Note: Descriptions are shown in the official language in which they were submitted.
WO 96/40831 r~ u_,~/{7u48
~ 21 96975
FLAVONOID ALDEHYDES AND USE IN PAINT
INT~ODUCTION
Field of thP InVP"rinn
This invention relates to flavonoid aldehydes as biocides in paints and wood
10 ~c~.v.llivc products ~or use in, and on the surfaces of, homes, boats, offices and
imdustrial buildings.
n . I~ U . UUl/d ~
Biocides are required in many paint and wood ,u~ V- Liv~, r ,. ", '-I;n -c to
1~ prevent microbial ~ l;n~ during shipment, storage or use. Biocides are also
required in these formulated products to help protect a substrate ~ubac~_.l.ly coated
with these formulated products from harmful Illi.luUl~;~A._..ls such as bacteria and
fungi and the like. Biocides used in paint products may be grouped into three major
classes: ,ulc~clva-iv~, mildewicides and a ' ' P~e~lva,iv~, are widely used
~0 in water-based paint systems to prevent in-can bacterial and fungal ~Pgr~ rinn durmg
storage and shipment. They are particularly useful in latex systems such as syntnetic
rubber, pulr~-,-y' , and natural rubber latexes. Mil-l.,w;,id,, are employed to
prevent ~ dl", of the dried paint films and underlying substrate by
Illi~.lUUll;.lll..~lll~. Antifoulant paints are used to prevent the growth of organisms on
25 the hulls of both commercial and pleasure boats. The attachment of such organisms
decreases the operatmg efficiency of the boats and increases their "~h,r~ - costs.
Mercurial-type biocides have been widely used as both ,UlC~ClVO.~iV~.~ and
~uild.w;.;de~ in paints. They have excellent L,~,lru~ ,l. c in both functions in many
situations. They offer fast kill time and can control high levels of bacterial
30 r U r ~ they are ha~ardous to handle and may present
cuvhullll~.l~l problems. Thus, their use may be limited to certain ~ppli~rin~c
Various IlUllllA,II.,Ulia.l ~ ,lv~l.iV~ and mildewicides have been hl~ ly
considered as substitutes for mercurial rnmrolln~l~
SUBSTITUTE SHEET (RULE 26)
WO 96/40831 PCT/US95/17048
2 1 (~6975
A wide variety of biocides have been tried as marine: r but the
has been dominated by r ~ of cuprous oxide and organotin
Cuprous oxide has been popular because it is efficient, relatively
~ l, and is specified in many military antifouling p~unt r as the
5 exclusive biocide. However, this chemical causes UpUlU ~;L~/ in the paint film,
which adversely affects efficiency, and it limits the paint colors which can be
formulated to those of a dark reddish brown. The use of organotin ~ l~ ' has
been growing in recent years; however, these l ' are more expensive than
cuprous oxide and also more difficult to l into punt r
o r they do not leach out completely during use so that when ships are
" ' 1, the disposal of the resulting I ' sand poses ~I rr
However, paint r ~ containing organotins yield uniform, tight films without
the 1~ UIJUIUalt~ problems associated with those formulated with cuprous oxide and
may be formulated in a wide variety of bright or light colors. For these latter
15 reasons, they are widely used on pleasure boats. Since both cuprous oxide andorganotin ~ r present technical or c~lv;lum.~ l problems, there is a need for
new and better antifoulant paint biocides.
Biocides also are employed as wood l,.~.valive products in order to prevent
~. ~. ;...,l;.~.~ of wood products that are exposed to conditions which promote
20 microbial growth and decay. For example, utility poles, cross ties, piling timbers,
freshly milled lumber and fence posts as well as wood chip piles used in pulp
r require the ~ . of biocides to stop or control fungal
In the past, two classes of biocides have been employed as wood
V~lfiv~. One class is oil-bome ~-~.v,.Li~. . (e.g., creosote and
25 p_~hlu~ ) while the second class is water-bome salts (e.g., mi~ctures of
inorganic ~ , ' such as copper, chromium, arsenic and zinc salts). The oil-
bome ~ vdfivl,.~ have been the most widely used biocides for wood ~ IV~ ;UI~.
