U.S. patent number RE39,033 [Application Number 10/141,079] was granted by the patent office on 2006-03-21 for potentiation of the microbicide 2-(thiocyanomethylthio)benzothiazole using an n-alkyl heterocyclic compound.
This patent grant is currently assigned to Buckman Laboratories International, Inc.. Invention is credited to Dean T. Didato, Daniel E. Glover, S. Rao Rayudu, Marilyn S. Whittemore.
United States Patent |
RE39,033 |
Whittemore , et al. |
March 21, 2006 |
Potentiation of the microbicide
2-(thiocyanomethylthio)benzothiazole using an N-alkyl heterocyclic
compound
Abstract
Microbicidal compositions are described. The compositions
comprise (a) 2-(Thiocyanomethylthio)benzothiazole (TCMTB) and (b)
an N-alkyl heterocyclic compound of the formula: ##STR00001## The
variable "n" ranges from 5 to 17, and the heterocyclic ring defined
by ##STR00002## is a substituted or unsubstituted ring having four
to eight members. Components (a) and (b) are present in a combined
amount effective to control the growth of at least one
microorganism. Methods for controlling the growth of microorganisms
on various substrates or in aqueous systems are also described.
Also described is the industrial application of the microbicidal
composition in the leather industry, the lumber industry, the
papermaking industry, the textile industry, the agricultural
industry, and the coating industry, as well as in aqueous
systems.
Inventors: |
Whittemore; Marilyn S.
(Germantown, TN), Glover; Daniel E. (Brighton, TN),
Rayudu; S. Rao (Germantown, TN), Didato; Dean T.
(Cordova, TN) |
Assignee: |
Buckman Laboratories International,
Inc. (Memphis, TN)
|
Family
ID: |
23811415 |
Appl.
No.: |
10/141,079 |
Filed: |
May 9, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
Reissue of: |
08456098 |
May 30, 1995 |
05693631 |
Dec 2, 1997 |
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Current U.S.
Class: |
514/183; 514/315;
504/100; 210/764; 210/612; 210/601; 514/231.2; 514/359; 514/408;
514/424; 8/159; 8/94.19R; 514/307 |
Current CPC
Class: |
A01N
43/78 (20130101); A01N 43/78 (20130101); A01N
43/84 (20130101); A01N 43/76 (20130101); A01N
43/60 (20130101); A01N 43/50 (20130101); A01N
43/46 (20130101); A01N 43/40 (20130101); A01N
43/36 (20130101); A01N 43/78 (20130101); A01N
2300/00 (20130101) |
Current International
Class: |
A01N
43/00 (20060101); A01N 43/40 (20060101); A01N
43/64 (20060101); A61K 31/535 (20060101) |
Field of
Search: |
;514/183,231.2,315,359,367,408,424,372 ;8/94.19R,159
;210/601,612,764 ;18/50 ;504/100 ;435/29,7.2,4 ;424/704 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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24 569 74 |
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Jun 1976 |
|
DE |
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43 09 690 |
|
Sep 1994 |
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DE |
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06-340504 |
|
Dec 1994 |
|
JP |
|
07-69315 |
|
Mar 1995 |
|
JP |
|
Primary Examiner: Lilling; Herbert J.
Attorney, Agent or Firm: Nixon Peabody LLP
Claims
The invention claimed is:
1. A microbicidal composition comprising: (a)
2-(Thiocyanomethylthio)benzothiazole (TCMTB) and (b) an N-alkyl
heterocyclic compound of the formula: ##STR00008## wherein n varies
from .[.5 to 17.]. .Iadd.9 to 15.Iaddend., the heterocyclic ring
defined by ##STR00009## is a substituted or unsubstituted ring
having four to eight members, and wherein the TCMTB (a) and the
N-alkyl heterocyclic compound (b) are present in a combined amount
effective to control the growth of at least one microorganism and
the .[.and the.]. amount of the N-alkyl heterocyclic compound (b)
present potentiates the microbicidal activity of the TCMTB (a).
2. A microbicidal composition according to claim 1, wherein .[.n
varies from 9 to 15, and.]. the heterocyclic ring is selected from
the group consisting of pyrrolidinyl, 2-pyrrolidinonyl, pyrrolinyl,
pyrazolidinyl, pyrazolinyl, pyrazolyl, imidazolidinyl,
imidazolinyl, imidazolyl, oxazolidinonyl, piperidinyl, piperazinyl,
morpholinyl, hexamethyleneiminyl, and heptamethyleneiminyl.
3. A microbicidal composition according to claim 1, wherein the
N-alkyl heterocyclic compound is selected from the group consisting
of N-dodecyl morpholine, N-dodecyl imidazole,
N-dodecyl-2,6-dimethyl-morpholine,
N-dodecyl-5-chloromethyl-2-oxazolidinone,
N-dodecyl-2-pyrrolidinone, N-dodecyl hexamethyleneimine, N-dodecyl
pyrrolidine, N-dodecyl-3-methyl-piperidine, N-dodecyl piperidine,
N-dodecyl-4-methyl-piperidine and
N-dodecyl-2-methyl-piperidine.
4. A microbicidal composition according to claim 1, wherein the
N-alkyl heterocyclic compound is N-dodecyl morpholine.
5. A microbicidal composition according to claim 4 wherein (a) and
(b) are present in a combined amount synergistically effective to
control the growth of at least one microorganism selected from
algae, fungi, and bacteria.
6. A microbicidal composition according to claim 1, wherein the
N-alkyl heterocyclic compound is N-dodecyl imidazole.
7. A microbicidal composition according to claim 6 wherein (a) and
(b) are present in a combined amount synergistically effective to
control the growth of at least one microorganism selected from
algae, fungi, and bacteria.
8. A microbicidal composition according to claim 1, wherein the
composition is an aqueous formulation.
9. A method for controlling the growth of microorganisms on a
substrate comprising the step of contacting a substrate susceptible
to the growth of microorganisms with (a)
2-(Thiocyanomethylthio)benzothiazole (TCMTB) and (b) an N-alkyl
heterocyclic compound of the formula: ##STR00010## wherein n .[.may
be from 5 to 17.]. .Iadd.varies from 9 to 15.Iaddend., the
heterocyclic ring defined by ##STR00011## is a substituted or
unsubstituted ring having four to eight members, and the TCMTB (a)
and the N-alkyl heterocyclic compound (b) are present in a combined
amount effective to control the growth of at least one
microorganism on the substrate and the amount of the N-alkyl
heterocyclic compound (b) present potentiates the microbicidal
activity of the TCMTB (a).
10. A method according to claim 9, wherein .[.n varies from 9 to
15, and.]. the heterocyclic ring is selected from the group
consisting of pyrrolidinyl, 2-pyrrolidinonyl, pyrrolinyl,
pyrazolidinyl, pyrazolinyl, pyrazolyl, imidazolidinyl,
imidazolinyl, imidazolyl, oxazolidinonyl, piperadinyl, piperazinyl,
morpholinyl, hexamethyleneiminyl, and heptamethyleneiminyl.
