Note: Descriptions are shown in the official language in which they were submitted.
1 332554
Background of the Invention
In an effort t~ protect the public from
unsanitary surfaces in-public restrooms, telephones and other
surfaces whic~ are contacted by the pub~ic, a number of
5 methods have been developed. Many people have an aversion
to using public restroom facilities or other objects for
< personal use which have been used previously by others.
With the increased concerns of Herpes simplex virus type 2
which is a persistent viral infection once contracted, the
community has become increasingly cautious about exposure.
One of the more commonly available methods for
protection is a disposable paper cover for the toilet
~;~ facility. The paper covers do not contain a germicide and
~.
; are not always available. -
A spray germicide for sanitizing surfaces which
is quick drying was disclosed in U.S. Patent No. 3,445,564
~` to Rirscher. The patent discloses a spray and other
alternative embodiments of a quick drying germicide. It
does not disclose effectiveness against Herpes or the
range of pathogens disclosed herein. Also, there is no
disclosure of use of the disappearing dye which provides a
visual assurance and confirmation of the area treated as
well as an indicator of germicidal destruction.
The use of a dye in a bacteriacidal solution was
disclosed in U.S. Patent No. 2,449,274 to Broll. The use
of the dye was in a bacteriacidal liquid to clean objects ;
such as tableware. The solution would change color upon
losing bacteriacidal strength. This was not a surface
spray application with a dye indicator.
; 30 There are a number of applications for spray
~, germicides such as the commercially available LysolR spray
~t by Lehn and Fink Products Division of Sterling Drug, Inc.
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1 332554
U.S. patent Nos. 3,282,776 and 4,201,764 are examples of surface
or surface and space spray combinations. None of the patents or
products include a dye which is used with the spray in any
manner.
Summary of the Invention
The invention in one aspect pertains to a disinfecting
composition consisting essentially of a lower alkyl alcohol
having 1 to 4 carbon atoms, an effective amount of a disinfecting
surfactant, a pH sensitive dye and alkali means for adjusting the
pH of the composition to produce a color in the liquid with the
dye so that upon neutralization the dye loses color.
The invention also comprehends a method for
disinfecting a surface prior to personal use which comprises
applying a thin layer of the above composition which has been
kept in a sufficiently airtight container with the dye visibly
noticeable in the layer on the surface and allowing the
composition to dry with the color of the dye disappearing upon
exposure in the atmosphere.
The invention also comprehends a biocide composition
for disinfecting a surface ~or personal use consisting
essentially of a lower alkyl alcohol having 1 to 4 carbon atoms,
an effective amount of a disinfecting surfactant, a pH sensitive
dye which changes color upon exposure to air and an alkali means
for adjusting the pH of the composition to produce a color in the
liquid with the dye so that upon neutralization the dye loses
color.
More particularly the invention provides a germicidal
composition with a disappearing dye which can be dispersed in a
fine spray or otherwise spread on a surface with the dye
indicating the delivery of the germicide. Although the dye is
added for color, it is thoroughly mixed and completely dispersed
~ in the system so that a coating or fine layer of the spray
;; imparts a visual colored composition which will also indicate an
active germicidal coating or layer. The germicide is effective
against Herpes simplex virus type 2 (HSV2) as well as bacteria
such as Staphylococcus aureus, Neisseria gonorrhoeae, enteric
bacteria Escherichia coli OllK58 (Pathogenic), Shigella
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1 332554
-2A-
sonnei and Salmonella typhimurium, and the yeast Candida
albicans. The dye will be quite noticeable initially on lighter
colored surfaces and therefore give a visual check as to the area
disinfected. However, with the presence of alkali means for
adjusting the pH of the composition, the dye quickly loses its
color when the composition dries.
Another aspect of the germicide is to provide one or
more detergents as germicides. The detergents are surface active
and attack the target pathogen but also, through the surface
active qualities, causes the composition to spread effectively on
the surface to be disinfected. The spreading aspect is
especially helpful when the composition is delivered in a spray
form because the individual droplets dispensed from any type of
spray device will spread to provide a more even layer before
drying.
The composition can be used in a spray form or applied
with an absorbent wipe in a layer which will dry rapidly or can
be wiped off the surface with toilet tissue.
The composition can be packaged in any type of airtight
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1 332554
container such as an aerosol spray or foil packet and can be made
in a convenient size which can be carried in a purse or pocket.
The composition can also be packaged in a larger size spray
dispenser for multiple applications for home or commercial use.
