Note: Descriptions are shown in the official language in which they were submitted.
2~9~5~
SMC 35834
--1--
BIOCID~ COMPOSITION
The present invention relates to compositions which are
useful as industrial biocides.
Industrial biocides are useful to prevent industrial
spoilage, in particular that caused by bacteria and fungi. Materials
which can be used as industrial biocides have antimicrobial
properties and particularly have antifungal and antibacterial
properties. Such materials are useful in the preservation of paints,
latices, adhesives, leather, wood, metal working fluids and cooling
water.
Many compounds have been disclosed which have
anti-microbial activity and some of these compounds are commercially
available and are used as industrial biocides. ~owever, whilst these
materials can provide use~ul protection, a need remains for materials
having improved characteristics.
In our European Patent Application Publication No 249328,
we disclose that certain cyclic compounds have useful antimicrobial
properties and can be used as biocides. We have now found that when
used with other compounds having antimicrobial properties a
beneficial effect can be achieved.
According to the present invention there is provided a
composltion which comprises A) at least one compound of the
formula (I)
/ \
B N - O~
\ I (I)
C = S
D
i ~ .
:~
`~ ~
.
2~6~3
-2- S 3~34
or a salt or complex thereof; and
B~ at least one other biologically active compound which is
(i) an agent affecting membrane pe~neability;
(ii) an isothiazolinone or an isothiazolothione; or
(iii) an aldehyde;
wherein
A is a nitrogen or carbon atom, which may be substituted7
B and D are, independently, oxygen or sulphur or a nitrogen or
carbon atom which may be substituted; or
A and/or B and/or B and/or D may be part of a ring syste~;
R is hydrogen, a hydrocarbyl group, a substituted h~drocarbyl
group, an acyl group, a substituted acyl group or a group
-COORl; and
Rl is a hydrocarbyl group
with the proviso that ~ and D ~re not both sulphur or both oxygen.
The bonds between the groups A and B and between the groups
B and D may be single or double bond~ but it will be appreciated that
there can be only one such double bond in the ring.
The group A, and optionally one or both of groups B and D
can be a group -CtR2)2-; a group -CR2~; a group ~ C-NR2; a group
-NR - or a group N-; where R is a hydrogen atom, a hydrocarbyl
group, a substituted hydrocarbyl group or two groups R2, together
with the carbon atom, or carbon atoms, to which they are attached
form a ring.
The groups A, B and D can form part of a Eurther r~ng
system but generally not more than two of the groups A, B ancl D form
part o a further ring system. The further ring system is typically
a hydrocarbon ring system containing five or six carbon atoms, for
example a cyclopentene, cyclohexane, cyclohexene, cyclohexadiene or
benzene ring. The further ring system, if present, typically
contains one or both of the groups A and B. I~ only the group A
forms part of a ring system, this may be a cyclohexane ring of the
type
- - . -
.
. , . ~ . . :
, , , ~ " ,
- . ... .
4 ~ ~
-3_ S 35834
CH ~ ~ CH2
\ /
CH2 C
/
CH2 --- C~2
where the group A is the carbon atom with the ~uo free valences,
which are linked to the group -NOR- and B respectively. If both A
and B form part of a ring system, the further ring is then fused to
S the azolethione rin~ system; for example as in
3-hydroxy-4,5,6,7-tetrahydrobenzothiazol-2~3H)-thione.
I~ many of the compounds used in the biocide compositions
of the present invention, the groups ~1 B and/or D are not part of a
fur~her ring system. Thus, if A, B and/or D is a carbon atom, or
substituted carbon atom, it may be, inter alia9 a group -CH=,
C(C2H5) ~ -C(C6~5)~, -C(C6~4Cl)~ -C(C~3)2 or C=N~. It
will be appreciated that in the foregoing, the group R2 is a hydrogen
atom, a methyl, ethyl, phenyl or chlorophenyl group. The group R2
typically is a hydrogen atom, a lower alkyl group, that is one
containing up to five carbon atoms, an aryl group or a subs~ituted
alkyl or substituted aryl group. If the group R2 iY a substituted
group, each substituent ls a hydrocarbonoxy group, a hydro~arbonthio
group, an acyl (that is a hydrocarboncarbonyl) group, an es~er (that
is an acyloxy) group, a halogen ~tom, a nitrile group or a hydroxy
group.
Preferred compounds are those in which the groups A and B
are both optiotlally substituted carbon atoms and the group D is a
sulphur atom or an optionally substituted nitrogen atom. l'he groups
A and B are preferably linked through a double bond as in the group
-CR2~CR2, in which the groups R2 may be the same or different. It is
preferred that D is a sulphur atom.