11. A method according to claim 9, wherein the N-alkyl heterocyclic
compound is selected from the group consisting of N-dodecyl
morpholine, N-dodecyl imidazole, N-dodecyl-2,6-dimethyl-morpholine,
N-dodecyl-5-chloromethyl-2-oxazolidinone,
N-dodecyl-2-pyrrolidinone, N-dodecyl hexamethyleneimine, N-dodecyl
pyrrolidine, N-dodecyl-3-methyl-piperidine, N-dodecyl piperiodine,
N-dodecyl-4-methyl-piperidine and
N-dodecyl-2-methyl-piperidine.
12. A method according to claim 9, wherein the N-alkyl heterocyclic
compound is N-dodecyl morpholine.
13. A .[.method according to claim 12 wherein (a) and (b).].
.Iadd.microbicidal composition, comprising 2-(Thiocyanomethylthio)
benzothiazole (TCMTB) and N-dodecyl morpholine, wherein the TCMTB
and N-dodecyl morpholine .Iaddend.are present in a combined amount
synergistically effective to control the growth of at least one
microorganism selected from algae, fungi, and bacteria.
14. A .[.method according to claim 13, wherein (a) and (b).].
.Iadd.microbicidal composition, comprising 2-(Thiocyanomethylthio)
benzothiazole (TCMTB) and N-dodecyl imidazole, wherein the TCMTB
and N-dodecyl imidazole .Iaddend.are present in a combined amount
synergistically effective to control the growth of at least one
microorganism selected from algae, fungi, and bacteria.
15. A method according to claim 9, wherein the N-alkyl heterocyclic
compound is N-dodecyl imidazole.
16. A method for controlling the growth of microorganisms on a hide
during a leather tanning process comprising the step of contacting
the leather with (a) 2-Thiocyanomethylthio)benzothiazole (TCMTB)
and (b) an N-alkyl heterocyclic compound of the formula:
##STR00012## wherein n .[.may be from 5 to 17.]. .Iadd.varies from
9 to 15.Iaddend., the heterocyclic ring defined by ##STR00013## is
a substituted or unsubstituted ring having four to eight members,
and wherein the TCMTB (a) and the N-alkyl heterocyclic compound (b)
are present in a combined amount effective to control the growth of
at least one microorganism on the hide and the amount of the
N-alkyl heterocyclic compound (b) present potentiates the
microbicidal activity of the TCMTB (a).
17. A method according to claim 16, wherein the N-alkyl
heterocyclic compound is N-dodecyl morpholine or N-dodecyl
imidazole and the microorganism is algae, fungi, or bacteria.
18. A liquor for use in a leather-tanning process comprising (a)
2-(Thiocyanomethylthio)benzothiazole (TCMTB) and (b) an N-alkyl
heterocyclic compound of the formula: ##STR00014## wherein n .[.may
be from 5 to 17.]. .Iadd.varies from 9 to 15.Iaddend., the
heterocyclic ring defined by ##STR00015## is a substituted or
unsubstituted ring having four to eight members, the TCMTB (a) and
the N-alkyl heterocyclic compound (b) are present in the liquor in
a combined amount effective to control the growth of at least one
microorganism on the leather, and the amount of the N-alkyl
heterocyclic compound (b) present potentiates the microbicidal
activity of the TCMTB (a); and the liquor is selected from a
pickling liquor, a chrome-tanning liquor, a vegetable-tanning
liquor, a post-tan washing liquor, a retanning liquor, a dye
liquor, and a fat liquor.
19. A liquor according to claim 18, wherein the N-alkyl
heterocyclic compound is N-dodecyl morpholine or N-dodecyl
imidazole and the microorganism is algae, fungi, or bacteria.
20. A method for controlling the growth of microorganisms on a
textile substrate in a textile manufacturing process comprising the
step of contacting the textile substrate with (a)
2-(Thiocyanomethylthio)benzothiazole (TCMTB) and (b) an N-alkyl
heterocyclic compound of the formula: ##STR00016## wherein n varies
from .[.5 to 17.]. .Iadd.9 to 15.Iaddend., the heterocyclic ring
defined by ##STR00017## is a substituted or unsubstituted ring
having four to eight members, and wherein the TCMTB (a) and the
N-alkyl heterocyclic compound (b) are present in a combined amount
effective to control the growth of at least one microorganism on
the textile substrate and the amount of the N-alkyl heterocyclic
compound (b) present potentiates the microbicidal activity of the
TCMTB (a).
21. A method according to claim 20, wherein the N-alkyl
heterocyclic compound is N-dodecyl morpholine, or N-dodecyl
imidazole and the microorganism is algae, fungi, or bacteria.
22. A method for controlling the growth of microorganisms on lumber
comprising the step of contacting the lumber with (a)
2-(Thiocyanomethylthio)benzothiazole (TCMTB) and (b) an N-alkyl
heterocyclic compound of the formula: ##STR00018## wherein n varies
from .[.5 to 17.]. .Iadd.9 to 15.Iaddend., the heterocyclic ring
defined by ##STR00019## is a substituted or unsubstituted ring
having four to eight members, and wherein the TCMTB (a) and the
N-alkyl heterocyclic compound (b) are present in a combined amount
effective to control the growth of at least one microorganism on
the lumber and the amount of the N-alkyl heterocyclic compound (b)
present potentiates the microbicidal activity of the TCMTB (a).
23. A method according to claim 22, wherein the N-alkyl
heterocyclic compound is N-dodecyl morpholine or N-dodecyl
imidazole and the microorganism is algae, fungi, or bacteria.
24. A method according to claim 22, wherein the contacting step
comprises dipping the lumber in a bath containing the TCMTB and the
N-alkyl heterocyclic compound.
25. A method according to claim 22, wherein the contacting step
comprises spraying an aqueous formulation of the TCMTB and the
N-alkyl heterocyclic compounds onto the lumber.
26. A method for controlling the growth of microorganisms on a seed
or plant comprising the step of contacting the seed or plant with
(a) 2-(Thiocyanomethylthio)benzothiazole (TCMTB) and (b) an N-alkyl
heterocyclic compound of the formula: ##STR00020## wherein n varies
from .[.5 to 17.]. .Iadd.9 to 15.Iaddend., the heterocyclic ring
defined by ##STR00021## is a substituted or unsubstituted ring
having four to eight members, and wherein the TCMTB (a) and the
N-alkyl heterocyclic compound (b) are present in a combined amount
effective to control the growth of at least one microorganism and
the amount of the N-alkyl heterocyclic compound (b) present
potentiates the microbicidal activity of the TCMTB (a).
27. A method according to claim 26, wherein the N-alkyl
heterocyclic compound is N-dodecyl morpholine or N-dodecyl
imidazole and the microorganism is algae, fungi, or bacteria.