However, products treated with these mixtures may have messy oily surfaces. Alsoboth creosote and ~ , ' ' have been objected to as being . ,.v;., 'ly
30 hazardous. The water-bome salts are also toxic chemicals which are dissolved in
water and injected into wood products where they become bound to or within the
wood. These salts have certain advantages over the oil-bome treatments. They leave
a cleaner surface that may be more readily painted. Also, their water soluble
~ WO96/40831 2 1 9 6975 ~ . ru48
provide savings in solvent costs. However, the use of chromium and
arsenic saits in particuiar presents e.,.;., I problems.
Mosses are a nuisance in hu~ lt~ i and landscape More
ih~ Li~ in certain geographic wnes favoring moss growth on surfaces frequented
5 by people (e.g., stairs, walkways, ioaths~ decks, patios), there is an ongoing concem
about slip and fall injuries. Particuiar interest has been expressed conceming the
safety of senior citizens in outdoor areas of nursing and r~-h~ ~tir~n centers where
the results of such siip and fail accidents are of more medicai (~ L ' '~/
orthopedic) concern. Products on the maricet for control of moss include zinc
10 chioride and ferric sulfate. Zinc chloride solutions have severai L~h.u.~6~. Tney
may cause injury to the respiratory tract; they are corrosive to the eyes resulting in
severe damage which may be followed by biindness; on skin contact they will
severely irritate or burn the skin; and upon swailowing, they are extremely corrosive
to the mouth and throat, where they burn the tissue, atld in sufficient quantities they
can cause death to the animai. Ferric sulfate is corrosive to the eye and is listed as
toxic to aquatic iife.
It therefore is of interest to develop biocides for use in paint and wood
ati~w which do not pose heaith and/or ~ ' hazards.
20 ~ Li~ ure
Antifouiing paint is disclosed in USPN 4,313,800. Miidew resistant paint
are disclosed in USPN 5,073,582. Skin diseases and contact sensitivity
in house painters using water-based paints, glues and putties is described in Fischer,
a 61., (1995) Contoc~ Denm~uids 32:39-45.
Sln~rM AR Y O F TllF DNn~E~rD10 N
The present invention provides ~ and methods for controliing
pathogenic organisms using flavonoid aidehydes in paint and wood ~lc~l~a~
The method includes the step of including an ~ , ~ agent in the paint or
30 wood ~JI~I~dti~ '- in an amount sufficient to control growth of target
pathogenic organisms. The growth modulating product has a formula shown in (1) below:
WO 96/40831 PCT/US95/17048
21 96~75
R4
s ~n~ R
wherein R represents -CH20H or -C~O; n is an integer from O to 3; each R,
, represents OH or an organic substituent containing from 1 to 10 carbon
10 atoms and from O to S 1,~ wherein the total number of carbon and
in all R! ' of said compound is no more than 15, and R~
represents hydrogen or an organic constituent containing from I to 10 carbon atoms.
These include natural compounds such as ~ d, coniferyl
aldehyde, and closely related . ' Also of interest are alpha substituted
IS aldehydes, such as ~-hexyl cinnamic aldehyde (HCA). The method finds use in
treating l ~ and ~ ~ ' crops for pathogenic organisms.
DE~ PrlON OF T~F. SPECl~IC El~RODTMl~ TS
Paint and wood ~ ..u~O substantially free of pathogenic organisms such
20 as fungi and bacteria are provided together with a method to biocontrol pathogen
~ r - on painted or treated surfaces using flavonoid aldehydes. By "biocontrol~
h intended control of pathogens via direct ~ ". - activity. A fungus andlor
bacteria colonizing ~ ' or coated substrate such as a roof and roof shingles,
barns, boat hulls, railroad ties, tree trunks, wooden ~jungle gyms,~ decks, docks,
2S wallcways, stairs, wooden patios, or wharves with brick surfaces, are contacted with a
paint or ,ul~l~ldti~ containing a flavonoid aldehyde. By "colonizing" is intended
association of a , ~ or insect with a surface or with a paint or ~ t;~
r... ~ ~;.. The flavonoid aldehydes can be isolated from a natural source, bewholly or partially synthetic, or be produced by ~l ' ~ techniques.
