SANITISING COMPOSITION

Abstract

The invention relates to methods and liquid compositions suitable for high level disinfection (“HLD”) of medical instruments at close to ambient temperatures. The methods and compositions are also effective for digesting biological contaminants, and, surprisingly, can be used to clean and disinfect concurrently. Both cleaning and disinfection efficacy meet the criteria for cleaning and disinfection of medical devices. Use of compositions according to the invention enable reprocessing of instruments more quickly with greater energy efficacy, and greater convenience than prior art reprocessing methods. Compositions according to the invention are also useful for simultaneous cleaning and disinfecting other surfaces, for example in hospitals, food preparation areas and the like, or fabrics and the like such as are treated in hospital laundries.

Claims

1. A method of cleaning a surface contaminated with an organic load while simultaneously achieving high level disinfection of the surface, wherein the method comprises the steps of: (i) providing a composition comprising an enzyme, an anionic hydrotrope, a biocide selected from the group of quaternary ammonium biocides, and an activity protector comprising a boron compound, (ii) diluting the composition by from 10:1 to 200:1, and (iii) treating the surface with the diluted composition, wherein the surface is the surface of a medical instrument, and wherein the aryl quaternary ammonium biocides is present at a concentration which, when diluted for use, is below the Minimum Inhibitory Concentration (“MIC”) of the quat to any challenge microorganism indicated in Therapeutic Goods Order (TGO54).

2. A method according to claim 1, wherein the composition is diluted by from 20:1 to 100:1.

3. A method according to claim 1, wherein the anionic hydrotrope is selected from the group consisting of water soluble anionic hydrotropes of formula: ##STR00003## wherein R.sup.1 and R.sup.2 are independently hydrogen or alkyl groups from 1 to six carbons, preferably from one to four carbons, and more preferably from one to two carbons.

4. A method according to claim 1, wherein the anionic hydrotrope is ##STR00004## wherein R.sup.1 and R.sup.2 are independently hydrogen or alkyl groups from 1 to six carbons, preferably from one to four carbons, and more preferably from one to two carbons.

5. A method according to claim 1, wherein the anionic hydrotrope is selected from the group consisting of alkali metal xylenesulphonates, alkali metal cumene sulphonates, other alkali metal alkylarylsulphonates and combinations thereof.

6. A method according to claim 1, wherein the boron compound is boronic acid.

7. A method according to claim 1, wherein the quaternary ammonium biocide is an aryl quaternary ammonium biocide.

8. A method according to claim 1, wherein the aryl quaternary ammonium biocide is benzalkonium halide.

9. A method according to claim 1, wherein the enzyme is a protease.

10. A method according to claim 1, wherein the composition further comprises a polyol.

11. A method according to claim 1, wherein the composition before dilution comprises a concentration of quaternary ammonium biocide of 2% w/w or more.

12. A method according to claim 1, wherein the composition before dilution comprises a concentration of quaternary ammonium biocide of 4% w/w or less.

13. A method according to claim 1, wherein the composition before dilution comprises a concentration of quaternary ammonium biocide of 2% w/w to 4% w/w.

14. A method according to claim 1, wherein the composition before dilution comprises a concentration of quaternary ammonium biocide of 2% w/w.

15. A method according to claim 1, wherein the composition before dilution comprises a concentration of quaternary ammonium biocide of 4% w/w.

16. A method according to claim 1, wherein the organic load includes a protein.

17. A method according to claim 1, wherein the surface is treated in a bath.

18. A method according to claim 1, wherein the High Level Disinfection is least a 6 log reduction of Mycobacteria and non-enveloped viruses.

19. A method according to claim 1, wherein the treating step is carried out at a temperature from 25° C. to 60° C.

Description

DETAILED DESCRIPTION OF THE INVENTION

Anionic Hydrotrope

[0053] An essential feature of the present invention is the inclusion of an anionic hydrotrope. A hydrotrope is a compound that solubilises hydrophobic compounds in aqueous solutions. Desirably the hydrotrope is selected from the group consisting of water soluble anionic hydrotropes of formula:

##STR00001##

And more preferably of the formula:

##STR00002##

and having no alkyl side chains greater than six carbons in length. In preferred hydrotropes R.sup.1 and R.sup.2 are independently alkyl groups from 1 to six carbons, preferably from one to four carbons, and more preferably from one to two carbons, although R.sup.1 or R.sup.2 may optionally be hydrogen. Very highly preferred hydrotropes are water soluble xylene sulphonate (R.sup.1 and R.sup.2 are methyl) and cumene sulphonate (R.sup.1 is isopropyl, R.sup.2 is hydrogen) salts.

