1
Evaluation
of
the
Toxicology
Database
On
Azadioxabicycooctane
FOR
REREGISTRATION
ELIGIBILITY
DECISION
(
RED)

September
28,
2005
2
1.0
HAZARD
CHARACTERIZATION
Azadioxabicyclooctane
consists
of
an
equilibrium
mixture
of
three
chemicals
(
I:
5­
hydroxymethoxymethyl­
1­
aza­
3,7­
dioxabicyclo(
3,3,0)
octane;
II:
5­
hydroxymethyl­
1­
aza­
3,7­
dioxabicyclo(
3,3,0)
octane;
III:
5­
hydroxypoly(
methylene­
oxy)
methyl­
1­
aza­
3,7­
dioxabicyclo(
3,3,0)
octane).
These
cannot
be
divided
into
components
for
individual
testing.
This
mixture
is
registered
for
control
of
slime­
forming
bacteria
and
fungi
at
the
following
use
sites:
oil
recovery
drilling
muds
and
packer
fluids;
industrial
adhesives
and
coatings;
resin/
latex/
polymer
emulsions;
metalworking
cutting
fluids;
latex
paints;
paper/
paper
products;
specialty
industrial
products;
and
wet­
end
additives
for
industrial
processing
chemicals
Azadioxabicyclooctane
has
a
moderate
order
of
acute
toxicity
via
the
oral
and
dermal
routes
of
exposure
(
Toxicity
Category
III)
and
a
moderate
to
severe
acute
toxicity
via
the
inhalation
route
(
Toxicity
Category
II).
Azadioxabicyclooctane
produces
severe
irritation
to
the
eyes
(
Toxicity
Category
I)
and
moderate
irritation
to
the
skin
(
Toxicity
Category
III).

Repeated
dose
toxicity
data
for
azadioxabicyclooctane
are
limited
to
a
90­
day
oral
toxicity
study
in
the
rat,
a
21­
day
dermal
toxicity
study
in
the
rabbit,
and
a
dermal
developmental
toxicity
study
in
the
rat.

In
a
90­
day
oral
toxicity
study
(
MRID
41641606
reformat
of
00109245,
supplemental
information
410801010),
Nuosept
®
95
(
Lot
No.
005­
267)
was
administered
to
20
CD1
Sprague­
Dawley
derived
rats/
sex/
dose
in
drinking
water
at
dose
levels
of
0,
125,
625,
or
3125
ppm
(
equivalent
to
0,
20,
100,
or
500
mg/
kg
bw/
day;
actual
intake
reported
was
0,
13,
62,
and
245
mg/
kg/
day).
Four
rats/
sex/
group
were
kept
for
a
4­
week
recovery
period
to
determine
reversibility
of
effects.
No
signs
of
toxicity
due
to
Nuosept
95
treatment
were
noted
in
mortality,
clinical
observations,
hematology,
clinical
chemistry,
ophthalmology,
gross
pathology,
or
histopathology.
Body
weight
and
body
weight
gain
were
lower
in
high­
dose
males
and
females
compared
to
their
respective
controls.
Although
food
consumption
was
reduced
in
the
intermediate
and
high­
dose
groups
in
both
males
and
females,
the
food
efficiency
was
unaffected.
Water
consumption
was
drastically
(
approximately
30­
50%)
reduced
in
high­
dose
males
and
females
throughout
the
entire
study
period.
Due
to
the
reduction
in
water
consumption,
the
intake
of
test
article
did
not
reach
predicted
levels.
Based
on
the
analytical
concentration
of
the
test
article
in
the
water
and
the
water
intake
data,
the
daily
doses
were
estimated
to
be
10.6,
56.5,
and
228.9
mg/
kg/
day
in
males
and
12.2,
64.3,
and
255.9
mg/
kg/
day
for
females.
The
LOAEL
is
56.5
and
64.3
mg/
kg/
day
in
males
and
females,
respectively,
based
on
the
reduction
in
water
consumption.
The
NOAEL
is
10.6
and
12.2
mg/
kg/
day
for
males
and
females,
respectively.
The
90­
day
study
is
classified
unacceptable/
guideline
but
is
upgradable.