Description of the Preferred Embodiments
The product is manufactured as a fluid which can be
packaged in a number of ways depending on the desired size of
container and method of delivery to the surface to be
disinfected. For example, the product may be packaged in a spray
dispensing unit for personal or home use in an appropriate size
of container. An aliphatic alcohol with high volatility is used
as the primary component by volume. The alcohol has
bacteriacidal characteristics and allows for the rapid drying of
the layer of disinfectant on the surface. For use in public
restrooms a rapidly drying composition would be most desirable
for use. Due to cost and availability, isopropanol is utilized
in the examples although other quick-drying alcohols could be
substituted. The isopropanol can be mixed with an amount of
water and the examples show about a 70% isopropanol mixture with
30% water.
-Disinfecting surfactant-detergent compounds are also
used in the composition. Surfactants are effective germicides
` and attack the membranes of the organisms. Also, the surfactants
are surface active reducing surface tension. This phenomena
causes the spreading over the surface of the disinfecting
composition providing a more effective distribution of the
germicide on the surface. It is thought that the surfactant has
two characteristics which contribute to this invention. Those
being germicidal activity including effectiveness over a wide
range of organisms including bacteria, virus and yeast and
providing a reduction of the surface tension of the composition
: to achieve effective spread and distribution of the germicide on
the surface to be disinfected. Two different types of
surfactants were tested separately and in combination for
` 35 efficacy. It was found that both sodium dodecyl sulfate and
`, octyl phenoxy polyethoxyethanol are effective germicide-
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1 332554
surfactants. There are other compounds which have the properties
of germicide-surfactants, and this invention is not limited to
those compounds shown in the examples.
The composition contains a dye which is dissolved and
dispersed in the composition. In the examples, the dye selected
is a pH sensitive blue dye which is colored at alkaline pH and
upon neutralization becomes colorless. The blue dye was chosen
for the examples because of a clean association by the public
with the blue color. Any other dye color could be used which
would be acceptable for a particular use. Other indicators which
have similar pH sensitivity can be used as shown, for instance in
the Table of "Indicators for Volumetric Work and pH
Determination" in the Merck Index, 10th Ed. (1983).
The composition has the pH adjusted with the addition
of an alkali such as sodium hydroxide so that the blue color is
deep and noticeable when applied, but losing color upon use. The
pH sensitive dye was chosen for use by the public because the
delivery of the alkaline composition in the environment causes
the color to change in a short period of time. The color change
is probably due to the neutralization of the composition from CO2
in the air and the surface on which it is sprayed. The
alkalinity of the material is adjusted carefully so that the
~ neutralization of the composition can produce a visual change
j from blue to clear within a short period of time. During the
neutralization of the composition and visible change of color,
the alcohol surfactant germicide is producing an effective kill
on organisms present. The disappearance of the color, the
killing of the organisms, and the drying of the germicide occur
'~ in rapid se~uence leaving a dry, germ-free surface for personal
use.
i It is also found that this composition does not leave a
c, film after drying so that the surface is not tacky and
undesirable for personal use. Also, the composition does not
-~ have an objectionable odor as found in phenolic type germicides.
Perfume additives may be used to provide a fragrance if desired
1 332554
--5--
but are not necessary to mask the odor of the germicidal
composition.
The delivery and method of use of the germicide can be
in various forms. There is a factor of the necessity of an
airtight container for a pH sensitive dye. If ambient air
is allowed to penetrate into the container, the liquid may be
neutralized and the color will disappear. Also, the highly
volatile alcohol will escape if the container is not sealed. The
disappearance of the color will not affect the strength of the
germicide detergent which have a long shelf life.
For personal use a small aerosol container which
delivers a fine spray is a practical packaging for the
composition. The pressure sealed container can have a
fluorinated hydrocarbon propellant of the freon type, although
for commercial packaging, a hydrocarbon propellant is preferred
to meet environmental standards. A propellant system to deliver
a fine spray is preferable because it will deliver a thin,
~ ,
rapidly spreading layer which will dry quickly. Atomizers or
other devices which deliver a denser spray may necessitate the
- 20 wiping of the composition with toilet paper or other wipe before
use because of a longer evaporation time for a dense spray.
Another preparation of the invention can be the saturation of a
woven wipe which is sealed in a foil or other airtight packaging.
The packet would be torn open for a one-time usage of the wipe
delivering the dye colored germicide to the surface. The rubbing
;of the wipe on the surface will promote evaporation of the
composition.
