U.S. patent number 3,852,441 [Application Number 05/341,191] was granted by the patent office on 1974-12-03 for synergistic mixtures of diphenylbismuth acetate and the zinc salt of 1-hydroxy-2-pyridine thione effect as antibacterial and antifungal agents.
This patent grant is currently assigned to The Procter & Gamble Company. Invention is credited to John A. Kooistra, Jr..
United States Patent |
3,852,441 |
Kooistra, Jr. |
December 3, 1974 |
**Please see images for:
( Certificate of Correction ) ** |
SYNERGISTIC MIXTURES OF DIPHENYLBISMUTH ACETATE AND THE ZINC SALT
OF 1-HYDROXY-2-PYRIDINE THIONE EFFECT AS ANTIBACTERIAL AND
ANTIFUNGAL AGENTS
Abstract
Mixtures of the heavy metal salts of 1-hydroxy-2-pyridinethione
and either diphenylbismuth acetate, bismuth trichloride, bismuth
formic iodide, or bismuth subgallate, provide synergistic
antibacterial and antifungal activity, especially against
Pseudomonas aeruginosa; compositions containing said mixture
exhibit this same antibacterial and antifungal effectiveness.
Inventors: |
Kooistra, Jr.; John A. (Forest
Park, OH) |
Assignee: |
The Procter & Gamble
Company (Cincinnati, OH)
|
Family
ID: |
26794097 |
Appl.
No.: |
05/341,191 |
Filed: |
March 14, 1973 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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98085 |
Dec 14, 1970 |
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Current U.S.
Class: |
514/188; 510/131;
510/319; 510/390; 510/500; 510/508; 510/492; 510/382; 510/133;
510/119; 546/7; 514/852 |
Current CPC
Class: |
C11D
3/48 (20130101); Y10S 514/852 (20130101) |
Current International
Class: |
C11D
1/00 (20060101); A01n 009/02 (); A01n 009/22 ();
A01n 011/001 () |
Field of
Search: |
;424/131,245,263,296 |
References Cited
[Referenced By]
U.S. Patent Documents
Other References
The Merck Index, 7th Ed.; 1960, pp. 155 and 157..
|
Primary Examiner: Goldberg; Jerome D.
Assistant Examiner: Robinson; Allen J.
Attorney, Agent or Firm: Aylor; Robert B. Hemingway; Ronald
L.
Parent Case Text
REFERENCE TO RELATED APPLICATION
This application is a continuation of my copending application Ser.
No. 98,085, filed Dec. 14, 1970, for "SYNERGISTIC MIXTURES OF
DIPHENYLBISMUTH ACETATE AND THE ZINC SALT OF
1-HYDROXY-2-PYRIDINETHIONE AND COMPOSITIONS CONTAINING SAME", now
abandoned.
Claims
What is claimed is:
1. An antibacterial and antifungal composition, active against
Pseudomonus aeruginosa and Escherichia coli bacteria, said
composition containing a mixture of:
A. zinc 1-hydroxy-2-pyridinethione; and
B. diphenylbismuth acetate in the ratio of (A) to (B) of from about
1:9 to about 9:1.
2. An antibacterial and antifungal composition containing a mixture
of:
A. zinc 1-hydroxy-2-pyridinethione; and
B. diphenylbismuth acetate in a ratio of (A) to (B) of from about
1:4 to about 4:1.
Description
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates to improved mixtures of antibacterial and
antifungal agents and compositions containing said mixtures. More
particularly, it relates to such mixtures which give improved
activity against more resistant bacteria such as Pseudomonas
aeruginosa.
Prior Art
U.S. Pat. No. 3,239,411, issued Mar. 8, 1966, teaches that various
organo-bismuth compounds, such as diphenylbismuth acetate, are
effective antibacterial and antifungal agents for use against a
wide variety of gram-positive and gram-negative organisms.
Similarly, the heavy metal salts of 1-hydroxy-2-pyridinethione are
known. See, e.g., U.S. Pat. No. 2,809,971. The use of zinc
pyridinethione as an antibacterial agent has been disclosed in
various soap, detergent and cosmetic preparations. See, e.g., U.S.
Pat. Nos. 3,412,033; 3,281,366; and 3,235,455.
SUMMARY OF THE INVENTION
The present invention is directed toward synergistic combinations
of the heavy metal salts of 1-hydroxy-2-pyridinethione and either
phenylbismuth acetate, bismuth trichloride, bismuth formic iodide,
or bismuth subgallate. This synergism is present in all proportions
of these classes of antibacterial compounds. It is most evident in
the preferred ratio of from 1:9 to 9:1, the most preferred ratio
being 80:20, and the most preferred mixture being diphenylbismuth
acetate:zinc 1-hydroxy-2-pyridinethione.
The synergism of these mixtures is evident against a wide variety
of bacteria, yeast and fungal species, but it is most impressive
against Pseudomonas aeruginosa (Ps.a.), which is extremely
difficult to inhibit. Ps.a. is currently a problem in many
hospitals due to its inherent resistance to most antibacterial
agents. At the same time, the mixtures will also display their
synergistic activity against the common organisms such as
Staphylococcus epidermidis (S.ep.), Streptococcus faecalis (St.
f.), and Escherichia coli (E.c.). Other bacteria which are
controlled by the mixture are disclosed hereinafter in the
examples.