In
a
21­
day
dermal
toxicity
study
(
MRID
41641605),
Nuosept
®
95/
Nuosept
®
C
(
Lot
number
and
purity
not
reported)
was
administered
directly
to
the
skin
of
New
Zealand
White
rabbits
(
5/
sex/
group)
at
doses
of
0,
100,
300,
and
1000
mg/
kg/
day,
6
hours/
day
for
5
days/
week
during
a
21­
day
period.
No
treatment­
related
effects
on
clinical
observations
and
mean
absolute
or
relative
organ
weights
were
observed.
Significant
findings
were
presented
for
food
consumption,
body
weight,
hematology,
and
clinical
chemistry
parameters,
however
these
findings
were
not
3
considered
to
be
treatment­
related.
Extensive
damage
to
the
treated
skin
was
observed
in
all
of
the
rabbits
in
the
high­
dose
group.
Numerous
microscopic
changes
in
the
skin
as
well
as
increases
of
neutrophils
and
platelets
were
observed
and
were
considered
to
be
secondary
to
tissue
repair.
The
dermal
LOAEL
is
100
mg/
kg/
day
(
based
on
severe
dermal
effects).
The
dermal
NOAEL
for
Nuosept
95/
Nuosept
C
could
not
be
determined
because
mild
skin
effects
were
observed
at
the
low
dose.
The
systemic
NOAEL
is
greater
than
or
equal
to
1000
mg/
kg/
day
and
the
LOAEL
is
greater
than
1000
mg/
kg/
day.
This
21­
day
dermal
toxicity
study
in
the
rabbit
is
classified
as
acceptable
(
non­
guideline).

In
a
dermal
developmental
toxicity
study
(
MRID
41537501,
reformat
of
41699001),
Nuosept
®
95
(
Batch
No.
006­
097­
597,
50.38%;)
in
distilled
water
was
applied
dermally
to
mated
female
Crl:
COBS
CD
(
SD)
BR
VAF+
rats
(
25/
dose)
at
dose
levels
of
0,
100,
300,
or
1000
mg/
kg/
day
from
days
6
through
15
of
gestation
(
gd
6­
15).
An
additional
25
mated
female
rats
were
administered
distilled
water
from
gd
6­
15.
There
were
no
treatment
related
effects
in
mortality,
clinical
signs,
weight
gain,
food
consumption,
gross
necropsy
parameters
and
cesarean
parameters.
There
was
clear
evidence
of
maternal
dermal
effects
in
all
treatment
groups.
The
low­
dose
group
had
well­
defined
erythema,
scabbing
that
obscured
some
of
the
treatment
site,
and
well­
defined
edema
in
most
animals
by
study
termination.
The
intermediate­
dose
group
displayed
moderate/
well­
defined
erythema.
Half
of
the
animals
had
scabbing
that
completely
obscured
the
treatment
site,
and
edema
that
was
well­
defined
by
study
termination.
The
highdose
group
exhibited
well­
defined
erythema,
scabbing
that
obscured
the
treatment
site
in
all
animals
by
gd
10,
and
moderate
to
severe
edema
that
extended
to
the
underlying
musculature.
The
LOAEL
for
maternal
dermal
toxicity
is
100
mg/
kg/
day
(
based
on
severe
dermal
irritation);
a
NOAEL
could
not
be
established.
The
systemic
maternal
toxicity
NOAEL
is
greater
than
or
equal
to
1000
mg/
kg/
day;
the
systemic
maternal
toxicity
LOAEL
is
greater
than
1000
mg/
kg/
day.
There
were
no
other
embryotoxic
or
fetotoxic
effects
observed
in
this
study.
The
developmental
NOAEL
is
greater
than
or
equal
to
1000
mg/
kg/
day;
the
developmental
LOAEL
is
greater
than
1000
mg/
kg/
day.
This
developmental
toxicity
study
in
the
rabbit
is
classified
as
unacceptable/
guideline
and
is
upgradable.