EXAMPLE I
A sample of the germicidal composition was prepared by
30 adding 400 mg of sodium dodecyl sulfate (SDS) and 400 mg
of octyl phenoxy polyethoxyethanol marketed as TritonTM X~100 a
product of Sigma Chemical Company to 100 ml of 70% by
volume isopropanol. 100 mg of blue dye thymophthalein was added.
~! The pH was adjusted with 0.05 ml of 12N NaOH
which produces a deep blue colored liquid when kept
,~ airtight. This also gave a pH of 12.53 to the
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liquid. When the composition is allowed to stand exposed
to air it becomes colorless and the pH drops to 8.8.
Tests were done on adjusting the pH with the blue dye
thymophthalein and it was found that the dark blue color
is present at about 11.27 pH. A lighter blue is present
at 11.01 pH and the liquid is clear at 10.95 pH and lower.
An initial dark blue was used in Example 1 to produce a
colored spray. ~owever, the NaOH added was not excessive
to prevent bleaching when the composition is exposed as a
spray or thin coating on a surface in the normal
atmosphere.
This composition was tested for germicide
effectiveness against Herpes simplex virus type 2 tHSV2),
Neisseria gonorrhoeae, Staphylococcus aureus, Escherichia
coli 011K58, Shigella sonnei, Salmonella tyPhimurium, and
Candida albicans. The composition was sprayed and dropped
on pathogen suspensions to test efficacy and dye color
disappearance.
In both the spray and drop tests 0.1 milliliters
(ml) of test pathogenic organisms containing approximately
lx10 organisms were placed on the surface of a sterile
plastic Petri dish. In the drop test 0.1 ml of the
composition of Example I was added by pipette to the
pathogen suspension in the Petri dish. The pathogen
suspension and drop of Example I were mixed and allowed to
i stand for twenty seconds. The blue color would disappear
before the twenty seconds elapsed. At the end of twenty
seconds the Petri plate was tilted and 0.1 ml of the test
` sample was removed. In the spray test the composition of
Example I was placed in an aerosol spray with a
~l fluorocarbon propellant. The 0.1 ml of the pathogenic
¦ organism suspension in the Petri plate was sprayed for two
seconds with Example I. It was determined that between
;~s 0.6 and 0.9 ml of liquid was delivered in the two second
spray. The spray spreads quite noticeably over the Petri
, plate and the blue color disappears in a s~ort time. The
î spray was allowed to mix on the Petri plate for twenty
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seconds. After twenty seconds the Petri plate was tilted
and 0.1 ml of the sample was removed for testing.
The test samples removed from the drop and spray
tests were diluted and plated on agar medium (casman media
for S. typhimurium, S. sonnei and E. coli; chocolate agar
supplemented with factor XV for N. gonorrheae, and sheep
blood agar for S. aureus and C. albicans~. HSV2 was added
to the first wells of a 96-well sterile microtiter tissue
culture plate, serially diluted and cultured with VERO
monkey kidney cells for five days. Phosphate buffered
saline was used as a diluent for all organisms except N.
~ gonorrheae and HSV2; phosphate buffered saline (PBS)
`~, containing 0.5% gelatin was used as a diluent for the N.
gonnorheae and minimal essential medium supplemented with
5% fetal calf serum and antibiotics was used for the HSV2.
Each test included a control (pathogenic organism +
diluent) and a quantitative titration of the pathogen to
determine the actual number of organisms in each test
suspension. All Petri and tissue culture plates were
incubated in 37C (5% CO2) incubators.
All Petri plates were observed the morning after
plating for colony forming units (CFU) and the number of
organisms present in the test suspensions were calculated.
The tissue culture plates containing VERO cells were
observed daily for virus specific cytopathic effects
(CPE). At the end of five days the last well in each
series of dilutions showing CPE was recorded and the titer
`t
of virus in the original test suspensions calculated.
Each assay involving HSV2 had a tissue culture control
~ 30 (VERO cells + media only1 and a virus control (VERO cells
,1 + HSV2 and no germicide).
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The results of the cidal activity of Example I
are shown in Table 1.
:
Table 1. The microcidal activity of Example I against the
different pathogens.
:~
No. Mean
of Percent
Pathogen tests killing Range Comments
S. aureus 6 >99* 99-99.9 Similar killing
by drop or spray.
10 C. albicans 6 >99 99-99.9 Similar killing
; by drop or spray.
N. ~onorrheae 5 >99 99-99.9 Similar killing
by drop or spray.
E. coli OllK58 3 >99 99-99.9 Similar killing
by drop or spray.