The method of the subject invention is carried out by adding an effective
palllu~_l. ~ ' ~' ~", amount of a compound of the invention to a paint or ~
r ' '- The compound preferably is added at the time of placing the
~ - in a shipping container, but can be added " 1~, prior to use of the
~ W096~4W31 21 96975 r~l,u.,,~,l,u4s
product. The amount of , ~~ ~ agent that is added depends to some e~tent
upon the paint or ~ ' ' and the specific: . ' ,, used and
therefore is empirically determined for best results. By "~ ' r '~ O ~ I~ is intended
controlling the growth of pathogens and can involve ~illing the pathogen and/or
5 slowing or arresting its ~
The cwmpounds of the present invention offer several advantages over those
currently in use. They possess good ~.t;,..;~.u~ l activity and are not ~ . ' '
with: . of w.... ' paint and wood ~.v~ti.., products. The
~ also are non-volatile, L~l~ul~ stable, thermally stable, and
10 depending upon their chemical structure, may be soluble in water and organic
solvents. r. generally they form no undesirable wlors in the paint and
wood ~.~.vd~ or in the resulting dried films. Still further, they are
wst , .~ with known biocides used in various paints and wood ~ iVC
products while having low or no toxicity toward humans and wildlife. The present15 invention also overwmes the problem of migration of previously available
u~ by bonding the bioactive wmpounds to the wood.
A preferred ~ dtiVi cwmpound is shown in formula (2) below:
R~
~ Q' (2)
Rl~
R2
25 wherein Rl 1l . CHO, R2 represents-OH or an organic substituent wntainingfrom l to 10 carbon atoms, R3 represents a methoxy group or organic substituent
wontaining from I to 10 catbon atoms, and R~ represents a hydrogen or an organicsubstituent wntaining from I to 10 carbon atoms. Of particular interest are havonoid
~ aldehydes, ~ u~ aromatic aldehydes. Examples of aromatic aldehydes of use
30 in the present invention are cilmamic aldehyde ((3) below):
WO 96/40831 2 1 9 6 9 7 5 PCT/US95/17048 ~
C h o
~ (3)
and conifcryl aldehyde ((4) below):
C ~3C~
C H0 (4)
flO~
Other compounds of interest include analogs of the compound of formula (I)
such as ' substituted at the alpha position with an alkyl, such as a hexyllS group, or a branched alkyl group such as an amyl group Generally the group at the
alpha position is from C-5 to C-10. Such compounds include alpha hexyl
d~ and alpha amyl lAPhyde. The chemical structure of alpha-
h_A~ ~ aldehyde (HCA) is shown in (5) (below).
CH~
(CH2)5 (5)
2S ~/
The Chemical Abstracts Service (CAS) name of HCA is 2-(~L~,IIJ~ ' jl....~) octanal
and the CAS Registry Number is [101-86-0]. The compound is also described by thechemical name of 2-hexyl-3-phenyl-2-propenal. The formula of the compound is
Cl5H~oO and the molecular weight is 216.3. HCA is a low to moderately volatile
compound, having a vapor pressure of 70 x 10' mm Hg at 25~C. Its parent
compound, cimlamic aldehyde, has a vapor pressure ~ 'y 40 times higher
(2970 ~ 10' mm Hg at 25~C). (R~ifPn~th W.G. (1995) Volatile .~ '
Cosmetics and Toiletries, 110: 85-93).
~ W096140831 21 96975 P~,l/u~
The arornatic and aliphatic aldehydes of the subject invention can be prepared
by various synthetic methods known to those skilled in the art. For example, sec, J.