28. A method according to claim 26, wherein the contacting step
comprises drenching the soil surrounding a seed or plant with an
aqueous formulation of the TCMTB and the N-alkyl heterocyclic
compound.
29. A method according to claim 26, wherein the contacting step
comprises spraying an aqueous formulation of the TCMTB and the
N-alkyl heterocyclic compounds onto the seed or plant.
30. A method for controlling the growth of microorganisms in an
aqueous system capable of supporting growth of a microorganism
comprising the step of treating the aqueous system with (a)
2-(Thiocyanomethylthio)benzothiazole (TCMTB) mad (b) an N-alkyl
heterocyclic compound of the formula: ##STR00022## wherein n varies
from .[.5 to 17.]. .Iadd.9 to 15.Iaddend., the heterocyclic ring
defined by ##STR00023## is a substituted or unsubstituted ring
having four to eight members, and wherein the TCMTB (a) and the
N-alkyl heterocyclic compound (b) are present in a combined amount
effective to control the growth of at least one microorganism and
the amount of the N-alkyl heterocyclic compound (b) present
potentiates the microbicidal activity of the TCMTB (a).
31. A method according to claim 30, wherein the N-alkyl
heterocyclic compound is N-dodecyl morpholine or N-dodecyl
imidazole and the microorganism is algae, fungi, or bacteria.
32. A method according to claim 30, wherein said aqueous system is
selected from the group consisting of a latex, a metal working
fluid, an aqueous emulsion, an aqueous detergent, cooling water,
and an aqueous resin formulation.
33. A method for controlling the growth of microorganisms on pulp
or paper in a papermaking process, comprising the step of
contacting the pulp or paper with (a)
2-(Thiocyanomethylthio)benzothiazole (TCMTB) and (b) an N-alkyl
heterocyclic compound of the formula: ##STR00024## wherein n varies
from .[.5 to 17.]. .Iadd.9 to 15.Iaddend., the heterocyclic ring
defined by ##STR00025## is a substituted or unsubstituted ring
having four to eight members, and wherein the TCMTB (a) and the
N-alkyl heterocyclic compound (b) are present in a combined amount
effective to control the growth of at least one microorganism and
the amount of the N-alkyl heterocyclic compound (b) present
potentiates the microbicidal activity of the TCMTB (a).
34. A method according to claim 33, wherein wet-lap pulp is
contacted by spraying an aqueous formulation of the TCMTB and the
N-alkyl heterocyclic compound onto the pulp after the pulp leaves a
press in the papermaking process.
35. A method according to claim 33, wherein wet-lap pulp is
contacted by mixing the TCMTB and an N-alkyl heterocyclic compound
into a pulp/white water mixture prior to reaching a formation wire
in a papermaking process.
36. A method according to claim 33, wherein the TCMTB and the
N-alkyl heterocyclic compound are incorporated into the body of the
paper.
37. A method according to claim 33, wherein contacting step is
accomplished by mixing the TCMTB and an N-alkyl heterocyclic
compound into a coating composition and the coating composition is
applied to the finished paper.
38. A method according to claim 33, wherein the N-alkyl
heterocyclic compound is N-dodecyl morpholine or N-dodecyl
imidazole and the microorganism is algae, fungi, or bacteria.
Description
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to compositions for controlling the
growth of microorganisms on a variety of substrates and in aqueous
systems. More particularly, the invention relates to combinations
of 2-(Thiocyanomethylthio)benzothiazole (TCMTB) with an N-alkyl
heterocyclic compound and the use of such combinations as
microbicides.
2. Background of the Invention
A large variety of commercial, industrial, agricultural, and wood
materials or products are subject to microbiologicl attack or
degradation which reduces or destroys their economic value.
Examples of such materials or products include surface coatings,
lumber, seeds, plants, leather and plastics. The various
temperatures at which such materials or products are manufactured,
stored, or used as well as their intrinsic characteristics make
them susceptible to growth, attack, and degradation by common
microorgansims such as algae, fungi, yeasts, and bacteria. These
microorganisms may be introduced during a manufacturing or other
industrial process, by exposure to air, tanks, pipes, equipment,
and humans. They can also be introduced while using a material or
product, for example, by multiple openings and reclosures of
packages or from stirring or removing material with contaminated
objects.
Aqueous systems are also highly subject to microbiological growth,
attack, and degradation. The aqueous system may be a fresh,
brackish or saltwater system. Exemplary aqueous systems include,
but are not limited to, latexes, surfactants, dispersants,
stabilizers, thickeners, adhesives, starches, waxes, proteins,
emulsifying agents, cellulose products, metal working fluids,
cooling water, waste water, aqueous emulsions, aqueous detergents,
coating compositions, paint compositions, and resins formulated in
aqueous solutions, emulsions or suspensions. These systems
frequently contain relatively large amounts of water and organic
material causing them to be environments well-suited for
microbiologic growth and thus attack and degradation.
Microbiological degradation of aqueous systems may manifest itself
as a variety of problems, such as loss of viscosity, gas formation,
objectionable odors, decreased pH, emulsion breaking, color change,
and gelling. Additionally, microbiological deterioration of aqueous
systems can cause fouling of the related water-handling system,
which may include cooling towers, pumps, heat exchangers, and
pipelines, heating systems, scrubbing systems, and other similar
systems.
Another objectionable phenomenon occurring in aqueous systems,
particularly in aqueous industrial process fluids, is slime
formation. Slime formation can occur in fresh, brackish or salt
water systems. Slime consists of matted deposits of microorganisms,
fibers and debris. It may be stringy, pasty, rubbery, tapioca-like,
or hard, and may have a characteristic undesirable odor that is
different from that of the aqueous system in which it formed. The
microorganisms involved in its formation are primarily different
species of spore-forming and nonspore-forming bacteria,
particularly capsulated forms of bacteria which secrete gelatinous
substances that envelop or encase the cells. Slime microorganisms
also include filamentous bacteria, filamentous fungi of the mold
type, yeast, and yeast-like organisms. Slime reduces yields in
production and causes plugging, bulking, and other problems in
industrial water systems.
Various chemicals known as industrial microbicides have been used
to prevent microbiological deterioration of industrial systems, raw
materials, and products. For instance,
2-(Thiocyanomethylthio)benzothiazole (TCMTB) is a well known,
effective microbicide available for such uses. The use of TCMTB as
an industrial microbicide has been described in U.S. Pat. Nos.
4,293,559, 4,866,081, 4,595,691, 4,944,892, 4,893,373, and
4,479,961. TCMTB is manufactured by Buckman Laboratories
International Inc., and sold as Busan.RTM. 30L product, Busan.RTM.
