymer may be derived from polymethylene polyphenyl isocyanate. 
     For example, R P2  may be a straight chain alkylene of formula —(CH 2 ) m —. In some cases, m is an integer from 2 to 10, for example from 2 to 8, for example from 4 to 8, for example, 6 (in other words, R P2  may be hexylene). 
     In other words, the lactam may be encapsulated in a polymer formed from polymethylene polyphenyl isocyanate and hexamethylenediamine. 
     In some cases, the polymer and/or encapsulate structure is selected and/or configured to permit controlled or triggered release. For example, the encapsulate may dissolve at a pre-determined rate under certain conditions. For example, the encapsulate may release in response to a trigger. The trigger may be, for example, the presence or a certain concentration of acid, base, a salt, an enzyme; or a non-chemical trigger such as ultrasound or light. 
     Suitably, the lactam is encapsulated to form particles whose average diameter is from about 10 nanometers to about 1000 microns, preferably from about 50 nanometers to about 100 microns, more preferably from about 2 to about 40 microns, even more preferably from about 4 to 15 microns. A particularly preferred range is from about 5 to 10 microns, for example 6 to 7 microns. The capsule distribution can be narrow, broad or multimodal. Multimodal distributions may be composed of different types of capsule chemistries. 
     The encapsulation process suitably is done in a carrier oil, which may be a ketone. For example, the carrier oil may be a C 5-20 alkyl ketone, for example a C 5-15 alkyl ketone, for example a C 5-10 alkyl ketone, for example a C 6-8 alkyl ketone, such as a C 7 alkyl ketone. The alkylketone may be branched or straight-chain. Preferably, it is straight chain. The oxo group of the alkyl ketone may be located at C2; in other words, the alkylketone may be an alkyl-2-one. A preferred carrier oil is 2-heptanone. 
     Levels of Lactam 
     The lactam is present at a level of from 0.0015 to 2.5 wt. %. This equates to from 15 to 25,000 ppm (parts per million). 
     The lactam is preferably present at a level of from 0.0015 to 1 wt. % (15 to 10,000 ppm). For example, the lactam may be suitably present at levels of 0.0015 to 0.5 wt. % (15 to 5,000 ppm), or even 0.05 to 0.5 wt. % (50 to 5,000 ppm), or even 0.05 to 0.1 wt. % (50 to 1,000 ppm). 
     Varnish 
     Post-print varnish is a thin protective layer on both sides of the banknote. It is applied as last step in the printing process. It smoothens the rough cotton surface and protects the printing including the security features. Studies reported varnished banknotes stay clean for a longer time increase circulation time. 
     A varnish may be considered a combination of liquid resins, solvents and additives (wax, adhesive, photo initiator, etc.), which is applied in the form of a continuous transparent ink layer. The varnish, once dry, makes the banknote surface impermeable to soiling. The first varnishes used on banknotes were so called solvent-based varnishes. The following types of varnish may be used:
         Water-based.   Ultraviolet drying (UV).   Double layer, which can be applied in two ways: i) two layers of waterbased varnish, and ii) an undercoat of water-based varnish with a top coat of UV varnish.       

     Water-based varnishes dry slowly under infrared (IR) and/or warm air. During the drying process, part of the varnish layer originally printed on the banknote evaporates, leaving a final varnish layer that is thinner than the original and almost invisible. UV varnishes are exposed to ultraviolet radiation which causes the particles in the varnish layer to bind and so dry very quickly, such that the thickness of the printed varnish layer is not reduced. 
     Preferably the varnish is a water-based varnish or an ultraviolet drying varnish. 
     Varnished Banknote 
     The varnish is applied to a banknote substrate. 
     Preferably the banknote comprises a material selected from: cellulosic substrate; polymer substrate; or plastic substrate. 
     Preferred cellulosic substrates are cotton, or cotton in a blend with flax, abaca (banana plant) or eucalyptus pulp. 
     Preferred polymer substrates include polypropylene, in particular biaxially orientated polypropylene (BOPP). 
     Further Ingredients 
     The varnish may further comprise standard varnish ingredients such as liquid resins, solvents, wax, adhesive, photo initiator. 
     Methods of Manufacture of Varnishing Banknotes 
     Preferred methods of manufacture for varnishing banknotes are the following options:
         a) Varnishing the substrate   b) Varnishing the banknote after printing (post-varnishing)   c) Varnishing with two layers, one applied to the substrate and the other to the banknote after printing       

