GB2404337A

GB2404337A - Two-part sterilant system

Info

Publication number: GB2404337A
Application number: GB0410203A
Authority: GB
Inventors: Bruce Philip Green
Original assignee: Individual
Current assignee: Individual
Priority date: 2003-07-29
Filing date: 2004-05-07
Publication date: 2005-02-02
Anticipated expiration: 2024-05-07
Also published as: GB2404337B; GB0410203D0

Abstract

A two-part sterilant system comprising a first part comprising a reagent in a carrier medium and a second part which is miscible with the first part and which comprises a second reagent in a carrier medium. The first reagent will react with the second reagent when mixed to provide a sterilising composition. The first reagent preferably comprises sodium chlorite or sodium chlorate. The second reagent may comprise a solution of citric acid, boric acid, and sorbic acid. The reagents react to form chlorine dioxide in situ. The first part is contained in a dispenser 2 and the second part is adsorbed or impregnated in at least one fabric member (18, Fig. 2) in a sealed container 20. The first part is preferably contained within a pump dispenser or trigger-operated dispenser and dispensed as a fluid, foam, paste, powder, spray or a gel. The fabric member may be a wipe or a wound dressing. Each of the fabric members may be provided in its own separate sealed sachet.

Description

- 1 2404337

STERILANT SYSTEM

FIELD OF THE INVENTION

The present invention relates to a two-part sterilant system, notably to a system for producing chlorine dioxide (ClO2).

The invention is particularly for use in sterilizing medical supplies and equipment, but it is not limited to these uses.

BACKGROUND TO THE INVENTION

Two-part sterilizing solutions are used in applications where the active sterilizing ingredient is unstable over time. The solution is therefore prepared in situ shortly before it is to be used. A particularly important sterilizing agent is chlorine dioxide, which may be formed from mixtures of various reagents including: chlorite and acid; chlorate, peroxide and acid; and chlorite, hypochlorite, and a suitable buffer. Chlorine dioxide has excellent sterilizing and bactericidal properties, and oral ingestion in man and animals has been shown to be relatively safe.

The cleaning of endoscopes and other medical equipment with suitable chlorine dioxide solutions is known from earlier patents in the name of the present inventor, for example, European Patent Number 0 785 719 and United States Patent Numbers 5,696,046 and 6,007,772, the contents of which are hereby incorporated by reference.

It is not always convenient to mix up batches of solutions for use in sterilizing equipment. For wiping down (rather than thoroughly cleaning inside and out) of endoscopes and 3960.a4 {2)vl.doc 06 May 2004 - 2 probes, wipes of alcohol, general-purpose detergent, or soapy water are generally used, but these are not as effective as chlorine dioxide. It is desirable to be able readily to make up small quantities of two-component sterilizing agents when desired and to be able to make such agents up in a form in which they may be readily handled for a particular application.

SUMMARY OF THE INVENTION

According to a first aspect of the present invention there is provided a two-part sterilant system comprising: (a) a first part comprising a first reagent in a carrier medium; and (b) a second part which is miscible with the first part and which comprises a second reagent in a carrier medium; wherein the first reagent and the second reagent will react when mixed to provide a sterilising composition; characterized in that the first part is contained in a pump dispenser whereby it may be dispensed as a fluid, and in that the second part is absorbed or impregnated in at least one fabric member in a sealed container.

The term "fluids is used herein to include liquids, foams, sprays, pastes, aerosols, powders, sole and gels. It is particularly preferred that the first part is dispensed as a; foam or a spray to facilitate its coverage of a desired area of the fabric member. Optionally, the dispenser may have a relatively large dispensing head, for supplying the fluid over all or a substantial part of a surface of the fabric member. For example, the dispensing head may take the form of a rose or sprinkler with a multitude of small orifices to 3960 ad (2)vl doc 06 May 2004 - 3 - spread the fluid over the fabric member.

The pump dispenser is preferably a trigger-operated dispenser, both for convenience and to facilitate the dispensing of metered quantities. However, other pump dispensers could be used, for example, a squeeze bottle with a suitable spray or foam nozzle. The invention will, for convenience, be described hereinafter with reference to the use of a trigger-operated dispenser, but it is to be understood that it is not limited to this embodiment.

By putting up the first part in a trigger-operated dispenser, small quantities may be readily dispensed without risk of spillage. Preferably the dispenser comprises a sprayer apparatus that provides the first part as a foam so that it is at least partly form-retaining and can be readily seen and manipulated. We have also found that providing the first part in a foam may have the beneficial effect of reducing the odour of chlorine dioxide when the wipe is activated. The invention will for convenience be described with reference to this preferred embodiment, but it will be understood that the invention is not limited to this embodiment.

The trigger sprayer may include a mixing chamber to facilitate mixing of the first part with air, for example as described in United States patent number 5,337,929.

The fabric members may be formed from any suitable fabrics, either woven or non-woven. They may be of natural or man made fibres, for example polyester, cotton, cellulose or mixtures thereof. Other suitable fabrics will be well known to those skilled in the textile or fabric arts.

3960.a42)vl.doc 06 May 2004 - 4 The fabric member may comprise a fabric wipe or cloth, or a gauze, pad, or other wound dressing material. Once prepared, the fabric member will have biocidal properties and may be used to dress wounds, ulcers, or the like while promoting a sterile local environment around the wound. For convenience, the invention will be described hereinafter with reference to the use of a fabric wipe, but it is to be understood that the invention is not limited to this embodiment.