: . , .: '
. ~ :
2~9~3
_4_ S 3583
The group R may be a hydrogen atom, an acyl group such as
benzoyl or acetyl, or an alkoxycarbonyl group such as an
ethoxycarbonyl group. If the group R is a substituted group the
substituent may be as disclosed for the group R2 or may be a
substituent which contains a further ring system of general formula
I, the two ring systems being linked through the group R, for example
as in the glu~aryl bis ester of the formula:
OCoc~2c~2cH2coo
I
N N
C~3 ~ \ ~ ~r CH3 ?
/c=Ss~c ~11
S S
Component (A) of the composition may be a salt or complex
of the compound of general fonmula ~. The salt or co~plex may be one
which contains any metal. The metal may be a transition metal, for
example a metal of group VIII, IB or IIB of the Periodic Table. Such
metals include iron, copper and zinc, particularly such metals in
their m~ximum possible valency s~ate.
:
All references herein to the Periodic Table are to the
Pe~iodic Table according to MendeleePf, as set out on the inside rear
cover of UGeneral and Inorganic Chemistry" by J.R.Partingon, Second
Edition publlshed by MacMillan and Co.Limited, I.ondon.
For convenience hereafter, the compounds of the general
formula I, and salts and complexes thereof wlll be referred to simply
as "Compound I".
A wide range of Compounds I can be used as component (A) in
~he compositions of the present invention. The Compounds I have
antl-microbial activity agalns~ a wide range of micro-organisms.
., ~
.
.',
~.
.~ . : ~ , , ~ . .
.
- - . ' ::
~069~3
_5_ S 35834
Compounds I which can be used in the compositions of the
present inventioD include:
3-hydroxy-4-methylthiazol-2(3H)-thione,
3-benzoyloxy-4-methylthiazol-2(3H)-thione,
3-hydroxy-4-phenylthiazol-2t3H)-thione,
3-hydroxy-4,5,6,7-tetrahydrobenzothiazol-2t3H)-thione,
3-acetoxy-4-methylthiazol-2(3H)-thione,
the glutaryl bis-ester of 3-hydroxy-4-methylthiazol-2(3H)-thione,
5,5-dimethyl-1-hydroxy-4-imino-3-phenylimid~æolidine-2-thione,
1-hydroxy-4-inino-3-phenyl-2-thiono-1,3-diaza~piro[4.5]decan~,
l-hydroxy-5-methyl-4-phenylimidazoline-2-thio~e,
3-ethoxycarbonyloxy_4-methylthiazol-2(3H)-thione,
4,5-dimethyl-3-hydroxythiazol-2(3EI)-thione,
~s,5-dimethyl-3-acetoxythiazol-2t3H)-thione,
4-ethyl-3-hydroxy-5-methylthiazol-2(3H)-thione,
4-ethyl-3-acetoxy-5-methylthiazol-2(3H)thione,
4-(4-chlorophenyl)-3-hydroxythiazol-2(3H)-thione,
3-hydroxy-S-methyl-4-phenylthiazol-2(3~)-thione,
3-ace~oxy-4-phenylthiazol-2(3H)-thione,
and th~ metal complexes and salts thereof. The metal complexes and
salts thereof includ~ ferric, cup~ic and zinc complexes and salts
such as
the zinc complex of 3-hydroxy-4-methylthiazol-2(3H)-thione, thP
ferric complex of 3-hydroxy-4-methylthiazol-2(3H)-thione,
the cupric complex of
l-hydroxy-4-imino-3-phenyl-2-thion-1,3-diazaspiro~4.S]decane,
the cupric complex of 4,5-dimethyl-3-hydroxythiazol-2(3H)~thione,
the zinc complex of 4,5-dimethyl-3-hyclroxythiazol-2(3H)-thione, and
the zinc complex of 4-ethyl-3-hydroxy-5-methylthiazol-2(3H)-thione.
We have obtained particularly useful results when component
(A) is the zinc complex or salt of 3-hydroxy-4-methylthiazol-2(3H)
-thione.
'
2~9~L~3
-6- S 35834
Component (B) (i) of the composition i5 an agsnt affecting
membrane permeability. Action on general membrane permeability is
discussed in ~Pharmaceutical Microbiology" edited by W.B.Hugo and
J.B.Russell, published in 1977 by Blackwell, Chapter 11, specifically
at pages 202 and 204. The d~scussion indicates that treatment of
bacterial cells with certain substances causes a leakage of a group
of characteristic chemical species. Po~assium ion is indicated to
the first substance to appear when such damage occurs. Materials
which cause such an effect can be used as component (B) (i) of the
composition of the present invention. Component (B) (i) may be a
phenol and o-phenylphenol has been found to give a useful effect.
Cationic disinfectants can be effective as an agent
affectinK membrane permeability. By disinfectan~ we mean a material
which is able to destroy pathogenic micro-organisms but not
necessarily resistant spores as discussed on page 701 of "Medical
Microbiology", 11th edition (1965) by R.Cruickshank, published by E
and S Livingstone Ltd. Effective cationic disinfectants for use in
the compositions of the present invention include blguanides and
quaternary ammonium compounds.