March, ed., Appendix B, Advanced Orgar~ic Chemistry: Rcacdors, ~' ' . and
Structure, 2nd Ed., McGraw-Hill, New York, 1977. t' ' ' ' ydu can be
S prepared a.~ '' " "y, for example, by oxidation of cinnamyl alcohol (Traynelis er
al., J. Am. Chcm. Soc. (1964) 86:298) or by ~ of styrene witb
.~h~ ' (Brit. patent 504,125). The subject aldehydes also can be
obtained by isolation from natural sources. For example, ~ 1. h~d., can be
isolated from ~ fungus, Stereum r~hr;l Birkinshaw et al., Biochcm.
10 J. (1957) 66:188.
HCA can be synthesiz_d as described, for example, in USPN 5,055,621. On
a laboratory scale, HCA can be synthesized by reaction of l,~..~W~ h~lc with octanal
under a nitrogen: . ' (aldol ' ) (Personal r- ~ ~ Eric
Walborsky, Firmenich Chemical r~ g Center, Port Newark, New Jersey).
15 The reaction is conducted in a stirred flask charged with methanol, 309 ppm
~'i,' .~' ~ , potassium hydroxide and ' '' ' ~d~ . Following tbe slow addition
of octanal, the reaction mixture is brought to a pH of 7.5-9.5 with acetic acid.Following c~..~.~l;u-- of methanol and wash of the reaction mixture with water, the
organic phasc is transferred to a distillation unit. A~ 'y 2û-24% of the pot
charge is removcd as l ' ' ' ~d~ and alights", with the remaining distillate
~: 'i ' h~ h~ uuh, aldehyde "heart cut." The "hcart cut" is subjectPd
to an additional r.r l:-~ - in which 1-5% (by weight) of the material is removedin ~light~ fracfions, dcpending upon odor cvaluation. The final product is a light
yellow oil having a specific gravity of 0.955-0.965 at 20~C, a refractive index of
1.548-1.562 at 20~C, a boiling point of 305~C at I atrn~r~rhPrP~ and a melting point
of 26~C.
HCA also can be obtained from Firmenich; their product is composed
principally of the (E)-cis isomer (93.8% maximum), and the (Z)-trans isomer (6%
maximum). Among minor ~ . is the self aldol ' product of
octanal (1-1.5% (Personal ~' June Burkhardt, Finnenich, Plainsboro,
New Jersey). The ~ ~.;ol product is stabilizcd with the addition of 0.04% 2, 6-
di-tert-butyl-p-cresol (butylatcd ~ dlUA~,h~ or BHT), which seNes as an anti-
o~cidant (Technical Data Sheet, IIw~ aldehyde 907600, Revision 853,
WO 96140831 2 1 q 6 9 7 5 PCT/llS95/17048 ~
Frmenich Inc., Plainsboro, New Jersey). HCA can be isolated from rice where it
has been reported to occur naturally. (Givaudan-Roure Index, Gi~ n,~
~'nTn~tinn Clifton, New Jersey, 1994, p. 89).
The compounds can be used by hlcul~l~Lil.g an effective paint ~lc~vdLiv~
5 amount of the compound into a paint or wood IJlc~ LilC. By "an effective paintamount~ is intended any amount which will prevent or control
d~ of the paint. In-can ~L'- I l;~n of paints is often caused by gram-
positive bacteria such as Pacillus cereus and S~ .'l aureus or gram-negativebacteria such as those of the r ~ or Y ~/~ classes. This 4c~
10 of the p~unt ingredients results in viscosity loss or generation of offensive odors.
Generally, paint p.~ ld~ J are employed in aqueous-based paint systems
such as latex systems. Solvent-based p~unts usually do not require a p.w~.~di.~
since the . ' ' will not support fungal and/or bacterial growth. In-
can ~JIC~.~tiV~ are fungicidal and/or I - ' ;; l-~ and their killing action preferably
15 is rapid to prevent production of enzymes by the ~ ., which are actually
the cause of the latex paint dewtruction.
When the present bioactive .~ . ' are employed as p~unt ~ Li~w~ it
is usually dwirable to add them to the paint ' ' in the same manner as other
ingredients are i r ' ~ It is preferred to . them as a substitute for
20 ' "~, all of the non-bioactive non-paint . in the p~unt r... ,....1 .l;, ...