30WB product, Busan.RTM. 1030 product, Busan.RTM. 1118 product as
well as other products. TCMTB has the following chemical structure:
##STR00003##
Despite the existence of such microbicides, industry is constantly
seeking more cost-effective technology which offers equal or better
protection at lower cost and lower concentration. The concentration
of conventional microbicides and the corresponding treatment costs
for such use, can be relatively high. Important factors in the
search for cost-effective microbicides include the duration of
microbicidal effect, the ease of use, and the effectiveness of the
microbicide per unit weight.
SUMMARY OF THE INVENTION
In view of industry's search for more cost effective microbicides,
the present invention offers an improvement over current products
or practices.
A first embodiment of the invention provides a microbicidal
composition. This compostion comprises (a)
2-(Thiocyanomethylthio)benzothiazole (TCMTB) as a microbicide, and
(b) an N-alkyl heterocyclic compound of the formula: ##STR00004##
For the N-alkyl heterocyclic compound, n may vary from 5 to 17 and
the heterocyclic ring defined by ##STR00005## may be a substituted
or unsubstituted ring having four to eight members. In the
microbicidal composition, the TCMTB (a) and the N-alkyl
heterocyclic compound (b) are present in a combined amount
effective to control the growth of at least one microorganism. The
combination of TCMTB with an N-alkyl heterocyclic compound achieves
superior microbicidal activity at lower concentrations and lower
cost than TCMTB alone against microbiological attack or degradation
such as discussed above.
Another embodiment of the present invention provides a method for
controlling the growth of a microorganism on a substrate. This
method comprises contacting the substrate with TCMTB and an N-alkyl
heterocyclic compound, having the above formula. The TCMTB and
N-alkyl heterocyclic compound are present in a combined amount
effective to control the growth of at least one microorganism on
the substrate.
Another embodiment of the invention provides a method for
controlling the growth of microorganisms in an aqueous system
capable of supporting growth of a microorganism. This method
comprises the step of treating the aqueous system with
2-(Thiocyanomethylthio)benzothiazole (TCMTB) and an N-alkyl
heterocyclic compound of the above formula, where the TCMTB and the
N-alkyl heterocyclic compound are present in a combined amount
effective to control the growth of at least one microorganism in
the aqueous system.
The combination of TCMTB and an N-alkyl heterocyclic compound
according to the invention is useful in preventing the
microbiological attack, degradation, or deterioration of various
types of raw materials and products such as leather, textiles,
pulp, paper and paperboard, coatings, lumber, as well as
agricultural products such as seeds and crops. Advantageously, the
combination may be used in various industrial processes used to
prepare or manufacture these products. Accordingly, additional
embodiments of the present invention employ the combination to
control the growth of microorganisms on or in such industrial
products, raw materials or processes.
The foregoing and other features and advantages of the present
invention will be made more apparent from the following detailed
description and preferred embodiments.
DETAILED DESCRIPTION OF THE INVENTION
In one embodiment, the present invention relates to a microbicidal
composition comprising 2-(Thiocyanomethylthio)benzothiazole (TCMTB)
as a microbicide and an N-alkyl heterocyclic compound. Mixtures of
N-alkyl heterocyclic compounds may be used. The TCMTB and the
N-alkyl heterocyclic compound are present in a combined amount
effective to control the growth of at least one microorganism.
The N-alkyl heterocyclic compounds employed in the present
invention have the following general formula: ##STR00006## The
variable "n" may vary from 5 to 17, and preferably from 9 to 15.
Most preferably, n is 11. The alkyl chain defined by
CH.sub.3C.sub.nH.sub.2n-- may be branched or unbranched. Branched
alkyl chains may lose some of their solubility in water or other
aqueous systems. Unbranched alkyl groups are generally
preferred.
The heterocyclic ring defined by ##STR00007## may have four to
eight members and is preferably a five-, six-, seven-, or
eight-member ring. Most preferably the heterocyclic ring is a
six-membered ring.
Although the heterocyclic ring always contains one nitrogen atom,
the remainder is generally a carbocycle. However, the ring may
contain one or more additional heteroatoms selected from N, O, or
S. The ring may be saturated or unsaturated. The ring may also have
common substituents such as alkyl groups, substituted alkyl groups,
alkenyl groups, substituted alkenyl groups, amino groups, an oxo
group to form a cyclic ketone, halogens, etc. The heterocyclic ring
may also be part of a multiple ring structure.
The heterocycles listed below exemplify substituted or
unsubstituted heterocyclic rings which may be used in the N-alkyl
heterocyclic compounds utilized in preferred embodiments of the
present invention. Examples of five-membered heterocyclic rings
include, but are not limited to, pyrrolidinyl, 2-pyrrolidinonyl,
pyrrolinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, imidazolidinyl,
imidazolinyl, imidazolyl and oxazolidinonyl. Six-membered rings
include, but are not limited to, piperidinyl, piperazinyl, and
morpholinyl. Seven- and eight-membered rings such as
hexamethyleneiminyl and heptamethyleneiminyl may also be used in
the present invention. One of ordinary skill will appreciate that
other heterocyclic rings may also be used.
N-alkyl heterocyclic compounds useful in the invention are
available either commercially from chemical supply houses or may be
prepared from starting materials using well-known literature
methods. U.S. Pat. No. 5,250,194 discloses exemplary methods and is
incorporated herein by reference.
U.S. Pat. No. 5,250,194 also describes N-dodecyl heterocyclic
compounds and their use as microbicides for aqueous systems to
inhibit the growth of microorganisms, the formation of slime in
aqueous systems, or the disfigurement or deterioration of
substances susceptible to microbiological growth. One example of an
N-alkyl heterocyclic compound useful as such a microbicide is
N-dodecyl morpholine (DDM). DDM is manufactured by BASF GmbH and by
Buckman Laboratories International Inc., Memphis, Tenn.
Preferred N-alkyl heterocyclic compounds for use in the present
invention include N-dodecyl morpholine, N-dodecyl imidazole,
N-dodecyl-2,6-dimethyl-morpholine,
N-dodecyl-5-chloromethyl-2-oxazolidinone,
N-dodecyl-2-pyrrolidinone, N-dodecyl hexamethyleneimine, N-dodecyl
pyrrolidine, N-dodecyl-3-methyl-piperidine, N-dodecyl piperidine,
N-dodecyl-4-methyl-piperidine and N-dodecyl-2-methyl-piperidine.
Most preferred of these compounds are N-dodecyl morpholine, (DDM),
and N-dodecyl imidazole, (DDI).
Depending on the application, microbicidal compositions according
to the present invention may be prepared in various forms known in
the art. For example, the composition may be prepared in liquid
form as an aqueous solution, dispersion, emulsion, or suspension, a
dispersion or suspension in a non-solvent, or as a solution by
dissolving the TCMTB and the N-alkyl heterocyclic compound in a
solvent or combination of solvents. Suitable solvents include, but
are not limited to, methyl ethers of glycols, M-pyrol or petroleum
distillates. The microbicidal composition may be prepared as a
concentrate for dilution prior to its intended use. Common
additives such as surfactants, emulsifiers, dispersants, and the
like may be used as known in the art to increase the solubility of
the TCMTB or N-alkyl heterocyclic compound in a liquid composition
or system, such as an aqueous composition or system. In many cases,
the biocidal composition of the invention may be solubilized by
simple agitation.
