Note: Descriptions are shown in the official language in which they were submitted.
CA 02297350 2000-02-02
Background of the Invention
Effective cleaning and disinfecting of working areas in medical, public and
household
environments is of major importance in the prevention of disease transmission.
With recent
increases in levels of concern about transmission risks of a number of
microbial agents, much
attention has been paid in recent years to the testing of widely accepted
cleaning agents for
actual effectiveness in respect of specific types of pathogenic microorganisms
where working
conditions do not facilitate prolonged txeatxnent under high temperatures.
It has been shown that several known bleaching agents for surface use in
hospital and laboratory
environments are effective against the HIV virus as well as Hepatitis B virus.
Thus for example
quarteraary ammonium salts, sodium hypochlorite and a variety of alcohols and
phenols were
found to have such anti viral activity in a research by Resnick et al., JAMA
April 11, 1986 -
Vol. 255, no. 14.
CA 02297350 2000-02-02
It is also known that microorganisms vary widely in their resistance to
chemical germicides.
Thus bacterial spores, mycobacteria and to a relatively lesser degree tubercle
bacilli, small or
non lipid viruses and vegetative fungi as well as asexual fungal spores,
vegetative bacteria and
medium sized or lipid containing viruses may withstand treatment with
disinfectants under
routine cleaning procedures.
Cleaning and bleaching procedures are of crucial importance in the
disinfection of medical
instruments as prior physical cleaning is a sine qua non for the effectiveness
of any sterilization
procedure.
It is of similar importance to ensure that disease does not spread via human
contact with public
or household surfaces and objects contaminated by pathogenic viruses,
bacteria, mycobacteria
or fungi. Untreated fruit and vegetables are yet another source of infection.
There is thus a widely recognized need for, and it would be highly
advantageous to have, a
bleaching-disinfecting preparation for hospital and laboratory surfaces and
for cleaning medical
instruments with a wide range germicidal action that combines fungicidal
activity with anti
viral, mycobactericidal and bactericidal effectiveness and that is applicable
in a suitable form
such as diluted liquid, ointment, dry powder or aerosol for general cleaning
and disinfecting in
the household or in public places.
Summary of the Invention
According to the present invention there is provided a wide range germicidal
action disinfecting
preparation consisting of a solution of of 7-13% sodium bromide w/v, 3-6%
peracetic acid w/v,
2-4% phthalic acid dipotassium salt 98°l° w/v, 3-5% acetanilide
w/v, 12-L6% isopropanol v/v,
64% water v/v, and optionally 1 % soap v/v for the disinfection of hospital
and laboratory
surfaces and medical environments. The inventive preparation significantly
increases the
CA 02297350 2000-02-02
prevention of disease transmission by cleaning and disinfecting of hospital
and laboratory
surfaces and equipment.
In accordance with another aspect of the invention the preparation may contain
a detergent that
enhances the disinfecting action of the preparation by removing organic
particles the presence of
which is known to sigiuficantly hilder decontamination.
W accordance with a further aspect of the invention the preparation may be
diluted for use in the
cleaning a.nd disinfecting of household equipment, furluslung and other
surfaces as well as
cleaning and disinfection of fruit and vegetables, and preservative for
flowers.
In accordance with yet another advantage of the present invention the
preparation may be
applied as an external disinfectant for human patients.
The inventive preparation may be prepared and used in liquid solutions, as
aerosol, as
humidifier in cleansilg tissues, as ointment with a suitable emulsifier or in
dry powder
formulation and it may be used on its own or in admixture with other
disinfectants or as an
addition to soaps or detergents.
In accordance with a further aspect of the invention a suitable catalytic
agent may be added to
activate the formulation and enhance the bleaching and disiifecting effect.
The inventive preparation acts effectively agaiist mycobacteria and a wide
number of viruses
such as polio, coxsackie, herpes, influenza, Hepatitis B, HIV and others while
at the same tine
destroying a variety of pathogenic bacteria such as E.coli, Pseudomonas
fluorescens, Shigella
dysenteriae, Clostridium botulinum, Staphylococcus aureus, and having an
enhanced fungicidal
4~~~/~f
Vii W.
CA 02297350 2000-02-02
Detailed Description of the Invention
The present invention relates to a preparation for bleaching and disinfecting
hospital and
laboratory surfaces and medical equipment, that is also applicable for
household sanitation and
skin disinfection at suitable low concentrations, consisting of a solution of
7-13% sodium
bromide w/v, 3-6% peracetic acid w/v, 2-4% phthalic acid dipotassium salt 9$%
w/v, 3-5%
acetanilide w/v, 12-16% isopropanol v/v, 64% water v/v, to which a detergent
is optionally
added.