     Varnishing Paper Banknotes (Post-Print) 
     Banknote paper is a porous material which readily absorbs damp, contaminant particles and microorganisms. Varnishing creates a layer protecting the banknote against surface soiling, enabling it to remain in circulation longer. Varnishing is currently used by numerous central banks worldwide, having become one of the solutions available to reduce the cost of cash by extending the lifetime of circulating banknotes. 
     Varnishing Polymer and Hybrid Substrate Banknotes (Pre- and Post-Printing) 
     In the case of polymer and hybrid substrates it is preferable to apply any pre-varnish coat for the ink to better adhere to the substrate. This is followed by a post-varnishing coat to reduce wear on the print during the banknote&#39;s circulating lifetime. These varnishes contribute to improving the mechanical and anti-soiling properties of these substrates with respect to conventional banknotes printed on cotton paper. 
     Varnishing protects against soiling. Nevertheless, the effectiveness of the anti-soiling protection depends both on the type of varnish used and the thickness of the varnish layer coating the banknote. In the case of thin layers, it is worth mentioning that if banknotes are in circulation for a long-time surface cracks may appear in the varnished surface, which can collect dirt and so cause dark lines to appear. One solution to this problem is to apply a double coat of varnish. 
     The invention will be further described with the following non-limiting examples. 
     EXAMPLES 
     Example 1—Preparation of Examples of Preferred Lactams 
     Preparation of 4-(4-chlorophenyl)-5-hydroxy-5-methylfuran-2(5H)-one 
     
       
         
         
             
             
         
       
     
     1-(4-Chlorophenyl)propan-2-one (40.00 g, 34.75 mL, 237.2 mmol), glyoxylic acid monohydrate (32.75 g, 355.8 mmol) and phosphoric acid (69.74 g, 711.7 mmol) were combined at room temperature before heating to 85° C. overnight. After cooling to room temperature, the mixture was poured into a mixture of water (500 mL) and ethyl acetate (500 mL). The layers were separated and the aqueous phase extracted with ethyl acetate (500 mL). The combined organic layers were washed with a 1:1 mixture of water and brine (2×500 mL), dried (MgSO 4 ) and concentrated under reduced pressure to yield 4-(4-chlorophenyl)-5-hydroxy-5-methylfuran-2(5H)-one (66.00 g, &gt;100% yield) as a brown oil. The material was used in the next step without further purification. 
     Preparation of 4-(4-chlorophenyl)-5-hydroxy-5-methyl-1H-pyrrol-2(5H)-one 
     
       
         
         
             
             
         
       
     
     4-(4-Chlorophenyl)-5-hydroxy-5-methylfuran-2(5H)-one (66.00 g, 293.8 mmol) was dissolved in thionyl chloride (196.8 g, 120.0 mL, 1654 mmol) and heated at 40° C. for 1 hour, then 80° C. for 2 hours. The mixture was concentrated under reduced pressure and azeotroped with 2-methyltetrahydrofuran (200 mL). The residue was diluted with 2-methyltetrahydrofuran (160 mL) and this solution added to a cooled stirring mixture of 28% ammonia in water (180 mL) in 2-methyltetrahydrofuran (20 mL) at 0° C. The mixture was warmed to room temperature and stirred overnight. Water (100 mL) and ethyl acetate (200 mL) were added and the layers separated. The aqueous phase was extracted with ethyl acetate (200 mL), and the combined organic extracts dried (MgSO 4 ) and concentrated under reduced pressure. Purification by dry flash column chromatography (5-60% ethyl acetate in heptane) yielded 4-(4-chlorophenyl)-5-hydroxy-5-methyl-1H-pyrrol-2(5H)-one (23.18 g, 35% yield) as a cream coloured solid. 
       1 H NMR (400 MHz, d 6 -DMSO) 8.55 (brs, 1H), 7.88-7.83 (m, 2H), 7.51-7.46 (m, 2H), 6.37 (d, 1H), 6.32 (s, 1H), 1.45 (s, 3H) 
     UPLC (Basic) 1.51/5.00 min, 100% purity, M+H +  224 
     MP 177° C. 
     Preparation of 4-(4-chlorophenyl)-5-methylene-1H-pyrrol-2(5H)-one 
     