By providing the second part absorbed in a fabric wipe, a sterilizing wipe may readily be prepared by applying the first part to the wipe. The user may fold the wipe or rub two halves together to facilitate mixing. The wipes are particularly useful for cleaning, disinfecting, and sterilising surfaces and equipment, notably in a medical environment.

The first part may include a coloured component so that a visual indication of the coverage of the wipe with the first part can be made.

In a preferred embodiment, at least one of the first and second parts is provided with an indicator reagent that 2S changes colour to show that sufficient mixing has taken place. Where the first part and the second part are of different pH, the indicator may be a pH-sensitive indicator.

Suitable indicators are well known to those skilled in the art, nonlimiting examples including: phenol red, litmus, thymol blue, pentamethoxy red, tropeolin oO, 2,4- dinitrophenol, methyl yellow, methyl orange, bromophenol blue, tetrabromophenol blue, alizarin sodium sulphonate, a 3960.a4 (2 jvl doc 06 May 2004 - 5 - naphthyl red, p-ethoxychrysoidine, bromocresol green, methyl red, bromocresol purple, chlorophenyl red, bromothymol blue, p-nitrophenol, azolitmin, neutral red, rosalic acid, cresol red, a-naphtholphthalein, tropeolin OOO, phenolphthalein, a naphtholbenzein, thymolphthalein, wile blue, alizarin yellow, diazo violet, tropeolin O. nitramine, Poirrer's blue, trinitrobenzoic acid, and mixtures thereof. It is preferred that the indicator is selected so that both parts are separately colourless and the colour develops when the two parts are mixed.

Alternatively, or additionally, one or more fluorescent additives may be included so that the mixture fluoresces to indicate mixing. Non-limiting examples of suitable fluorescing agents include: 4-methylumbelliferone, 3,6- dihydroxanthone, quinine, thioflavin, 1-napthol, harmine, coumarin, acridine orange, cotarmine, and mixtures thereof.

The indicator (colour change or fluorescent) may be included in either part. Preferred proportions by weight are about 0.1 to 10%, notably about 0.5 to 2%.

The carrier mediums may be fluids such as liquids or sols, or they may be more form-retaining or viscous compositions such as gels or pastes. It is preferred that at least one reagent is present in an aqueous fluid, although other additives may of course be present. Preferably both reagents are put up in aqueous fluids.

The trigger-operated dispenser may be a conventional atomiser or foamer, or other manual pump in which the contents are expelled manually by operation of the trigger by the user.

3960.a4 {2Jvl.doc 06 May 2004 - 6 Alternatively, the dispenser may contain a propellant to dispense the contents when operation of the trigger opens a valve, as is well known in applications such as shaving foam canisters and the like. Suitable dispensers will be well known to those skilled in the art.

The preferred sterilizing agent is chlorine dioxide, which may be formed from suitable known reagents. In a preferred embodiment one reagent is a chlorite (notably sodium chlorite) and the other is an acid, preferably with a buffer.

Suitable acids include lactic acid, citric acid, boric acid, phosphoric acid, acetic acid, sorbic acid, ascorbic acid, hydrochloric acid or mixtures thereof. In a preferred embodiment a mixture of acids is used, notably a mixture of citric, sorbic and boric acids.

A particularly preferred system is as described in EP 0 785 719, with the corrosion inhibitors optionally not included, and with other additives as desired for particular applications. In addition to suitable indicators, optional additives include foam-promoting agents or stabilizers, humectants, essential oils and fragrances. Other sterilising agents may also be employed; for example chlorine or oxygen.

Chlorine may be produced by reaction between a hypochlorite such as sodium hypochlorite, and a suitable acid or buffer.

Oxygen may be produced by reaction between a peroxide and a catalyst such as catalase, optionally in the presence of a buffer. For convenience hereinafter, the invention will be described with reference to chlorine dioxide as the sterilizing agent.

Suitable foam promoters will be well known to those skilled 3960. ad (2) vl doc 06 May 2004 l - 7 in the art. Non-limiting examples include: sodium laureth sulphate, ammonium lauryl sulphate, cocamide DEA, cocamidopropyl betaine, sodium lauryl sarcosinate, cocamidopropylamine oxide, monoethanolamine lauryl sulphate, cocamidopropyl hydroxysultaine, cocoyl sarcosinate. Anionic, cationic, non-ionic and amphoteric surfactants may be employed depending on the chemistry of the reagents. The foam promoters are selected to provide a stable foam structure. The foam promoter may comprise from about 0.1 to 50% by weight of the first part, notably from about 1 to 10%, preferably from about 3 to 6%.

Suitable foam stabilizers well known to those skilled in the art may also be used, in proportions similar to those for the foam-promoters. Nonlimiting examples include: alkanolamides, for example monoethanolamides and diethanolamides, amine oxides, betaines, protein hydrolyeates and cellulose derivatives such as carboxymethylcellulose.

In a preferred embodiment, a humectant is included in at least one of the first and second parts. Humectants serve to reduce the rate of evaporation of components and improve product feel if direct skin contact is involved. We have found that the use of a humectant reduces the volatility of chlorine dioxide, which reduces the odour of chlorine dioxide and prolongs the life of the activated mixture. Non-limiting examples of suitable humectants include sodium lactate and polyols, for example glycerine, sorbitol, propylene glycol, diethylene glycol and ethylene glycol. The humectant may be present in any desired amount, particularly from about 0.1 to 50% by weight, notably from about 0.5 to 10%, preferably from about 1 to 3%.