The actual amount of ~).WCI~ used varies depending upon many p~m~-t~
Generally, it is preferred to employ from about 0.5 to 5 volume % of a compound of
formula (I) as~part of a total paint r ~ ~ for this purpose.
The compounds of the subject invention also find use for the prevention and/or
25 killing of mildew (-..ilJ~ ;Ji~). Accordingly, the term "effective ' ' ..;ci~hl
amount" is intended to include any amount which will kill or control the growth of
mildew-causing ~ ,, Mildew or mold causing . ~ vary
according to the exposure e..vh.,n..l~.,.. A ,, ' ' pullulans is the most
commonly found species in temperate and colder climates. Tropical and subtropical
30 conditions favor the growth of, u,, of the classes A..,
,~spergillus and Penicillium as well as the alg~ ri~ . UL6LLU.. virides. The effective
' ' . ;.,;J~I amount is varied empirically based up changew in the parameters of the
h.~' ' and the substrate having the . li ' of the subject invention
WO96140831 2 1 9 6 9 75 PCT/US95117048
~ l therein. Generaily, it is preferred to employ from about 0.5 to 5
volume % of a compound shown in formula (I) as part of a totai paint r .. . 1- ...
Another use for the subject compounds is as an antifoulant. An effective
antifouiant amount of one or more of the compounds of formuia (1) is ~ , '
5 into a hull coating ' ~ By the term "effective antifouling amount" is
intended any amount which wiil prevent or control fouiing on the huii. Fouling
organisms include plant forms such as aigae and animai forms such as those of the
classes Anthropeda, ('oPIP~.t~'~ and Mollusca. The green aigae L.t~.l , ' is
the organism most often found on the hulls of large ships. The effective antifouiing
10 amount will vary because of changes in the parameters of the cn~;-- and the
substrate on which it is applied to the huiis. Generaiiy, it is preferred to empioy
from about 0.5 to 5 volume % of the active moiety shown in formula (1) as part of a
totai paint ' ' for this purpose.
An additionai use of the subject r ~ " is as a wood l~G~~ .,. An
lS effective wood-preserving amount of one or more of these compounds is
into a wood treatment product. The term "effective wood ~ amount~ is
intended to include an amount of the compound which prevents or controls
~1. ~" .- 1 ~ ;.... of the wood product to which it is applied. Wood products not in water
are subject to two forms of fungai attack, surface attack (e.g., soft rot) and internai
20 attack (e.g., white and brown rots). Fungi imperfecti and A ~ are the major
cause of soft rot and the P ' ,y~tt.~ class of fungi is the major cause of internai
attacic. White rots attack the lignin and brown rots attack the cellulose. The
commonly l~nown dr,v rot is a brown rot. Also, wood products exposed to seawaterare attacked by marine organisms such as Pholads, 7'eredo, and Lin~noria tripunctata.
25 The effective amount of compound employed in this appiication is empiricaily
determined based upon parameters which include the specific preserving
employed, the type of wood product to be protected, and the type of .,..~i-, the
wood product is exposed to. Generaily, it is preferred to employ from about 0.5 to 5
volume % of the active moiety shown in formula (I) as part of a totai wood
The biocides of the present invention can be added to the wood products by
dther pressure or , ~UIG ' . ~, '' If pressure ~ ~ is employed,
air, hydrostatic pressure or vacuum methods, or ~- 5. ~ . thereof, can be used.
WO 96140831 2 1 q 6 9 7 5 PCT/US95117048 ~
If . ~ l, " of wood is desired, dipping, spraying, brushing or the
lilce can be used.
The bioactive . ' of the present invention can be either added directly
to cellulosic materials such as the wood products in a preformed state, or the
. ' of formula (1) can be added to, for example, bound to a cellulose binding
protein. In this latter case, the bioactive compounds bond to the pul.~l.~ide
structure of the cellulosic material (e.g., wood, paper and the like) upon contact.