It has also been found that the above-disclosed combination of
antibacterial agents possesses synergistic antibacterial properties
whereby a lasting antibacterial and antifungal effect is obtained
particularly in soap and non-soap synthetic detergent compositions,
and other products where an antibacterial action is desired. Such
compositions containing the synergistic combination of the present
invention in amounts of from about 0.2% to about 3%, preferably
about 1%, are particularly effective.
The term "soap" as used herein is meant to designate alkali metal
soaps such as the sodium and potassium salts of the higher fatty
acids of naturally occurring plant or animal esters, e.g., palm
oil, coconut oil, babassu oil, soybean oil, castor oil, tallow,
whale and fish oils, grease and lard and mixtures thereof. Sodium
and potassium soaps can be made by direct saponification of the
fats and oils or by the neutralization of the fatty acids which are
prepared in a separate manufacturing process. Examples of suitable
soaps are the sodium, potassium, ammonium and alkylolammonium salts
of higher fatty acids (C.sub.10 - C.sub.20). Particularly useful
are the sodium and potassium salts of the mixtures of fatty acids
derived from coconut oil and tallow, i.e., sodium or potassium
tallow and coconut soap.
Anionic synthetic detergents which can be used with the
antibacterial combinations of the present invention can be broadly
defined as the water-soluble salts, including the alkali metal,
ammonium and substituted ammonium salts, of organic sulfuric
reaction products having in their molecular structure an alkyl
radical containing from about 8 to about 22 carbon atoms and a
radical selected from the group consisting of sulfonic acid and
sulfuric acid ester radicals.
Important examples of the synthetic detergents which can be used
with the compositions of the present invention are the following:
alkali metal (e.g., sodium and potassium) ammonium and substituted
ammonium (e.g., lower alkyl ammonium) salts of the following: alkyl
sulfates, especially those obtained by sulfating the higher
alcohols produced by reducing the glycerides of tallow or coconut
oil; random paraffin sulfonates, in which the alkyl group contains
from about 8 to about 22 carbon atoms, prepared by treating random
paraffin hydrocarbons in sulfur dioxide and chlorine in the
presence of light followed by treating with a base; branched or
linear alkyl benzene sulfonates, in which the alkyl group contains
from about 8 to about 18 carbon atoms, preferably from about 10 to
about 14 carbon atoms, especially those of the types described in
U.S. Pat. No. 2,220,099, and 2,477,383; sodium alkyl glyceryl ether
sulfonates, especially those ethers of the higher alcohols derived
from tallow and coconut oil; coconut oil fatty acid monoglyceride
sulfates and sulfonates; sulfuric acid esters of the reaction
product of one mole of a higher fatty alcohol (e.g., tallow or
coconut alcohols) and from about 1 to about 6, preferably about 3
moles of ethylene oxide; alkyl phenol ethylene oxide ether sulfates
with about 4 units of ethylene oxide per molecule and in which the
alkyl radicals contain about 9 carbn atoms; the reaction product of
fatty acids esterified with isethionic acid and neutralized with
sodium hydroxide where, for example, the fatty acids are derived
from coconut oil; fatty acid amides of the methyl taurine in which
the fatty acids, for example, are derived from coconut oil;
sulfonated olefins of U.S. Pat. No. 3,332,880; and other known
others the art, a number being specifically set forth in U.S. Pat.
Nos. 2,486,921, 2,486,922 and 2,396,278.
The nonionic synthetic detergents which can be used with the
antibacterial combinations of the present invention may be broadly
defined as compounds produced by the condensation of alkylene oxide
groups (hydrophilic in nature) with an organic hydrophobic compound
which may be aliphatic or alkyl-aromatic in nature. The length of
the hydrophilic or polyoxyalkylene radical which is condensed with
any particular hydrophobic group can be readily adjusted to yield a
water-soluble compound having the desired degree of balance between
hydrophilic and hydrophobic elements.
For example, a well-known class of nonionic synthetic detergents is
made available on the market under the trade name of "Pluronic."
These compounds are formed by condensing ethylene oxide with a
hydrophobic base formed by the condensation of propylene oxide with
propylene glycol. The hydrophobic portion of the molecule which, of
course, exhibits water insolubility has a molecular weight of from
about 1,500 to about 1,800. The addition of polyoxyethylene
radicals to this hydrophobic portion tends to increase the water
solubility of the molecule as a whole and the liquid character of
the products is retained up to the point where polyoxyethylene
content is about 50% of the total weight of the condensation
product.
Other suitable nonionic synthetic detergents include:
1. The polyethylene oxide condensates of alkyl phenols, e.g., the
condensation products of alkyl phenols having an alkyl group
containing from about 6 to 12 carbon atoms in either a straight
chain or branched chain configuration, with ethylene oxide, the
said ethylene oxide being present in amounts equal to 10 to 60
moles of ethylene oxide per mole of alkyl phenol. The alkyl
substituent in such compounds may be derived from polymerized
propylene, diisobutylene, octane, or nonane, for example.
2. Those derived from the condensation of ethylene oxide with the
product resulting from the reaction of propylene oxide and ethylene
diamine -- products which may be varied in composition depending
upon the balance between the hydrophobic and hydrophilic elements
which is desired. For example, compounds containing from about 40
to about 80% polyoxyethylene by weight and having a molecular
weight of from about 5,000 to about 11,000 resulting from the
reaction of ethylene oxide groups with a hydrophobic base
constituted of the reaction product of ethylene diamine and excess
propylene oxide, said base having a molecular weight of the order
of 2,500 to 3,000, are satisfactory.
