AU2013270577B2 - Microbicidal composition - Google Patents

Microbicidal composition Download PDF

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AU2013270577B2
AU2013270577B2 AU2013270577A AU2013270577A AU2013270577B2 AU 2013270577 B2 AU2013270577 B2 AU 2013270577B2 AU 2013270577 A AU2013270577 A AU 2013270577A AU 2013270577 A AU2013270577 A AU 2013270577A AU 2013270577 B2 AU2013270577 B2 AU 2013270577B2
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atcc
methyl
ppm
benzisothiazolin
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Beverly Jean El A'mma
Beat Heer
Richard Levy
Kiran Pareek
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Rohm and Haas Co
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Abstract

Synergistic microbicidal compositions containing N-methyl-1,2 benzisothiazolin- 3-one.

Description

1 The present application is a divisional of Australian parent application no. 2008203011, the disclosure of which is included herein by reference. This invention relates to a synergistic combination of selected microbicides having greater activity than would be observed for the individual microbicides. In some cases, commercial microbicides cannot provide effective control of microorganisms, even at high use concentrations, due to weak activity against certain types of microorganisms, e.g., those resistant to some microbicides, or due to aggressive environmental conditions. Combinations of different microbicides are sometimes used to provide overall control of microorganisms in a particular end use environment. For example, U.S. Pat. App. Pub. No. 2007/0078118 discloses synergistic combinations of N-methyl-1,2-benzisothiazolin-3-one (MBIT) with other biocides. However, there is a need for additional combinations of microbicides having enhanced activity against various strains of microorganisms to provide effective control of the microorganisms. Moreover, there is a need for combinations containing lower levels of individual microbicides for environmental and economic benefit. The problem addressed by this invention is to provide such additional combinations of microbicides. STATEMENT OF THE INVENTION The present invention is directed to a microbicidal composition comprising: (a) N-methyl-1,2-benzisothiazolin-3-one; and (b) a microbicide comprising 2 ethylhexyl glycerin. In another embodiment of the invention, the weight ratio of 2 ethylhexyl glycerin to N-methyl-1,2-benzisothiazolin-3-one is from 1:0.017 to 1:2.3.
2 DETAILED DESCRIPTION OF THE INVENTION As used herein, the following terms have the designated definitions, unless the context clearly indicates otherwise. "MBIT" is N-methyl-1,2 benzisothiazolin-3-one. The term "microbicide" refers to a compound capable of 5 killing, inhibiting the growth of or controlling the growth of microorganisms at a locus; microbicides include bactericides, fungicides and algaecides. The term "microorganism" includes, for example, fungi (such as yeast and mold), bacteria and algae. The term "locus" refers to an industrial system or product subject to contamination by microorganisms. The following abbreviations are used 0 throughout the specification: ppm = parts per million by weight (weight/weight), mL = milliliter, ATCC = American Type Culture Collection, MBC = minimum biocidal concentration, and MIC = minimum inhibitory concentration. Unless otherwise specified, temperatures are in degrees centigrade (*C), and references to percentages (%) are by weight. Amounts of organic microbicides are given on 5 an active ingredient basis in ppm (w/w). The compositions of the present invention unexpectedly have been found to provide enhanced microbicidal efficacy at a combined active ingredient level lower than that of the individual microbicides. Additional microbicides beyond those listed in the claims may be present in the composition. 0 In one embodiment of the invention, the antimicrobial composition comprises N-methyl-I,2-benzisothiazolin-3-one and hexahydro-1,3,5-tris(2 hydroxyethyl)-s-triazine. Preferably, a weight ratio of hexahydro-1,3,5-tris(2 hydroxyethyl)-s-triazine to N-methyl-i,2-benzisothiazolin-3-one is from 1:0.0014 to 1:150, more preferably from 1:0.012 to 1:150. 5 In one embodiment of the invention, the antimicrobial composition comprises N-methyl-1,2-benzisothiazolin-3-one and 4,4-dimethyloxazolidine. Preferably, a weight ratio of 4,4-dimethyloxazolidine to N-methyl-1,2 benzisothiazolin-3-one is from 1:0.0011 to 1:3.5. In one embodiment of the invention, the antimicrobial composition 0 comprises N-methyl-1,2-benzisothiazolin-3-one and sodium hydroxymethylglycinate. Preferably, a weight ratio of sodium hydroxymethylglycinate to N-methyl-1, 2-benzisothiazolin- 3-one is from 1:0.0005 3 to 1:2.3, more preferably from 1:0.14 to 1:2.3, more preferably from 1:0.29 to 1:2.3. In one embodiment of the invention, the antimicrobial composition comprises N-methyl-1,2-benzisothiazolin-3-one and myristamidopropyl PG-dimonium chloride phosphate. Preferably, a weight ratio of myristamidopropyl PG-dimonium chloride phosphate to N-methyl-1,2-benzisothiazoln-3-one is from 1:0.0011 to 1:150. In one embodiment of the invention, the antimicrobial composition comprises N-methyl-1,2-benzisothiazolin-3-one and octanoic acid. Preferably, a weight ratio of octanoic acid to N-methyl-1,2-benzisothiazolin-3-one is from 1:0.0046 to 1:0.68, more preferably from 1:0.029 to 1:0.68. In one embodiment of the invention, the antimicrobial composition comprises N-methyl-1,2-benzisothiazolin-3-one and 2-ethylhexyl glycerin. Preferably, a weight ratio of 2-ethylhexyl glycerin to N-methyl-1,2-benzisothiazolin-3-one is from 1:0.017 to 1:2.3. In one embodiment of the invention, the antimicrobial composition comprises N-methyl-1,2-benzisothiazolin-3-one and glyceryl monocaprylate. Preferably, a weight ratio of glyceryl monocaprylate to N-methyl-1,2-benzisothiazolin-3-one is from 1:0.0004 to 1:75, more preferably from 1:0.012 to 1:75. In one embodiment of the invention, the antimicrobial composition comprises N-methyl-1,2-benzisothiazolin-3-one and glyceryl monocaprate. Preferably, a weight ratio of glyceryl monocaprate to N-methyl-1,2-benzisothiazolin-3-one is from 1:0.0031 to 1:8.8, more preferably from 1:0.017 to 1:8.8. In one embodiment of the invention, the antimicrobial composition comprises N-methyl-1,2-benzisothiazolin-3-one and 7o-ethyldihydro-1H, 3H, 5H-oxazolo (3,4 C) oxazole. Preferably, a weight ratio of 7a-ethyldihydro-1H, 3H, 5H-oxazolo (3,4-C) oxazole to N-methyl-1,2-benzisothiazolin-3-one is from 1:0.0003 to 1:7.1. In one embodiment of the invention, the antimicrobial composition comprises N-methyl-1,2-benzisothiazolin-3-one and 2-(thiocyanomethylthio) 4 benzothiazole. Preferably, a weight ratio of 2-thiocyanomethylthio) benzothiazole to N-methyl-1,2-benzisothiazolin-3-one is from 1:0.063 to 1:143. The microbicides in the composition of this invention may be used "as is" or may first be formulated with a solvent or a solid carrier. Suitable solvents 5 include, for example, water; glycols, such as ethylene glycol, propylene glycol, diethylene glycol, dipropylene glycol, polyethylene glycol, and polypropylene glycol; glycol ethers; alcohols, such as methanol, ethanol, propanol, phenethyl alcohol and phenoxypropanol; ketones, such as acetone and methyl ethyl ketone; esters, such as ethyl acetate, butyl acetate, triacetyl citrate, and glycerol triacetate; carbonates, such as propylene carbonate and dimethyl carbonate; and mixtures thereof. It is preferred that the solvent is selected from water, glycols, glycol ethers, esters and mixtures thereof. Suitable solid carriers include, for example, cyclodextrin, silicas, diatomaceous earth, waxes, cellulosic materials, alkali and alkaline earth (e.g., sodium, magnesium, potassium) metal salts (e.g., 5 chloride, nitrate, bromide, sulfate) and charcoal. When a microbicide component is formulated in a solvent, the formulation may optionally contain surfactants. When such formulations contain surfactants, they are generally in the form of emulsive concentrates, emulsions, microemulsive concentrates, or microemulsions. Emulsive concentrates form 3 emulsions upon the addition of a sufficient amount of water. Microemulsive concentrates form microemulsions upon the addition of a sufficient amount of water. Such emulsive and microemulsive concentrates are generally well known in the art; it is preferred that such formulations are free of surfactants. U.S. Patent No. 5,444,078 may be consulted for further general and specific details on 5 the preparation of various microemulsions and microemulsive concentrates. A microbicide component also can be formulated in the form of a dispersion. The solvent component of the dispersion can be an organic solvent or water, preferably water. Such dispersions can contain adjuvants, for example, co-solvents, thickeners, anti-freeze agents, dispersants, fillers, pigments, 0 surfactants, biodispersants, sulfosuccinates, terpenes, furanones, polycations, stabilizers, scale inhibitors and anti-corrosion additives.