There
are
no
data
on
the
reproductive
toxicity
of
azadioxabicyclooctane.
There
are
also
no
carcinogenicity
nor
chronic
toxicity
data
for
azadioxabicyclooctane.

Azadioxabicyclooctane
has
been
tested
for
mutagenic
activity
in
several
assays,
including
a
bacterial
DNA
damage/
repair
assay
(
MRID
93050019;
93050020),
dominant
lethal
assay
(
MRID
00088953),
unscheduled
DNA
synthesis
assay
(
MRID
93050021;
42711001;
41642401),
micronucleus
assay
(
41728601),
in
vitro
cell
transformation
assay
(
MRID
93050022),
and
the
Ames
assay
(
MRID
93050017;
93050018).
In
the
Ames
Salmonella
test
,
azadioxabicyclooctane
was
negative
in
all
strains
tested,
but
deficiencies
in
the
studies
were
identified
and
these
deficiencies
must
be
addressed
to
upgrade
the
studies.
Azadioxabicyclooctane
was
also
negative
in
the
micronucleus
assay,
but
was
positive
in
one
unscheduled
DNA
synthesis
assay
(
93050021)
and
the
bacterial
DNA
damage/
repair
assay.
Several
tests
(
MRID
numbers
93050022
through
93050025)
were
conducted
with
C3H/
10T1/
2
cell
cultures
examining
cell
transformation
in
vitro.
Although
results
were
largely
negative,
these
studies
collectively
did
not
properly
identify
the
test
material
being
examined.
In
fact,
only
one
study
(
MRID
93050018)
properly
characterized
the
4
test
material.
Thus,
although
the
data
suggest
largely
negative
responses,
the
lack
of
test
article
characterization
(
especially
in
light
of
positive
responses
observed
in
two
studies)
points
to
the
need
for
proper
test
article
characterization
before
an
adequate
conclusion
can
be
made
about
the
mutagenicity
of
azadioxabicyclooctane.

From
the
available
repeat
dose
toxicity
studies,
there
was
no
evidence
of
neurotoxicity
of
azadioxabicyclooctane.

2.0
REQUIREMENTS
Insert
Table
3.0
DATA
GAPS
The
ADTC
determined
that
there
are
several
data
gaps
in
the
hazard
database
for
azadioxabicyclooctane
based
on
the
current
use
patterns.
Due
to
the
potential
for
inhalation
exposure
in
the
occupational
setting,
a
repeated
dose
(
90­
day)
inhalation
toxicity
study
should
be
performed
with
azadioxabicyclooctane.
The
oral
study
used
for
selection
of
the
inhalation
endpoint
does
not
adequately
characterize
potential
inhalation
hazard
and
was
used
with
added
uncertainty
factors
(
10x)
in
the
absence
of
inhalation
data.
A
route­
specific
study
would
reduce
or
eliminate
this
uncertainty.
An
acceptable
dermal
sensitization
study
must
also
be
submitted.

To
address
data
requirements
for
indirect
food
uses
of
antimicrobials,
the
Agency
has
established
an
interim
two­
tiered
system
for
toxicology
testing
requirements
in
order
to
complete
a
risk
assessment
for
indirect
food
uses
of
antimicrobial
pesticide
chemicals
for
which
the
FDA
has
established
a
food
additive
regulation
that
specifically
states
that
the
use
is
"
safe",
but
for
which
the
Agency
requires
its
own
assessment.
Tier
I
toxicology
data
requirements
would
apply
to
all
indirect
food
additives
that
result
in
residue
concentrations
in
food
ranging
from
0­
200ppb.
The
requirements
would
consist
of
an
acute
toxicity
testing
battery,
a
subchronic
toxicity
study
in
the
rodent,
a
developmental
toxicity
study
in
the
rodent
(
rat),
and
a
mutagenicity
testing
battery.
Each
of
these
data
requirements
has
been
fulfilled
for
AZA.
The
Agency
also
conducts
a
literature
search
and
can
also
conduct
a
Structure
Activity
Relationship
analysis
(
SAR)
if
appropriate.