3. The condensation product of aliphatic alcohols having from 8 to
18 carbon atoms, in either straight chain or branched chain
configuration, with ethylene oxide, e.g., a coconut alcohol
ethylene oxide condensate having from 10 to 30 moles of ethylene
oxide per mole of coconut alcohol, the coconut alcohol fraction
having from 10 to 14 carbon atoms.
4. Long chain tertiary amine oxides corresponding to the following
general formula, R.sub. 1 R.sub. 2 R.sub. 3 N.fwdarw.O, wherein
R.sub.1 contains an alkyl, alkenyl or monohydroxy alkyl radical of
from about 8 to about 18 carbon atoms from 0 to about 10 ethylene
oxide moieties, and from 0 to 1 glyceryl moiety, and R.sub.2 and
R.sub.3 contain from 1 to about 3 carbon atoms and from 0 to about
1 hydroxy group, e.g., methyl, ethyl, propyl, hydroxy ethyl, or
hydroxy propyl radicals. The arrow in the formula is a conventional
representation of a semi-polar bond. Examples of amine oxides
suitable for use in this invention include dimethyldodecylamine
oxide, oleyldi(2-hydroxyethyl) amine oxide, dimethyloctylamine
oxide, dimethyldecylamine oxide, dimethyltetradccylamine oxide,
3,6,9-trioxaheptadecyldiethylamine oxide,
di(2-hydroxyethyl)-tetradecylamine oxide,
2-dodecoxyethyldimethylamine oxide,
3-dodecoxy-2-hydroxypropyldi(3-hydroxypropyl)amine oxide,
dimethylhexadecylamine oxide.
5. Long chain tertiary phosphine oxides corresponding to the
following general formula RR'R"P.fwdarw.O, wherein R contains an
alkyl, alkenyl or monohydroxyalkyl radical ranging from 8 to 18
carbon atoms in chain length, from 0 to about 10 ethylene oxide
moieties and from 0 to 1 glyceryl moiety and R' and R" are each
alkyl or monohydroxyalkyl groups containing from 1 to 3 carbon
atoms. The arrow in the formula is a conventional representation of
a semipolar bond. Examples of suitable phosphine oxides are:
dodecyldimethylphosphine oxide,
tetradecyldimethylphosphine oxide,
tetradecylmethylethylphosphine oxide,
3,6,9 -trioxaoctadecyldimethylphosphine oxide,
cetyldimethylphosphine oxide,
3-dodecoxy-2-hydroxypropyldi(2-hydroxyethyl)phosphine oxide,
stearyldimethylphosphine oxide,
cetylethylpropylphosphine oxide,
oleyldiethylphosphine oxide,
dodecyldiethylphosphine oxide,
tetradecyldiethylphosphine oxide,
dodecyldipropylphosphine oxide,
dodecyldi(hydroxymethyl)phosphine oxide,
dodecyldi(2-hydroxyethyl)phosphine oxide,
tetradecylmethyl-2hydroxypropylphosphine oxide,
oleyldimethylphosphine oxide,
2-hydroxydodecyldimethylphosphine oxide.
6. Long chain dialkyl sulfoxides containing one short chain alkyl
or hydroxy alkyl radical of 1 to about 3 carbon atoms (usually
methyl) and one long hydrophobic chain which contains alkyl,
alkenyl, hydroxy alkyl, or keto alkyl radicals containing from
about 8 to about 20 carbon atoms, from 0 to about 10 ethylene oxide
moieties and from 0 to 1 glyceryl moiety. Examples include:
octadecyl methyl sulfoxide, 2-ketotridecyl methyl sulfoxide,
3,6,9-trioxaoctadecyl 2-hydroxyethyl sulfoxide,
dodecyl methyl sulfoxide,
oleyl 3-hydroxy propyl sulfoxide,
tetradecyl methyl sulfoxide,
3-methoxytridecyl methyl sulfoxide
3-hydroxytridecyl methyl sulfoxide,
3-hydroxy-4-dodecoxybutyl methyl sulfoxide.
The zwitterionic synthetic detergents useful with the antibacterial
agents of the present invention can be broadly described as
derivatives of aliphatic quaternary ammonium, phosphonium, and
sulfonium compounds, in which the aliphatic radicals can be
straight chain or branched, and wherein one of the aliphatic
substituents contains from about 8 to 18 carbon atoms and one
contains an anionic water solubilizing group, e.g., carboxy,
sulfonate, sulfate, phosphate, or phosphonate. A general formula
for these compounds is: ##SPC1##
wherein R.sup.2 contains an alkyl, alkenyl, or hydroxy alkyl
radical of from about 8 to about 18 carbon atoms, from 0 to about
10 ethylene oxide moieties and from 0 to 1 glyceryl moiety; Y is
selected from the group consisting of nitrogen, phosphorous, and
sulfur atoms; R.sup.3 is an alkyl or monohydroxyalkyl group
containing 1 to about 3 carbon atoms; x is 1 when Y is a sulfur
atom and 2 when Y is a nitrogen or phosphorous atom, R.sup.4 is an
alkylene or hydroxyalkylene of from 1 to about 4 carbon atoms and Z
is a radical selected from the group consisting of carboxylate,
sulfonate, sulfate, phosphonate, and phosphate groups.