5 When both microbicides are each first formulated with a solvent, the solvent used for the first microbicide may be the same as or different from the solvent used to formulate the other commercial microbicide, although water is preferred for most industrial biocide applications. It is preferred that the two 5 solvents are miscible. Those skilled in the art will recognize that the microbicide components of the present invention may be added to a locus sequentially, simultaneously, or may be combined before being added to the locus. It is preferred that the first microbicide and the second microbicide component be added to a locus D simultaneously or sequentially. When the microbicides are added simultaneously or sequentially, each individual component may contain adjuvants, such as, for example, solvent, thickeners, anti-freeze agents, colorants, sequestrants (such as ethylenediamine-tetraacetic acid, ethylenediaminedisuccinic acid, iminodisuccinic acid and salts thereof), .5 dispersants, surfactants, biodispersants, sulfosuccinates, terpenes, furanones, polycations, stabilizers, scale inhibitors and anti-corrosion additives. The microbicidal compositions of the present invention can be used to inhibit the growth of microorganisms or higher forms of aquatic life (such as protozoans, invertebrates, bryozoans, dinoflagellates, crustaceans, mollusks, etc.) ) by introducing a microbicidally effective amount of the compositions onto, into, or at a locus subject to microbial attack. Suitable loci include, for example: industrial process water; electrocoat deposition systems,; cooling towers; air washers; gas scrubbers; mineral slurries; wastewater treatment; ornamental fountains; reverse osmosis filtration; ultrafiltration; ballast water; evaporative 5 condensers; heat exchangers; pulp and paper processing fluids and additives; starch; plastics; emulsions; dispersions; paints; latices; coatings, such as varnishes; construction products, such as mastics, caulks, and sealants; construction adhesives, such as ceramic adhesives, carpet backing adhesives, and laminating adhesives; industrial or consumer adhesives; photographic chemicals; 0 printing fluids; household products, such as bathroom and kitchen cleaners and sanitary wipes; cosmetics; toiletries; shampoos; soaps; detergents; industrial cleaners; floor polishes; laundry rinse water; metalworking fluids; conveyor 6 lubricants; hydraulic fluids; leather and leather products; textiles; textile products; wood and wood products, such as plywood, chipboard, wallboard, flakeboard, laminated beams, oriented strandboard, hardboard, and particleboard; petroleum processing fluids; fuel; oilfield fluids, such as injection 5 water, fracture fluids, and drilling muds; agriculture adjuvant preservation; surfactant preservation; medical devices; diagnostic reagent preservation; food preservation, such as plastic or paper food wrap; food, beverage, and industrial process pasteurizers; toilet bowls; recreational water; pools; and spas. Preferably, the microbicidal compositions of the present invention are used 0 to inhibit the growth of microorganisms at a locus selected from one or more of mineral slurries, pulp and paper processing fluids and additives, starch, emulsions, dispersions, paints, latices, coatings. construction adhesives, such as ceramic adhesives, carpet backing adhesives, photographic chemicals, printing fluids, household products such as bathroom and kitchen cleaners and sanitary 5 wipes, cosmetics, toiletries, shampoos, soaps, detergents, industrial cleaners, floor polishes, laundry rinse water, metal working fluids, textile products, wood and wood products, agriculture adjuvant preservation, surfactant preservation, diagnostic reagent preservation, food preservation, and food, beverage, and industrial process pasteurizers. o The specific amount of the composition of this invention necessary to inhibit or control the growth of microorganisms and higher aquatic life forms in a locus depends upon the particular locus to be protected. Typically, the amount of the composition of the present invention to control the growth of microorganisms in a locus is sufficient if it provides from 0.1 to 1,000 ppm of the isothiazoline 15 ingredient of the composition in the locus. It is preferred that the isothiazolone ingredients of the composition be present in the locus in an amount of at least 0.5 ppm, more preferably at least 4 ppm and most preferably at least 10 ppm. It is preferred that the isothiazolone ingredients of the composition be present in the locus in'an amount of no more than 1000 ppm, more preferably no more than 500 0 ppm, and most preferably no more than 200 ppm.