       
         
         
             
             
         
       
     
     To a cooled solution of 4-(4-chlorophenyl)-5-hydroxy-5-methyl-1H-pyrrol-2(5H)-one (10.00 g, 44.51 mmol) in dry dichloromethane (100 mL) at 0° C. was added a solution of boron trifluoride diethyl etherate (8.213 g, 7.142 mL, 57.87 mmol) in dry dichloromethane (45 mL) over 15 minutes. The mixture was stirred at 0° C., before slowly warming to room temperature and stirring for 2 hours. The reaction was quenched with ice-water (100 mL) and the layers separated. The aqueous layer was extracted with dichloromethane (100 mL), and the combined organic layers washed with a 1:1 mixture of water and saturated aqueous sodium hydrogen carbonate solution (100 mL), dried (MgSO 4 ) and filtered. Silica was added to the filtrate and the mixture stirred for 10 minutes before filtering through a plug of silica, washing through with dichloromethane followed by a 3:1 mixture of dichloromethane:diethyl ether. Fractions containing the desired product were combined and concentrated under reduced pressure. Upon concentration a precipitate formed, which was collected by filtration, washing with diethyl ether, to yield 4-(4-chlorophenyl)-5-methylene-1H-pyrrol-2(5H)-one (5.25 g, 57% yield) as a cream coloured solid. 
       1 H NMR (400 MHz, d 6 -DMSO) 10.10 (s, 1H), 7.54-7.47 (m, 4H), 6.36 (s, 1H), 5.04 (t, 1H), 4.85 (s, 1H) 
     UPLC (Basic) 1.87/5.00 min, 100% purity, M+H +  206 
     MP 182° C. 
     Preparation of 5-hydroxy-5-methyl-4-(p-tolyl)furan-2(5H)-one 
     
       
         
         
             
             
         
       
     
     1-(p-Tolyl)propan-2-one (25.00 g, 24.00 mL, 168.7 mmol), glyoxylic acid monohydrate (23.29 g, 253.0 mmol) and phosphoric acid (49.60 g, 506.1 mmol) were combined at room temperature before heating at 90° C. overnight. After cooling to room temperature, the mixture was poured into a stirring mixture of ice-water (400 mL) and ethyl acetate (400 mL). The layers were separated and the organic phase washed with water (100 mL), dried (MgSO 4 ) and concentrated under reduced pressure. The mixture was azeotroped with 2-methyltetrahydrofuran (50 mL) to yield 5-hydroxy-5-methyl-4-(p-tolyl)furan-2(5H)-one (16.50 g, 48% yield) as a brown solid. 
       1 H NMR (400 MHz, d 6 -DMSO) 7.86 (s, 1H), 7.75 (d, 2H), 7.28 (d, 2H), 6.59 (s, 1H), 2.32 (s, 3H), 1.61 (s, 3H) 
     Preparation of 5-hydroxy-5-methyl-4-(p-tolyl)-1H-pyrrol-2(5H)-one 
     
       
         
         
             
             
         
       
     