3960.a42Jvl.doc 06 May 2004 - 8 Where one of the reagents is basic or oxidising, for example sodium chlorite, it is particularly preferred that this reagent is provided in the trigger dispenser rather than in the wipe, because such reagents may react with the fabric over time. Preferably the optional humectant is included in the first part, with the sodium chlorite or other first reagent.

The first and/or second part may further include a biocide to ensure that, in the event of poor mixing of the parts, a biocidal effect is still present. The first and/or second part may also include a preservative.

Equal weights of the first part and the second part may provide, when mixed, a sterilizing composition having a pH of from 1.0 to 10.5, but it is preferred that the composition has a pH of from 4.5 to 6.5 as this may result in a more

stable compound.

A plurality of fabric members may be provided in a single resealable container, for example a canister with a lid, or a resealable sachet. In a preferred embodiment, each fabric member is provided in its own sachet which may be factory sealed and disposed of after use. In a particularly preferred embodiment, each sealed sachet contains a single fabric wipe.

Other aspects and benefits of the invention will appear in

the following specification, drawings and claims.

3960.a4 (2Jvl.doc 06 May 2004 - 9 -

BRIEF DESCRIPTION OF THE DRAWINGS

The invention will now be further described, by way of example, with reference to the following drawings in which: Figure 1 shows a trigger sprayer for use in a sterilant system in accordance with an embodiment of the present invention; Figures 2 and 4 show an canister of sterilizing wipes, respectively open and closed, for use in a sterilant system in accordance with an embodiment of the present invention; Figure 3 shows an alternative trigger sprayer for use in a sterilant system in accordance with an alternative embodiment of the invention; and Figure 5 illustrates a sterilant system in accordance with a further embodiment of the invention.

DETAILED DESCRIPTION

The trigger sprayer shown in Figure 1 is of a construction well known per se. The sprayer comprises a body 2 connected to a sprayer head 4 by an internally screw-threaded connector ring 14. A spray nozzle 10 in the head is connected to an aqueous liquid 12 by means of a dip tube 6. A user dispenses the liquid 12 through the nozzle 10 by operation of a trigger 8. Rotation of the nozzle allows the user to dispense the fluid as either a spray of fine droplets or as a foam.

3960.a42)vl.doc 06 May 2004 - 10 In the present example, the liquid 12 (first part) comprises 0.75% of a first reagent (sodium chlorite), 3.0% foam promoter (Cocamidopropyl Betaine). The remainder is deionised water. In this specification, all parts are by weight unless otherwise indicated. Operation of the trigger 8 dispenses the first part 12 as a foam.

An alternative design of pump dispenser 2 is illustrated in Figure 3. The trigger 8 is formed integrally with the nozzle 10. Depressing the trigger 8 dispenses a portion of the fluid contents as a foam (referred to as the 'Activator Foam' because it activates the sterilizing powers of a sterilizing wipe). A protective cap 26 is provided to cover the nozzle and trigger 8 when not in use.

Turning now to Figures 2 and 4, a sealable container 20 is also of a construction well known per se. The container 20 is a hollow cylinder fitted with a cap 22. The container 20 contains a roll of interleaved fabric sheets 18. In this example, the fabric sheets 18 are to be used as sterilizing wipes, but it will be understood that the sheets 18 could also be used for other applications such as biocidal wound- dressings.

The cap 22 has a central opening through which a tip of the central wipe 18 is disposed. By pulling the central wipe 18, a user may remove this wipe from the container, leaving the next wipe in its place. A stopper 16 is provided on the cap 22 for releasably sealing the container 20.

In this example, the wipes 18 are impregnated with an aqueous acid solution (second part). In this example, the acid 3960.a4 (2Jvl don 06 May 2004 - 11 solution comprises 0.5% citric acid, 0.05% sorbic acid, 0.05% boric acid. The solution also comprises 0.35% of a buffer (trisodium phosphate). The solution also comprises 0.25% Trisodium Citrate, 1.0% glycerine, 0.1% Benzotriazole, 0.1% Sodium Molybdate and 0. 3% Sodium Nitrate. The remainder is deionised water.

The pump dispenser 2 and container 20 together comprise the sterilant system. To prepare a sterilizing wipe, a user removes an impregnated wipe 18 from the container 20, and applies a portion of foam from the sprayer 2 to the wipe 18.

To facilitate mixing of the reagents in the foam and the wipe, the user may fold the wipe in half and crush or rub the folded wipe before opening it out. Preferably, one of the components is provided with a pH-sensitive indicator which changes colour or becomes coloured when adequate mixing has occurred, thereby indicating that sufficient C1O2 has been generated in the wipe.

Once the sterilising wipe has been prepared, it may be used for a number of applications, including wiping surfaces and sterilizing medical equipment such as endoscopes.

In the preferred embodiment illustrated in Figure 5, each sterilizing wipe 18 is provided in its own sealed container 20, in this example a sachet. The sterilant system comprises a box 28 of sterilizing wipes 18 in individual sachets 20.

Each sachet 20 may be factory sealed and may be disposed of after the wipe has been removed. The foam pump dispenser 2 is also provided in the box 28 of sterilizing wipe sachets 20.