Paper products can be treated to make a ' '~ LollL paper, cardboard boxes or
the like using the flavonoid compound - pc ly ~ binding conjugate. A
sufficient portion of a cellulose binding domain up to the full length cellulose can be
used when the target p~ ~h.uide is a cellulose. The preparation of cellulose
binding domains is described in U.S. Patent Nos. 5,340,731; 5,202,247 and
5,166,317. Bindir,g proteins from scaffold proteins also can be used. See Shoseyev
et al. (PCT application EP/0594/04132). The conjugate can be prepared with or.
without a cleavable bond using methods known to those sl~lled in the art.
Paint and wood ~ di~ products which may contain the biocidal
of the invention as ~ h~ and l ~ ..h,;d~,i, include such as latex
and solvent interior and exterior punts, coatingS for new and existing
structures. Other p~unt products include industrial finishing products such as interior
20 and exterior coatings and marine antifouling paints.
The following examples are offered by way of illustration and not by way of
limitation.
F.
n~ ntl M~thn~i~
The chemicals used in the examples given below were obtained from the
following sources: cinnamic aldêhyde, Spectrum Chemical Company, N.J.; coniferyl aldehyde, APIN Chemical, U.K.; Tween 80 and sodium l,;~. Spectrum
Chemical Company, Gardena, Ca. 1'~ are given as the, of
the indicated solution before dilution.
~ WO 96/40831 2 1 9 6 q 7 5 PCT/US95/17048
E3~
E~al and ~ P~PllPnry Test of C
The bioactive . , ' are tested for fungal and algal repellency.
~ ~irmcrl~r slides are used as the substrates in both cases. One half of each slide is
S coated on one side with a film of polymer containing the active agent to be tested,
while the other half of the slide is not treated with the active agent. The total slide is
exposed to the challenge of either fungi or algae with the expectation that the half of
the slide containing the active agent will prevent growth of the challenging organism,
while the untreated half will not.
In the fungal test, the slide is placed on the surface of an agar plate which isseeded with fungi. After incubation for about 14 days, the slide is examined forextent of growth or lack thereof on the treated surface of the slide. Since leaching of
the active agent would create an ~. .l. -.,.1,1. zone-of ' ' outside the perimeter
of the treated surface of the slide, no growth on the treated surface along with a
small, or no, zone of inhibition is the desired result.
In the algal test, the slide is immersed in a nutrient broth which has been
inoculated with the alg~ r~ After an incubation for 30 days
(under light) and a water rinse, the slide is exannined . ~ ~ly and the extent
of algal attachment is noted. Total lack of attachment on the treated surface is the
desired result. Additional i r~ is obtained by comparing the extent of growth
of alg~ throughout the broth. Significant leaching of the active agent from the
treated surface would inhibit growth in the broth as well as on tbe treated surface.
Example 2
Paint ~ ,;de TPC~C
The test procedure is followed exactly from the following published
procedure: R.A. Zabel and W.E. Horner, Joumal of Coanngs Technology, 53, 33-
3',7, (1981), except that the organism A.., ' " pullulans M30-4 is used, isolated
from mildewed exterior latex paint. Duplicates are run in each case. Two separate
30 tests are run for slightly different time periods.
WO 96/40831 2 1 9 6 9 7 5 PCTIUS95/17048 ~
E~
Wood 1'~ ,.,.\ T
This aspect of the present invention extends tne utility of the above described
~ ' ~ ' , ' to the wood ~ Liu~ area. Two general techniques are
S shown. In one, a solution of the compound is applied to the wood direc~dy. In the
second, a solution of compound bound to a cellulose binding protein is applied to the
wocd.
Eight (about 2.5 cm x 2.5 cm x 0.5 cm) weighed pine wood blocks ~ placed
in a dish and a 5-6 mm Hg vacuum applied for 30 minutes. Two solutions are
prepared, one containing a compound of formula (1) and the second a compound of
formula (I) bound to a cellulose binding protein. While still under vacuum, eachsolution is added to a dish and the wood blocks are submerged in one of the two
solutions. After soaking for 30 minutes the wood blocks are removed from the
solutions, excess compound is wiped off and the blocks are heated at 75~ C. for 16
hours. After cooling to room i r ' the blocks are weighed and analyzed to
determine the ;' ' of compound throughout the wood chip.