     5-Hydroxy-5-methyl-4-(p-tolyl)furan-2(5H)-one (16.50 g, 80.80 mmol) was dissolved in thionyl chloride (48.06 g, 29.47 mL, 404.0 mmol) and heated at 50° C. for 1 hour, before heating at reflux for 1 hour. After cooling to room temperature, the mixture was concentrated under reduced pressure and azeotroped with 2-methyltetra-hydrofuran (2×50 mL). The residue was diluted with 2-methyltetrahydrofuran (60 mL) and this solution added to a cooled stirring mixture of 28% ammonia in water (55 mL, 808.0 mol) in 2-methyltetrahydrofuran (10 mL) at 0° C. The mixture was warmed to room temperature and stirred overnight. 2-Methyltetrahydrofuran was removed under reduced pressure, and the residue diluted with water (200 mL) and diethyl ether (100 mL) and the mixture stirred for 20 minutes at room temperature. The solids were collected by filtration and stirred in water (100 mL) and diethyl ether (50 mL) at room temperature for 10 minutes. The solids were collected by filtration and washed with water, diethyl ether and dried under vacuum at 50° C. to yield 5-hydroxy-5-methyl-4-(p-tolyl)-1H-pyrrol-2(5H)-one (10.49 g, 31% yield) as a light beige solid. 
       1 H NMR (400 MHz, d 6 -DMSO) 8.44 (brs, 1H), 7.73 (d, 2H), 7.21 (d, 2H), 6.24 (s, 2H), 2.29 (s, 3H), 1.45 (s, 3H) 
       13 C NMR (400 MHz, d 6 -DMSO) 170.4 (s, 1C), 161.1 (s, 1C), 139.8 (s, 1C), 129.7 (s, 2C), 128.9 (s, 1C), 128.2 (s, 2C), 119.1 (s, 1C), 87.8 (s, 1C), 26.7 (s, 1C), 21.5 (s, 1C) 
     UPLC (Basic) 1.41/5.00 min, 100% purity, M+H +  204 
     MP 178° C. Decomposition 
     Preparation of 5-methylene-4-(p-tolyl)-1H-pyrrol-2(5H)-one 
     
       
         
         
             
             
         
       
     
     To a cooled solution of 5-hydroxy-5-methyl-4-(p-tolyl)-1H-pyrrol-2(5H)-one (8.68 g, 42.7 mmol) in dry dichloromethane (87 mL) at 0° C. was added a solution of boron trifluoride diethyl etherate (6.85 g, 5.96 mL, 55.5 mmol) in dry dichloromethane (40 mL) over 15 minutes. After 1 hour the mixture was allowed to slowly warm to room temperature. After a further 3 hours, the reaction was diluted with dichloromethane (50 mL) and ice-water (100 mL) and stirred for 10 minutes. The layers were separated and the organic layer washed with water (100 mL), a 1:1 mixture of water and saturated aqueous sodium hydrogen carbonate solution (100 mL) and brine (100 mL) and the organic layer filtered through Celite, washing with dichloromethane. Any excess water was removed by pipette before drying the filtrate (MgSO 4 ) and concentrating under reduced pressure to a brown solid. The solids were stirred in hot dichloromethane (120 mL) for 15 minutes before slowly cooling to room temperature and then 0° C. The solids were collected by filtration to yield 5-methylene-4-(p-tolyl)-1H-pyrrol-2(5H)-one (3.87 g, 49% yield) as a yellow solid. Silica was added to the filtrate and the mixture stirred for 10 minutes before filtering through a plug of silica, washing through with dichloromethane and then a 4:1 mixture of dichloromethane:diethyl ether. The filtrate was concentrated under reduced pressure to yield 5-methylene-4-(p-tolyl)-1H-pyrrol-2(5H)-one (0.58 g, 7%) as a yellow solid. Total yield of 5-methylene-4-(p-tolyl)-1H-pyrrol-2(5H)-one (4.45 g, 56% yield). 
       1 H NMR (400 MHz, d 6 -DMSO) 10.11 (brs, 1H), 7.35 (d, 2H), 7.25 (d, 2H), 6.25 (s, 1H), 5.01 (s, 1H), 4.85 (s, 1H), 2.31 (s, 3H) 
     UPLC (Basic) 1.83/5.00 min, 100% purity, M+H +  186 
     MP 200° C. Decomposition 
     Example 2 —Pseudomonas, Staphylococcus  and  Candida  Static Biofilm Growth Inhibition of as Measured by Viability on Paper (Cellulosic) and Polymer Substrates 
     The lactam used in these experiments was 4-(4-chlorophenyl)-5-methylene-pyrrol-2-one and is given the code 488. The structure is:— 
     