Tier
II
studies
would
be
triggered
by
the
presence
of
significant
(
i.
e.
=
200ppb)
residues
in
food
or
evidence
of
significant
toxicity
from
the
Tier
I
data
set,
which
may
include
developmental
/
reproductive,
or
other
systemic
toxicity
such
as
presence
of
neoplastic
growth
or
significant
target
organ
toxicity.
In
such
cases,
chronic
toxicity
and
carcinogenicity
testing
would
be
required
in
addition
to
the
above
Tier
I
studies.

For
AZA,
the
Agency
will
hold
in
reserve
a
two­
generation
reproduction
toxicity
study
in
the
rat
5
and
a
subchronic
toxicity
study
in
the
non­
rodent
which
would
become
data
requirements
pending
the
evaluation
of
the
Tier
1
data
to
adequately
address
risk
from
indirect
food
uses
of
azadioxabicyclooctane
Separately,
risks
from
the
metalworking
fluid
use
of
azadioxabicyclooctane
cannot
be
adequately
characterized
without
conduct
of
chronic
toxicity
studies
in
the
rat
and
mouse
as
well
as
carcinogenicity
studies
in
these
two
species.
Metalworking
fluid
use
is
considered
a
high
exposure
antimicrobial
use
pattern
and
requires
chronic
toxicity
and
carcinogenicity
data.

4.0
HAZARD
ASSESSMENT
4.1
Acute
Toxicity
Adequacy
of
database
for
Acute
Toxicity:
The
acute
toxicity
database
for
azadioxabicyclooctane
is
considered
incomplete.
An
acceptable
dermal
sensitization
study
must
be
submitted.

The
acute
oral
toxicity
of
azadioxabicyclooctane
is
low,
with
a
calculated
median
lethal
dose
of
1940
mg/
kg
from
a
study
in
the
rat
(
MRID
41641601).
Pale
kidneys,
enlarged
spleen,
and
livers
with
focal
hemosiderosis
were
observed
in
this
study.
The
acute
dermal
toxicity
of
azadioxabicyclooctane
is
also
low,
with
a
dermal
median
lethal
dose
of
greater
than
2000
mg/
kg
(
MRID
41671801).
Dermal
effects
were
noted,
but
no
systemic
toxicity.
The
acute
inhalation
toxicity
study
(
MRID
42650901)
showed
a
median
lethal
dose
range
of
between
0.441
mg/
L
and
0.819
mg/
L,
with
epistaxis,
labored
breathing,
rales,
and
rhinorrhea
in
all
dose
groups.
Corneal
opacity
was
observed
in
the
primary
eye
irritation
study
(
MRID
41641602)
resulting
in
a
Toxicity
Category
I
classification.
Moderate
dermal
irritation
effects
were
noted
in
the
primary
dermal
irritation
study
(
MRID
41641603),
leading
to
a
Toxicity
category
III
classification.

The
acute
toxicity
data
for
azadioxabicyclooctane
is
summarized
in
Table
1.