Biguanides which may be used in the compositions of the
present invention contain at least one biguanide unit of the formula
(II) _
-NC-C-NH C-NH- (II)
11 11
NH NH
.
Typically the biguanide contains at least two units of the
formula (II) which are linked by a bridging group which contains at
least one methylene group. The bridging group may include a
polymethylene chain which may optionally be interrupted by hetero
atoms such as oxygen, sulphur or nitrogen. The bridging group may
include one or more cyclic nuclei which may be saturated or
unsaturated. It is generally preferred that the bridging group is
such that there are at leas~ three, and especially at least four,
.
''~: ' ' ' ' ' ~
-- . . ::
. . ~ .
2~9~5~
-7 S 3~34
carbon atoms directly interposed between two adjacent units of the
fonmula (II). In general it is preferred that there are not more
than ten carbon atoms, especially not more than eight carbon atoms,
interposed betw~en two adjacent units of the formula (II).
The biguanide units May be terminated by any suitable group
which may be a hydrocarbyl or substituted hydrocarbyl group or which
may be an amine group or an amine hydrochloride group or by a group
-NH-C-NH-CN
Il
NH
If the terminating group is a hydrocarbyl group this may be an alkyl,
cycloalkyl or aryl group or may be an alkyl, cycloalkyl or ,aryl group
or may be a combination thereof as ln an aralkyl group. If the
.erminating group is a substituted hydrocarbyl group, the substltuent
can be any substituent which does not have an undesirable adverse
effect on the microbial activity of the biguanide compound and
typically is a hydrocarbonoxy group, a hydrocarboncarbonyl (than is
an acyl) group, an ester (that is an acyloxy) group, a halogen atom
or a nitrile group and there may be more than one substituent, for
example more than one halogen atom.
A suitable biguanide is a material which contains two units
of ~he formula (II) and in which the the units are linked by a
; polymethylene group, particularly a hexamethylene group. The
terminating groups may be 4-chlorophenyl groups, for example as in
the compound oE formula (III):-
H H El H H H
~1 1 1 1 i I ~
Cl ~ ~ N-C-N-C-N-~CH~)6-N-C-N-C-N~ ~ Cl (III)
~ ~v
NH NH NH NH
The compound of formula (II) is available as a
chlorhexidine salt.
::
.~ ~
2~9~3
-8 S 3S834
The biguanide may alternatively be a polymeric biguanide,
for example a linear pol~neric biguanide which has a recurring
polymer unit represented by the formula
X-NH-C-NH-C-NH-Y-NH-C-NH-C-NH-
Il 11 11 11
NH NH NH NH
wherein X and Y may be the same or different and represent bridgin8
groups in which together the total number of carbon atoms directly
interposed bet~een the pairs of nitrogen atoms linked by X and Y is
at least 9 and not more than 17.
The bridging groups X and Y may consist of polymethylene
chains, optionally interrupted by hetero atoms, for example, oxygen,
sulphur or nitrogen. X and Y may also incorporate cyclic nuclei
~hich may be saturated or unsaturated, in which case the number of
carbon ato~s directly interposed between the pairs of nitrogen atoms
linked by X and Y is taken as including that segment of the cyclic
group, or groups, which is the shortest. Thus, the number of carbon
a~oms directly lnterposed between the nitrogen atoms in the group
~ .
-NH-CH2-O CH2 N~l
is 4 and not 8.
The preferred polymeric biguanide for use in the present
inventiorl is poly(hexamethylene biguanide), in ~hich X and Y both
represent the -(CH2)6- group.
Polymeric biguanides may be prepared by the reaction of a
bisdicyanidiamide having the fonmula
,:
.
20~9~
9- S 3583
CN-NH-C-NH-X-NH C-NH-CN
Il 1i
NH NN
with a diamine H2N-Y-NH2, wherein X and Y have the meanings defined
previously herein;
5or by reaction between a diamine salt of dicyanimide having
the formuLa
:~ + +
(H3N-X-NH3)(N(CN)2)2
with a diamine ~2N-Y-NH2 wherein X and Y have the meanings defined
~; previously herein. These methods of preparation are d~scribed in UK
lOPatent Specifications Nos. 702,268 and 1152243 respectively, and any
of the polymeric biguanides described therein may be used as
component (B)(i) in the composition of the present invention.
The polymer chains are terminated either by an amino
~; hydrochloride grnup or by an -NH-C-NH-CN group, and the terminating
11 ,
NH
`
groups may be the same or different on each polymer chain.
Typically, the polymerlc biguanides are obtained as
mixtures of polymers in which the polymer chains are of different
lengths, the number of individual biguanide units, i.e.