[0054] Examples of suitable anionic hydrophobic compounds include sodium xylenesulphonate (“SXS”), and sodium cumene sulphonate (“SCS”). However other suitable anionic hydrotropes include sodium-2-ethyl hexylsulphate, phosphate ester of oxyethylated phenol, amine alkylaryl sulphonate, linear alkyl naphthalene sulphonate, sodium dihexyl sulphosuccinate, and sodium dodecylbenzene sulphonate.

[0055] Desirably the anionic hydrotrope is present in a concentration sufficient that the quaternary ammonium biocide is effective in use to provide “High Level” disinfection (as herein defined) of the bath in the presence of the at least one enzyme and of a typical proteinaceous load in the bath. The ratio of anionic hydrotrope to quat is at least 2:1, more preferably 5:1.

Activity Protector

[0056] Desirably an “activity protector” is present and, is selected from (1) compositions known to be effective in stabilizing enzymes in liquid aqueous solutions, including enzyme stabilizing compounds and systems, (2) selected “micelle inhibitors”, and mixtures of (1) and (2). In preferred embodiments of the invention the “activity protector” is an enzyme stabilizer and more particularly is a suitable concentration of boron anions. Other reversible enzyme inhibitors could be suitable in this application, for example phenyl boronic acid and similar compounds described in EP 0707642A1. Desirably these are solvated in a polyol and may be combined with enzyme stabilizing synergists or adjuvants forming an enzyme stabilizing system. Preferred “micelle inhibitors” include species known to modify as well as to inhibit micelle formation and may be selected from water miscible solvents such as C1 C6 alkanols, C1 C6 diols, C2 C24 alkylene glycol ethers, alkylene glycol alkyl ethers, and mixtures thereof. A highly preferred micelle inhibitor is di-(propylene glycol) methyl ether (“DPM”) and analogues thereof which modify micelle formation. It is especially preferred to combine the use of borate ions with DPM which has been found by the present inventor synergistically to enhance the biocidal activity protection conferred on the quat. biocide without irreversibly denaturing the enzyme.

Quat

[0057] It is highly preferred that the quat biocide is an aryl quat compound, preferably benzalkonium halide. Other biocidal quaternary compounds could be used.

Enzymes

[0058] It is well known that enzymes may become denatured in storage, in the presence of other enzymes, and/or in the presence of antagonistic anions such as for example anionic surfactants, quaternary ammonium compounds and detergency “builders”. A number of enzyme stabilizing systems have been developed and are well known in the enzyme formulation art. An example of an “enzyme stabilizing system” is a boron compound (e.g. boric acid) which in the past has been used alone or with selected other adjuvants and or synergists (e.g. polyfunctional amino compounds, antioxidants, etc.) to protect proteolytic and other enzymes in storage and in various products. It has been theorised that an enzyme stabilizing system such as boron and calcium form intramolecular bonds which effectively cross-link or staple an enzyme molecule so as to hold it in its active spatial configuration. Enzyme stabilizers have not hitherto been used to protect the biocidal activity of a quat. biocide. The present invention is based on the surprising discovery that at least some enzyme stabilizing systems are effective in protecting the biocidal activity of quat. biocides in the presence of protein.

[0059] The present invention also includes an “activity protector” of the kind discussed in our patent specification PCT/AU01/00381, e.g. boron in a ratio to quat. biocide chosen to substantially to minimise the Minimum Inhibitory Concentration (“MIC”) of quat. biocide in the presence of the enzymes in the formulation and at a given level of protein load. MIC is a measure of the minimum concentration of the biocide which succeeds in preventing bacterial growth in a culture during a specified time period, for example 24 hrs. Details of the MIC test are shown in Bailey & Scott “Diagnostic Microbiology”, 8.sup.th edition, 1990 at page 177. The TGA tests are specified at TGO 54 annexed. MIC tests referred to herein are conducted over 24 hrs.