Examples include:
4-[N,N-di(2-hydroxyethyl)-N-octadecylammonio]-butane-1-carboxylate;
5-[S-3-hydroxypropyl-S-hexadecylsulfonio]
3-hydroxypentane-1-sulfate;
3-[P,P-diethyl-p-3,6,9-trioxatetradecoxylphosphonio]-2-hydroxy-propane-1-ph
osphate;
3-[N,N-dipropyl-N-3-dodecoxy-2-hydroxypropylammonio]-propane-1-phosphonate;
3-(N,N-dimethyl-N-hexadecylammonio)propane-N,N-dimethyl-N-hexadecylammonio)
propane-1-sulfonate;
3-(N,N-dimethyl-N-hexadecylammonio)-2-hydroxypropane-1-sulfonate;
4-[N,N-di(2-hydroxyethyl)-N-(2-hydroxydodecyl)ammonio]-butane-1-carboxylate
;
3-[S-ethyl-S-(3-dodecoxy-2-hydroxypropyl)sulfonio]-propane-1-phosphate;
3-[P,P-dimethyl-P-dodecylphosphonio]-propane1-phosphonate; and
5-[N,N-di(3-hydroxypropyl)-N-hexadecylammonio]2-hydroxy-pentane-1-sulfate.
The amphoteric synthetic detergents useful in the present invention
can be broadly described as derivatives of aliphatic secondary and
tertiary amines in which the aliphatic radical can be straight
chain or branched and wherein one of the aliphatic substituents
contains from about 8 to about 18 carbon atoms and one contains an
anionic water solubilizing group, e.g., carboxy, sulfonate,
sulfate, phosphate, or phosphonate. Examples of compounds, falling
within this definition are sodium 3-dodecylaminopropionate, sodium
3-dodecylaminopropane sulfonate, dodecyl-.beta.-alanine,
N-alkyltaurines such as the one prepared by reacting dodecylamine
with sodium isethionate according to the teaching of U.S. Pat. NO.
2,658,072, N-higher alkyl aspartic acids such as those produced
according to the teaching of U.S. Pat. No. 2,438,091, and the
products sold under the trade name "Miranol" and described in U.S.
Pat. No. 2,528,378.
Detergent formulations containing the antibacterial compositions of
the present invention can also contain from about 0 to about 90%,
preferably from about 10 to about 90% of water-soluble alkaline
detergency builder salts, either of the organic or inorganic types.
Examples of such builder salts can be found in U.S. Pat. No.
3,336,233, issued Aug. 15, 1967, column 9, lines 29-66, which is
incorporated herein by reference.
The detergent formulations can also contain any of the usual
adjuvants, diluents, and additives, for example, perfumes,
anti-tarnishing agents, anti-redeposition agents, dyes,
fluorescers, suds builders, suds depressors and the like without
detracting from the advantageous properties of the antibacterial
compositions of the present invention.
Examples of diluents which may be incorporated into a synthetic
detergent bar in amounts of up to about 80% of the bar include
soaps, especially heavy metal insoluble soaps, (metallic salts of
higher fatty acids); starches such as cornstarch; and clays such as
china clay or fuller's earth. Other diluents include inorganic
salts such as sodium and potassium chlorides and sulfates. Such
diluents add bulk to the bar and improve its cosmetic properties
without impairing its detergent or cohesive properties.
In compositions the antibacterial mixture may tend to react slowly
to give the phenylbismuth di(pyridinethione-N-oxide) of the
copending application of John D. Curry, application Ser. No.
98,086, filed concurrently herewith now abandoned. Therefore, the
shelf life of some compositions containing the mixture of
diphenylbismuth acetate and zinc 1-hydroxy-2-pyridinethione may be
limited insofar as stability is concerned. The mixture of
diphenylbismuth acetate and zinc 1-hydroxy-2-pyridinethione is
stable for an indeterminate period of time in buffered solutions
such as "pHisoderm." The mixture is also effective when slurried in
water or dissolved in an organic solvent. Care, however, should be
taken when using the mixture in an organic solvent, since zinc
1-hydroxy-2-pyridinethione when solubilized may have increased
toxicity to mammals. This same care should be taken when zinc
1-hydroxy-2-pyridinethione is mixed in alkaline solutions with
detergents and other salts containing a sodium ion. Under such
circumstances, the zinc 1-hydroxy-2-pyridinethione can be changed
to the soluble sodium 1-hydroxy-2-pyridinethione which will display
mammalian toxicity.
These precautions as to mammalian toxicity, of course, are
irrelevant where contact with mammals is kept to a minimum.
The following examples illustrate the invention.
EXAMPLE I
Diphenylbismuth acetate and 1-hydroxy-2-pyridinethione were tested
to determine the minimal inhibitory concentration (MIC) in agar for
the two materials and their mixtures against Staphylococcus
epidermidis, Streptococcus faecalis, Pseudomonas aeruginosa, and
Escherichia coli.
10,000 ppm stock solutions of diphenylbismuth acetate and
1-hydroxy-2-pyridinethione were prepared in dimethyl formamide. The
sanitizers were combined in the desired test ratios, mixed
thoroughly and added directly to 19 to 20 ml of molten trypticase
soy agar to give a series of graded concentrations for each ratio.