3960.a4 (2)vl.doc 06 May 2004 - 12

EXPERIMENTAL RESULTS

Experiment 1 Sterilant wipes in accordance with one embodiment of the invention were tested and compared with conventional wipes saturated with isopropanol (IPA), a general-purpose detergent, and sterile deionised water.

The test method to evaluate effectiveness of the wipes in killing/removing test organisms dried onto test surfaces, involved the following steps.

1. Mark out a six inch (30.5 cm) square test area on the test surface.

2. Inoculate the test surface with 0.5 ml of test organism suspension.

3. Spread the inoculum over the test area using a plastic spreader.

4. Allow the inoculum to dry (about 30 minutes).

5. Don a pair of disposable plastic gloves.

6. Prepare a ClO2 wipe in accordance with the invention, using a prescribed mixing time.

7. Wipe the test area for the prescribed wiping time.

8. Place the wipe in 10 ml of universal neutralizer in a 3960.a42)vl.doc 06 May 2004 - 13 Universal bottle (Test Suspension A). Vortex stir to release organisms.

9. Wipe the entire test area with a cotton-tipped swab (thoroughly/10 times).

10. Dip the swab into 10 ml of universal neutralizer in a Universal bottle after each sampling of the test area and rotate the swab against the inner wall of the bottle to release organisms (Test Suspension B).

11. Prepare 5 serial deci-dilutions of Test Suspension A and Test Suspension B in diluent.

12. Inoculate 0.5 ml of each dilution onto a culture plate and spread using a plastic spreader. Incubate the plates and do a viable count.

13. Calculate log1O reductions achieved from the difference in the initial inoculum and the number of test organisms recovered after disinfection with a C1O2 wipe.

Test variables were as follows.

Test Surface A flat stainless steel instrument tray.

Test Organism 3960 a4 (2)vl.doc 06 May 2004 ] - 14 Spores of Bacillus subtilis var.niger NCTC 10073 freshly prepared by the method of Beeby & Whitehouse.

Inoculum The test surface was inoculated with 1 X 108 spores.

Suspending Fluid Sterile deionised water.

Disinfectant Concentrations 1. 200 ppm C1O2 (notional) 2. 300 ppm C1O2 (notional).

Mixing Times + 30 seconds.

Wiping Times + 30 + 60 seconds.

Controls 1. 1% Hospec general purpose neutral liquid detergent (Young's Detergents)/Kimcare Medical Wipes (Kimberly-Clark).

2. Sterets Alcowipe: 70% IPA (Seton Prebbles Ltd).

3. Sterile deionised water: Kimcare Medical Wipes (Kimberly-Clark).

3960.a4 (2)vl.doc 06 May 2004 Results are given in Table 1.

Exp. Disinfectant/ Mixing Wiping VC VC No. Detergent time time Surface Wipe (seconds) (seconds) 1 200 ppm C102 15 15 177 143 2 200 ppm C1O2 15 30 36 14 3 200 ppm C1O2 15 60 10 8 4 200 ppm C1O2 30 15 800 300 _.__ _ 200 ppm C1O2 _ 30 240 27 6 200 ppm C1O2 30 60 29 26

_ __

7 300 ppm C1O2 15 15 1240 330 8 300 ppm C1O2 15 30 530 250 300 ppm C1O2 15 60 160 140 300 ppm C1O2 30 15 1450 900 11 300 ppm C1O2 30 30 30 70 12 300 ppm C1O2 30 60 20 10 5 13 1% Hospec 60 7.3x10 4.3x10 14 70% IPA 60 1.9x104 3. 7x104 Deionised H2O 60 2.0x10 5 3.0xl 05

TABLE 1

VC = Viable Count Interpretation of Results 1. Washing/wiping with water, neutral detergent (1% Hospec), or alcohol (70% IPA) were ineffective 2. For the notional 200 ppm C1O2 wipes the best results were obtained with a mixing time of 15 seconds and a wiping 3960. ad (2)vl. don 06 May 2004 time of 60 seconds.

3. For the notional 300 ppm C1O2 wipes the best results were obtained with a mixing time of 30 seconds and a wiping time of 60 seconds.

4. Results for 200 ppm ClO2 (notional) were surprisingly better than results for 300 ppm (notional), except for mixing times of 30 seconds combined with wiping times of at least 30 seconds.

5. A wiping time of 60 seconds achieved better results than a wiping time of 30 seconds, which in turn achieved better results than a wiping time of 15 seconds.

6. Both C1O2 concentrations achieved good results after a wiping time of 60 seconds. The test surface was inoculated with 1 x 103 spores. After using the ClO2 wipes, surface counts were reduced to 10 and 29 (200 ppm ClO2) and to 160 and 20 (300 ppm C102).

7. A wipe containing 200 or 300 ppm may be useful, as may mixing times of 15 or 30 seconds (or, clearly, any intermediate times). However, it is preferred that wiping times longer than 15 seconds are employed.

These results were obtained using bacterial spores. It is to be expected that a vegetative bacterium such as MRSA will be much more sensitive, so that lower ClO2 concentrations and/or i shorter mixing or wiping times may be effective against such bacteria.

3960.a4 (2)vl doc 06 May 2004 17 Further experiments (2-4) were carried out using 41 gem spunlace sheets comprised of 50.5% wood pulp and 49.5% PET.