The preparation according to the invention effectively disinfects and prevents
disease
transmission for a wide range of microorganisms including a wide number of
viruses such as
polio, coxsackie, herpes, influenza, Hepatitis B, HIV and others while at the
same rime
destroying mycobacteria and a variety of pathogenic bacteria such as E.coli,
Pseudomonas
fluorescens, Salmonella, Shigella dysenteriae, Clostridium botulinum,
Staphylococcus aureus,
and having an enhanced fungicidal effect.
In accordance with another embodiment of the invention, the preparation also
contains
detergents that enable the elimination of organic matter from work surfaces
and equipment.
The inventive preparation has the additional advantage that elimination of
organic matter highly
facilitates disinfection whereby the combination has a synergetic effect.
The preparation is suitable for household and skin disinfection use at a
dilution of 1:10.
In accordance with the invention the disinfectant preparation may be prepared
as a liquid
aqueous or inert solution or it may be formulated as an ointment with a
suitable emulsifier or it
CA 02297350 2000-02-02
may be prepared for use in an aerosol container or as a liquid in saturated
cleansing tissues or
contained within a solid soap preparation.
It is envisaged that the inventive preparation may be applied as a pesticide
at high
concentrations and it may be added to water as a preservative for cut flowers
at low, household
concentrations.
The inventive preparation may comprise a catalytic component to enhance the
bleaching and
germicidal action.
It will be appreciated by those versed in the art that the invention has been
described
hereinabove in respect of a preferred embodiment and many variations,
modifications and
improvements may be made that still remain within the ambit of the invention
as described and
claimed.
Laboratory testing
Efficacy testing of the preparation in accordance with the invention was
carried out by the
appropriate AOAC official methods as shown below:
I. MYCOBACTE~CTDAL, TESTS
Materials and Methode
The tests were earned out with a formulation consisting of 7% sodium bromide
w/v, 3%
peracetic acid w/v, 3% phthalic acid, dipotassium salt 98% w/v, 4% acetanilide
w/v, 15%
isopropanol v/v, 64% water v/v, 1 % soap v/v.
Mycobacterium terrae ATCC 15755: M. terrae was in 7H9 broth (Difco) containing
glycerol
but no antibiotics. The bacterial suspension was centrifuged at 2,500 rpm for
15 minutes and the
pellet resuspended to give approximately 2.5 x 10g cells/mL (No. 8 McFarland
Standard).
Sterile Saline: The diluent and rinse used for the mycobactericidal test was
0.85% sterile saline.
CA 02297350 2000-02-02
Organic Load: Fetal bovine serum was used as the organic load. It was added to
the test
microbial suspension at a final concentration of 5%.
The test involved drying a microbial suspension on a hard surface carrier and
covering the dried
inoculum with the use dilution of the disinfectant for the specified contact
time and temperature.
At the end of the contact time, a diluent/rinse was used for recovering the
inoculum from the
carrier and the eluent was passed through a membrane filter (0.45 pm pore
diameter) to capture
the test organism. The filters were then placed on plates with an appropriate
recovery medium
and incubated to allow viable organisms to form visible colonies. The numbers
of colony
forming units were recorded and the level of inactivation of the test organism
was calculated by
comparison to controls.
The testing used glass vials as the hard surface carriers. The organism used
for the
mycobactericidal tests was Mycobacterium terrae suspended in fetal bovine
serum at a final
nnnnan+r~tinn n~ C°/ Txxrn nHcaminol fnrmWofinsxa xx~a~~
to°.°~tvd ai a vv~ntact iu v° of ~~v m~~'ute.°~
vvuvvaaaa uwvai v~ r i v. i rr v vxavuuvux ivmaauaua.avu.~ rr v
and 30 minutes. After rinsing and filtering, the filter was placed on 7H11
agar and incubated at
37C for a total of 4 weeks. The plates were monitored and counted (CFU) at
weekly intervals.
Control carriers were used in the same manner as test carriers except sterile
saline was applied
tn ~tn ~rio~ jir~vvultia~ii lx'istead of fine vh°vii'ii~c''~1
formulator. The number of teJt Va111eJ 1'~~U.J ~.
w mu iws
Three control carries were incorporated in each test. The results are reported
as loglo reductions
in viability with reference to the mean titre on the control carriers. For the
product to be
considered mycobactericidal, it was expected to reduce the viable titre of all
the test organisms
by a minimum of 6 loglo (at least 1 million fold) under the conditions of this
test.
Carrier Inoculation: Carrier inoculation involved placing 10 ~.L (between 10~
to 10g colony
forming units) of the test microbial suspension on the inside bottom surface
of a glass vial and
allowing the inoculum to dry.
Ra r~rl tr
The results for this phase of the tests are shown in Table 1.