7 EXAMPLES Materials and Methods The synergism of the combination of the present invention was 5 demonstrated by testing a wide range of concentrations and ratios of the compounds. One measure of synergism is the industrially accepted method described by Kull, F.C.; Eisman, P.C.; Sylwestrowicz, H.D. and Mayer, R.L., in Applied Microbiology 9:538-541 (1961), using the ratio determined by the formula: 0 Qa/QA + Qb/QB = Synergy Index ("SI") wherein: QA = concentration of compound A (first component) in ppm, acting alone, which produced an end point (MIC of Compound A). 5 Qa = concentration of compound A in ppm, in the mixture, which produced an end point. QB = concentration of compound B (second component) in ppm, acting alone, which produced an end point (MIC of Compound B). Qb = concentration of compound B in ppm, in the mixture, which produced 3 an end point. When the sum of Qa/QA and Qb/QB is greater than one, antagonism is indicated. When the sum is equal to one, additivity is indicated, and when less than one, synergism is demonstrated. The lower the SI, the greater the synergy 5 shown by that particular mixture. The minimum inhibitory concentration (MIC) of a microbicide is the lowest concentration tested under a specific set of conditions that prevents the growth of added microorganisms. Synergy tests were conducted using standard microtiter plate assays with media designed for optimal growth of the test microorganism. Minimal salt 0 medium supplemented with 0.2% glucose and 0.1% yeast extract (M9GY medium) was used for testing bacteria; Potato Dextrose Broth (PDB medium) was used for testing yeast and mold. In this method, a wide range of combinations of microbicides was tested by conducting high resolution MIC 8 assays in the presence of various concentrations of MBIT. High resolution MICs were determined by adding varying amounts of microbicide to one column of a microtitre plate and doing subsequent ten-fold dilutions using an automated liquid handling system to obtain a series of endpoints ranging from 2 ppm to 5 10,000 ppm active ingredient. The synergy of the combinations of the present invention was determined against several microorganisms, as described in the Tables below. The bacteria were used at a concentration of about 5 x 106 bacteria per mL and the yeast and mold at 5 x 105 fungi per mL. These microorganisms are representative of 3 natural contaminants in many consumer and industrial applications. The plates were visually evaluated for microbial growth (turbidity) to determine the MIC after various incubation times at 250 C (yeast and mold) or 30* C (bacteria). The test results for demonstration of synergy of the MBIT combinations of the present invention are shown below in Tables 1 through 10. In each test, Second Component (B) was MBIT and the First Component (A) was the other commercial microbicide. Each table shows the specific combinations of MBIT and the other component; results against the microorganisms tested with incubation times; the end-point activity in ppm measured by the MIC for MBIT alone (QB), for the other component alone (QA), for MBIT in the mixture (Qb) and for the other component in the mixture (Qa); the calculated SI value; and the range of synergistic ratios for each combination tested (other component/MBIT or
A/B).
9 Table: 1 Ca: ppm Al of BIOBAN GK(hexahydro-1,3,5-tris(2-hydroxyethyl)-s-triazine) Cb: ppm Al of MBIT(N-methyl-1,2-benzisothiazolin-3-one) Ratio:Ca:Cb 5 ---------------- - -- -____--_---- Test Organisms Contact Ca Cb S.I. Ca:Cb Time A. niger 3 days 425 ATCC # 16404 - 37.5 110 18.8 0.76 1:0.1709 200 18.8 0.97 1:0.0940 300 18.8 1.21 1:0.0627 300 9.4 0.96 1:0.0313 425 9.4 1.25 1:0.0221 300 4.7 0.83 1:0.0157 425 4.7 1.13 1:0.0111 300 2.4 0.77 1:0.0080 425 2.4 1.06 1:0.0056 300 1.2 0.74 1:0.0040 425 1.2 1.03 1:0.0028 7 days 650 150 42.5 75 0.57 1:1.7647 65 75 0.60 1:1.1538 110 75 0.67 1:0.6818 200 75 0.81 1:0.3750 300 75 0.96 1:0.2500 400 75 1.12 1:0.1875 425 18.8 0.78 1:0.0442 525 18.8 0.93 1:0.0358 650 9.4 1.06 1:0.0145 525 4,7 0.84 1:0.0090 650 4.7 1.03 1:0.0072 525 2.4 0.82 1:0.0046 650 2.4 1.02 1:0.0037 650 1.2 1.01 1:0.0018 C.albicans 48 hrs 875 ATCC# 10231 30 87.5 