Thorough rotation of plates assured adequate mixing of the
sanitizers before agar solidification. Appropriate control plates
containing concentrations of each sanitizer alone as well as plain
agar were included. All plates remained at room temperature
overnight.
0.1 ml of a diluted 18 hour broth culture of each organism was
added directly onto a sterile one-half inch diameter filter disc.
The Staphylococcus epidermidis was used at a dilution of 1:1000;
the Streptococcus faecalis was used at a dilution of 1:10; the
Pseudomonas aeruginosa was used at a dilution of 1:10,000; and the
Escherichia coli was used at a dilution of 1:10,000. Sufficient
discs were prepared in this manner to inoculate each plate with all
organisms. Discs containing bacteria were placed on the agar
surfaces, marked in numbered sectors, for approximately 30 minutes
and then removed. Plates were incubated 48 hours at 37.degree.C.
and then examined for growth.
The following table summarizes the ratios of diphenylbismuth
acetate (PBA) to 1-hydroxy-2-pyridinethione (ZPT) tested, the
minimal inhibitory concentrations in parts per million of said
mixtures and the ratio sum for each mixture. Where the ratio sum is
greater than 1, there is antagonism, and where the ratio sum is
less than 1, there is synergism, assuming a straight line
relationship for activity vs. concentration. The results clearly
show synergism for all ratios of the two antibacterial agents.
Table I ______________________________________ Synergism of Several
Ratios of PBA and ZPT Against Four Organisms (MIC Test in Agar)
______________________________________ Organism PBA:ZPT MIC (ppm)
Ratio Sum* ______________________________________ Staphylococcus
100:0 8.0 1.000 epidermidis 80:20 6.0 0.900 60:40 4.0 0.700 40:60
1.0 0.200 20:80 1.0 0.225 0:100 4.0 1.000 Streptococcus 100:0 10.0
1.000 faecalis 80:20 2.0 0.240 60:40 0.5 0.070 40:60 0.5 0.080
20:80 1.0 1.180 0:100 5.0 1.000 Pseudomonas 100:0 >100 1.000
aeruginosa 80:20 20 0.227 60:40 20 0.253 40:60 20 0.280 20:80 20
0.307 0:100 60 1.000 Escherichia 100:0 >20 1.000 coli 80:20 8.0
0.720 60:40 6.0 0.780 40:60 4.0 0.680 20:80 4.0 0.840 0:100 4.0
1.000 ______________________________________ *Ratio sum> 1 =
antagonism Ratio sum < 1 = synergism Calculations according to:
Zwart Voorspuiji, A. and Nass, C. Arch. Intern Pharmacodynamie
109:211-28, 1957.
The minimal inhibitory concentrations (MIC) of 80:20 ratios of
PBA:ZPT against several other organisms were determined according
to the method described in Example I. Diluted broth cultures of the
organisms were added directly to agar surfaces with Kline antigen
microdroppers instead of using the filter paper discs previously
described. Results were as follows:
Organism MIC (ppm) ______________________________________
Staphylococcus aureus 0.2 Streptococcus species 0.1 Sarcina lutea
0.8 Diphtheroid species 0.4 Serratia marcescens 8.0 Klebsiella
pneumoniae 8.0 Enterobacter aerogenes 8.0 Corynebacterium acnes*
1.0 Pityrosporum ovale** 10.0 Aspergellus niger*** 30.0 Microsporum
gypseum*** 1.0 ______________________________________ *Brain heart
infusion agar + 1% glucose used as growth medium (anaerobic)
**Trypticase soy agar + 0.1% of a 95:5 ratio of oleic:palmitic acid
used as growth medium ***Modified gradient plate technique of Hunt
and Sardham used (Applied Microbiology 17 2:329-30, 1969)
These results emphasize the broad spectrum of antimicrobial
activity present in the 80:20 PBA:ZPT sanitizer system.
EXAMPLE II
The following test method was used in this example: Cornified
epidermis was obtained from the plantar surfaces of human
volunteers by cutting sheets of uniform thickness (0.4 mm.) with a
dermitome. Discs, 6 mm. in diameter, were cut from the sheets with
a number 2 cork borer. A 2.5% stock solution of "pHisoderm" and
10,000 parts per million stock solutions of diphenylbismuth acetate
(PBA) and zinc 1-hydroxy-2-pyridinethione (ZPT) in
dimethylformamide (DMF) were prepared. Small, screwcapped vials
held 20 ml. of the "pHisoderm" solution containing either the
sanitizers along or in combination in concentrations ranging from
0.5% to 2% (125 to 300 ppm). A control was also included.
The required number of discs were added to the vials which then
were mechanically agitated for 10 minutes in a 50.degree.C. water
bath. The vial contents were then transferred to a small beaker
where the washing liquid was removed by suction and three 30 ml.
distilled water rinses were applied with suction removal of the
rinse water. The discs were then dried for 2 hours on paper towels
covered with a plexiglass shield to prevent air contamination. The
dry discs were placed on the surface of 15 ml. of solidified
trypticase soy agar (TSA) in a Petri dish (no more than two
discs/dish). A 1:10,000 dilution of each of the test organisms in
TSA at 50.degree.C. was prepared. Each Petri dish was overlaid with
10 ml. of agar seeded with one of the organisms. After agar
solidification, the dishes were refrigerated overnight to allow
diffusion of any sanitizer away from the disc into the medium. The
following day, the dishes were placed in a 37.degree.C. incubator
for 24 hours.