       
         
         
             
             
         
       
     
     Lactam was incorporated into ultraviolet drying varnish (UV Guard, Gleitsmann security) to a final concentration of 7, 100 and 275 mg/l (ppm). This equates to 0.0007 wt. %, 0.01 wt. % and 0.0275 wt. % respectively. 
     The varnish was printed onto paper (cellulosic) and polymer banknote substrate using an IGT proofer (IGT testing systems). 
       P. aeruginosa  (PA01),  S. aureus  (Newman&#39;s strain) were cultured overnight at 37° C. on TSA plates.  C. albicans  3153A was cultured for 2 days at 30° C. on MEA plates. Prior to use in tests, colonies of each organism were added to 20 mL of brain-heart infusion broth (BHI) containing 5 mL of glass beads and homogenised for 30 seconds. The optical density of each organism was measured and adjusted to give 1×10 7  cfu/mL. Substrates where cut into 9 cm 2  d discs and placed in the well of 6-well plates. Inoculum (0.3 ml) was placed directly on polymer and paper substrates (with and without lactam) and incubated overnight in appropriate broth. Textiles were subsequently removed, washed in PBS and metabolism assessed by AlamarBlue on a plate reader. Data were presented as percentage of no-lactam control. 
     SEM Sample Preparation 
     Fixative was prepared as described (Erlandsen, Kristich, Dunny, Wells,  J. Histochem Cytochem,  2004), using 2% para-formaldehyde, 2% gluteraldehyde and 0.15M Sodium Cacodylate and 0.15% Alcian Blue, pH 7.4. The fixative was applied (enough to cover the biofilms) to the wells containing the biofilms on suitable substrates for 2 hours, this varied between 2 and 22 hours. 
     
       
         
           
               
               
               
               
               
             
               
                   
               
               
                 Component 
                 5 ml 
                 10 ml 
                 15 ml 
                 20 ml 
               
               
                   
               
             
            
               
                  8% Para-formaldehyde 
                  1.25 ml 
                  2.5 ml 
                  3.75 ml 
                   5 ml 
               
               
                 0.3M Sodium Cacodylate 
                   2.5 ml 
                    5 ml 
                   7.5 ml 
                   10 ml 
               
               
                 25% Gluteraldehyde 
                   0.4 ml 
                  0.8 ml 
                   1.2 ml 
                  1.6 ml 
               
               
                 Distilled water 
                  0.85 ml 
                  1.7 ml 
                  2.55 ml 
                  3.4 ml 
               
               
                 Alcian Blue 
                 0.0075 g 
                 0.015 g 
                 0.0225 g 
                 0.03 g 
               
               
                   
               
            
           
         
       
     
     Paraformaldehyde was prepared at 60° C. with 80 ml distilled water and 8 g of paraformaldehyde using a hot plate and magnetic stirrer. NaOH was added drop by drop until solution cleared and was adjusted to pH 7.2 with HCl. 
     Following fixation, the fixative solution was removed and 0.15M Sodium Cacodylate buffer was added to the samples. The samples were then stored in the fridge until processing. Samples were washed 3×5 mins with 300 ml fresh buffer to remove any remaining glutaraldehyde. Subsequently, a solution of 1% Osmium tetroxide (OsO4) was prepared 1:1 with 0.15M Sodium Cacodylate buffer, and added to the samples before incubating for 1 hour at room temperature. Samples were rinsed with distilled water 3×10 mins. 
     0.5% aqueous Uranyl acetate was then added to the samples before incubation in the dark for 30 mins at room temperature. 
     Samples were then dehydrated in an ascending ethanol series: 
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                 Alcohol 
                 Time 
               
               
                   
                   
               
             
            
               
                   
                 30% alcohol 
                 2 × 5 mins 
               
               
                   
                 50% alcohol 
                 2 × 5 mins 
               
               
                   
                 70% alcohol 
                 2 × 5 mins 
               
               
                   