The sheets' dimensions were 160 mm x 180 mm x 0.36 mm. In each experiment the wipes each contained 3 ml of Solution A (formulated as set forth below), made by treating a canister of 50 wipes with 150 ml of Solution A. Each wipe was activated with 1.5 ml of Solution B (formulated as set forth below) from a foam dispenser.

Solution A (Wipe) Formulation: Ingredients Actual % Tolerance w/w 1 Citric acid 0.50% +/ C.A.S. 77-92-9 0.60-0.40% 2 Sorbic acid 0.005% +/ C.A.S. 0.006 0.004% 3 Boric acid 0.005% +/ C.A.S. 10043-35-3 0.006 0.004% 4 Trisodium citrate 0.25% +/ C.A.S. 68-04-02 0.30-0.20% Trisodium phosphate 0.35% +/_ _ _ C.A.S. 10101-89-0 0.45-0.25% 6 Glycerin 1.00% +/ C.A.S. 56-81-5 1.10-0.90% 7 Benzotriazole 0.10% +/ C.A.S. 95-14-7 0.15-0.05% 8 Sodium molybdate 0.10% +/ C.A.S. 10102-40-6 0.15-0.05% 9 Sodium nitrate 0.20% +/ C.A.S. 7631-99-4 0.25-0.15% Preservative (Paramotol) 0.15% +/ C.A.S. 0.20-0.10% 11 Deionised water Balance Balance C.A.S. 7732-18-5 Solution B (Foam) 3960.a4(2)vl doc 06 May 2004 - 18 Formulation: Ingredients Actual % Tolerance 1 Sodium chlorite (25% solution) ow/w75% 0.85 0.65% 2 Cocamidopropyl betaine 3.00% +/ 3.10 2.90%

_

3 Indicator/colour solution 0.60% +/ (Indicator is cosmetic yellow, 0.07 No. 5, cl 19140 at 1% solution - 0. 50% 0.6%) 4 Preservative 0.15% +j (Euxyl K 100) 0.20 0.10%

_

Deionised Water (Purified) 95.50% +/ C.A.S. 7732-18-5 Balance Experiment 2 A study was carried out to compare the effectiveness of (a) ClO2 wipes in accordance with the invention (b) a 70% IPA wipe (c) a neutral detergent wipe and (d) a water wipe in removing and/or killing (1) B. subtilis spores, and (2) P. aeruginosa cells dried onto the insertion tube of a flexible endoscope.

Wipes were prepared fresh as required by squirting foam onto a wipe and then scrunching the wipe with the fingers to mix the reagents to form ClO2.

3960.a4 (2)vl.doc 06 May 2004 - 19 EXPERIMENT 2 Test organisms B. subtilis NCTC 10073 spores A suspension containing approximately 108 spores/ ml was prepared by the method of Beeby & Whitehouse. A 1 in 10 dilution in sterile distilled water was prepared to produce a suspension containing approximately 107 spores/ ml.

P. aeruginosa NCTC 6749 A culture containing approximately 108 cells/ ml was prepared by inoculating a tube of nutrient broth and incubating for 18 h at 37 C. Insertion tube used in Experiment 2 The insertion tube was 1 metre long, in good condition, with clear markings. The test site used was the 10 cm section between the 30 and 40 markings.

Test Method 1. Immerse a cotton-tipped swab into a suspension of spores or vegetative cells.

2. Inoculate entire surface area of test site with the suspension. Repeat several times. Regarding B. subtilis spores, assume that (1) the volume of inoculum = 0.1 ml, and (2) the mortality rate on drying out is zero. Hence the viable count of the inoculum = approximately 106 spores.

Regarding P. aeruginosa cells, assume that (1) the volume of 3960.a42)vl.doc 06 May 2004 l - 20 inoculum = 0.1 ml, and (2) the mortality rate on drying out is 1 log. Hence the viable count of the inoculum = approximately 106 cells.

3. Place inoculated insertion tube across the top of an empty discard jar with the 10 cm test site resting over the centre of the jar. Allow inoculum to dry out (approximately minutes).

4. Don pair of disposable plastic gloves.

5. Prepare a Wipe: ClO2 (scrunch time = 15 see), IPA, Hospec or water.

6. Wipe test site for the prescribed wipe time (30 see) as follows: Wrap wipe loosely around the insertion tube and then wipe up and down the test site repeatedly.

7. Place the wipe in 20 ml of universal neutralizer in a Universal bottle. Vortex stir to release recovered spores/ cells (Test Suspension A).

8. Swab entire test site with a cotton-tipped swab. Dip swab into 10 ml of universal neutralizer in a Universal bottle and rotate swab against the inner wall of the bottle to release recovered spores/ cells. Repeat 10 times then break off cotton-tip of swab and leave in the neutralizer.

Vortex stir to release recovered spores/ cells (Test Suspension B).

9. Prepare 5 serial deci-dilutions of Test Suspension A and Test Suspension B in diluent.

10. Inoculate 0.5 ml of each dilution onto a culture plate and spread using a plastic spreader. Incubate plates. Viable count.

11. Calculate logy reductions achieved from the difference in the number of spores or cells inoculated onto the test site (approximately 106) and the number recovered after cleaning and/or disinfection.

3960 ad (2)vl.doc 06 May 2004 À 21 Wipes used in Experiment 2 1. C1O2 Wipe (scrunch time = 15 seconds).