The diameter of any clear zones of bacterial growth inhibition were
measured in mm. including the diameter of the disc. A clear zone is
indicative of sanitizer retention on excised stratum corneum after
the washing and rinsing process. Synergism of two compounds is
evident if a zone produced by a combination is greater than the
zone produced by the same concentration of both compounds tested
separately.
The diameters of zones around discs washed in the stated
concentrations of either PBA, ZPT, or combinations of these agents
are presented in Table II for two test organisms -- Escherichia
coli and Pseudomonas aeruginosa. The presence or absence of
synergism is also indicated.
Table II ______________________________________ Synergism of
PBA:ZPT Combinations Against Two Gram-negative Bacteria in the
Human Corneum Disc Test ______________________________________ %
Concentration in 2.5% pHisoderm Test Organisms PBA ZPT Escherichia
Pseudomonas coli aeruginosa ______________________________________
0.5 -- 10* 0 1.0 -- 12 0 1.5 -- 12 0 2.0 -- 12 0 -- 0.5 8 0 -- 1.0
9 0 -- 1.5 9 0 -- 2.0 11.5 0 0.5 0.5 23 + 11.5 + 1.0 0.5 24 + 20 +
0.5 1.0 18 + 11.5 + 1.0 1.0 23 + 14.5 + 1.5 0.5 22 + 13.5 + 0.5 1.5
21 + 15 + "pHisoderm" control 0 0
______________________________________ *All values are total zone
diameters in mm. (including 6 mm. disc) +Synergism
As can be seen, synergism was realized with every combination of
PBA and ZPT tested against both organisms, but the most pronounced
effect was found with Pseudomonas aeruginosa.
EXAMPLE III
The following test method was utilized:
PBA and ZPT were prepared in "pHisoderm" in the following
concentrations:
0.01% PBA:
0.01% zpt;
0.0075% pba + 0.0025% zpt;
0.005% pba + 0.005% zpt; and
0.0025% PBA + 0.0075% ZPT.
These concentrations were prepared by adding the dry sanitizer
powders directly to "pHisoderm" and mixing thoroughly with a
mechanical mixer. The final product for a given test was
distributed in 12 small plastic bottles for use by each test
subject at home and two larger bottles for laboratory
handwashings.
Groups of 12 subjects were employed in the testing of each product.
They were chosen at random from a large group of available subjects
who use non-sanitizer products exclusively for personal hygiene.
During the three-day testing period, each subject used only the
assigned product for hand cleansing. Three regimented washings were
required daily in the laboratory as follows:
a. The hands were moistened under 100.degree.F. running tap
water.
b. The product was squeezed from a plastic bottle into the palm in
an amount approximately the size of a 25 cent piece.
c. The product was distributed over the hands and lather worked for
90 seconds.
d. The hands were then rinsed under running tap water for 30
seconds.
Ad libitum washings were done at home. On day 1, the test product
was given to the panelists for home use and they completed the
three standard washes for that day. The same schedule was followed
the second day and on the morning of the third day, quantitative
determinations of the bacterial hand flora were made as
follows:
a. Four successive handwashings were performed as described above
using a "blank" bar soap containing no antibacterial agents.
b. A fifth standardized washing was done in a basin containing 1
liter of sterile distilled water with careful rinsing in this
basin.
c. Aliquots were taken from the thoroughly mixed basin contents,
added to 30 ml. of sterile distilled water in a membrane funnel,
and passed through sterile membrane filters with vacuum
suction.
d. The filters were incubated on pads saturated with 2X
concentrated trypticase soy broth plus 10% horse serum and 1% Tween
80 in plastic dishes for 48 hours at 37.degree.C.
e. Colonies appearing on the filters were counted and these numbers
multiplied by the appropriate dilution factor to determine the
number of bacteria/liter in the fifth basin.
The numbers of colonies for each panelist were converted to
logarithms to minimize unusually high or low counts. These values
were averaged to determine the mean log of the fifth basin
bacterial count. Log percent reduction for each subject was
determined using the log of 1.3 .times. 10.sup.6, or 6.1184, as an
average for the fifth basin bacterial count obtained from more than
500 handwashings by nonantibacterial soap users. These values were
then averaged to obtain the mean log percent reduction.
Table III ______________________________________ Synergism of
Ratios of PBA:ZPT in Handwashing Tests
______________________________________ Sanitizer Concentrations
Mean Log in "Phisoderm"* Mean Log % Reduction
______________________________________ 0.01% PBA 5.7942 52.60 0.01%
ZPT 5.7764 0.0075% PBA + 0.0025% ZPT 5.3790 81.78 0.005% PBA +
0.005% ZPT 5.5987 69.78 0.0025% PBA + 0.0075% ZPT 5.6258 67.83
______________________________________ *Two-day exposures
As can be seen from the above table, the mixture of antibacterial
agents gave synergistically better performance than the individual
components. This test confirms in vivo the synergism demonstrated
in the previous in vitro tests.