15 0.60 1:0.1714 110 15 0.63 1:0.1364 200 15 0.73 1:0.0750 300 15 0.84 1:0.0500 425 15 0.99 1:0.0353 110 7.5 0.38 1:0.0682 200 7.5 0.48 1:0.0375 300 7.5 0.59 1:0.0250 425 7.5 0.74 1:0.0176 525 7.5 0.85 1:0.0143 650 7.5 0.99 1:0.0115 425 3.75 0.61 1:0.0088 525 3.75 0.73 1:0.0071 650 3.75 0.87 1:0.0058 10 875 3.75 1.13 1:0.0043 650 1.86 0.80 1:0.0029 875 1.86 1.06 1:0.0021 650 0.94 0.77 1:0.0014 875 0.94 1.03 1:0.0011 72 hrs 875 - 30 87.5 15 0.60 1:0.1714 110 15 0.63 1:0.1364 200 15 0.73 1:0.0750 300 15 0.84 1:0.0500 425 15 0.99 1:0.0353 425 7.5 0.74 1:0.0176 525 7.5 0.85 1:0.0143 650 7.5 0.99 1:0.0115 875 1.86 1.06 1:0.0021 875 0.94 1.03 1:0.0011 Ps. aeruginosa 24hrs 55 ATCC#9027 - 75 32.5 37.5 1.09 1:1.1538 43.75 18.75 1.05 1:0.4286 43.75 9.4 0.92 1:0.2149 43.75 4.6 0.86 1:0.1051 48hrs 55 - - 125 26.25 75 1.08 1:2.8571 32.5 37.5 0.89 1:1.1538 43.75 37.5 1.10 1:0.8571 55 4.6 1.04 1:0.0836 Saureus 24hrs 55 ATCC#6538 - 15 15 7.5 0.77 1:0.5000 32.5 3.8 0.84 1:0.1169 43.75 3.8 1.05 1:0.0869 32.5 1.8 0.71 1:0.0554 43.75 1.8 0.92 1:0.0411 55 1.8 1.12 1:0.0327 55 0.94 1.06 1:0.0171 48hrs 55 - 30 0.1 15 0.50 1:150.0000 1.5 15 0.53 1:10.0000 2.625 15 0.55 1:5.7143 4.375 15 0.58 1:3.4286 5.5 15 0.60 1:2.7273 10 15 0.68 1:1.5000 15 15 0.77 1:1.0000 21.25 15 0.89 1:0.7059 26.25 15 0.98 1:0.5714 32.5 15 1.09 1:0.4615 21.25 7.5 0.64 1:0.3529 26.25 7.5 0.73 1:0.2857 11 32.5 7.5 0.84 1:0.2308 43.75 7.5 1.05 1:0.1714 32.5 3.8 0.72 1:0.1169 43.75 3'8 0.92 1:0.0869 55 3.8 1.13 1:0.0691 32.5 18 0.65 1:0.0554 55 18 1.06 1:0.0327 43.75 0.94 0.83 1:0.0215 55 0.94 L.3 1:0.0171 Table: 2 Ca: ppm AI of BIOBAN 1135 (4,4-dimethyloxazolidine) Cb: ppm Al of MBIT(N-methyl-1,2-benzisothiazolin-3-one) 5 Ratio:Ca:Cb Test Organisms Contact Ca Cb S.I. Ca:Cb Time A. niger 3 days 525 ATCC # 16404 - 37.5 200 18.8 0.88 1:0.0940 300 18.8 1.07 1:0.0627 300 9.4 0.82 1:0.0313 425 9.4 1.06 1:0.0221 425 4.7 0.93 1:0.0111 525 4.7 1.13 1:0.0090 425 2.4 0.87 1:0.0056 525 2.4 1.06 1:0.0046 425 1.2 0.84 1:0.0028 525 1.2 1.03 1:0.0023 7 days 875 - 150 525 75 1.1 1:0.1429 650 37.5 0.99 1:0.0577 875 9.4 1.06 1:0.0107 650 4.7 0.77 1:0.0072 875 4.7 1.03 1:0.0054 650 2.4 0.76 1:0.0037 875 2.4 1.02 1:0.0027 875 1.2 1.01 1:0.0014 C alhicans 48 hrs 1100 - ATCC # 10231 30 425 15 0.89 1:0.0353 525 15 0.98 1:0.0286 650 15 1.09 1:0.0231 650 7.5 0.84 1:0.0115 875 7.5 1.05 1:0.0086 875 3.75 0.92 1:0.0043 1100 3.75 1.13 1:0.0034 875 1.86 0.86 1:0.0021 1100 1.86 1.06 1:0.0017 875 0.94 0.83 1:0.0011 12 1100 0.94 1.03 1:0.0009 72hrs 1100 30 425 15 0.89 1:0.0353 525 15 0.98 1:0.0286 650 15 1.09 1:0.0231 875 7.5 1.05 1:0.0086 875 3.75 0.92 1:0.0043 1100 3.75 1.13 1:0.0034 875 1.86 0.86 1:0.0021 1100 1.86 1.06 1:0.0017 875 0.94 0.83 1:0.0011 1100 0.94 1.03 1:0.0009 Ps. aeruginosa 24hrs 100 ATCC#9027 - 75 55 37.5 1.05 1:0.6818 55 9.4 0.68 1:0.1709 100 9.4 1.13 1:0.0940 55 4.6 0.61 1:0.0836 100 4.6 1.06 1:0.0460 48hrs 100 125 21.25 75 0.81 1:3.5294 26.25 75 0.86 1:2.8571 32.5 75 0.93 1:2.3077 43.75 75 1.04 1:1.7143 55 37.5 0.85 1:0.6818 100 9.4 1.08 1:0.0940 100 4.6 1.04 1:0.0460 S.aureus 24hrs 55 ATCC#6538 30 21.25 15 0.89 1:0.7059 26.25 15 0.98 1:0.5714 32.5 15 1.09 1:0.4615 26.25 7.5 0.73 1:0.2857 32.5 7.5 0.84 1:0.2308 43.75 7.5 1.05 1:0.1714 43.75 3.8 0.92 1:0.0869 55 3.8 1.13 1:0.0691 55 1.8 1.06 1:0.0327 55 0.94 1.03 1:0,0171 48hrs 55 30 21.25 15 0.89 1:0.7059 26.25 15 0.98 1:0.5714 32.5 15 1.09 1:0.4615 26.25 7.5 0.73 1:0.2857 32.5 7.5 0.84 1:0.2308 43.75 7.5 1.05 1:0.1714 43.75 3.8 0.92 1:0.0869 55 3.8 1.13 1:0.0691 55 1.8 1.06 1:0.0327 13 55 0.94 | 1.03 | 1:0.0171 Table: 3 Ca: ppm AI of SUTTOCIDE A ( sodium hydroxymethylglycinate) 5 Cb: ppm Al of MBIT(N-methyl-I.,2-benzisothiazolin-3-one) Ratio:Ca:Cb Test Organisms Contact Ca Cb S.I. Ca:Cb Time A. niger 3 days 875 ATCC # 16404 - 75 425 18.8 0.74 1:0.0442 525 18.8 0.85 1:0.0358 650 18.8 0.99 1:0.0289 425 9.4 0.61 1:0.0221 525 9.4 0.73 1:0.0179 650 9.4 0.87 1:0.0145 875 9.4 1.13 1:0.0107 650 4.7 0.81 1:0.0072 875 4.7 1.06 1:0.0054 525 2.4 0.63 1:0.0046 650 2.4 0.77 1:0.0037 875 2.4 1.03 1:0.0027 650 1.2 0.76 1:0.0018 875 1.2 1.02 1:0.0014 7 days 1100 - 150 110 75 0.60 1:0.6818 200 75 0.68 1:0.3750 300 75 0.77 1:0.2500 425 75 0.89 1:0.1765 