                 90% alcohol 
                 2 × 5 mins 
               
               
                   
                 Absolute alcohol 
                 4 × 5 mins 
               
               
                   
                 Dried absolute alcohol 
                 2 × 5 mins 
               
               
                   
                   
               
            
           
         
       
     
     Samples were transferred from the original 24-well plate into a petri dish of hexamethyldisilazane (HMDS) for 5 mins, then to a second dish for 5 mins before being placed in a new 24-well plate lined with filter paper. The plate was then placed in a desiccator overnight to allow evaporation and drying or samples. 
     After sputter coating with gold-palladium in an argon filled chamber, samples were viewed under a JEOL JSM-6400 scanning electron microscope and images were assembled using the Photoshop software. 
     Table 2 and  FIGS.  1  &amp;  2    show the effect of reduction of microorganism  P. aeruginosa  on treatment of cellulosic (paper) and polymer substrates with lactam varnish, using a varnish control and varnish with lactam incorporated 
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Reduction of microorganism  P .  aeruginosa  on  
               
               
                 treatment of cellulosic (paper) and polymer 
               
               
                 substrates with lactam varnish 
               
            
           
           
               
               
               
               
            
               
                   
                   
                 Paper 
                 Polymer 
               
               
                   
                   
                 substrate 
                 substrate 
               
               
                   
                   
                 Mean 
                 Mean 
               
               
                   
                   
               
               
                   
                 Varnish + 275 ppm lactam 
                 38.50 
                 36.92 
               
               
                   
                 Varnish + 100 ppm lactam 
                 89.31 
                 40.81 
               
               
                   
                 Varnish + 7 ppm lactam 
                 94.46 
                 92.93 
               
               
                   
                 Varnish Control 
                 92.52 
                 95.53 
               
               
                   
                   
               
            
           
         
       
     
     Table 3 and  FIGS.  3  &amp;  4    show the effect of reduction of microorganism  C. albicans  on treatment of cellulosic (paper) and polymer substrates with lactam varnish, using a varnish control and varnish with lactam incorporated 
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 Reduction of microorganism  C .  albicans  on  
               
               
                 treatment of cellulosic (paper) and polymer  
               
               
                 substrates with lactam varnish 
               
            
           
           
               
               
               
            
               
                   
                 Paper 
                 Polymer 
               
               
                   
                 substrate 
                 substrate 
               
               
                   
                 Mean 
                 Mean 
               
               
                   
               
            
           
           
               
               
               
            
               
                 Varnish + 275 ppm lactam 
                 38.24 
                 38.61 
               
               
                 Varnish + 100 ppm lactam 
                 78.72 
                 50.74 
               
               
                 Varnish + 7 ppm lactam 
                 104.96 
                 84.88 
               
               
                 Varnish Control 
                 104.62 
                 90.44 
               
               
                   
               
            
           
         
       
     
     Table 4 and  FIGS.  5  &amp;  6    show the effect of reduction of microorganism  S. aureus  on treatment of cellulosic (paper) and polymer substrates with lactam varnish, using a varnish control and varnish with lactam incorporated 
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 Reduction of microorganism  S .  aureus  on  
               
               
                 treatment of cellulosic (paper) and polymer 
               
               
                 substrates with lactam varnish 
               
            
           
           
               
               
               
            
               
                   
                 Paper  
                 Polymer 
               
               
                   
                 substrate 
                 substrate 
               
               
                   
                 Mean 
                 Mean 
               
               
                   
               
            
           
           
               
               
               
            
               
                 Varnish + 275 ppm lactam 
                 51.45 
                 63.83 
               
               
                 Varnish + 100 ppm lactam 
                 72.84 
                 75.89 
               
               
                 Varnish + 7 ppm lactam 
                 86.60 
                 92.63 
               
               
                 Varnish Control 
                 97.77 
                 101.75 
               
               
                   
               
            
           
         
       
     
     It can be seen from the experimental data that the lactam varnished substrates have reduced microorganism levels, particularly for the polymer substrate varnished with lactam and particularly for varnishes comprising greater than 15 ppm lactam.