2. 70% IPA wipe: Azowipe (Vernon Carus).

3. Hospec wipe: Kimberley Clark Medical Wipe immersed in 1% Hospec and then squeezed to remove excess solution.

4. Water wipe: Kimberley Clark Medical Wipe immersed in sterile water and then squeezed to remove excess water.

EXPERIMENT 2 - RESULTS Exp Test organism Disinfectant/ Scrunch Wipe Viable Count (0.5 detergent time time ml) (see) (see) Surface Wipe 1 B. sub tills C1O2 15 30 0 2 ClO2 (repeat) 15 30 0 3 70% IPA 30 5.0 x 102 2.7 x 103 4 1% Hospec 30 1.5 x 102 2.6 x 103 Water 30 3.0 x 101 2.5 x 103 6 P. aeruginosa C1O2 15 30 0 0 7 C1O2 (repeat) 15 30 0 9 70% IPA 30 0 9 1% Hospec 30 6. 2 x 103 8.0 x 104 Water 30 2.5 x 104 1. 5 x 105

Table 2

3960.a4 (2Jvl doc 06 May 2004

_

Exp Test organism Disinfectant/ Total spores/ cells detergent recovered Surface1 Wipe2 1 B. sub tills C1O2 i 2 C1O2 (repeat) 0 0 3 70% IPA 1.0 x 10 1.0 x 102: 4 1% Hospec 3.0 x 10 1.0 x 10 Water 6.0 x 10 1.0 x 10 6 P. aeruginosa C1O2 0 0

_

7 C1O2 (repeat) 0 0 8 70% IPA 4.0 x 101 0 _ 1% Hospec 1.2 x 105 3.2 x 106 Water 5.0 x 10 6.0 x 10

Table 3

1 Viable count in Table 1 x 20 (0.5 ml of 10 ml neutralizer plated out).

2 Viable count in Table 1 x 40 (0.5 ml of 20 ml neutralizer plated out).

EXPERIMENT 2 - CONCLUSIONS

1. C1O2 wipes were completely effective against both B. subtilis spores and P. aeruginosa cells. No spores or cells were recovered in duplicate experiments.

2. IPA wipes exhibited good activity against P. aeruginosa cells but did not eliminate all of the test cells - 40 viable cells were recovered from the test site on the insertion tube.

3960. ad (2) vl.doc 06 May 2004 -l, 1' 3. IPA wipes were ineffectiveagainst B. subtilis spores.

IPA proved less effective than 1% Hospec or water which may be attributable to the coagulant properties of alcohol (fixing spores on the test site).

4. Wipes saturated with 1% Hospec were ineffective against either B. subtilis spores or P. aeruginosa cells.

5. Wipes saturated with water were ineffective against either B. subtilis spores or P. aeruginosa cells.

Experiment 3 EVALUATION OF THE EFFECTIVENESS OF C1O2 WIPES IN KILLING/ REMOVING METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS (MRSA)

DRIED ONTO A STAINLESS STEEL TEST SURFACE

Test Method The following test method was used to evaluate the effectiveness of C102 Wipes in killing/ removing test organisms dried onto test surfaces. The test method involves the following steps: 1. Mark out an 18 inch (457.2 mm) square on the test surface.

2. Inoculate test surface with 4.5 ml of test organism I suspension.

3. Spread inoculum over 18 inch (457.2 mm) square test area using a plastic spreader.

4. Allow inoculum to dry (30-60 minutes).

5. Don pair of disposable plastic gloves.

6. Prepare a C1O2 Wipe using the prescribed scrunch time (15 seconds).

3 960 ad 2 vl. doc 06 May 2004 Ed 7. Wipe test area for the prescribed wipe time (30 seconds).

8. Place the ClO2 Wipe in 20 ml of universal neutralizer in a universal bottle. Vortex stir to release organisms. (Test Suspension A).

9. Swab entire test area with a cotton-tipped swab. Dip swab into 10 ml of universal neutraliser in a universal bottle and rotate cotton-tip against the inner wall of the bottle to release organisms. Repeat 10 times. Finally, snap off cotton-tip into the neutralizer. Vortex stir to release organisms. (Test Suspension B).

10. Prepare 5 serial deci-dilutions of Test Suspension A and Test Suspension B in diluent.

11. Inoculate 0.5 ml of each dilution onto a culture plate and spread using a plastic spreader. Incubate plates. Viable count.

12. Calculate logic reductions achieved from the difference in the initial inoculum and the number of test organisms recovered after cleaning/ disinfection with a Cl02 Wipe.

13. Repeat above using control wipes (70% IPA, It Hospec & sterile water).

Variables selected in Experiment 3 Test surface A flat stainless steel laboratory bench.

Test organism Methicillin Resistant Staphylococcus aureus (MRSA): a clinical isolate from the Royal Preston Hospital.

3960.a42}vl.doc 06 May 2004 - 25 Inoculum The test surface was inoculated with >109 bacterial cells: 4.5 ml of an overnight culture in Nutrient Broth.

Suspending fluid Nutrient Broth Scrunch time seconds Wipe time seconds Controls 1. 70% IPA wipe: Azowipe (Vernon Carus).

2. 1% Hospec general purpose neutral liquid detergent (Young's Detergents) / Kimcare Medical Wipe (Kimberly-Clark).

The wipe was immersed in 1% Hospec and then squeezed with the fingers to remove excess fluid.