20 -X-NH-C-NH-C-NH-
11 11
NH NH
and -Y-NH-C-NH-C-NH-
11 11
, NH NH
. . ~ .,
.
2~9~3
10- S 35~34
together being from 3 to about 80.
In the case of the preferred poly(hexa~lethylene biguanide)
having the formula (IV)
(CH2)6-NH-C-NH-C-NH 1--
NH N~ (IV)
rl
the value of n is in the range from 6 to 15, the average molecular
weight of the polymer mixture being from about 1100 to about 3300.
The biguanides are used as salts with suitable inorganic or
organic acids, for example as the hydrochloride salts or the acetate
or gluconate.
The component (B)(i) may be a quaternary a~monium salt of
the general formula ~V):-
[NR3R4RSR6] E ~V)
where
R3 is a hydrocarbyl group or substituted hydrocarbyl group;
R4, RS and R6 may be the same or different and are a hydrogen
atom or a hydrocarbyl group or a substituted hydrocarbyl
group; and
E is an anion havlng a valency and being in an a~ount: to giYe
a neutral compound;
wherein
R3, R4, R5 and R6 tog~ther contain at least six aliphatic carbon
atoms or two or more of R4, R5 and R6, together with the nitrogen
atom to which they are attached, form one or more heterocyclic rings.
Preferred quaternary ammonium salts are those in ~hich R3
is a group containing at least six allphatic carbon atoms. If any of
the groups R3, R4, RS or R6 is a substltuted hydrocarbyl group the
subs~ituent may be a hydrocarbonoxy group, an acyl group, an acyloxy
group, a nltrile group, a hydroxy group or a halogen atom.
"
-
2~6~3
-11- S 35834
The hydrocarbyl group may be an alkyl, cycloalkyl or aryl
group or a mixture of such groups as in an alkaryl group. The
quaternary ammonium compound may one in wh:ich not more than two the
groups R3, R4, R5 and R6 are alkyl groups containing at least six
carbon atoms. In a compound of the foregoing type the remaining
groups are preferably hydrogen, lower alkyl (as previously defined
herein) or a group con~aining an aromatic ring such as a benzyl
group. Alternatively, the residual groups may fonm a heterocyclic
ring, for example a pyridine ring or a nitrogen containing adamantane
structure.
The anion E is preferably a mono-valent anion such as a
halide anion, a bisulphate (HSO4) anion or an alkylsu].phate anion.
Quaternary a~monium compounds which may be used as
component (B)(i) in the compogition of the present invention include
diethyldodecylbenzyl ammonium chloride;
dimethyloctadecyl-(dimethylbenzyl)am~onium chloride;
dimethyldidecylammonium chloride; dimethyldidodecylammonium chloride;
; trimethyl-tetradecylam~tonium chloride; benzyldimethyl(C12-C18
alkyl)a~ttonium chloride; dichlorobenzyldimethyldodecylammonium
chloride; hexadecylpyridiniu~ chloride; hexadecylpyridinium bromide;
hexadecyltrimethylammonium bromide; dodecylpyridinium chloride; !'
dodecylpyridinium bisulphate;
benzyldodecyl-bistbeta-hydroxyethyl)~ttmonium chloride;
dodecylbenzyltrimethylammonium chloride; benzyldimethyltC12-C18
alkyl~ ammonium chloride; benzyldimethyl (C12-C16 alkyl) ammoniu~
chloride; trimethyl ~C12-C16 alkyl) ammonium bromide;
dodecyldimethylethyl ammonium ethylsulphate;
dodecyldimethyl (l-naphthylmethyl)ammonium chloride;
hexadecyldinethylbenzyl al~nonium chloride; dodecyldimethylbenzyl
an~tonium chloride and
l-t3-chloroallyl)-3,5,7-triaza-1-azonia-adamantane chloride. The
quaternary ammonium salt may be benzyldimethyl(C12-C16)alkyl ammonium
chloride.
2~694L~3
-12- S 35~34
Component (B)(ii) of the composition is an lsothiazolinone
or an isothia~olothione.
The isothia~olinone or isothiazolothione derivstive which
is component (B)(ii) of the composition is typically a compound of
the general fonmula:
R8 \ ¦ ¦ :
/ C
; ¦¦ N - R7
C
. R9 S
wherein:
2 is an oxygen or sulphur atom;
R7 is a hydrogen atom, a substituted or unsubstituted
hydrocarbyl:group, a substituted or unsubstituted
hydrocarbylthio group, a substituted or unsubstituted
hydrocarbyloxy group, a carbamoyI group or a cation;
R8 i8 a hydrogen atom, a halogen atom, a cyano group, or a
~ubstituted or unsubstituted hydrocarbyl group;
R9 is a hydrogen atom, a halogen atom, a cyano group, or a
substituted or unsubstituted hydrocarbyl group; or
R~ and R9, together with the carbon atoms to which they are attached,
form a five- or six-membered ring, which may optionally be
substituted.