Table
1.
Acute
Toxicity
Profile
for
Azadioxabicyclooctane
Guideline
Number
Study
Type/
Test
substance
(%
a.
i.)
MRID
Number/
Citation
Results
Toxicity
Category
870.1100
(
§
81­
1)
Acute
Oral­
Rat
Nuosept
®
95
(
50%
a.
i.)
MRID
41641601
LD50
=
1940
mg/
kg/
day
III
870.1200
(
§
81­
2)
Acute
Dermal­
Rabbit
Nuosept
®
95
(
50%
a.
i.)
MRID
41671801
LD50
>
2000
mg
/
kg
III
870.1300
(
§
81­
3)
Acute
Inhalation­
Rat
Nuosept
®
95
(
50%
a.
i.)
MRID
42650901
LC50
 
0.441
mg/
L
II
6
Table
1.
Acute
Toxicity
Profile
for
Azadioxabicyclooctane
Guideline
Number
Study
Type/
Test
substance
(%
a.
i.)
MRID
Number/
Citation
Results
Toxicity
Category
870.2400
(
§
81­
4)
Primary
Eye
Irritation­
Rabbit
Nuosept
®
95
(
50%
a.
i.)
MRID
41641602
Corrosive
I
870.2500
(
§
81­
5)
Primary
Dermal
Irritation­
Rabbit
Nuosept
®
95
(
50%
a.
i.)
MRID
41641603
Moderate
irritant
III
870.2600
(
§
81­
6)
Dermal
Sensitization
NA
Assumed
Sensitizer
no
data
available
4.2
Subchronic
Toxicity
Adequacy
of
database
for
Subchronic
Toxicity:
Subchronic
toxicity
data
for
azadioxabicyclooctane
are
limited
to
a
90­
day
oral
toxicity
study
in
the
rat
and
a
21­
day
dermal
toxicity
study
in
the
rat.

870.3100
90­
day
Oral
Toxicity
Study
(
rat)

In
a
90­
day
oral
toxicity
study
(
MRID
41641606
reformat
of
00109245,
supplemental
information
410801010),
Nuosept
®
95
(
Lot
No.
005­
267)
was
administered
to
20
CD1
Sprague­
Dawley
derived
rats/
sex/
dose
in
drinking
water
at
dose
levels
of
0,
125,
625,
or
3125
ppm
(
equivalent
to
0,
20,
100,
or
500
mg/
kg
bw/
day;
actual
intake
reported
was
0,
13,
62,
and
245
mg/
kg/
day).
Four
rats/
sex/
group
were
kept
for
a
4­
week
recovery
period
to
determine
reversibility
of
effects.

No
signs
of
toxicity
due
to
Nuosept
95
treatment
were
noted
in
mortality,
clinical
observations,
hematology,
clinical
chemistry,
ophthalmology,
gross
pathology,
or
histopathology.
Body
weight
and
body
weight
gain
were
lower
in
high­
dose
males
and
females
compared
to
their
respective
controls.
Although
food
consumption
was
reduced
in
the
intermediate
and
high­
dose
groups
in
both
males
and
females,
the
food
efficiency
was
unaffected.
Water
consumption
was
drastically
(
approximately
30­
50%)
reduced
in
high­
dose
males
and
females
throughout
the
entire
study
period.
The
water
consumption
in
these
high­
dose
rats
recovered
after
the
test
article
was
removed
from
the
drinking
water.
Water
consumption
in
the
intermediate­
dose
group
was
slightly
(
approximately
10­
20%)
lower
than
controls
with
statistical
significance
reached
sporadically.
These
rats
resumed
their
consumption
of
water
during
the
recovery
period.
Due
to
the
reduction
in
water
consumption,
the
intake
of
test
article
did
not
reach
predicted
levels.
Based
on
the
analytical
concentration
of
the
test
article
in
the
water
and
the
water
intake
data,
the
daily
doses
were
estimated
to
be
0,
10.6,
56.5,
and
228.9
mg/
kg/
day
in
males
and
0,
12.2,
64.3,
7
and
255.9
mg/
kg/
day
for
females.

The
LOAEL
is
56.5
and
64.3
mg/
kg/
day
in
males
and
females,
respectively,
based
on
the
reduction
in
water
consumption.
The
NOAEL
is
10.6
and
12.2
mg/
kg/
day
for
males
and
females,
respectively.