Examp]e 4
Wood Rnt T.~ct
The test procedure is based upon the following published procedure: H.P.
Sutter, l -' r J ' ;UiUiiU~ Bullenn, 14 (3), 95-99 (1978). The organisms
employed are r: , ' u puteana ATCC 36336 and Lendnus lepide~s ATCC 12653
(a ~ Vt~, ~ fungus). Duplicates are run in each case. The growth of brown
rot ( " ' d ~ ' ,) fungi on pine blocks after 25 days at 28~ C. is evaluated as:
Growth Key
0 - no growth
1 - slight growth
2 - moderate growth
3 - heavy growth
4 - very heavy growth
12
~ WO 91i/40831 2 1 9 6 9 7 5 PCr/US9S/17048
Four (~Z.5 cm x 2.5 cm ~-0.5 cm) print wood blocks are surface treatçd with
either a wood ~ ti~, comprising a test ~ of a compound according
to formula (I) or a commercial wood ~ ,. The wood blocks are brush
coated on all surfaces and in some cases multiple coats are applied. Pressure
5 Dent is not used. These present examples correspond to what a consumer would
do to apply a wood ~ . The amount of biocide applied to each wood block
is calculated by ' ~ the weight increase of each wood block after treatmentand calculating the biocide present in the weight increase.
Example S
~ 111' ' ' ' ' TPCtj~
p- r '~ ~ e ~ I are tes~ed in a standard Minimum Inhibitory
~'. (MIC) tçst against 8 different bacteria and 8 different fungi. Also
tested in this MIC test are Tween 80 (2%), NaHCO3 (6%) and Tween 80 (2%) plus
15 NaHCO3 (6%) ac blanks.
13
WO 96/40831 2 1 9 6 9 7 5 PCT/llS95/17048 ~
The organisms included in the MIC test are:
S ~
aeruginosa ATCC 9028;
2 - F ' acruginosa ~pyrithione resistant);
3 F 'I Ul~aLic~ aerogenes ATCC 13048;
4 - 5~ 7 aureus ATCC 6538;
5 - F. 7~ syriugac ATCC 19310;
6 - Fs ' ~ ~ rt~ ATCC 11355;
7 - Y~ vesiculoria ATCC 11551;
8 - Y~ ' piwseoli ATCC 19315; and
EY~:
1 - Aspergillus niger ATCC i6404;
2 - Tr~ ,.S~ ~ ATCC 9533;
3 - Candiara albicarrs ATCC 10231;
4 - Y ~ , oryzac ATCC 34393;
S - Fuseariurn o~ysporum ATCC 15643;
6 - Glomerella augulata ATCC 10593;
7 - Actcrnaria solani ATCC 11078;
8 - r ~ - solani ATCC 28268.
Example 6
B~yuJ~h~ ~ 'e Activity of Fl-vr nnitl Al~i~hyde
Bryophy~a (moss) bioassay studies were carried out as follows:
fifteen 9 cm plastic petri dishes were lined with Whatman filter paper discs (7.5 cm).
Three ml water were pipetted onto each filter paper disc. Mosses were placed in
groups of five; each moss section was about 3.5 cm x 3.5 cm. Two ml of test
solution were sprayed as a fine mist (Gilmour sprayer) from a distance of 10 cm. and
the petri dishes placed on a table at ambient room i , and observed at 24, 48
and 60 hours. The area of desiccation was measured at each .' v.~li.,n time point.
See Table I (below) for results.
In a second set of ~A~ ~ ', the above pror edure was repeated with
Dic~n moss using a single ~ IA~ of ciMamic aldehyde (2%) in a vehicle
of 2% Tween 80 and 6% NaHCO3 as compared to the individual ~ 'J~ of the
'( ~ See Table 2 for results.