3. Sterile deionised water / Kimcare Medical Wipe (Kimberly-Clark). The wipe was immersed in water and then squeezed with the fingers to remove excess fluid.

3960.a4 (2)vl.doc 06 May 2004 - 26 Results Exp Disinfectant/ Mixing Wiping Viable Count detergent time time Surface Wipe (see) (see) 1 C1O2 15 30 0 0 2 C1O2 (repeat) 15 30 0 0 3 70% IPA 30 5.5 x 104 4 1% Hospec 30 5.5 x 104 6.0 x 104 Deionised H2O 30 5.7 x 104 5.9 x 104

Table 4

Exp Disinfectant Mixing Wiping Total number of / detergent time time organisms recovered (see) (see) Surface1 wipe2 _.. _ 1 C1O2 15 30 0 0 2 C1O2 15 30 0 (repeat) 6 2 3 70% IPA. 1.1 x 10 3.6 x 10 4 1% Hospec 30 1.1 x 106 2.4 x 10 Deionised 30 1.1 x 10b 2.4 x 10 H2O

Table 5

1 Viable Count in Table 1 x 20 (0.5 ml of 10 ml neutralizer plated out).

2 Viable Count in Table 1 x 40 (0.5 ml of 20 ml neutralizer plated out).

3960 ad (2)vl.doc 06 May 2004 t 27 Interpretation of results 1. Wiping with a ClO2 Wipe for 30 seconds was completely effective. No test organisms were recovered from either the test surface or the wipes in duplicate experiments.

2. Wiping the test surface with a 70% IPA wipe (Azowipe) for 30 seconds was ineffective. This could be due to: (a) an exposure time of 30 seconds was not long enough to kill the MRSA (b) the IPA evaporated off the test surface before the minimum exposure time required to kill the MRSA] (c) the volume of IPA on the wipe was insufficient to deal with the >109 MRSA dried onto the 18 inch test surface (d) a combination of the above.

3. Only 360 test organisms were recovered from the Azowipe. This could be due to: (a) entrapment of test organisms in the fibres (b) incomplete/ slow neutralization of the residual IPA on the wipe by the neutralizer (c) a combination of the above 4. Wipes saturated with either 1% Hospec or sterile water were ineffective. L Experiment 4 This experiment was carried out to evaluate the effectiveness of ClO2 Wipes in killing/ removing spores of Bacillus subtilis var. niger NCTC 10073 dried out for 24 h at room 3960.a4 {2)vl.doc 06 May 2004 - 28 temperature on a stainless steel test surface.

Test Method 1. Mark out a 12 inch (304.8 mm) square on the test surface.

2. Inoculate test surface with 1.0 ml of aqueous spore suspension.

3. Spread inoculum over 12 inch (304.8 mm) square test area using a plastic spreader.

4. Allow inoculum to dry out naturally at room temperature for 24 h. 5. Don pair of disposable plastic gloves.

6. Prepare a ClO2 Wipe using the prescribed scrunch time (15 seconds).

7. Wipe test area for the prescribed wipe time (30 seconds).

9. Swab entire test area with a cotton-tipped swab. Dip swab into 10 ml of universal neutralizer in a universal bottle and rotate cotton-tip against the inner wall of the bottle to release organisms. Repeat 10 times. Finally, snap off cotton-tip into the neutralizer. Vortex stir to release organisms. (Test Suspension B).

11. Inoculate 0.5 ml of each dilution onto a culture plate and spread using a plastic spreader.

12. Repeat above using a control wipe (a Medical Wipe saturated with sterile water).

3960.a4 (2)vl.doc 06 May 2004 - 29 13. Incubate plates. Viable count.

14. Calculate logy reductions achieved using the C102 Wipe from the difference in viable count obtained using the C102 Wipe and the control wipe.

Variables selected in Experiment 4 Test surface A flat stainless steel instrument tray.

Test organism Bacillus subtilis var. tiger NCTC 10073. A spore suspension was prepared by the method of Beeby & Whitehouse.

Inoculum The test surface was inoculated with (a) 106 spores, and (b) 103 spores.

Suspending fluid Deionised water.

Drying time The inoculated instrument tray was allowed to dry out naturally at room temperature for 24 h in a dark cupboard.

3960. ad (2) vl doc 06 May 2004 - 30 Scrunch time seconds.

Wipe time seconds.

Control 1. Sterile deionised water / Kimcare Medical Wipe (Kimberly-Clark). The wipe was immersed in water and then squeezed with the fingers to remove excess fluid.

Results Exp. Inoculum Disinfectant Mixing Wiping Viable Count (spores) / detergent time time Surface Wipe 6 (see) (see) 1 10 C1O2 15 30 0 0 2 1Ob Water 15 30 2.0 x 2.1 3 1o8 C1O2 15 30 4.8 x 1.3 4 1Oa Water 15 30 6.6 x 1.9

Table 6

3960.a4 (2)vl.doc 06 May 2004 - 31 Exp. Inoculum Disinfectant Mixing Wipe Total number of (spores) / detergent time time spores (see) (see) recovered 6 -- Surface1 Wipe2 1 10 C1O2 15 30 0 0 2 10 Water 15 30 4.0 x 8.4 x 8 103 103 3 10 ClOz 15 30 9.6 x 5.2 x 103 103 4 108 Water 15 30 1.3 x 7.6 x lo6 loot

Table 7

1 Viable Count in Table 1 x 20 (0.5 ml of 10 ml neutraliser plated out).

2 Viable Count in Table 1 x 40 (0.5 ml of 20 ml neutraliser plated out).

Interpretation of results 1. Spores dried out for 24 h at room temperature on a stainless steel test surface were not easy to dislodge using a Medical Wipe saturated with deionised water. With the 106 inoculum the recovery was 4.0-8.4 x 103 spores leaving 2-3 log1O spores on the surface (assuming no mortality) . With the 108 inoculum the recovery was 1.3 -7.6 x 106 spores leaving 1 2 logy spores on the surface.