Microbicidal compositions of the present invention may also be
prepared in solid form, for example as a powder or tablet, using
means known in the art. In a preferred method of preparation, a
liquid product containing TCMTB is deposited on a carrier such as
diatomaceous earth or kaolin and mixed with an N-alkyl heterocyclic
compound in the form of a liquid or solution to form a powder or
tablet.
The TCMTB and the N-alkyl heterocyclic compound may be combined in
a single composition. Alternatively, the TCMTB and the N-alkyl
heterocyclic compound may be employed as separate components such
that combined amount for the intended use is effective to control
the growth of at least one microorganism.
As mentioned above, a microbicidal composition of the invention
demonstrates an unexpected enhanced microbicidal effect between the
respective components, TCMTB and an N-alkyl heterocyclic compound.
That is, the combination of TCMTB and an N-alkyl heterocyclic
compound achieves superior microbicidal activity at lower
concentrations to control the growth of microorganisms as compared
to the microbicidal capability of TCMTB alone. Thus, the N-alkyl
heterocyclic compound potentiates, or even synergistically
enhances, the microbicidal effect of the TCMTB. Such a superior
effect presents a distinct economic advantage and increases the
microbicide's effectiveness per unit weight.
According to the present invention, control of the growth of a
microorganism on a substrate or in an aqueous system means control
to, at, or below a desired level and for a desired period of time
for the particular substrate or system. This can vary from the
complete prevention or inhibition of microbiological growth to
control at a certain desired level and for a desired time. The
combination of TCMTB and N-alkyl heterocyclic compound described
here can, in many cases, reduce the total microbiological count to
undetectable limits and maintain the count at that level for a
significant period of time. Accordingly, the combination may be
used to preserve a substrate or system.
The effective amount or percentage of the combination of TCMTB and
an N-alkyl heterocyclic compound necessary to achieve the desired
result will vary somewhat depending on the substrate or aqueous
system to be protected, the conditions for microbial growth, and
the degree of protection desired. For a particular application, the
amount of choice may be determined by routine testing of various
amounts prior to treatment of the entire affected substrate or
system. In general, an effective amount used on a substrate ranges
from about 0.0001% to about 4% (w/w); preferably about 0.0001% to
about 0.2%. With aqueous systems, an effective amount may range
from about 0.5 to about 5000 parts per million, more preferably
from about 5 to about 1000 parts per million of the aqueous system,
and most preferably from, about 10 to about 25 parts per million.
Similar amounts effectively control slime formation. For slime
control, effective amounts preferably range from about 1 to about
200 parts per million, and more preferably, from about 1 to about
25 parts per million of the aqueous system.
In a preferred embodiment, combinations of TCMTB and an N-alkyl
heterocyclic compound are those combinations having a weight ratio
of TCMTB to N-alkyl heterocyclic compound from about 99:1 to about
1:99. More preferably the weight ratio is from about 60:10 to about
10:60, and most preferably, from about 50:50 to about 25:75. The
weight ratio may vary depending on the intended use, the
microorganism encountered as well as the particular material,
product, or system to which the combination according to the
invention is applied.
The combination of TCMTB and an N-alkyl heterocyclic compound may
be applied in a variety of industrial uses and processes for
microorganism control. The combination may be used in place of and
in the same manner as other microbicides traditionally used in the
particular industry. As discussed above, such industries include,
but are not limited to the leather industry, the lumber industry,
the papermaking industry, the textile industry, the agricultural
industry, and the coating industry. The combination of TCMTB and an
N-alkyl heterocyclic compound may also be used with aqueous systems
such as those previously discussed which are subject to
microbiological attack and degradation. The problems caused by
microbiological attack and deterioration in these various
applications has been described above. The use of the combination
of TCMTB and an N-alkyl heterocyclic compound according to the
invention to control the growth of microorganisms in particular
exemplary applications is described below.
The invention also relates to a method for controlling the growth
of microorganisms on various substrates. The method comprises the
step of contacting a substrate susceptible to microbiological
growth or attack with TCMTB and an N-alkyl heterocyclic compound,
as described above. The TCMTB and N-alkyl heterocyclic compound are
present in a combined amount effective to control the growth of at
least one microorganism on the substrate. Preferably, the method
may be used to eliminate or prevent substantially all
microbiological growth on the substrate. As discussed above, the
TCMTB and the N-alkyl heterocyclic compound may be applied together
or as separate compositions. Preferred applications of this general
method are discussed below.
In the leather industry, the combination of TCMTB and an N-alkyl
heterocyclic compound may be used to control the growth of
microorganisms on a hide during a tanning process. To achieve this
control, the hide is contacted with a combined amount of TCMTB and
an N-alkyl heterocyclic compound effective to control the growth of
at least one microorganism on the hide. The combination of the
TCMTB and the N-alkyl heterocyclic compound may be used in the
tanning process in similar amounts and manner similar to that used
to apply other microbicides used in the tanning industry. The type
of hide may be any type of hide or skin that is tanned, for example
cowhide, snake skin, alligator skin, sheep skin, and the like. The
amount used, to some extent, will depend on the degree of
microbiological resistance required and may be readily determined
by one skilled in the art.
A typical tanning process comprises a number of stages, including,
but not limited to, a pickling stage, a chrome-tanning stage, a
vegetable-tanning stage, a post-tan washing stage, a retanning
stage, a dyeing stage, and a fatliquoring stage. The combination of
TCMTB and an N-alkyl heterocyclic compound may be used during all
process stages in the tanning process in addition to those stages
where a known microbiological problem is occurring. In each stage,
the combination of TCMTB and an N-alkyl heterocyclic compound may
be a component of the appropriate tanning liquor applied to the
hide undergoing tanning.
Incorporating the TCMTB and an N-alkyl heterocyclic compound in a
tanning liquor protects the hide from microbiological deterioration
during the tanning process. Preferably, the combination is
uniformly dispersed, e.g., under agitation, into an appropriate
liquor to be used in a tanning process. Typical tanning liquors
include, for example, a pickling liquor, a chrome-tanning liquor, a
vegetable-tanning liquor, a post-tan washing liquor, a retanning
liquor, a dye liquor, and a fatliquor. This method of application
ensures that the combination applied to the hides protects against
microbiological attack, deterioration, or other microbiological
degradation.
In a somewhat analogous nature, the combination of the invention
may also be employed to control the growth of microorganisms on a
textile substrate in a textile manufacturing process. Contacting
the textile substrate with a combination of TCMTB and an N-alkyl
heterocyclic compound according to the invention effectively
controls the growth of a microorganism on the textile substrate. In
a textile process, the combination may be used in similar amounts
and a manner similar to other microbicides commonly used in such
processes. As one of ordinary skill would appreciate, particular
amounts generally depend on the textile substrate and the degree of
microbiological resistance required.