[0060] In the present case in which an enzyme is present in addition to the quat. biocide and in which it is desired to retain the enzymatic activity of the enzyme as well as the biocidal activity of the quat, biocide then the quantity of “activity protector” required will need to be greater than that required merely to protect the enzyme and will need to be sufficient both to stabilise the enzyme and protect the biocidal activity of the quat. biocide. Moreover, as the composition is anticipated to come into contact with an external proteinaceous load (from contaminants in the surgical instruments bath) then the “activity protector” concentration will need to be greater still.

[0061] The inventor discovered that boron surprisingly protects a quaternary biocide from deactivation by a protein in such a way and to such an extent that the MIC of the biocide is not increased in the presence of a protein. In preferred embodiments of the invention the MIC is dramatically reduced, for example, more than halved notwithstanding the presence of up to 2 wt. % based on the weight of solution, of protein. This allows the formulation of a wide range of new and useful compositions which remain effective as disinfectants or antibacterials in circumstances in which the prior art would be significantly less effective or not effective at all.

[0062] The invention also enables storage-stable liquid biocidally effective compositions to be prepared with a lower concentration of quat. biocide and at much lower cost. By “shelf stable” is meant that the composition retains at least 50% of its biocidal efficacy after 12 months storage in a sealed container at 18-25° C. Preferred embodiments of the invention retain better than 98% biocidal efficacy under these conditions.

[0063] Without wishing to be bound by theory, the inventor speculates that polymeric borate ions associate with the cationic quat. biocide, thus protecting the quat biocide from combining with proteins. When the formulation is diluted the polymeric ions become unstable and release the quat biocide for disinfection. Alternatively, it may be that the biocidal activity of the quat. biocide significantly relates to denaturing proteins of cell membranes and that boron complexes with charged groups of non-living proteins and prevents wasting quat. on denaturing non-living proteins. However, as enzymes are structurally quite different from quat. biocides, and as the complete mechanism by which quat. biocides kill bacteria is also uncertain, it was not previously predictable that any enzyme stabilizer would be effective in maintaining the biocidal activity of a quat. biocide (an enzyme antagonist). The mechanism by which the activity of the quat biocide is maintained may be different from that whereby the enzyme is stabilised.

Examples of the Invention

[0064] Several formulations with varying concentration of hydrotropes, various groups of commercially available proteases and quaternary amine (biocidal active) in accordance with the invention were prepared as shown in Table 1 annexed hereto. Some of the formulations are with or without non-ionic surfactants

[0065] The formulations of Table 1 are identified by designations 126-8 to 126-20 and all are examples of multi-enzyme cleaning and sanitising products according to the invention for use in manual baths and AER medical instrument reprocessors. The preferred use concentration is between 5 mL/L and 20 mL/L and at temperatures from 25° C. to 60° C. (maximum temperature to which flexible endoscopes could be exposed).

[0066] The cleaning efficacy and foaming properties of the composition was then tested and compared with formulations identified as 84-0, 84-2, 84-4 and 84-10 made in accordance with PCT/AU01/00381 and shown in Table 2 annexed hereto.

[0067] Tables 3, 4 and 5 annexed hereto demonstrate the cleaning efficacy of the formulations of Table 1. PF-126 formulations were diluted with distilled water at 25° C. (Table 3), hot water at 50° C. (table 4) or at 40° C. (Table 5) (to concentrations 2 mL/L, 5 mL/L and 20 mL/L) in glass beakers. The temperature of the solutions were maintained in a water bath for the duration of testing. A test soil was introduced in the form of a Browne load check strip at the same time as a stopwatch was started. The test strips were monitored over time to identify how long it took the test soil to be completely removed from the Browne load check strip.

[0068] All samples in Table 2 (in accord with PCT/AU01/00381) failed to remove the test soil even at 50° C. and a concentration of 20 mL/L within 60 minutes.

[0069] Formulations according to the present invention on the other hand as shown in Tables 4 & 5, removed the test soil within commercially acceptable times even at static conditions.

[0070] The static cleaning used in the above experiments is the worst case scenario. With agitation/mixing of the solutions simulating the agitations encountered in AERS and washer disinfectors the cleaning efficacy speeds up markedly: for example, formulation 126-8 cleans Browne Load Check at [25° C. 5 ml/1] in 31 minutes at static conditions, in 7 minutes at AER agitations and less than 4 minutes in an orbital shaker mimicking the agitations of washer disinfectors. The formulation 84-0—the best performing out of the prior art formulations—could not clean Browne strip at 20 ml/l 50° C. in 60 minutes even in an orbital shaker.