;: :
:, -
2~9~3
-13- S 35834
Preferably component (B)(ii) is at least one
isothiazolinone derivative, ~hat is a compound in which Z is an
oxygen atom. If the groups R7, R8 and R9 are, or contain,
substituted hydrocarbyl groups, the substituents may be as disclosed
for the group R2 and are typically halogen, alkoxy or alkylthio,
particularly those in which the alkyl groups contain l to 4 carbon
atoms. If R7 is a carbamoyl group, this of th~ general type -CONHR10
where R10 is a hydrogen atom or a hydrocarbyl group, which may be
substituted. It is generally preferred that the group R7 is a
hydrogen atom or a lower alkyl group (as previously defined herein).
R7 is especially hydrogen or a methyl group.
R~ and Rg ~ay, together with the carbon atoms to which they
are attached, form a flve- or six-membered ring, which may be
substituted, the substituents typically being halogen, alkyl, alkoxy,
or alkylthio gro~ps. The ring thus ob~ained may contain a heteroatom,
for example a nitrogen atom but in general R8 and R9 fon~ a
hydrocarbon ring such as a benzane, cyclopentene or cyclohexene ring.
Alternatively, R8 and R9 are separate groups and one or both o R8
and R9 can be a hydrogen atom. It is generally preferred that at
least one of R8 and R9 is other than a hydrogen atom and is,
particul~rly, a halogen atom, for example chlorine or a lower alkyl
; group.
Isothia~olinone compound (Z is oxygen) which can be used as
component (B)(ii) of the mixture include 2-methylisothiazolin-3-one
(R7 is methyl, R8 and R9 are both hydrogen);
5-chloro-2-methylisothiazolin-3-one (~7 ls methyl, R8 is hydrogen and
R is chlorine); mi~turqs of the foregoing two compounds;
4,5-dichloro-2-methylisothiazolin-3-one ~R7 is methyl and R8 and R9
are both chlorine); 1,2-benzisothiazolin-3-one (R7 is hydrogen and R8
and R9, together with the carbon atoms to which they are attached,
~ form a benzene ring); 4,5-trimethylene-4-isothiazolln-3-one (R7 is
; hydrogen and R8 and R9 togethcr with the csrbon atoms to which ~hey
are attached, ~orm a cyclopentene ring); and
2-methyl-4,5-trimethylene-4-isothiazolin-3-one (R7 is methyl and R~
and R9 together with the carbon atoms to which they are attached,
form a cyclopentene ring).
.
: . .
~ S 3583
In component tB)(ii). if R7 is a cation this may he a
cation having a valency of more than one but is particularly a
monovalent cation such as an alkali metal, an amlne or quaternary
ammonium cation.
Component (B)(iii) of the composition is an aldehyde.
Suitable aldehydes are fonmaldehyde and glutersldehyde.
Useful results have been achieved when component (B) is
o-phenylphenol, chlorhexidine hydrochloride salt, poly(hexamethylene
biguanide) hydrochloride salt, (C12-C16) al~yldimethyLbenz~lammonium
chloride, a mixture of quaternary a~monium chlorides including
(Cl2-Cl6)alkyldimethylbenzyl, octyldecyldimethyl, dioctyldimethyl and
didecyl dimethyl ammonium chloride,
5-chloro-2-methylisothia~olin-3-one, 1,2-benzisothiazolin-3-one,
2-methyl-4,5-trinethylene-~-isothiazolin-3-one, Eormaldehyde or
glutaraldehyde.
:,
Useful compositions in accordance with the present
invention comprise
A) the zi~c complex or salt of
3-hydro~y-4-methylthiazol-2(3H)-thione; and
B) o-phenylphenol,chlorhexidine hydrochloride salt,
poly(hexamethylene biguAnide) hydrochloride salt, (Cl~-Cl6)
alkyldimethylbenzylammonium chloride, a mlxture of quaternary
ammonium chlorides including (C12-C16)alkyldimethylbenzYl,
octyldecyldimethyl, dioctyldimethyl and didecyl dimethyl ammonium
chloride, 5-chloro-2-methylisothiazolin-3-one;
1,2-benzisothiazolin-3-one;
2-methyl-4,5-trlmethylene-4-isothiazolin-3-one, :Eormaldehyde or
glutaraldehyde.
The relative proportions of components (A) and (B) are
dependent on the particular componen~s, especially component ~B).
The antimicrobial activity of materials which can be used in the
compositions of the present invention varies quite widely, this being
particularly so in respect oE materials which can be used as
component (B), some of which are effective at a concentration of less
2 ~ 5 3
-15- S 3583
than lo ppm by ~rei~ht whilst other materials may be used in amounts
as high as 0.1% by weight to achieve an effect. Hence, depending on
the particular combination of (A) and (B), the relative proportions
thereof, by weight, may be from lO0:1 to 1:50.