CA 02297350 2000-02-02
Table 1. The germicidal activity of the Formulation against li~ycobacterium
terrae:
i Experiment Contact time Average I Average I Loge Reduction i
I I CFU~'Control CFU~'Test I
i i i I Carrier
i i I Carrier I
1 1 i 20 ( 4.0x106 i 0 i >6.6 I
1 2 ( 30 i 4.4x10 i 0 i >6.6 I
, , i ,
i 3 i 20 1 4.3x106 1 0 i >6.6 i
C''n~r~~l"e;n~.
~/V iV11iJ1V 1.
The formulation in its diluted form was proved as mycobactericidal
T_T_, FTTNC''TT_C''TT~AT. TFCT$
The tests were carried out with a formulation consisting of 7% sodium bromide
w/v, 3%
peracetic acid w/v, 3% phthalic acid, dipotassium salt 98% w/v, 4% acetanilide
w/v, 15%
isopropanol v/v, 64% water v/v, 1% soap v/v wherein two of the ingredients
have a
~~nnrat,trotinn lnixrnr ~hae~ tha in~rcanti~ra fnr~»~~t~n,~,
VV11VV11416~1.1V11 1V ~~ V1 1(1.. 411V 111 ~ V11 1 ~ V 1V1 11 V 1.
Trichophyton mentagrophytes was used as the test organisem.
Rdatcariale Rr TllFcathnr~e
1~1(1.W1141.1J W 1~1V411V~4U
Sterile Saline: The diluent and rinse used for the fungicidal test was 0.85%
sterile saline.
Organic Load: Fetal bovine serum was used as the organic load. It was added to
the test fungal
suspension at a final concentration of 5%.
The ~,3uantitative Carrier Test:. The test involves drying a microbal
suspension on a hard
surface carrier and covering the dried inoculum with the use-dilution of the
disinfectant for the
specified contact time and temperature, At the end of the contact time, a
diluent / rinse was used
for specified contact time and temperature. At the end of the contact time, a
diluent / rinse was
used for recoveritlg the inoculum from the carrier and the eluent was passed
through a
membrane filter {0.45 ~Im pore diameter) to capture the test organism. The
filters were then
placed on plates with an appropriate recovery medium and itlcubated to allow
viable organisms
to form visible colonies. The numbers of colony forming units were recorded
and the level of
inactivation of the test orgatusm was calculated iti comparison to controls.
The testing used glass vials as the hard surface carriers.
CA 02297350 2000-02-02
The organism used for the fungicidal tests was a cordial suspension of
Trichophyton
mentagrophytes suspended in fetal bovine serum at a final concentration of S%.
One chemical
formulation was tested at a contact tune of 10 minutes. After rinsing a.nd
filtering, the filter was
placed on Mycobiotic agar and incubated at 28 ° c for a total of 10
days. The plates were
monitored and counted (CFU) daily. Control carriers were used in the same
manner as test
carries except sterile saline was applied to the dried inoculum instead of the
chemical
formulation. The number of test carriers was 5. Three control carriers were
incorporated in each
test. The results are reported as loglo reductions in viability in reference
with the control carries.
For the product to be considered fungicidal, it was expected to reduce the
viable titre of all the
test organisms by a minimum of 5 logio under the conditions of this test.
Carrier W ovulation: Carrier inoculation involved placing 10 L (between 105 -
106 colony forming
units) of the test fungal suspension on the inside bottom surface of a glass
vial and allowing the
inoculum to dry.
The results of the fungicidal testing are shown in Table 2.
Table 2. The fungicidal activity of the formulation against Trichophyton
mentagrophytes
Experiment Date of Expt Contact Time Average GFU/Test Loge, I
~ ~ (minutes) ~ CFU/Control~ Average ~ Reduction
Carner I Carrier
I I I Sep 29, 1997 I I0 I 7.3xI05 I 0 I 5.9 I
I 2 I Oct 2, 1997 I IO I 5.9xI05 I (3 I 5.8 I
(-''nn~lncinn
The formulation in its diluted form proved fungicidal.
rrr wr~r turner ~r~crrc
111. ~ uw vlir~ ray 1 a.iV 1 V
Poliovirus type 1 (Sabin~ was used for this test i11 accordance with the
Canadian General
C+.~nr~nrrlo Rn~~d tv~t f~~' ~'~ar~alcldvs.
V l.~,ylui4 <iJ tJ
Ma Pri al c Rr MPt nrl c
wt___~ ~3.._...
TY1P a~7~ent ~d d~l~vni used for the vii~uc~dul test YvU.s ERSS.