14
~ WO 96/40831 2 1 9 ~ 9 7 5 PCT/U595/17048
~ - Table1
l~ryophyta (Moss)
Percent of Desiccation (over time)
Moss r ~ o24 hrs. 48 hrs. 60 hrs.
Dicranum Fl 0 15 40 90
F2 0 10 40 80
F3 0 15 50 85
F4 0 5 15 25
F5 0 6 18 26
Spagnum
(Bog Moss) Fl 0 20 60 90
F2 0 25 55 75
F3 0 20 60 85
F4 0 8 12 20
F5 0 9 12 22
Wood~nd Fl 0 20 60 85
F2 0 10 40 70
F3 0 20 50 80
F4 0 10 15 20
FS 0 12 18 28
Fl C~uLunic ~dehyde (2%) in 2% Tween 80, 6% NaHC03.
F2 Saponin (10~ Brix) 0.86 ml conc. diluted in 50 ml water.
F3 rFl + F21.
F4 -CONr~ROL H~O.
F5 2% Tween 80,6% NaHC03.
. Table2
3S Moss
PercPnt ~l~cci~tjnn ~t 60 hn~
Percent
r. ~Pci~tinn
Cinnamic aldehyde (2%) 70
T80(2%) 10
NaHC03 (6%) 20
T80 + NaHC03 (2% + 6%) 25
Fl 90
F4 10
WO 96/40831 2 1 9 6 q 7 5 PCT/lJS95il7048 ~
The most effective ' ' tested was 2% cinnamic aldehyde in 2%
Tween 80 and 6% NaHCO3; moss so treated showed 90% desiccation (Dicran~m
and Bog Moss) and 85% desiccation (Woodland Moss) at o0 hours. Cinnamic
aldehyde (2 %) in water produced 70% desiccation of Dicramlm Moss at 60 hours.
S Saponin (1:60in H2O) 10~ Brix caused 80%, 75% and 70% desiccation in
Dicrcruvn, Bog Moss and Woodland Moss, .~L~I~. The ' ~ ' of
cinnamic aldehyde a%) in vehicle and saponin (1:60 in HlO) 10~ Brix was more
effective than saponin (1:60 in H2O) alone, but less effective than 2% cinnamic
aldehyde in vehicle.
Exam~Le 8
Formula~ion,,~ Interios Latex Flat Paint
A latex paint ' ' ' is mixed according to the p~u~ulLu.." in Table 3,
below.
Table 3
IS Latex Paint r~
Ingredient Pounds Gallons
Water 297.18 35.63
Propylene Glycol 35.00 4.05
Cellosize ER~1400 3.5 0.30
Kathon LX 1.5% 1.7 0.20
Drewplus L-422 2.0 0.27
Tanol 731 7.0 0.76
Triton N-101 2.0 0.23
AMP-95 2.8 0.36
Ti-Pure R-931 200.0 6.6
Optiwhite 75,0 4.09
Duramite 96.97 4.31
Ucar 379 265.50 29.35
Texanol 13.0 1.65
2.5% Cellosize ER-4400 100.0 11.92
Totals: 1103.7S 100.00
~ WO96140831 21 9 69 75 r~ u48
Cinnamic aldehydc is substitutcd for }Cathon and propylene glycol in various
One gallon batches are produced and stored in sealed one gallon paint pails.
At 30, 60, 120 and 180 days, lids are removed and the paint compared to
S batches. Inspection for molds, skimming and settling is conducted. Test paints also
are compared to ' flat latex on wood surfaces over the same time frame.
Tnc~tinnc are made for mold and general coating d v ;~
All I ' ' and patent A~ mentioned in this ~ are
indicative of the level of sldll of those sldlled in the art to which this invention
10 pertains. All l ' ' and patent -~ are herein ~ by
reference to the same extent as if each individual publication or patent application was
and individually indicated to be r ' ~ by reference.
The invention now having boen fully described, it will be apparcnt to one of
ordinary sldll in the art that many changes and . .I;~ - can be made thereto
lS without departing from the spirit or scope of the appended claims.