2. C1O2 Wipes were effective in killing/ removing spores dried out for 24 h at room temperature on the stainless steel test surface. With the 106 inoculum, no spores were recovered 3960 a4 (2)vl.doc 06 May 2004 - 32 from either the surface or wipe which represents a 3-4 log1O reduction on both the surface and wipe. With the 103 inoculum, a 2- 3 log1O reduction of spores was achieved on the surface and a 3-4 log1O reduction on the wipe.

Thus, the invention provides a sterilant system which can be prepared in situ and which provides bactericidal, fungicidal, virucidal, and sporicidal fabrics. The system is particularly useful for sterilizing wipes and for the dressing of wounds and ulcers.

3960.a42)vl.doc 06 May 2004 - 33

Claims

1. A two-part sterilant system comprising: (a) a first part comprising a first reagent in a carrier medium; and (b) a second part which is miscible with the first part and which comprises a second reagent in a carrier medium; wherein the first reagent and the second reagent will react when mixed to provide a sterilizing composition; characterized in that the first part is contained in a pump dispenser whereby it may be dispensed as a fluid, and in that the second part is absorbed or impregnated in at least one fabric member in a sealed container.

2. A sterilant system according to claim 1, wherein the first part includes a foam promoter and is contained in a trigger-operated foam dispenser.

3. A sterilant system according to claim 1 or claim 2, wherein the at least one fabric member comprises a plurality of fabric wipes, each of which is provided in its own separate sealed sachet.

4. A sterilant system according to any preceding claim, wherein at least one of the first part and the second part includes an indicator reagent that changes colour when the parts are mixed together.

5. A sterilant system according to claim 4, wherein the first part and the second part have a different pH and wherein the indicator reagent changes colour in response to a change in pH when the parts are mixed.

3960.a4 (2)vl.doc 06 May 2004 - 34

6. A sterilant system according to any preceding claim, wherein one of the first part and the second part comprises a solution containing sodium chlorite or sodium chlorate and the other comprises an acidic solution

7. A sterilant system according to claim 6, wherein the acidic solution comprises a solution of citric acid, sorbic acid and boric acid.

8. A sterilant system according to any preceding claim, wherein the first part further comprises from 0.1 to 50% w/w of at least one foam promoter.

9. A sterilant system according to claim c9, wherein said foam promoter comprises from 3 to 6% w/w.

10. A sterilant system according to any preceding claim, wherein one of the first part and the second part further comprises from 0.1 to 50% w/w of a humectant.

11. A sterilant system according to claim 10, wherein said humectant comprises from 1 to 3% w/w.

12. A sterilant system according to any preceding claim, wherein when equal weights of the first part and the second part are mixed they provide a sterilizing composition having a pH of from 4.5 to 6.5.

13. A sterilant system according to claim 6, wherein the first part comprises said solution of sodium chlorite or sodium chlorate.

3960 a4 (2)vl doc 06 May 2004 - 35

14. A sterilant system according to claim 13, wherein said first part comprises a solution of sodium chlorite and said acidic solution comprises an aqueous mixture of citric acid, sorbic acid, boric acid, and a buffer.

15. A sterilant system according to claim 1, wherein one of the first reagent and the second reagent comprises chlorite and the other comprises hypochlorite.

16. A sterilant system according to claim 1, wherein one of the first reagent and the second reagent comprises chlorate and the other comprises a peroxide and an acid.

17. A two-part sterilant system comprising: (a) a first part comprising a first reagent in a carrier medium; and (b) a second part which is miscible with the first part and which comprises a second reagent in a carrier medium; wherein the first reagent and the second reagent will react when mixed to provide a sterilizing composition; characterized in that the first part is contained in a trigger-operated dispenser whereby it may be dispensed as a foam, a spray, a paste, a powder or a gel, and in that the second part is absorbed or impregnated in at least one fabric wipe in a sealed container.

18. A two-part wound-dressing system comprising: (a) a first part comprising a first reagent in a carrier medium; and (b) a second part which is miscible with the first part and which comprises a second reagent in a carrier medium; 3960.a4 (2)vl.doc 06 May 2004 - 36 wherein the first reagent and the second reagent will react when mixed to provide a sterilizing composition; characterized in that the first part is contained in a dispenser whereby it may be dispensed as a fluid, and in that the second part is absorbed or impregnated in at least one fabric wound dressing in a sealed container.

19. A wound-dressing system according to claim 18, wherein said dispenser is a trigger dispenser which will dispense the first part as a spray or foam and wherein the second part is absorbed or impregnated in a plurality of fabric wound dressings each of which is provided in its own separate sealed sachet.

20. A sterilant system substantially as herein described with reference to the drawings.

3960.a4 (2Jvl.doc 06 May 2004