The
90­
day
study
is
classified
unacceptable/
guideline
but
is
upgradable.
If
the
histopathology
report
is
reformatted
and
re­
submitted
for
review,
conclusions
regarding
organ
weight
changes
and/
or
enzyme
changes
can
be
made
when
an
understandable
histopathology
report
is
submitted.
There
were
several
deficiencies
in
the
submitted
histopathology
report:
(
1)
several
animal
numbers
were
referenced
for
which
there
were
no
histopathology
reports
(
animal
#'
s
430
and
higher);
(
2)
incidence
of
lesions
in
control
animals
that
appeared
higher
than
in
treated
animals,
such
as
individual
cell
necrosis
of
the
liver,
hemosiderosis
of
the
spleen,
and
chronic
respiratory
disease.
If
an
acceptable
report
is
re­
submitted,
the
study
may
then
be
considered
for
upgrade.
This
can
be
done,
as
the
week
4
results
for
hematology
can
be
discarded
based
on
faulty
methodology
(
guideline
requires
only
a
13
week
measurement),
and
a
recovery
group
is
not
part
of
the
guideline
requirement,
therefore
all
the
deficiencies
from
that
examination
can
be
disregarded
and
the
conclusions
set
aside
for
that
group.
Information
on
the
characterization
of
the
test
article
also
needs
to
be
provided.
Azadioxabicyclooctane
is
an
equilibrium
mixture
of
three
isomers,
and
information
needs
to
be
provided
on
the
percentages
of
these
three
isomers
in
the
mixture
used
in
this
study.

21­
day
Dermal
Toxicity
Study
In
a
21­
day
dermal
toxicity
study
(
MRID
41641605),
Nuosept
®
95/
Nuosept
®
C
(
Lot
number
and
purity
not
reported)
was
administered
directly
to
the
skin
of
New
Zealand
White
rabbits
(
5/
sex/
group)
at
doses
of
0,
100,
300,
and
1000
mg/
kg/
day,
6
hours/
day
for
5
days/
week
during
a
21­
day
period.

No
treatment­
related
effects
on
clinical
observations
and
mean
absolute
or
relative
organ
weights
were
observed.
Significant
findings
were
presented
for
food
consumption,
body
weight,
hematology,
and
clinical
chemistry
parameters.
These
findings
were
not
considered
to
be
treatment­
related,
however,
because
of
the
following
reasons:
(
1)
body
weights
and
food
consumption
were
reduced
by
week
3
due
to
the
extensive
skin
damage
and
decreased
mobility
in
the
high­
dose
group;
(
2)
statistical
changes
occurred
sporadically;
and
(
3)
a
dose­
dependent
relationship
was
not
observed.
Extensive
damage
to
the
treated
skin
was
observed
in
all
of
the
rabbits
in
the
high­
dose
group.
Numerous
microscopic
changes
in
the
skin
as
well
as
increases
of
neutrophils
and
platelets
were
observed
and
were
considered
to
be
secondary
to
tissue
repair.

The
dermal
LOAEL
is
100
mg/
kg/
day
(
based
on
severe
dermal
effects).
The
dermal
NOAEL
for
Nuosept
95/
Nuosept
C
could
not
be
determined
because
skin
effects
were
observed
at
the
low
dose.
The
systemic
NOAEL
is
greater
than
or
equal
to
 
1000
mg/
kg/
day.
The
systemic
LOAEL
is
greater
than
1000
mg/
kg/
day.

This
21­
day
dermal
toxicity
study
in
the
rabbit
is
classified
acceptable
(
non­
guideline).
8
Although
the
study
does
not
satisfy
the
guideline
requirement
for
a
21/
28­
day
dermal
toxicity
study
(
OPPTS
870.3200
[_
82­
2];
OECD
410),
no
systemic
effects
were
observed
rats
exposed
to
Nuosept95/
Nuosept
C
at
concentrations
up
to
and
including
a
limit
dose.
Information
needs
to
be
provided
on
the
composition
of
the
test
article.
Azadioxabicyclooctane
is
an
equilibrium
mixture
of
three
isomers
and
information
should
be
provided
on
the
percentages
of
these
three
isomers
in
the
mixture.
To
date,
the
registrant
has
not
supplied
this
information.