The composition of the present invention may be prepared by
mixing components (A) and (B) together using any suitable technique
depending on the physical state of the components and their
solubility. Thus, A and B may both be solids and be mixed together
using any suitable solids mixing technique. Alternatively, A and B
may be both be liquids and be mutually miscible. A and B may both be
soluble in the same solvent and be mixed together us solutions in
said solvent. A further alternative is to suspend one component
which is a solid in a liquid medium which is the other component or
is a solution o the other component in a liquid medit~n in which only
lS one of A and B is soluble.
The mixtures of the present invention provide improved
anti-microbial ac~ivity allowing lower concentrat~ons of component
(A) to be used.
The compositions of the present invention provide good wet
state preservation making the compositions advantageous for use as a
cutting fluid preservative and also in cooling water applications.
Wood and leather preservation is another advantageous field of
application of the compositions. The compositions o~ the present
invention can also be incorporated into paint7 as paint film
ftmgicide and many of the compositions can be used without the
addition oP a further active component.
The compositions oE the present invention may consist only
of components (A) and (B). However, typically the composition
comprises the components (A) and (B) as a solution, suspension or
emulsion in a suitable liquid medit~n such as water. The composition
may comprise a suspension or emulslon of the components ~A) and (B),
or a solution thereof, in a liquid medium in ~hich o~e, or both, of
the components is inso1uble.
,
. ~
,: ~
.; . - . .~ ~:
2 0 ~ 3
-16- S 35~34
The composition may be incorporated into a medium to be
protected against micro-organisms using any suitable ~ixing
technique. Thc composition is incorporated into the medium to be
protected in an amount to provide from 0.00002 to 5% by weight of the
composition of (A) and (B) relative to the total compositiont more
preferably from 0.00005 to lZ by weight of (A) and (B). It will be
appreciated that the quantity of the composition of (A) and (B)
required will be dependent on various factors such as the medium to
be protected, the micro-organisms against which protection is desired
and the extent of protection required.
If the composition is being used to preserve a solid
substrate such as leather or wood, the composition may be applied
directly to the substrate or may be incorporated into a coating
composition such as a paint, varnish or lacquer which is then applied
to the substrate. Alternatively, the solid material may be
impregnated with the composition of the present invention.
The compositions of the present invention can be used for
the treatment o$ various media to inhibit the growth of
micro-organisms.
Thus, as a further aspect of the present invention there i9
pro~ided a method for inhibiting ~he growth of micro-organisms on, or
in, a medium which comprise~ treating the medium with a composition
o ~A) and (B) as hereinbefore defined.
The composition can be used in conditions in which
micro-organisms grow and cause problems such as, for example, in
AqUeOus environments including cooling water systems, paper mill
liquors, metal working fluid~s geological drilling lubricants,
polymer emulsions, and emulsion paints. The composition can also be
used to impregnate solid materials such as wood or leather or can be
coated onto the surfaces thereof directly or incorporated into a
paint, varnish or lacquer.
- .
~, - . - : ,
20~9~3
17- S 35834
As a Eurther aspect the present invention provides a
material which is susceptible to attack by micro-organisms and which
cont~ins a composition of (A) and (B) in sn amount sufficient to at
least reduce attack by micro-organisms.
Further aspects of the present invention are described in
the following illustrative examples.
In the fcllowing exarnples, the products obtained were
subjected to microbiocidal evaluation. The microbiological testing
was effected, as follows:
~_
,
Soil was taken from a depth o up to 100 mm, the top 25 mm
being discarded. To each gramme of soll (wet weight) was added 2 cm3
of sterile saline (0.85% w/v sodium chlorlde). The resulting mixture
was stirred for 30 minutes and allowed to settle for five minutes.
The liquid was separated from large particulate matter using a syphon.
20 cm3 of the resulting liquid was added to malt broth to
give a final volume of 1 dm3.
,
Evaluati~n .~ L~
lO cm3 of the soil/broth mixture prepared as described were
placed into 2S cm3 bottles. To the contents of each bottle were
added quantities of one or more bioc:Ldes to gi~e an ll x 11 array
containing different relative proportions of biocide or biocide
mixture. The bottles were vortex mixed for 1-3 seconds and incubated
for one weelc at 25C without agitation.
::~ ` , ,
~, .. , ; ' ' ' ~
2~6~3
-18- S 35~34
The bottles were inspected and any containing visible
bacterial or ungal growth were recorded and the contents thereof
discarded. 10 mm3 aliquot samples were removed from the remalning
bottles and these were placed on malt agar plates, allowed to dry and
incubated at 25C for two to three days. The presence or absence of
growth was recorded and the bottles providing the aliquots giving
growth were discarded.