The step of contacting the textile substrate with the combination
of TCMTB and an N-alkyl heterocyclic compound may be accomplished
using means known in the textile art. To control microbiological
growth, a textile process generally dips the textile substrate into
a bath containing a microbicide, alone or with other chemicals used
to treat the textile substrate. Alternatively, the textile
substrate may be sprayed with a formulation containing a
microbicide. In the bath or the spray, the combination of TCMTB and
N-alkyl heterocyclic compound according to the invention are
present in a combined amount effective to control the growth of at
least one microorganism on the textile substrate. Preferably, the
bath and the spray are aqueous-based compositions.
To preserve the value of its raw materials and products, the lumber
industry also must control the growth of microorganisms in order to
prevent microbiological degradation. The combination of TCMTB and
an N-alkyl heterocyclic compound according to the invention is
effective to control the growth of microorganisms on lumber.
The combination of TCMTB and an N-alkyl heterocyclic compound may
be used to protect the lumber in similar amounts and a similar
manner employed for other microbicides used in the lumber industry.
Contacting lumber with an effective amount of the combination may
be accomplished, for example, by spraying the lumber with an
aqueous formulation containing the combination of TCMTB and an
N-alkyl heterocyclic compound, by dipping the lumber into a dip
bath containing the combination, or other means known in the art.
Dipping the lumber in an aqueous bath is preferred.
The TCMTB and the N-alkyl heterocyclic compound are preferably
uniformly dispersed in a bath (for example, by agitation) prior to
the dipping of the lumber into the bath. In general, the lumber is
dipped into the bath, raised, allowed to drip dry, and then air
dried. The dip time will depend, as is known in the art, on a
variety of factors such as the degree of microbiological resistance
desired, the moisture content of the lumber, type and density of
the wood, etc. Pressure may be applied to promote penetration of
the combination into the lumber being treated. Applying a vacuum to
the upper surface of the lumber may also be used to degas the
lumber and promote increased wetting of the lumber by a bath
containing the microbicidal combination.
The combination of TCMTB and an N-alkyl heterocyclic compound
according to the invention also has uses in the agricultural
industry. To control the growth of microorganisms on a seed or
plant, the seed or plant may be contacted with TCMTB and an N-alkyl
heterocyclic compound in a combined amount effective to control the
growth of at least one microorganism on the seed or plant. This
contacting step may be accomplished using means and amounts known
in the agricultural industry for other microbicides. For example,
the seed or plant may be sprayed with an aqueous formulation
containing the combination of TCMTB and N-alkyl heterocyclic
compound, or dipped into a bath containing the combination. After
being sprayed or dipped, the seed or plant is generally dried by
means known in the art such as drip drying, heated drying, or air
drying. For plants or crops, the combination may also be applied
using a soil drench. Soil drenching is particularly advantageous
when the microorganisms of concern inhabit the soil surrounding the
plant.
Yet another aspect of the present invention is a method for
controlling the growth of microorganisms in an aqueous system
capable of supporting such growth. The aqueous system is treated
with TCMTB and an N-alkyl heterocyclic compound such that the TCMTB
and N-alkyl heterocyclic compound are present in a combined amount
effective to control the growth of at least one microorganism in
the aqueous system. This includes controlling, and preferably
preventing, slime formation in the aqueous system.
Examples of various aqueous systems include, but are not limited
to, latexes, surfactants, dispersants, stabilizers, thickeners,
adhesives, starches, waxes, proteins, emulsifying agents, cellulose
products, aqueous emulsions, aqueous detergents, coating
compositions, paint compositions, alum compositions, and resins
formulated in aqueous solutions, emulsions or suspensions. The
combination may also be employed in aqueous systems used in
industrial processes such as metal working fluids, cooling waters
(both intake cooling water and effluent cooling water), and waste
waters including waste waters or sanitation waters undergoing
treatment of the waste in the water, e.g. sewage treatment.
As with the other uses discussed above, the combination of the
invention may be used in the same amounts and in the same manner as
microbicides traditionally used in these various aqueous systems.
The combination not only protects the aqueous system prior to use
or when stored, but in many cases protects the aqueous system when
in use or in appropriate applications even after the aqueous system
has dried. When used in a paint formulation for example, the
combination not only protects the paint in the can, but also the
paint film after being applied to a substrate.
Another embodiment of the present invention is a method for
controlling the growth of microorganisms on paper or in a
papermaking process, e.g., in a pulp or paper slurry and on a
finished paper product such as paper board. The paper, pulp, or
slurry is contacted with TCMTB and an N-alkyl heterocyclic compound
in a combined amount effective to control the growth of at least
one microorganism on the paper, the pulp or in a slurry. The
contacting step is accomplished using means and amounts known in
the papermaking art.
According to this aspect of the invention, for example, a forming
web on a papermaking machine (or a wet-lap pulp) may be contacted
with the combination of TCMTB and an N-alkyl heterocyclic compound
by spraying an aqueous dispersion containing the TCMTB and N-alkyl
heterocyclic compound onto the pulp after the pulp leaves the
presses in a papermaking process. Or, the TCMTB and the N-alkyl
heterocyclic compound may be incorporated into a bath used at the
wet or size press and the web contacted by nipping the web to
incorporate the combination into the web with any other agents
applied at the press. Alternatively, the pulp may be contacted by
mixing the TCMTB and N-alkyl heterocyclic compound into the
pulp/white water mixture, preferably prior to the pulp reaching the
formation wire.
When treating paper (which includes paperboard and other cellulosic
products or substrates), the TCMTB and N-alkyl heterocyclic
compound may be added into pulp slurries in the headbox, in the
substrate forming solution, or in the white water system to treat
the water system itself or for incorporation into the body of the
paper. Alternatively, as with other known microbicides, the
combination of TCMTB and an N-alkyl heterocyclic compound according
to the invention may be mixed into a coating used to coat the
finished paper.
The activity of the combinations described above has been confirmed
using standard laboratory techniques as discussed below. In many
cases, the N-alkyl heterocyclic compound potentiates, or even
synergistically enhances, the microbicidal affect of the TCMTB.
The following examples are intended to illustrate, not limit, the
present invention.
EXAMPLE 1
FUNGICIDAL ACTIVITY OF TCMTB AND AN N-ALKYL HETEROCYCLIC
COMPOUND
The fungicidal effects of combinations use of TCMTB and N-alkyl
heterocyclic compounds against fungi were measured by the binary
dilution method. Combinations of TCMTB and N-dodecyl morpholine
(DDM) were tested against a mixture of fungal species isolated from
molded chrome-tanned leather. Busan.RTM. 30L product, a 30% TCMTB
formulation, and BL 2180 product, a 60% formulation of DDM, were
used in these tests. Both the Busan.RTM. 30L product and BL 2180
product are available from Buckman Laboratories Inc., Memphis,
Tenn.