[0071] Table 7 shows the proteolytic activity of the preferred formulations compared to formulations without a quaternary biocide. It can be seen rather surprisingly and unexpectedly that the proteolytic activity of the formulations containing a quaternary biocide is substantially higher.

[0072] Table 8 shows the biocidal activity of preferred formulations against S.aureus ATCC 6538 and P.aeruginosa ATCC 15442. It can be seen that the biocidal activity is retained even at a high dilution factor of 1:1000.

[0073] Table 9 shows stability data for some preferred formulations. Each formulation is tested for proteolytic activity when first made and is then stored at 25° C. and 45° C. After 220 days in storage, the proteolytic activities of the formulations were retested. Storage at 45° C. for 220 days is equivalent to storage for 700 days (about 2 years) at 25° C. It is generally recognised in the art that loss of up to 50% of proteolytic activity on storage is acceptable.

[0074] Examples of some formulations that combine proteases and quats, proteases and hydrotropes, and quats and hydrotropes are shown in Table 10. Formulation 126-8 is in accordance with the invention. The other formulations are either not stable (hazy) or exhibit unacceptably poor bactericidal efficacy (greater than 40 minutes as per the test protocol in the table) or unacceptably poor cleaning (greater than 30 minutes as per the test protocol in the table).

Biocidal Efficacy

[0075] Compositions 126-8, 126-9, 126-13, 126-14 were evaluated for biocidal efficacy as per EN 1276 (biocidal) and EN 14348 (turbeculocidal). Table 6 annexed summarises the treatment envelop (conc, temp, time) required to achieve HLD for these formulations. As expected, Mycobacteria (TB) presented the greatest challenge. The increased concentration of QUATs improved the bactericidal efficacy. At the same time, the effect of water hardness was not as detrimental to quat activity as a person skilled in the art would expect indicating that enzymes might be symbiotic with quat in achieving high levels of kill.

[0076] A sample of the product diluted with hard water is added to a test suspension of mycobacteria in a solution of an interfering substance. The mixture is maintained at one of the temperatures and the contact times specified. At the end of this contact time, an aliquot is taken; the bactericidal and/or the bacteriostatic action in this portion is immediately neutralized or suppressed by a validated method.

Test Conditions

[0077] Test organism(s) Mycobacterium terrae (ATCC 15755)

[0078] Test temperature(s) 40° C., 45° C., 50° C.

[0079] Contact time(s) 5 min-30 min

[0080] Product diluent(s) 0 ppm and 300 ppm hard water

[0081] Interfering substance(s) Clean conditions=0.3 g/L bovine serum albumin

[0082] Dirty conditions=3 g/L bovine serum albumin+3 mL/L erythrocytes

Controls and Validations

[0083] All controls and validations were within the basic limits (EN 14348).

Results

[0084] See Table 6 annexed.

[0085] When used at 40° C., P126-4 is bactericidal within 30 minutes.

[0086] When used at 45° C., P126-4 is bactericidal within 15 minutes.

[0087] When used at 50° C., P126-4 is bactericidal within 5 minutes.

[0088] Using soft water (preferably RO water or distilled water) for dilution is recommended. It is of note that most potable water supplies have water hardness of below 50 ppm.

[0089] Since mycobacteria are regarded as one of the greatest challenges in the hierarchy of pathogens (inferior only to only endobacterial spores) the above results indicate that the formulation is capable to disinfecting instruments to High Level Disinfection as per TGO54. Similar results have been obtained with formulas PF-126-5, -6, -7, -8, -9 & -13, -14.

[0090] As will be apparent to those skilled in the art from the teaching hereof compositions according to the invention may be modified to provide lower levels of disinfection such as “Intermediate Level Disinfection”, “Hospital Grade Disinfection”, Safe to Handle” Disinfection” or as a Sanitiser if the intended use and applicable standards permit. Compositions according to the invention are also useful for cleaning and/or disinfection of other surfaces in hospitals, medical and dental practices, nursing homes or the like—for example chamber pots, trays, instrument transport trolleys and other large equipment—and for cleaning and/or disinfection food preparation areas, food utensils, dispensing equipment, cool rooms and the like, or fabrics and the like such as are treated in hospital laundries.