111V V (.L1
CA 02297350 2000-02-02
A formulation consisting of 7% sodium bromide w/v, 3% peracetic acid w/v, 3%
phthalic acid,
dipotassium salt 98% wlv, 4% acetanilide wlv, 15% isopropanol v/v, 64% water
v/v, 1 % soap
vlv (formula # 1 ) and a preferred embodiment of the inventive formulation,
consisting of a
solution of 10% sodium bromide w/v, 5% peracetic acid w/v, 3% phthalic acid
dipotassiutn salt
98% w/v, 4% acetaiulide w/v, 15% isopropanol v/v, 64% water v/v, and 1% soap
(formula #2)
were used as virucidal agents in these tests.
Polio Sabin 1 virus already available in the laboratory was grown on Vero
cells. Cells were
maintained by passage in minimum essential medium (MEM) from Gibco BRL Cat
#12360-038
with the appropriate supplements and 10% fetal bovine serum (FBS) in 25cm2
flasks in a C02
incubator at 37°C. Two hundred microliter of the viral suspension was
inoculated onto a
confluent monolayer washed three times with 10 mL of Earls Balanced Salt
Solution (EBSS)
from Gibco BRL Cat #450-1 100EL to remove serum. The virus was allowed to
spread by
gentle rocking of the flask. The flask was kept at 37°C for 50 min. to
allow for virus adsorption.
Supplemented MEM without serum was added and incubated for 24 hours. When 75%
Cytopathic effect (CPE) was observed. The virus was separated from the cells
by three rapid
freeze thawing followed by gentle centrifugation at 1500 rpm for 15 min. The
supernatant which
contained the virus was dispensed in aliquots of 200~L and stored at -
80°C. The virus titre was
determined using a plaque assay method.
Organic Load: Fetal bovine serum was used as the organic load in this study.
It was added to
the test virus suspension at a final concentration of 5%.
The testing used flat stainless steel discs as the hard surface carriers. The
organism used for the
virucide tests was polio type 1 (Sabin) suspended in fetal bovine serum at a
final concentration
of 5%. Carrier inoculation involved placing 10~L (between 10g-lO6 plaque
forming units) of the
testvirus suspension on the center of each stainless steel carrier and
allowing the inoculum to
dry for 1 hour.
CA 02297350 2000-02-02
The dried itioculum was covered with the use dilution of the disinfectant for
the specified
contact tune and temperature. At the end of the contact time, EBSS was used as
neutralizer/eluent for recovering the reaction mixture from the carrier. The
eluent was serially
diluted and plaque assay done to obtain titre of virus remaining.
Each formulation ~~las tested at a contact time of 10 minutes. Control
carriers were used in the
same manner as test carriers except sterile EBSS was applied to the dried
inoculum instead of
the chemical formulation. The number of test carriers was 5. Three control
carriers were
incorporated ul each. The results are reported as loglo reductions in
viability in reference with
the control carriers. For the formulation to be considered virucidal, it was
expected to reduce
the viable titre of all the test organisms by a minimum of 4 loglo under the
conditions of this test.
Results
The results of virucidal tests are showTn in Tables 3 and ~. Table ~ shows
that formula #1 was
not virucidal, it had little or no activity against poliovirus at a contact
time of 10 minutes at
20=1°C.
Table 3. The germicidal activity of the formulation #1 against Polio type 1
(Sabin)
ExperimentDate of Contact Average PFU/ControlPFU/Test AverageLogy
Expt Time Carrier Garner I Reduction
minutes ~
I
1 i Sep 9, 199710 1.X105 1.8X104 j 1
~
I 2 ~ Sep 22, 10 7.2XI04 I.9XI04
j 1997
I I I I I I I
The polivirus was retested with formula #2 which increased the concentration
of two of the
ingredients. Formula #2 is a preferred embodiment of the inventive formula
that consists of a
CA 02297350 2000-02-02
solution of 10% sodium bromide w/v, 5% peracetic acid w/v, 3% phthalic acid
dipotassium salt
98% w/v, 4% acetanilide w/v, 15% isopropanol v/v, 64% water v/v, and 1% soap
v/v. The test
methods used were identical of those describcd above. Formula #2 ~.~.ras able
to reduce the ~~:~us
titre by more than 4 loglo (Table 4).
Table 4. The germicidal activity of the inventive formulation against
Poliovirus type 1
(Sabin)
ExperimentDate of Contact Average PFU/ControlPFU/Test AverageLogo
I I Expt Time Carrier I Carrier I Reduction
I minutes I
I
1 I Nov 24, 10 I 6.8X104 0 4.83
I
I
I 1997 I I I
2 ~ Nov 25, ~ 10 i 1.25X10 I 0 i 5.10 I
1~~
7,4 f'nn~l_ycin_n_
Formula #2 proved virucidal.