525 75 0.98 1:0.1429 650 75 1.09 1:0.1154 425 37.5 0.64 1:0.0882 525 37.5 0.73 1:0.0714 650 37.5 0.84 1:0.0577 875 37.5 1.05 1:0.0429 525 18.8 0.6 1:0.0358 650 18.8 0.72 1:0.0289 875 18.8 0.92 1:0.0215 1100 18.8 1.13 1:0.0171 525 9.4 0.54 1:0.0179 650 9.4 0.65 1:0.0145 875 9.4 0.86 1:0.0107 1100 9.4 1.06 1:0.0085 650 4.7 0.62 1:0.0072 875 4.7 0.83 1:0.0054 1100 4.7 1.03 1:0.0043 650 2.4 0.61 1:0.0037 875 2.4 0.81 1:0.0027 14 1100 2.4 1.02 1:0.0022 650 1.2 0.60 1:0.0018 875 1.2 0.80 1:0.0014 1100 1.2 1.01 1:0.0011 Calbicans 48 hrs 3000 ATCC# 10231 30 525 15 0.68 1:0.0286 650 15 0.72 1:0.0231 875 15 0.79 1:0.0171 1100 15 0.87 1:0.0136 2000 15 1.17 1:0.0075 525 7.5 0.43 1:0.0143 650 7.5 0.47 1:0.0115 875 7.5 0.54 1:0.0086 1100 7.5 0.62 1:0.0068 2000 7.5 0.92 1:0.0038 3000 7.5 1.25 1:0.0025 875 3.75 0.42 1:0.0043 1100 3.75 0.49 1:0.0034 2000 3.75 0.79 1:0.0019 3000 3.75 1.13 1:0.0013 2000 1.86 0.73 1:0.0009 3000 1.86 1.06 1:0.0006 2000 0.94 0.70 1:0.0005 3000 0.94 1.03 1:0.0003 72 hrs 3000 - 30 525 15 0.68 1:0.0286 650 15 0.72 1:0.0231 875 15 0.79 1:0.0171 1100 15 0.87 1:0.0136 2000 15 1.17 1:0.0075 3000 15 1.5 1:0.0050 525 7.5 0.43 1:0.0143 650 7.5 0.47 1:0.01,15 875 7.5 0.54 1:0.0086 1100 7.5 0.62 1:0.0068 2000 7.5 0.92 1:0.0038 3000 7.5 1.25 1:0.0025 875 3.75 0.42 1:0.0043 1100 3.75 0.49 1:0.0034 2000 3.75 0.79 1:0.0019 3000 3.75 1.13 1:0.0013 2000 1.86 0.73 1:0.0009 3000 1.86 1.06 1:0.0006 2000 0.94 0.70 1:0.0005 3000 0.94 1.03 1:0.0003 Ps. aeruginosa 24hrs 87.5 ATCC#9027 - 75 65 9.4 0.87 1:0.1446 87.5 9.4 1.13 1:0.1074 48 hrs 110 - 15 125 87.5 9.4 0.87 1:0.1074 110 9.4 1.08 1:0.0855 110 4.6 1.04 1:0.0418 S.aureus 24hrs 110 ATCC#6538 - 15 - 30 7.5 0.77 1:0.2500 42.5 7.5 89 1:0.1765 52.5 7.5 98 1:0.1429 65 7.5 1.09 1:0.1154 52.5 3.8 0.73 1:0.0724 65 3.8 0.84 1:0.0585 87.5 3.8 1.05 1:0.0434 110 0.94 1.06 1:0.0085 48hr8 110
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30 6.5 15 0.56 1:2.3077 8.75 15 0.58 1:1.7143 11 15 0.6 1:1.3636 20 15 0.68 1:0.7500 30 15 0.77 1:0.5000 42.5 15 0.89 1:0.3529 52.5 15 0.98 1:0.2857 65 15 1.09 1:0.2308 42.5 7.5 0.64 1:0.1765 52.5 7.5 0.73 1:0.1429 65 7.5 0.84 1:0.1154 87.5 7.5 1.05 1:0.0857 65 3.8 0.72 1:0.0585 110 1.8 1.06 1:0.0164 110 0.94 1.03 1:0.0085 Table:4 Ca: ppm Al of Phospholipid PTM( myristamidopropyl PG-dimonium chloride phosphate ) Cb: ppm Al of MBIT(N-methyl-1,2-benzisothiazolin-3-one) Ratio:Ca:Cb Test Organisms Contact Ca Cb .1. Ca:Cb Time A. niger 3 days 1100 ATCC # 16404 - 37.5 - 425 18.8 0.89 1:0.0442 525 18.8 0.98 1:0.0358 650 18.8 1.09 1:0.0289 525 9.4 0.73 1:0.0179 650 9.4 0.84 1:0.0145 875 9.4 1.05 1:0.0107 1100 2.4 1.06 1:0.0022 1100 1.2 1.03 1:0.0011 7 days 1100 16 150 2 75 0.50 1:37.5000 20 75 0.52 1:3.7500 200 75 0.68 1:0.3750 525 75 0.98 1:0.1429 650 75 1.09 1:0,1154 20 37.5 0.27 1:1.8750 52.5 37.5 0.3 1:0.7143 87.5 37.5 0.33 1:0.4286 200 37.5 0.43 1:0.1875 425 37.5 0.64 1:0.0882 650 37.5 0.84 1:0.0577 875 37.5 1.05 1:0.0429 650 18.8 0.72 1:0.0289 875 18.8 0.92 1:0.0215 1100 18.8 1.13 1:0.0171 650 9.4 0.65 1:0.0145 875 9.4 0.86 1:0.0107 1100 9.4 1.06 1:0.0085 1100 4.7 1.03 1:0.0043 1100 2.4 1.02 1:0.0022 1100 1.2 1.01 1:0.0011 Calbicans 48 hrs 1100 ATCC# 10231 - 30 525 15 0.98 1:0.0286 650 15 1.09 1:0.0231 525 7.5 0.73 1:0.0143 650 7.5 0.84 1:0.0115 875 7.5 1.05 1:0.0086 650 3.75 0.72 1:0.0058 875 3.75 0.92 1:0.0043 1100 3.75 1.13 1:0.0034 875 1.86 0.86 1:0.0021 1100 1.86 1.06 1:0.0017 875 0.94 0.83 1:0.0011 1100 0.94 1.03 1:0.0009 72 hrs 1100 - 60 2 30 0.50 1:15.0000 20 30 0.52 1:1.5000 110 30 0.60 1:0.2727 200 30 0.68 1:0.1500 300 30 0.77 1:0.1000 425 30 0.89 1:0.0706 525 30 0.98 1:0.0571 650 30 1.09 1:0.0462 525 15 0.73 1:0.0286 650 15 0.84 1:0.0231 525 7.5 0.6 1:0.0143 650 7.5 0.72 1:0.0115 875 7.5 0.92 1:0.0086 650 3.75 0.65 1:0.0058 17 875 3.75 0.86 1:0.0043 1100 3.75 1.06 1:0.0034 875 1.86 0.83 1:0.0021 1100 1.86 1.03 1:0.0017 875 0.94 0.81 1:0.0011 1100 0.94 1.02 1:0.0009 Ps. aeruginosa 24hrs 425 ATCC#9027 75 425 4.6 1.06 1:0.0108 48hrs 525 125 525 4.6 1.04 1:0.0088 S.aureus 24hrs 10 ATCG#6538 - 15 2.625 7.5 0.76 1:2.8571 3.25 7.5 0.83 1:2.3077 4.375 7.5 0.94 1:1.7143 5.5 7.5 1.05 1:1.3636 4.375 3.8 0.69 1:0.8686 5.5 3.8 0.80 1:0.6909 10 3.8 1.25 