TABLE-US-00001 TABLE 1 Example Formulations Ingredient/Composition ID 126-8 126-9 126-10 126-11 126-12 126-13 126-14 126-15 126-16 126-17 126-18 126-19 126-20 DPM.sup.7 4 1 8 4 4 4 4 4 4 4 4 4 4 Sodium Cumene Sulphonate.sup.8 10 10 10 10 10 10 10 10 10 Sodium xylene sulphonate 12 Potassium xylene sulphonate 8 Sodium toluene sulphonate 8 Sodium salt dodecylbenzene 8 sulphonic acid Boron as boric acid 2 2 1 1 1 2 2 1 0.8 2 2 2 2 Serine Protease 3 3 3 3 2 2 2 3 3 3 (Savinase 16 L.sup.9) Cysteine Protease Papain 2 2 Metalloprotease 2 Endopeptidase Trypsin Amylase Termamyl 300 L 1 1 1 1 1 1 1 1 1 1 Teric GN9 1 1 1 1 1 0 1 1 1 Guardiquat 1450 2 4 2 4 2 2 2 (as 100% active).sup.10 Barquat MB-80 2 2 2 2 (benzalkonium chloride) Carboquat ™ MW-50 3 2 Didecyl Dimethyl Ammonium Carbonate Cold potable water qc qc qc qc pH (1:100 dilution) 8.2 9 7.8 8.6 9.3 6.9 9.2 9.3 8.1 7.7 8.4 8.2 9.0 .sup.7Diproplene Glycol methyl ether e.g. “Dowanol DPM” ex Dow Chemicals .sup.8Sodium Cumene Sulphonate e.g. ex Stepan .sup.9Savinase, Lipolase, Puradex and Termamyl ex Novazyme .sup.10Quats ex Albright and Wilson.

TABLE-US-00002 TABLE 2 Formulation number and corresponding composition for formulations based on compositions in accord with PCT/AU01/00381 84-0 84-2 84-4 84-6 84-10 Ingredient (w/v) % (w/v) % (w/v) % (w/v) % (w/v) % DPM 4 4 4 4 4 Propylene glycol 15 15 15 15 15 Teric 168 6 6 6 6 6 4Na-EDTA 1 1 1 1 1 Sulfamic acid 3 3 3 3 3 Genamin LAP 100D 10 10 10 10 10 Barquat MB-80 1 3 5 7 11 Savinase Ultra 10 10 10 10 10 16XL-NF Distilled water 50 48 46 44 40

TABLE-US-00003 TABLE 3 Cleaning times (in minutes) for full digestion of Browne load check strip soil at 25° C. Concentration PF-126-8 PF-126-9 PF-126-13 PF-126-14  2 mL/L >60 >60 >60 >60  5 mL/L 31 27 34 30 20 mL/L 17 16 12 19

TABLE-US-00004 TABLE 4 Cleaning times (in minutes) for full digestion of Browne load check strip soil at 50° C. Concentration PF-126-8 PF-126-9 PF-126-13 PF-126-14  2 mL/L 35 36 43 46  5 mL/L 16 19 26 24 20 mL/L 9 9 9 9

TABLE-US-00005 TABLE 5 Cleaning times (in minutes) for full digestion of Browne load check strip soil at 40° C. Concentration PF-126-8 PF-126-9 PF-126-13 PF-126-14  2 mL/L 43 43 50 52  5 mL/L 21 21 21 21 20 mL/L 9.5 9.5 9.5 9.5

TABLE-US-00006 TABLE 6 Biocidal Efficacy Results 126-8 126-9 126-13 126-14 Dilution Temp Diluent Pass at (min) 10 mL/L 40° C.  0 ppm 30 20 >30 20 10 mL/L 40° C. 300 ppm 30 20 30 30 20 mL/L 40° C.  0 ppm 20 15 20 15 20 mL/L 40° C. 300 ppm 20 15 20 15 10 mL/L 45° C.  0 ppm 15 15 15 10 10 mL/L 45° C. 300 ppm 15 10 15 10 20 mL/L 45° C.  0 ppm 10 10 10 10 20 mL/L 45° C. 300 ppm 10 5 10 5 10 mL/L 50° C.  0 ppm 5 5 5 5 10 mL/L 50° C. 300 ppm 5 5 5 5 20 mL/L 50° C.  0 ppm 5 5 5 5 20 mL/L 50° C. 300 ppm 5 5 5 5