Five separate handwashing tests of 1% PBA:ZPT (80:20) in
"Phisoderm" have shown the following results:
Log % Reduction Mean Log ______________________________________
99.2 4.0236 98.35 4.3360 98.12 4.3933 99.03 4.1060 97.3 4.5490
______________________________________
These results demonstrate the skin degerming effectiveness of
PBA:ZPT under conditions of actual use by human subjects.
EXAMPLE IV
Using the test procedure of Example I with Kline antigen
microdroppers for delivery of the inoculum to the agar surfaces,
the minimal inhibitory concentration was determined for an 80:20
mixture of PBA:ZPT in a water slurry containing no solvent against
the following indicated organisms:
Table IV ______________________________________ Organism MIC (ppm)
______________________________________ Staphylococcus aureus 0.5
Escherichia coli 6 Pseudomonas aeruginosa 40 Serratia marcescens 8
Klebsiella pneumoniae 8 Enterobacter aerogenes 10
______________________________________
EXAMPLE V
Shampoo Compositions ______________________________________
Composition No. 1 No. 2 No. 3
______________________________________ Sulfated coconut fatty 23 23
alcohol -- sodium salt Sodium stearate 8.7 8.7 Sodium alkyl
glyceryl 28.3 ether sulfonate.sup.1 Sodium acyl 5.0
sarcosinate.sup.2 Sodium sulfate 0.8 0.8 2.6 Sodium chloride 6.6
Trisodium phosphate 2.1 2.1 Diethanolamide of 2.0 coconut fatty
acids Acetylated lanolin 1.0 1.0 1.0 Phenyl mercuric 0.113 0.113
0.0035 acetate.sup.3 Perfume 1.0 1.0 0.4 80:20 mixture of 0.5 1.0
1.5 PBA:ZPT Water (make up balance) pH 7.7 7.7 7.4
______________________________________ .sup.1 Alkyl radicals
derived from fatty alcohol, 25.3% from coconut and 3% from tallow.
.sup.2 Acyl radicals derived from coconut fatty acids. .sup.3 31/2%
solution in oleic acid in No. 1 and No. 2.
Composition No. 4 Percent ______________________________________
Dimethyl coconut.sup.1 amine oxide 8.0 Disodium lauryl
beta-iminodipropionate 5.0 Sodium coconut.sup.1 sulfate 4.0
Coconut.sup.2 diethanolamide 1.0 80:20 mixture of PBA:ZPT 1.0
Perfume 0.5 Color Less than 0.01 Citric acid to adjust pH to 7.5
Water Balance ______________________________________ .sup.1 The
alkyl radical is derived from middle cut coconut alcohol and has
approximately the following chain length composition: 2% C.sub.10,
66 C.sub.12, 23% C.sub.14, and 9% C.sub.16. .sup.2 Wholecut.
COMPOSITION NO. 5
A shampoo composition is obtained by uniformly mixing together the
following ingredients:
4% triethanolamine salt of the sulfated condensation product of 3
moles of ethynene oxide and one mole of coconut oil fatty alcohol
having the following chain length distribution: 2% C.sub.10, 66%
C.sub. 12, 23% C.sub.14, and 9% C.sub.16.
10% triethanolamine N-acyl sarcosinate, the acyl radicals being
derived from coconut oil fatty acids and having the following chain
length distribution: 10% C.sub.8.sub.-10, 48% C.sub.12, 21%
C.sub.14, 10% C.sub.16, and 11% C.sub.18. 3% monoethanolamide of
coconut oil fatty acids having the following chain length
distribution: 16% C.sub.6.sub.-10, 48% C.sub.12, 17% C.sub.14, 9%
C.sub.16, and 10% C.sub.18.
1% 80:20 mixture of PBA:ZPT.
3% diethanolamide of coconut oil fatty acids having the following
chain length distribution: 16% C.sub.6.sub.-10, 48% C.sub.12, 17%
C.sub.14, 9% C.sub.16, and 10% C.sub.18.
0.65% ethylenediamine tetraacetic acid (as its partial
triethanolammonium salt)
7% ethanol
0.75% methyl cellulose, a 2% solution of which has a viscosity of
4000 cps. at 68.degree.F. and a gel point of 140.degree.F.
0.15% sodium sulfite
0.007% phenyl mercuric acetate
0.75% perfume
balance, water.
Shampoo formulations containing 80:20 mixtures of PBA:ZPT are
desirable since they will control dandruff.
EXAMPLE VI
Personal Use Detergent Lotion
______________________________________ Composition No. 6 Percent
______________________________________ Potassium coconut.sup.1
glyceryl ether 3.0 sulfonate (about 23% diglyceryl and the balance
substantially all monoglyceryl Sodium coconut.sup.1 glyceryl ether
4.0 sulfonate (diglyceryl and monoglyceryl content as above Sodium
tallow glyceryl ether 3.0 sulfonate (diglyceryl and monoglyceryl
content as above; the tallow alkyl radicals correspond to those of
sub- stantially saturated tallow alcohols containing approximately
2% C.sub.14, 32% C.sub.16, and 60% C.sub.18 Coconut.sup.1
dimethylamine oxide 5.0 Sodium salt of sulfated condensation 2.0
product of one mole of nonylphenol with 4 moles ethylene oxide
Potassium pyrophosphate 1.0 80:20 mixture of PBA:ZPT 2.0 Sodium
chloride (in addition to 3.0 that from detergents) Sodium toluene
sulfonate 2.5 Sodium carboxymethylcellulose 0.3 (degree of
substitution 0.65-0.95; viscosity of 1% soln., 1000-2800 cps. at
25.degree.C.) Acrylamide polymer (contains 5-10 0.1 mole percent
acrylic acid radicals; monomer is less than 0.05%; viscosity of a
0.5% soln. is about 10-15 centipoises at 25.degree.C.) Salts
(sodium and potassium chloride 1-2 and sulfate from detergents)
Water Balance ______________________________________ .sup.1 Coconut
indicates alkyl radicals corresponding to those of middle cut
coconut fatty alcohol containing approximately 2% C.sub.10, 66%
C.sub.12, 23% C.sub.14, and 9% C.sub.16.