The original Trichoderma culture used in the testing was isolated
and cultured from a molded sample of chrome-tanned or wet blue
leather. From a suspension of Trichoderma viride spores, 0.1 ml of
this suspension was added to each of 200 standard sized test tubes.
Each test tube contained 4.5 ml of nutrient salts broth. The test
tubes were then agitated to ensure proper mixing of the fungal
spores into the nutrient broth. Once inoculated and treated at
various concentrations and combinations, the test tubes were
incubated at 28.degree. C. for 7 days. The lowest concentration of
each chemical compound or combination of compounds that prevented
the growth of the fungi in the broth, i.e., the minimum inhibitory
concentration, was taken as the endpoint.
Synergism was demonstrated using the method devised by Kull, E. C.,
Eisman, P. C., Sylwestrwicz, H. D., and Mayer, R. L., Applied
Microbiology, 1961, pp.538-541. Kull et al. apply the following
summation (synergism index) to determine whether a synergistic
effect was achieved: QA/Qa+QB/Qb In this formula QA, Qa, QB, and Qb
have the following values:
Qa=Concentration of compound A in parts per million, acting alone,
which produced an end point. This represents the minimum inhibitory
concentration (MIC) for compound A.
Qb=Concentration of compound B in parts per million, acting alone,
which produced an end point. This represents the minimum inhibitory
concentration (MIC) for compound B.
QA=Concentration of compound A in parts per million, in the
mixture, which produced an end point.
QB=Concentration of compound B in parts per million, in the
mixture, which produced an end point.
When the synergism index is greater than one, antagonism is
indicated and when the index is equal to one, additivity is
indicated. When the index is less than one, synergism exits.
Table 1 below shows the synergism indices as calculated using the
Kull method for TCMTB (compound A), dodecyl morpholine (DDM,
compound B) and TCMTB/DDM combinations against a mixture of fungal
species isolated from molded chrome-tanned leather. The most
effective combination tested was found to be 2 ppm Busan.RTM. 30L
and 6 ppm of BL 2180 product, a 60% DDM formulation.
TABLE-US-00001 TABLE 1 A: Busan .RTM. B: Dodecyl 30L Morpholine
Synergism (30% TCMTB) (60% DDM) Index 4* 0* 1 3 2 0.75 2 6 0.5 2 10
0.5 1.75 60 0.4375 0 80 1 *ppm of formulated preparation
EXAMPLE 2
BACTERICIDAL ACTIVITY OF TCMTB AND AN N-ALKYL HETEROCYCLIC
COMPOUND
The bactericidal effects of the combined use of TCMTB and N-alkyl
heterocyclic compounds against bacteria were measured using an 18
Hour Stokes Broth Test. Combinations of TCMTB and N-dodecyl
morpholine (DDM) and of TCMTB and N-dodecyl imidazole (DDI) were
tested against the bacteria Sphaerotilus Natans, ATCC 15291. In
this test, compound A was Busan.RTM. 1030 product, a 30% TCMTB
formulation, and compound B was either DDM (99%) or DDI (99%). The
Busan.RTM. 30L product is available from Buckman Laboratories,
Memphis, Tenn.
The Stokes broth substrate used to evaluate effectiveness of
microbicidal compositions against Sphaerotilus natans was prepared
by adding the following to one liter of water:
TABLE-US-00002 Compound g/L Peptone 1.0 Glucose 1.0
MgSO.sub.4.7H.sub.2O 0.2 CaCl.sub.2 0.05 FeCl.sub.3.6H.sub.2O
0.01
The broth was adjusted to pH 7.0 with sodium hydroxide. Forty gram
portions were then added to 180 mL Pyrex milk dilution bottles
fitted with Escher rubber stoppers and then sterilized. Each of the
following substances was then added to each bottle in the order
listed. 1. Sterile distilled water as required in each individual
case to bring the total weight of the contents of each bottle to 50
g after all subsequent additions specified below (including
inoculation with the aqueous suspension of test organism) have been
made. 2. Solution of toxicant or control agent to be evaluated in
such individual volumes as to give the concentration desired in
parts per million by weight. 3. One mL of a 24-hour old S. natans
culture grown in Stokes broth.
After the inoculant suspension of the test organism was added to
the bottles they were incubated at 28.degree. C. for 18 hours.
After this incubation, a 1 mL of sample was withdrawn from each
bottle, diluted, plated on Stokes agar, and incubated for 72-96 hr.
at 28.degree.-30.degree. C. Stokes agar was prepared like the broth
with 12.5 g agar added per liter. The number of colonies on each
plate was determined and converted to the count per milliliter of
substrate.
From these data, the percentage kills were calculated. The
difference between the count for the control substrate (with no
toxicant) and the count obtained from the substrate containing
toxicant was divided by the count for the control substrate to give
the fraction killed, which was then converted to "percentage
killed" by multiplying by 100. The results are shown in Tables 2
and 3. A percentage kill of 90 percent or higher represents an
extremely useful bactericidal composition. Synergism indices were
calculated for some combinations using the Kull method as described
in Example 1. Others may be calculated from the data presented.
TABLE-US-00003 TABLE 2 A: Busan .RTM. 1030 B: Dodecyl Synergism
(30% TCMTB) Morpholine CFU/mL % Kill Index 0* 0 3.80 .times.
10.sup.6 -- 1 0 1.13 .times. 10.sup.6 70.3 2 0 1.09 .times.
10.sup.6 71.3 5 0 7.82 .times. 10.sup.5 79.4 10 0 <10.sup.3
>99.9 20 0 <10.sup.3 >99.9 0 1 1.58 .times. 10.sup.6 58.4
0 2 1.24 .times. 10.sup.6 67.4 0 5 8.51 .times. 10.sup.5 77.6 0 10
<10.sup.3 >99.9 0 20 <10.sup.3 >99.9 1 1 1.02 .times.
10.sup.6 73.2 1 2 6.33 .times. 10.sup.5 83.3 1 5 2.17 .times.
10.sup.4 99.4 0.60 1 10 <10.sup.3 >99.9 2 1 1.14 .times.
10.sup.3 97.0 0.30 2 2 1.85 .times. 10.sup.5 95.1 0.40 2 5 6.41
.times. 10.sup.3 83.1 2 10 <10.sup.3 >99.9 5 1 1.23 .times.
10.sup.3 96.8 0.60 5 2 8.74 .times. 10.sup.4 97.7 0.70 5 5
<10.sup.3 >99.9 1.00 5 10 <10.sup.3 >99.9 10 1
<10.sup.3 >99.9 10 2 <10.sup.3 >99.9 10 5 <10.sup.3
>99.9 10 10 <10.sup.3 >99.9 *ppm of formulated
preparation
TABLE-US-00004 TABLE 3 A: Busan .RTM. 1030 B: Dodecyl Synergism
(30% TCMTB) Morpholine CFU/mL % Kill Index 0* 0 5.05 .times.