Immediately after removing the 10 mm3 aliquot samples, 200
mm portions of freshly prepared Soil Organism Sample were added to
the remaining bottles. After a fur~her week, lO mm3 aliquot samples
were removed, placed on malt agar plates and evaluated for growth.
The procedure described was con~inued until 3 portions,
each of 200 mm3, of freshly prepared Soil Organism Sample had been
added to the bottles containing the initial soillbroth mixture.
Lsamples l to lO
A number of compo~nds and mixtures were tested in the
manner described. The concentrations of biocide, or biocide mixture,
which prevented any viable organisms gi~ing rise to growth in the
fourth 10 mm3 aliquot placed on a malt a~ar plate were noted as
providing effective control of the soil organis~s.
The results obtained were used to plot the variation of FLC
with the relative proportions of the biocide components. These
results are set out in accompanying Figures l to 11, wherein
Figure 1 is a plot of the variation of E'LC using ZHMT ~the
zinc complex or salt of 3-hydroxy-4-methylthiazol-2(3H)-thione) or
1,2-benzisothiazolin-3 one (BLT) or mixtures thereof;
Figure 2 is a plot of the variation of FLC using ZE~T or
formaldehyde or mixtures thereof;
-- .
~6~3
-l9- S 3583
Figure 3 is a plot of the variation of FLC using ZHMT or
1,2-dibromo-2,4-dicyanstobutane (T38) or mixtures thereof;
Figure 4 is a plot of the variati.on of FLC using Z~T or
glutaraldehyde or mixtures thereof;
Figure 5 is a plot of the variation of FLC using ZHMT or
chlorhexidine or mixtures thereof;
Figure 6 is a plot of the variation of PLC using ZHMT or
2-methyl-4,5-trimethylene-4-isothiazolin-3-one (MTI) or mi.~tures
thereof;
Figure 7 is a plot of the variation of FLC using ZE~T or
5-chloro-2 methylisothiazolin-3-one (CIT~ or mixtures thereoE;
Figure 8 is a plot of ~he variation of FLC using Z~MT or
; (G12-C16)alkyldimethyl-ben~yl~mmonium chloride (VCL) or mixtures
thereof;
Figure 9 i9 a plot of the variation of FLC using ZHMT or
o-phenylphenol or mixtures thereof;
~ .
Pigure lO is a plot of the variation of PLC using ZEMT or a
mixture of quaternary ammonium chlorides (available as Bardac 205M
(205) from Lonza Inc) or mixtures thereof;
Pigure ll is a plot of the variation of FLC using Z}~T or a
po].~(hexamethylenebiguanide) hydrochloride having an average
molecular weight in the range 1850 to 3300 (PHB) or mixtures of ZHMT
and PHB.
In Figures 1 to 11, the concentration of the biocides are
given as a proportion of the fractional lethal concentration (FLC)
where
PLC = Concentration of compound in mixture which controls ~rowth
Concentration of compound alone which controls growth.
,
2~69~3
-20~ S 35834
In Figures 1 to 11, the dotted straight line represents the
relationshlp if an additive effect is achieved. The 8011d line
represents the relationship as d0termlned experimentally. In Figure
3, using a mi~ture which is not in accordance with the present
invention, the dotted and soLid lines coincide showing that this
mixture gives an additive effect.
It should be noted that not all o~ the biocides used with
Z~MT were effective under the test conditions described even at the
highest concentration tested and with these materi~ls a value of one
on the axis of the graph represents a nu~erical extrapolation to a
point where the compound alone gives control.
From the plots in Figures l to 11, it is possible to
determine the minimum value of the FLC of the mixtures used and the
proportions of the two components which are required to attain this
minim~lm value. This information is se~ out in the following Table
together with results using further mixtures which ar~ not
represented in Figures 1 to 11 including mixtures, not in accordance
with the present invention, which did not give a minimum value of
FLC.