EXAMPLE VII
Cream Shampoo ______________________________________ Composition
Percent ______________________________________ Sodium coconut
glyceryl ether 14.8 sulfonate (about 29% diglyceryl and the balance
substantially Sodium tallow glyceryl ether sulfonate 2.0 (about 28%
diglyceryl and the balance substantially monoglyceryl) Sodium
chloride 6.7 Sodium sulfate 3.5 Sodium N-lauroyl sarcosinate 3.0
80:20 mixture of PBA:ZPT 2.0 coconut.sup.1 diethanolamine 0.5
Acetylated lanolin 1.0 Perfume 0.4 Water Balance
______________________________________ .sup.1 2% C.sub.10, 66%
C.sub.12, 23% C.sub.14, and 9% C.sub.16.
A milled toilet detergent bar is prepared in accordance with
methods known and used in the art and having the following
composition:
Percent Sodium alkyl glyceryl ether sulfonate 8.0 (alkyl group
derived from the middle- cut.sup.1 of alcohols obtained by
catalytic reduction of coconut oil) Potassium alkyl sulfate (alkyl
group derived from the middle cut.sup.1 of 20.0 alcohols obtained
by catalytic reduction of coconut oil) Magnesium soap of 80:20
tallow: 17.0 coconut fatty acids Inorganic salts (sodium and
potassium 32.0 chlorides and sulfates) 80:20 mixture of PBA:ZPT 1.0
balance Water and minors to 100 .sup.1 Middle-cut coconut alcohols
having a chain length distribution substantially as follows: 2%
C.sub.10, 66% C.sub.12, 23% C.sub.14 and 9% C.sub.16.
This bar cleans well and exhibits good odor reducing properties
evidencing antibacterial effectiveness. It reduces the number of
bacteria on the skin and does not discolor significantly.
A granular built synthetic detergent composition having the
following formulation can be prepared and the antibacterial
compositions of the present invention can be incorporated
therein.
______________________________________ Percent Sodium
dodecylbenzene sulfonate 17.5 Sodium tripolyphosphate 50.0 Sodium
sulfate 14.0 Sodium silicate (SiO.sub.2 :Na.sub.2 O=2:1) 7.0 80:20
mixture PBA:ZPT 3.0 balance Water and minors to 100
______________________________________
This composition, in addition to performing well in its cleaning
capacity, imparts considerable antibacterial and antifungal
activity to fabrics cleansed in its solution.
Substantially equivalent results are obtained, i.e., good cleaning
and good odor reducing properties when the sodium dodecylbenzene
sulfonate of Example IV is replaced, on an equal weight basis, by
the following:
sodium tallow alkyl sulfate;
potassium coconut alkyl glyceryl ether sulfonate;
sodium salt of randomly sulfonated paraffin containing an average
of 15.2 carbon atoms;
ammonium tridecyl sulfate;
condensation product of octyl phenol with 15 moles of ethylene
oxide per mole of octyl phenol;
dimethyldodecylamine oxide;
dodecyldimethylphosphine oxide;
tetradecyl methyl sulfoxide;
3-(N,N-dimethyl-N-hexadecylammonio)propane-1-sulfonate;
3-dodecylaminopropionate; and
dodecyl-.beta.-alanine.
The invention has been described above in conjunction with toilet
and laundry detergents. It will be obvious to those skilled in the
art, however, that the antibacterial compositions of the present
invention can also be beneficially employed in such products as
shampoos, foot powders, antiseptic ointments, cosmetic products and
the like.
A fabric softening composition having the following formula gives
good antibacterial and antifungal protection.
______________________________________ Percent Dialkyl
dimethylammonium chloride 6 (alkyl groups are derived from tallow)
Alkyl phenol polyethyleneoxide 1.5 (alkyl groups average C.sub.15
and 20 moles of ethyleneoxide) Color and perfume 0.3 80:20 mixture
of PBA:ZPT 1.0 Water balance
______________________________________
When in any of the above examples I-VII either bismuth trichloride,
bismuth formic iodide, or bismuth subgallate are substituted for
the diphenylbismuth acetate, and/or the cadmium, cupric, ferrous,
ferric, manganese, mercuric, silver, antimonous, cobaltous, lead,
bismuth, auric, mercurous, arsenic, or nickel salts of
1-hydroxy-2-pyridinethione are substituted for the zinc
1-hydroxy-2-pyridinethione, substantially equivalent results are
obtained in that the mixtures exhibit a synergistic antibacterial
and antifungal activity.
* * * * *