TABLE-US-00007 TABLE 7 Proteolytic activity Proteolytic Activity Increase in Assayed Formulation (Au/ml) Proteolytic Activity PF-126-8 (with quat) 0.476 67% PF-126-8 (without quat) 0.317 PF-126-9 (with quat) 0.484 70% PF-126-9 (without quat) 0.341 PF-126-13 (with quat) 0.468 65% PF-126-13 (without quat) 0.303

TABLE-US-00008 TABLE 8 Biocidal activity of preferred formulations against S. aureus ATCC 6538 and P. aeruginosa ATCC 15442 126-8 126-9 126-13 126-14 Dilution Temp Diluent Pass at (min) S. aureus ATCC 6538 1 mL/L 40° C.  0 ppm 20 25 20 20 (1:1000) 1 mL/L 40° C. 300 ppm 35 40 40 40 (1:1000) 5 mL/L (1:200) 40° C. 300 ppm 20 20 20 20 5 mL/L (1:200) 40° C.  0 ppm 15 15 15 15 P. aeruginosa ATCC15442 1 mL/L 40° C.  0 ppm 30 30 30 30 (1:1000) 1 mL/L 40° C. 300 ppm 45 45 45 45 (1:1000) 5 mL/L (1:200) 40° C. 300 ppm 25 30 25 25 5 mL/L (1:200) 40° C.  0 ppm 20 20 20 20

TABLE-US-00009 TABLE 9 Shelf life stability of preferred formulations: Proteolytic Activity (Au/ml) Storage 0 220 Loss on Formulations Temperature Days Days* storage PF-126-8  25° C.  0.320 0.31 3.1% 45° C. 0.36 0.32 9.9% PF-126-9  25° C. 0.34 0.32 5.3% 45° C. 0.36 0.28 21.3%  PF-126-13 25° C. 0.32 0.32 1.2% 45° C. 0.33 0.29 12.4%  PF-126-14 25° C. 0.33 0.32 0.6% 45° C. 0.33 0.28 16.2%  *220 Days at 45° C. is equivalent to storage for about 2 years at 25° C.

TABLE-US-00010 TABLE 10 Examples of prior art formulations: Ingredient/Composition ID 126-8 126-81 126-82 126-83 126-84 126-85 126-86 126-87 126-88 Includes: P = Protease; Q = quat; P + Q + H P + Q; P; No Q; Q + H; Q; No H; H + Q; H; No Q; P + H; P; No H; H = hydrotrope No H No H No P No P No P No P No Q No Q DPM.sup.[7] 4 4 4 4 4 4 4 4 4 Sodium Cumene Sulphonate.sup.[8] 10 0 0 10 0 10 10 10 0 Boron as boric acid 2 2 2 2 2 2 2 2 2 Serine Protease (Savinase 16 L.sup.[9]) 3 3 3 0 0 0 0 3 3 Cysteine Protease Papain Metalloprotease Endopeptidase Trypsin Amylase Termamyl 300 L 1 1 1 1 1 1 1 1 1 Teric GN9 1 1 1 1 1 1 1 1 1 Guardiquat 1450 (as 100% active).sup.[10] 2 2 0 2 2 2 0 0 0 Cold potable water qc qc qc qc qc qc qc qc qc Cleaning efficacy expressed as time 15 >120 25 >120 >120 50 80 15 50 (in minutes) to clean Browne STF Load Check at 40° C., static conditions Pass/fail cleaning test P F P F F F F P F Bactericidal properties expressed as time 10 20 >180 20 20 40 >180 >180 >180 (min) required to pass suspension test as per EN1278 against P. aeruginosa ATCC15442. 1:100 dilution, 40° C. Pass as HLD? P P F P P P F F F Appearance clear hazy hazy clear clear clear clear clear clear liquid liquid liquid liquid liquid liquid liquid liquid liquid Loss of protease activity on 30 days 6 14 15 na na na na 9 20 storage at 45° C. (%) .sup.[7]Diproplene Glycol methyl ether e.g. “Dowanol DPM” ex Dow Chemicals .sup.[8]Sodium Cumene Sulphonate e.g. ex Stepan .sup.[9]Savinase, Lipolase, Puradex and Termamyl ex Novazyme .sup.[10]Quats ex Albright and Wilson.