1:0.3800 4.375 1.8 0.56 1:0.4114 5.5 1.8 0.67 1:0.3273 10 1.8 1.12 1:0.1800 10 0.94 1.06 1:0.0940 48hrs 10 30 0.1 15 0.51 1:150.0000 0.5 15 0.55 1:30.0000 1 15 0.6 1:15.0000 1.5 15 0.65 1:10.0000 2.125 15 0.71 1:7.0588 2.625 15 0.76 1:5.7143 3.25 15 0.83 1:4.6154 4.375 15 0.94 1:3.4286 5.5 15 1.05 1:2.7273 4.375 7.5 0.69 1:1.7143 5.5 7.5 0.80 1:1.3636 10 7.5 1.25 1:0.7500 4.375 3.8 0.56 1:0.8686 5.5 3.8 0.68 1:0.6909 10 3.8 1.13 1:0.3800 5.5 1.8 0.61 1:0.3273 10 1.8 1.06 1:0.1800 10 0.94 L03 1:0.0940 18 Table: 5 Ca: ppm Al of Caprylic acid (octanoic acid) Cb: ppm AI of MBIT(N-methyl-1,2-benzisothiazolin-3-one) Ratio:Ca:Cb Test Organisms Contact Ca Cb S.I. Ca:Cb Time A. niger 3 days 650 ATCC # 16404 75 425 18.8 0.9 1:0.0442 525 18.8 1.06 1:0.0358 525 9.4 0.93 1:0.0179 650 9.4 1.13 1:0.0145 525 4.7 0.87 1:0.0090 525 2.4 0.84 1:0.0046 650 2.4 1.03 1:0.0037 650 1.2 102 1:0.0018 7 days 875 150 110 75 0.63 1:0.6818 200 75 0.73 1:0.3750 300 75 0.84 1:0.2500 425 75 0.99 1:0.1765 650 37.5 0.99 1:0.0577 425 18.8 0.61 1:0.0442 650 18.8 0.87 1:0.0289 875 18.8 1.13 1:0.0215 875 9.4 1.06 1:0.0107 875 4.7 1.03 1:0,0054 875 2.4 1.02 1:0.0027 875 1.2 1.01 1:0.0014 C.albicans 48 hrs 65 ATCC# 10231 - 30 42.5 7.5 0.9 1:0.1765 52.5 7.5 1.06 1:0.1429 65 1.86 1.06 1:0.0286 65 0.94 1.03 1:0.0145 72 hrs 87.5 30 42.5 15 0.99 1:0.3529 42.5 7.5 0.74 1:0.1765 52.5 7.5 0.85 1:0.1429 65 7.5 0.99 1:0.1154 65 3.75 0.87 1:0.0577 87.5 3.75 1.13 1:0.0429 87.5 1.86 1.06 1:0.0213 87 5 0.94 1.03 1:0.0107 Ps. aeruginosa 48 hrs >11000 ATCC#9027 - 125 8750 75 1.4 1:0.0086 11000 37.5 1.3 1:0.0034 19 11000 18.75 1.15 1:0.0017 11000 9.4 1.08 1:0.0009 11000 4.6 1.04 1:0.0004 S.aureus 24hrs 425 ATCC#6538 - 15 425 0.94 1.06 1:0.0022 48hrs 425 - 30 425 1.8 1.06 1:0.0042 J 425 0.94 1.03 1:0.0022 Table: 6 Ca: ppm AI of SENSIVA SC 50( 3[(2-ethylhexyl)oxy]1,2-propanediol) (2 ethylhexyl glycerin) 5 Cb: ppm Al of MBIT(N-methyl-1,2-benzisothiazolin-3-one) Ratio:Ca:Cb Test Organisms Contact Ca Cb S.I. Ca:Cb Time A. niger 3 days 875 ATCC # 16404 - 37.5 650 9.4 0.99 1:0.0145 650 4.7 0.87 1:0.0072 875 4.7 1.13 1:0.0054 875 2.4 1.06 1:0.0027 875 1.2 1.03 1:0.0014 7 days 1100 - 150 87.5 75 0.58 1:0.8571 110 75 0.60 1:0.6818 200 75 0.68 1:0.3750 300 75 0.77 1:0.2500 425 75 0.89 1:0.1765 525 75 0.98 1:0.1429 650 75 1.09 1:0.1154 650 37.5 0.84 1:0.0577 1100 9.4 1.06 1:0.0085 1100 4.7 1.03 1:0.0043 1100 2.4 1.02 1:0.0022 1100 1.2 1.01 1:0.0011 C.albicans 48 hrs 2000 - ATCC# 10231 - 30 6.5 15 0.50 1:2.3077 11 15 0.51 1:1.3636 110 15 0.56 1:0.1364 200 15 0.60 1:0.0750 300 15 0.65 1:0.0500 650 15 0.83 1:0,0231 875 15 0.94 1:0.0171 1100 15 1.05 1:0.0136 2000 1.86 1.06 1:0.0009 20 2000 0.94 1.03 1:0.0005 72hrs 2000 30 425 15 0.71 1:0.0353 525 15 0.76 1:0.0286 650 15 0.83 1:0.0231 875 15 0.94 1:0.0171 1100 15 1.05 1:0.0136 2000 1.86 1.06 1:0.0009 2000 0.94 1.03 1:0.0005 Ps. aeruginosa 24hrs 11000 ATCC#9027 75 8750 37.5 1.3 1:0.0043 48hrs 11000 125 4250 75 0.99 1:0.0176 ~aureus 24hrs 2000 ATCC#6538 30 8.75 15 0.5 1:1.7143 20 15 0.51 1:0.7500 42.5 15 0.52 1:0.3529 65 15 0.53 1:0.2308 110 15 0.56 1:0.1364 200 15 0.6 1:0.0750 425 15 0.71 1:0.0353 525 15 0.76 1:0.0286 650 15 0.83 1:0.0231 875 15 0.94 1:0.0171 1100 15 1.05 1:0.0136 2000 1.8 1.06 1:0.0009 2000 0.94 1.03 1:0.0005 48hrs 2000 - 30 8.75 15 0.50 1:1.7143 20 15 0.51 1:0.7500 42.5 15 0.52 1:0.3529 65 15 0.53 1:0.2308 110 15 0.56 1:0.1364 200 15 0.60 1:0.0750 425 15 0.71 1:0.0353 525 15 0.76 1:0.0286 650 15 0.83 1:0.0231 875 15 0.94 1:0.0171 1100 15 1.05 1:0.0136 2000 1.8 1.06 1:0.0009 -- _-_---------------- _ 2000 0.94 | 1.03 1:0.0005 21 Table: 7 Ca: ppm AI of CAPMUL MCM C8 (glyceryl monocaprylate) Cb: ppm Al of MBIT(N-methyl- 1, 2-benzisothiazolin- 3-one) Ratio:Ca,:Cb 5 ------------ Test Organisms Contact Ca Cb S.I Ca:Cb Time A. niger 3 days 4250 ATCC # 16404 - 75 2000 37.5 0.97 1:0.0188 3000 37.5 1.21 1:0.0125 2000 18.8 0.72 1:0.0094 3000 18.8 0.96 1:0.0063 4250 18.8 1.25 1:0.0044 2000 9.4 0.60 1:0.0047 3000 9.4 0.83 1:0.0031 4250 9.4 1.13 1:0.0022 2000 4.7 0.53 1:0.0024 3000 4.7 0.77 1:0.0016 4250 4.7 L06 1:0.0011 2000 2.4 0.5 1:0.0012 3000 2.4 0.74 1:0.0008 4250 2.4 1.03 1:0.0006 3000 1.2 0.72 1:0.0004 4250 1.2 1.02 1:0.0003 7 days 4250 - 