S. sonnei 3 >99 99-99.9 Similar killing
by drop or spray.
S. tYphimurium 3 >99 99-99.9 Similar kill.ing
~ by drop or spray.
`~ 20 HSV2 5 99 99-99 Similar killing
-~ by drop or spray;
may be >than 99%
killing but
toxicity of
Example I to VERO
-- cells made lower
dilutions
; unreadable.
.j
*Percent killing was determined by dividing the total number
of viable organisms in suspensions exposed to Example~I by
the total number of viable organisms in suspensions exposed
to phosphate buffered saline and multiplying by 100.
.
`-` 1 332554
g
Example I was compared to Lyso ~ for strength of
killing the organisms HSV2, S. aureaus, N. qonorrheae. E.
coli OllK58, S. typhimurium, S. sonnei and C. albicans,
the complete range of pathogens tested previously. Tne
test was conducted for spray delivery. Example I was
found to compare with the percentage kill equivalent to
Lysol in a side by side test.
The composition was diluted to determine
efficacy. Distilled water was chosen as the dilutant
because PBS caused a cloudy liquid and additional alcohol
would affect germicidal properties. There was a
noticeable decrease in surface tension reduction
decreasing the spreading of the composition with a tenfold
dilution with water. After a 100-fold dilution with
water, no reduction in surface tension was seen. The
killing effect of the dilutions of this example are shown
in Table 2.
I Table 2. The microcidal effect of different dilutions of
¦ Example I against different pathogens.
1 20 No. _ Percent of killing at dilution
t of
Pathogen tests ` 0 10-fold100-fold 1000-fold
S. aureus 3 >99/>99/>99* 37~90/99 8/54/90 20/75/ND**
C. albicans 3 >99/~99/>99 40/13/95 0/53/25 0/0/ND
N.
gonorrheae 3 >99/>99/>99 99/72/90 0/74/92 15/0/79
~SV2 2 9g/99 99/99 0/7 ~0/7
*Results of replicate experiments are separated by slant
(/) bars.
**ND = not done.
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Example II
Another embodiment of the composition can be
made using SDS as the only surfactant-germicide. The
composition was prepared as described in Example I
omitting the addition of Triton X-100 and using 400 mg of
SDS as the only surfactant-germicide. Testing the cidal
activity of Example II on the organisms listed in Table 3
below was carried out as previously outlined in Example I.
Table 3. The microcidal effect of Example II.
,; -- . . _ .
No.
~-~ of % Killings
Organism Tests Spray Drop
. .,
~ HSV2 2 99 99
`~ N.~ orrheae 2 >99 >99
S. aureas 2 >99 >99
C. albicans 2 >99 >99
,
,, . - :.
Example III
,~ An alternate embodiment of the composition can
be made using Triton X-100 as the only surfactant-
germicide. The composition was prepared as described in
Example I omitting SDS and adding 400 mg of Triton X-100
as the only surfactant-germicide. Testing on the
organisms listed in Table 4 below was carried as
previously outlined in Example I.
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Table 4. The microcidal effect of Example III.
No.
Example of % Killings
5 Organism Tests Spray Drop
HSV2 2 99 99
N. gonorrheae 3 ~99 ~99
S. aureas 3 >99 ~99
C. albicans 3 799 ~99
.
EXAMPLE IV
A formula for use with a hydrocarbon propellant is
shown in the following Table 5.
Table 5. Biocide and Hydrocarbon Propellant
- Ingredient Weight %
Isopropanol 51.62
Dionized Water 27.52
r Triton X-100 0.38
20 TEA-Lauryl Sulfate0.38
` Thymolphthalein 0.10
- Aeropin-70 Propellant20.00
~ .
In this example, Triethanol Amine Lauryl Sulfate was
used. The composition is adjusted to pH 11.5 with 10~ NaOH. The
` Aeropin-70 is a hydrocarbon propellant of the following
composition ln Table 6.
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1 33255~
-12-
Table 6. Hydrocarbon Propellant
Liquid Volume ~ Molecular % Weight %
Propane 51.05 54.79 47.92
Isobutane 19.81 17.84 20.61
N-butane 29.14 27.32 31.47
The propellant system used can be any environmentally
acceptable spray that disperses the biocide in a fine layer.
As seen from the results of testing on a wide range of
pathogens, a composition containing one or more surfactant-
germicide provides very effective killing strength used as a drop
or spray. Various other surfactants could be substituted as the
i- surfactant-germicide and it is not the intent of the invention to
limit the compounds used.
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