10.sup.6 -- 0.2 0 2.63 .times. 10.sup.6 47.9 0.5 0 1.97 .times.
10.sup.6 61.0 1 0 1.17 .times. 10.sup.6 76.8 2 0 1.06 .times.
10.sup.6 79.0 5 0 8.20 .times. 10.sup.3 83.8 10 0 <10.sup.3
>99.9 20 0 <10.sup.3 >99.9 0 1 1.26 .times. 10.sup.6 75.1
0 2 6.80 .times. 10.sup.3 86.5 0 5 <10.sup.3 >99.9 0 10
<10.sup.3 >99.9 0 20 <10.sup.3 >99.9 0 50 <10.sup.3
>99.9 0 100 <10.sup.3 >99.9 0.2 2 1.04 .times. 10.sup.6
79.4 0.2 5 <10.sup.3 >99.9 1.02 0.2 10 <10.sup.3 >99.9
0.2 20 <10.sup.3 >99.9 0.5 1 8.40 .times. 10.sup.3 83.4 0.5 2
5.00 .times. 10.sup.5 90.1 0.45 0.5 5 <10.sup.3 >99.9 0.5 10
<10.sup.3 >99.9 0.5 20 <10.sup.3 >99.9 1 1 3.80 .times.
10.sup.3 92.5 0.30 1 2 7.70 .times. 10.sup.5 84.8 1 5 1.25 .times.
10.sup.6 75.2 1 10 <10.sup.3 >99.9 1 20 <10.sup.3 >99.9
2 1 3.50 .times. 10.sup.5 93.1 0.40 2 2 6.50 .times. 10.sup.5 87.1
2 5 <10.sup.3 >99.9 2 10 <10.sup.3 >99.9 *ppm of
formulated preparation
EXAMPLE 3
BACTERICIDAL ACTIVITY OF TCMTB AND AN N-ALKYL HETEROCYCLIC
COMPOUND
The bactericidal effects of the combined use of TCMTB and N-alkyl
heterocyclic compounds against bacteria were measured using an 28
Day API Test. Combinations of TCMTB (compound A) and N-dodecyl
morpholine (DDM, compound B) and of TCMTB (compound A) and
N-dodecyl imidazole (DDI, compound B) were tested against the
bateria Desulfovibrio desulfuricans, ATCC 7757. Busan.RTM. 1030
product, a 30% TCMTB formulation, DDM (99%), and DDI (99%) were
used in this testing. The Busan.RTM. 30L product is available from
Buckman Laboratories, Memphis, Tenn.
The medium used was a modified API Broth having the following
composition:
TABLE-US-00005 Compound g/L Yeast extract 5.0 Ascorbic acid 0.1
Sodium lactate 5.2 MgSO.sub.4.7H.sub.2O 0.2 K.sub.2HPO.sub.4 0.6
Ferrous ammonium sulfate 0.1 NaCl 10.0
The compounds were dissolved in 1.0 L of deionized water to form
the medium. The pH of the medium was adjusted to a final pH of 6.9
with 1N sodium hydroxide prior to sterilization.
The medium was sterilized at 121.degree. C. for 20 minutes and then
allowed to cool to room temperature. The medium is then inoculated
by adding 10 ml per liter of a 24-hr. broth culture of
Desulfovibrio desulfuricans.
From stock solutions of the chemicals to be tested, sufficient
amounts of the compounds were added to 1 oz. glass bottles so that
when a bottle was completely filled, the desired concentration of
the chemical was present. The bottles were then completely filled
with the inoculated API broth (10 mL innoculum per liter medium) in
a manner such as to exclude all air and provide an anaerobic state.
The bottles were capped tightly and inverted gently to mix the
contents and, at the same time, to make sure no air bubbles were
present.
Incubation was carried out at 37.degree. C. for 28 days. Growth was
indicated by blackening of the culture medium, whereas those
bottles having sufficient chemical to inhibit growth would remain
clear. Observations of growth were made at the end of 7 days, 14
days, 21 days and 28 days. Results were reported using a scale of 0
to 4, where 0=no growth and 4=heavy growth.
The results are shown in Tables 4 and 5. The minimum inhibitory
concentration (MIC) was defined as the lowest concentration tested
which gave no growth in the bottle (clear liquid). Synergism
indices were calculated for some combinations using the Kull method
as described in Example 1. Others may be calculated from the data
presented.
TABLE-US-00006 TABLE 4 A: Busan .RTM. 1030 B: Dodecyl Day Day Day
Day Synergism (30% TCMTB) Morpholine 7 14 21 28 Index 0* 0 4 4 4 4
10 0 4 4 4 4 20 0 0 2 3 4 40 0 0 0 0 0 80 0 0 0 0 0 160 0 0 0 0 0 0
10 4 4 4 4 0 20 4 4 4 4 0 40 2 3 4 4 0 80 0 1 3 4 0 160 0 0 2 3 0
320 0 0 0 0 5 10 4 4 4 4 5 20 4 4 4 4 5 40 4 4 4 4 5 80 0 2 4 4 5
160 0 0 0 0 0.63 10 10 4 4 4 4 10 20 2 3 4 4 10 40 1 2 3 4 10 80 0
0 0 0 0.50 10 160 0 0 0 0 20 10 4 4 4 4 20 20 0 0 0 0 0.56 20 40 0
0 0 0 20 80 0 0 0 0 20 160 0 0 0 0 40 10 0 0 0 0 40 20 0 0 0 0 40
40 0 0 0 0 40 80 0 0 0 0 40 160 0 0 0 0 *ppm of formulated
preparation
TABLE-US-00007 TABLE 5 A: Busan .RTM. 1030 B: Dodecyl Day Day Day
Day Synergism (30% TCMTB) Morpholine 7 14 21 28 Index 0* 0 4 4 4 4
10 0 4 4 4 4 20 0 0 2 3 4 40 0 0 0 0 0 80 0 0 0 0 0 160 0 0 0 0 0 0
1 4 4 4 4 0 2 4 4 4 4 0 4 4 4 4 4 0 8 2 4 4 4 0 16 0 0 0 0 0 32 0 0
0 0 5 2 4 4 4 4 5 4 4 4 4 4 5 8 0 0 0 0 0.63 5 16 0 0 0 0 10 1 4 4
4 4 10 2 4 4 4 4 10 4 0 2 3 4 10 8 0 0 0 0 0.75 10 16 0 0 0 0 10 32
0 0 0 0 20 1 4 4 4 4 20 2 0 2 3 4 20 4 0 0 0 0 0.75 20 8 0 0 0 0 20
16 0 0 0 0 40 1 0 0 2 4 40 2 0 0 0 0 40 4 0 0 0 0 40 8 0 0 0 0 40
16 0 0 0 0 *ppm of formulated preparation
* * * * *