150 1100 75 0.76 1:0.0682 2000 75 0.97 1:0.0375 3000 75 1.21 1:0.0250 2000 37.5 0.72 1:0.0188 3000 37.5 0.96 1:0.0125 4250 37.5 1.25 1:0.0088 2000 18.8 0.60 1:0.0094 3000 18.8 0.83 1:0.0063 4250 18.8 1.13 1:0.0044 2000 9.4 0.53 1:0.0047 3000 9.4 0.77 1:0.0031 4250 9.4 1.06 1:0.0022 2000 4.7 0.50 1:0.0024 3000 4.7 0.74 1:0.0016 4250 4.7 1.03 1:0.0011 2000 2.4 0.49 1:0.0012 3000 2.4 0.72 1:0.0008 - ----------------- 4250 2.4 1.02 1:0.0006 Calbicans 48 hrs 200 ATCC # 10231 - 30 0.2 15 0.50 1:75.0000 2 15 0.51 1:7.5000 20 15 0.60 1:0.7500 42.5 15 0.71 1:0.3529 65 15 0.83 1:0.2308 22 87.5 15 0.94 1:0.1714 110 15 1.05 1:0.1364 200 1.86 1.06 1:0.0093 200 0.94 1.03 1:0.0047 72 hrs 200 30 0.2 15 0.50 1:75.0000 2 15 0.51 1:7.5000 20 15 0.6 1:0.7500 42.5 15 0.71 1:0.3529 65 15 0.83 1:0.2308 87.5 15 0.94 1:0.1714 110 15 1.05 1:0.1364 200 1.86 1.06 1:0.0093 200 0.94 1.03 1:0.0047 Ps. aeruginosa 48 hrs >11000 ATCC#9027 - 125 2000 75 0.78 1:0.0375 3000 75 0.87 1:0.0250 -_-_-------------------_ 4250 75 0.99 1:0.0176 Saureus 24hrs 650 - ATCO#6538 30 0.2 15 0.50 1:75.0000 2 15 0.50 1:7.5000 6.5 15 0.51 1:2.3077 11 15 0.52 1:1.3636 20 15 0.53 1:0.7500 30 15 0.55 1:0.5000 42.5 15 0.57 1:0.3529 52.5 15 0.58 1:0.2857 65 15 0.60 1:0.2308 87.5 15 0.63 1:0.1714 110 15 0.67 1:0.1364 200 15 0.81 1:0.0750 300 15 0.96 1:0.0500 425 15 1.15 1:0.0353 425 7.5 0.9 1:0.0176 525 7.5 1.06 1:0.0143 48hrs 875 30 0.2 15 0.50 1:75.0000 2 15 0.50 1:7.5000 6.5 15 0.51 1:2.3077 11 15 0.51 1:1.3636 20 15 0.52 1:0.7500 30 15 0.53 1:0.5000 42.5 15 0.55 1:0.3529 52.5 15 0.56 1:0.2857 65 15 0.57 1:0.2308 87.5 15 0.60 1:0.1714 110 15 0.63 1:0.1364 200 15 0.73 1:0.0750 23 300 15 0.84 1:0.0500 425 15 0.99 1:0.0353 650 7.5 0.99 1:0.0115 875 1.8 1.06 1:0.0021 A____ __ 875 0.94 1.03 1:0.0011 Table:8 Ca: ppm Al of CapmulMCM C10 (Mono/digyceride of caprice acid (Glyceryl Monocaprate)) 5 Cb: ppm AI of MBIT(N-methyl-1,2-benzisothiazolin-3-one) Ratio:Ca:Cb Test Organisms Contact Ca Cb S.I. Ca:Cb Time A. niger 3 days 2000 ATCC# 16404 - 75 4.25 37.5 0.50 1:8.8235 6.5 37.5 0.50 1:5.7692 11 37.5 0.51 1:3.4091 30 37.5 0.52 1:1.2500 65 37.5 0.53 1:0.5769 110 37.5 0.56 1:0.3409 200 37.5 0.60 1:0.1875 300 37.5 0.65 1:0.1250 425 37.5 0.71 1:0.0882 525 37.5 0.76 1:0.0714 650 37.5 0.83 1:0.0577 875 37.5 0.94 1:0.0429 1100 37.5 1.05 1:0.0341 1100 18.8 0.8 1:0.0171 2000 4.7 1.06 1:0.0024 2000 2.4 1.03 1:0.0012 2000 1.2 1.02 1:0.0006 7 days 2000 - 150 525 75 0.76 1:0.1429 650 75 0.83 1:0.1154 875 75 0.94 1:0.0857 1100 75 1.05 1:0.0682 1100 37.5 0.8 1:0.0341 1100 18.8 0.68 1:0.0171 2000 9.4 1.06 1:0.0047 2000 4.7 1.03 1:0.0024 2000 2.4 1.02 1:0.0012 2000 1.2 1.01 1:0.0006 C.albicans 48 hrs 425 ATCC# 10231 - 30 87.5 15 0.71 1:0.1714 110 15 0.76 1:0.1364 200 15 0.97 1:0.0750 300 15 1.21 1:0.0500 24 200 7.5 0.72 1:0.0375 300 7.5 0.96 1:0.0250 425 7.5 1.25 1:0.0176 200 3.75 0.60 1:0.0188 300 3.75 0.83 1:0.0125 425 3.75 1.13 1:0.0088 200 1.86 0.53 1:0.0093 300 1.86 0.77 1:0.0062 425 1.86 1.06 1:0.0044 200 0.94 0.50 1:0.0047 300 0.94 0.74 1:0.0031 425 0.94 1.03 1:00022 72 hrs 425 30 87.5 15 0.71 1:0.1714 110 15 0.76 1:0,1364 200 15 0.97 1:0.0750 300 15 1.21 1:0,0500 200 7.5 0.72 1:0,0375 300 7.5 0.96 1:0.0250 425 7.5 1.25 1:0.0176 200 3.75 0.60 1:0.0188 300 3.75 0.83 1:0.0125 425 3.75 1.13 1:0.0088 200 1.86 0.53 1:0.0093 300 1.86 0.77 1:0.0062 425 1.86 1.06 1:0.0044 200 0.94 0.50 1:0.0047 300 0.94 0.74 1:0.0031 425 0.94 1.03 1:0.0022 Ps. aeruginosa 24hrs 11000 ATCO#9027 - 75 37.5 18.75 9.4 4.6 48 hirs 11000 125 - 75 37.5 18.75 - 9.4 - 4.6 _ _ S.aureus 24hrs 200 ATCC#6538 - 30 5.25 15 0.53 1:2.8571 11 15 0.56 1:1.3636 20 15 0.60 1:0.7500 30 15 0.65 1:0.5000 42.5 15 0.71 1:0.3529 52.5 15 0.76 1:0.2857 65 15 0.83 1:0.2308 87.5 15 0.94 1:0.1714 110 15 1.05 1:0.1364 48hrs 200 - 30 - 65 15 0.83 1:0.2308 87.5 15 0.94 1:0.1714 __ _ _ _ _ _ _ _ _ _ _ _ _ _ 110 --- 15 1--L.5 ....I....1:0. 