`
2 ~ 3
-21- S 35834
TABLE
. _ _ ~
¦ Example I Z~MT ta) I Blocide _ ¦ Total
I or Comp ¦ Amount I Type ( c ) I Amount ¦ FLC
¦Ex. ¦ (ppm) (b) ¦ (c) ¦ (ppm) (b) ¦ (d)
__ l _ .. _ l l _ l _
¦ A ¦ 50 I NIL ¦0 ¦ 1.0
¦ B ¦ 0 ¦ BIT ¦63 ¦ 1.0
¦ 1 ¦ 20 I BIT I22 1 0.75
¦ C ¦ 0 ¦ FA ¦1600 ¦ 1.0
¦ 2 ¦12.5 I FA ¦400 ¦ 0.5
¦ D ¦12.5 ¦ GA ¦2000 ¦ 1.0~ 1
¦ 3 ¦ 15 I GA ¦1200 1 0.75 ~ ¦
¦ E ¦ 0.0 ¦ CH ¦6000 ¦ 1.0
12.5 I C~ 11200 1 0.45
F ¦12.5 ¦ MTI I 60.0 ¦ 1.0
¦ 5 ¦17.5 ¦ MTI ¦ 36 ¦ 0.8
¦ G I Q.0 I CIT I 2.4 ¦ 1.0
1 6 117.5 I CIT ¦ 1~2 ¦ 0.85
¦ H ¦ 0.0 j VCL ¦ 350 ¦ 1.0
1 7 ¦ 15 ¦ VCL ¦ 140 ¦ 0.7
I I ¦ 0.0 1 OPP ¦ 300 ¦ 1.0
1 8 120.0 1 OPP I 105 1 0.75
; I J I 0 1 205 1 125 1 1.0
9 112.5 1 205 1 37.5 1 0.55
I K ¦12.5 ¦ PHB I 200 ¦ 1.0+
¦10 ¦ 15 ¦ P~B I 120 1 0.75+ ¦
¦ L ¦ 0 ¦ T38 ¦ 100 ¦ 1.0++ ¦
¦ M ¦ 30 ¦ T38 1 70 ¦ 1.3++ ¦
¦ N ¦ ltO ¦ T38 ¦ 10 ¦ 0.9+
¦ 0 ¦ 0.0 I BR0 ¦ 49.2 ¦ 1.0
¦ P ¦62.5 ¦ BR0 ¦ 49.2 ¦ 2.25
I Q ¦ 75 ¦ BR0 ¦ 24.6 ¦ 2.0
¦ R ¦ 0 ¦ DS ¦1000 ¦ 1++
¦ S ¦62.5 ¦ DS ¦1000 1 2.25t+ ¦
I r 1 75 I DS I500 1 2.0
¦ U ¦ 0 ¦ DA l1000 ¦ 1.0
I V 187.5 ¦ DA ¦800 ¦ 2.55-~
~~ ~ . I
~0~5~
-22- S 35834
Notes to Table
a) ZEMT is the zinc complex or salt of
3-hydroxy-4-methylthiazol-2(3H)-thione and can be prepared
as described in Example 2 of EP-A-249328.
(b) Amoun~ is the concentration of ~he active componen~ in
parts per million w/v of the liquid mixture.
tc) BIT is 1,2-benzisothiazoline-3-one
FA is formaldehyde
GA is glutaraldehyde
CH is chlorhexidine
MTI is 2-methyl-4,5-trimethylene-4-isothiazolin-3-one
CIT is 5-chloro-2-methylisothiazolin-3-one
;~ VCL is (C12-C16)alkyldimethylbenzyl ammoni~ chloride
OPP ls o-phenylphenol
205 is a mixture of quaternary ammonium chlorides available
as Bardac 205M from Lonza Inc.
PHB is poly(hexamethylene biguanide)hydrochloride having an
average molecular weight in the range 1850 to 3300
T38 is 1,2-dibromo-Z,4-dicyanatobutane
BR0 is 5-bromo-5-nitro-1,3-dioxan
DS is Z,3,5,6-tetrachloro-4-methylsulphonylpyridine
DA is
3-(2-chlorobenzyl)-2,2-dimethyl-4-(triazol-1-yl)butan-3-ol
(d) PLC is the fractional lethal concentration and i9 the
concentration which prevents growth i~ ~he "Evaluation of
microbial activity" test as described herein and for any
compound i8 given by the relation~hip:-
FLC - Concentxation of compound in mixture wh~ch prevents ~rowth
Concentration of compound alone which prevents growth
.
:~ :
2~69~3
-23- S 3583
+ At the maximum level used, thsse compounds alone did not
prevent growth under the test conditions described and were
effective to prevent growth only in combination with a
quantity of ZXMT. With these compounds, FLC is calculated
as follows:-
(Concn. of com~ound in mixture preventin~ ~rowth) x [l-~FLC of Z~MT
Highest concn of compound used at highest concn
of other
compo~:md) ]
Hence, if the highest concentration of a co~pound which
prevents growth is 1000 ppm in the presence of 10 ppm of Z~MT, the
~: relationship is
(Concn of compound in mixture prev nt n~ ~rowth) x (1-10)
1000 50
that is 0.8 (Concentration of compound in mixture preventl~rowth)
1000
.
++ At the maximum level used, these compound alone did not
prevent ~rowth under the ~est conditions described and were
; effective to preven~ growth only in combination with a
quantity of ZHMT but did not give a minimum value of FLCo
With these compounds, PLC is determined using the highest
concentration of the particular compound used, for example
a compound alone does not give control at 1600 ppm but in a
mixture with ZHMT control i8 achieved with 40 ppm of ZHMT
and 1~00 ppm of the compound, the FLC for this mixture is
40 ~ 1200 = 0.8 -~ 0.75 ~ 1.55
1600
.