1364_ Table:9 Ca: ppm AI of BIOBAN1246 (7a -ethyldihydro-1H, 3H, 5H-oxazolo (3,4-C) oxazole) 5 Cb: ppm Al of MBIT(N-methyl-1,2-benzisothiazolin-3-one) Ratio:Ca:Cb Test Organisms Contact Ca Cb S.I. Ca:Cb Time A. niger 3 days 650 ATCC # 16404 - 37.5 200 18.8 0.81 1:0.0940 300 18.8 0.96 1:0.0627 425 18.8 1.16 1:0.0442 425 9.4 0.90 1:0.0221 525 9.4 1.06 1:0.0179 425 4.7 0.78 1:0.0111 525 4.7 0.93 1:0.0090 650 4.7 1.13 1:0.0072 425 2.4 0.72 1:0,0056 525 2.4 0.87 1:0.0046 650 2.4 1,06 1:0.0037 425 1.2 0.69 1:0.0028 525 1.2 0.84 1:0.0023 650 1.2 1.03 1:0.0018 7 days 875 - 150 425 75 0.99 1:0,1765 650 37.5 0.99 1:0.0577 525 18.8 0.73 1:0.0358 650 18.8 0.87 1:0.0289 875 18.8 1.13 1:0.0215 875 9.4 1.06 1:0.0107 875 4.7 1.03 1:0.0054 875 2.5 1.02 1:0.0029 875 1.2 1.01 1:0.0014 C.albicans 48 hrs 3000 ATCC# 10231 - 30 875 15 0.79 1:0.0171 1100 15 0.87 1:0.0136 2000 15 1.17 1:0.0075 875 7.5 0.54 1:0.0086 1100 7.5 0.62 1:0.0068 2000 7.5 0.92 1:0.0038 26 3000 7.5 1.25 1:0.0025 2000 3.75 0.79 1:0.0019 3000 3.75 1.13 1:0.0013 2000 1.86 0.73 1:0.0009 3000 1.86 1.06 1:0.0006 3000 0.94 1.03 1:0.0003 72 hrs 4250 30 875 15 0.71 1:0.0171 1100 15 0.76 1:0.0136 2000 15 0.97 1:0.0075 3000 15 1.21 1:0.0050 875 7.5 0.46 1:0.0086 1100 7.5 0.51 1:0.0068 2000 7.5 0.72 1:0.0038 3000 7.5 0.96 1:0.0025 4250 7.5 1.25 1:0.0018 2000 3.75 0.6 1:0.0019 3000 3.75 0.83 1:0.0013 4250 3.75 1.13 1:0.0009 2000 1.86 0.53 1:0.0009 3000 0.94 0.74 1:0.0003 4250 0.94 1.03 1:0.0002 Ps. aeruginosa 24hrs 100 ATCC#9027 - 75 26.25 75 1.26 1:2.8571 43.75 37.5 0.94 1:0.8571 55 37.5 1.05 1:0.6818 55 18.75 0.80 1:0.3409 100 18.75 1.25 1:0.1875 55 9.4 0.68 1:0,1709 100 9.4 1.13 1:0.0940 100 4.6 1.06 1:0.0460 48 hrs 150 125 26.25 75 0.78 1:2.8571 32.5 75 0.82 1:2.3077 43.75 75 0.89 1:1.7143 55 75 0.97 1:1.3636 100 75 1.27 1:0.7500 55 37.5 0.67 1:0.6818 100 37.5 0.97 1:0.3750 150 37.5 1.3 1:0.2500 100 18.75 0.82 1:0.1875 150 18.75 1.15 1:0.1250 150 9.4 1.08 1:0.0627 150 4.6 1.04 1:0.0307 S.aureus 24hrs 55 ATCC#6538 - 30 2.125 15 0.54 1:7.0588 5.5 15 0.60 1:2.7273 10 15 0.68 1:1.5000 27 26.25 15 0.98 1:0.5714 32.5 15 1.09 1:0.4615 21.25 7.5 0.64 1:0.3529 26.25 7.5 0.73 1:0.2857 32.5 7.5 0.84 1:0.2308 43.75 7.5 1.05 1:0,1714 32.5 3.8 0.72 1:0.1169 43.75 3.8 0.92 1:0.0869 55 3.8 1.13 1:0.0691 55 1.8 1.06 1:0.0327 55 0.94 1.03 1:0.0171 48hrs 55 - 30 2.125 15 0.54 1:7.0588 5.5 15 0.60 1:2.7273 10 15 0.68 1:15000 26.25 15 0.98 1:0.5714 32.5 15 1.09 1:0.4615 21.25 7.5 0.64 1:0.3529 26.25 7.5 0.73 1:0.2857 32.5 7.5 0.84 1:0.2308 43.75 7.5 1.05 1:0.1714 32.5 3.8 0.72 1:0.1169 43.75 3.8 0.92 1:0.0869 55 3.8 1.13 1:0.0691 55 1.8 1.06 1:0.0327 _ _ _ _ _ _ _ ...... 5.55 0.94 1 .03 1:0.0171 Table:10 Ca: ppm Al of TCMTB (2-(thiocyanomethylthio) benzothiazole) Cb: ppm Al of MBIT(N-methyl-1,2-benzisothiazoin-3-one) Ratio:Ca:Cb Test Organisms Contact Ca Cb S.I. Ca:Cb Time A. niger 3 days 3 ATCC # 16404 37.5 3 2.4 1.06 1:0.8000 3 1.2 1.03 1:0.4000 7 days 8.75 - - 150 0.525 75 0.56 1:142.8571 0.65 75 0.57 1:115.3846 0.875 75 0.60 1:85.7143 1.1 75 0.63 1:68.1818 2 75 0.73 1:37,5000 3 75 0.84 1:25.0000 4.25 75 0.99 1:17.6471 8.75 9.4 1.06 1:1.0743 6.5 2.4 0.76 1:0.3692 8.75 2.4 1.02 1:0.2743 28 8.75 1.2 1.01 1:0.1371 Calbicans 48 hrs 4.25 - ATCC# 10231 - 30 4.25 1.86 1.06 1:0.4376 4.25 0.94 1.03 1:0.2212 72 hrs 5.25 60 1.1 30 0.71 1:27.2727 2 30 0.88 1:15.0000 3 30 1.07 1:10.0000 4.25 15 1.06 1:3.5294 4.25 7.5 0.93 1:1.7647 5.25 3.75 1.06 1:0.7143 4.25 1.86 0.84 1:0.4376 5.25 1.86 1.03 1:0.3543 4.25 0.94 0.83 1:0.2212 ------ 5.25 0.94 1.02 1:0.1790 Ps. aeruginosa 48 hrs 212.5 ATCC#9027 - 125 21.25 75 0.70 1:3.5294 26.25 75 0.72 1:2.8571 43.75 75 0.81 1:1.7143 55 75 0.86 1:1.3636 100 75 1.07 1:0.7500 150 37.5 1.01 1:0.2500 150 18.75 0.86 1:0.1250 212.5 18.75 1.15 1:0.0882 150 9.4 0.78 1:0.0627 212.5 9.4 1.08 1:0.0442 212.5 4.6 1.04 1:0,0216 S.aureus 24hrs 26.25 ATCC#6538 - 15 26.25 0.94 106 1:0.0358 The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment 5 or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of -the common general knowledge in the field of endeavour to which this specification relates. Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

Claims (2)

1. A microbicidal composition comprising: (a) N-methyl-1,2-benzisothiazolin-3-one; and (b) a microbicide comprising 2-ethylhexyl glycerin.
2. The microbicidal composition according to claim 1 wherein the weight ratio of 2-ethylhexyl glycerin to N-methyl-1,2-benzisothiazolin-3-one is from 1:0.017 to 1:2.3.
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