4.3
Prenatal
Developmental
Toxicity
Adequacy
of
database
for
Prenatal
Developmental
Toxicity:
There
is
only
one
developmental
toxicity
study
(
MRID
41537501,
reformat
of
41699001)
in
rats
available
for
this
chemical
conducted
by
the
dermal
route.

Developmental
Toxicity
 
Rat
In
a
developmental
toxicity
study
(
MRID
41537501,
reformat
of
41699001),
Nuosept
95
(
Batch
No.
006­
097­
597,
50.38%
a.
i.)
in
distilled
water
was
applied
as
a
neat
material
to
mated
female
Crl:
COBS
CD
(
SD)
BR
VAF+
rats
(
25/
dose)
at
dose
levels
of
0,
100,
300,
or
1000
mg/
kg/
day
from
days
6
through
15
of
gestation
(
gd
6­
15).
An
additional
25
mated
female
rats
were
administered
distilled
water
from
gd
6­
15.

There
were
no
treatment­
related
effects
in
mortality,
food
consumption,
or
cesarean
parameters.
There
was
clear
evidence
of
maternal
dermal
effects
in
all
treatment
groups.
The
low­
dose
group
had
well­
defined
erythema
and
scabbing
that
obscured
some
of
the
treatment
site,
and
welldefined
edema
in
most
animals
by
study
termination.
The
intermediate­
dose
group
displayed
moderate/
well­
defined
erythema.
Half
of
the
animals
had
scabbing
that
completely
obscured
the
treatment
site,
and
edema
that
was
well­
defined
by
study
termination.
The
high­
dose
group
exhibited
well­
defined
erythema,
scabbing
that
obscured
the
treatment
site
in
all
animals
by
gd
10,
and
moderate
to
severe
edema
that
extended
to
the
underlying
musculature.

Systemic
effects
were
shown
by
a
slight
dose­
related
reduction
in
mean
bodyweight
gain
statistically
lower
than
controls
in
the
1000
mg/
kg/
day
group
(
p<
0.5)
during
the
gd
7­
14
period.
Enlargement
of
lymph
nodes
were
observed
at
autopsy
in
the
300
(
1
animal)
and
1000
(
2
animals)
mg/
kg/
day
dose
groups.
There
were
no
adverse
effects
on
litter
parameters
or
embryofoetal
development.

The
LOAEL
for
maternal
dermal
toxicity
is
100
mg/
kg/
day
(
based
on
dermal
irritation);
a
NOAEL
could
not
be
established.
The
systemic
maternal
toxicity
NOAEL
is
300
mg/
kg/
day;
the
maternal
toxicity
LOAEL
is
1000
mg/
kg/
day
(
based
on
decreased
body
weight
gain).
The
developmental
NOAEL
is
 
greater
than
or
equal
to
1000
mg/
kg/
day;
the
developmental
LOAEL
is
greater
than
1000
mg/
kg/
day.

This
developmental
toxicity
study
in
the
rabbit
is
classified
as
unacceptable/
guideline
and
is
upgradable.
The
study
report
contains
several
minor
deficiencies,
including
lack
of
information
9
on
the
homogeneity,
concentration,
or
stability
of
the
test
material
in
the
vehicle
and
historical
control
data.
However,
there
is
useful
information
concerning
the
dermal
toxicity
of
the
chemical
in
this
study,
and
testing
up
to
a
dermal
limit
dose
showed
no
systemic
effects.
Although
the
dosing
duration
was
not
considered
long
enough
for
a
dermal
developmental
toxicity
study
(
dosing
should
begin
on
day
0
for
dermal
studies),
the
lack
of
a
systemic
effect
at
the
limit
dose
is
supported
by
the
submitted
21/
28
day
dermal
toxicity
study,
which
also
found
no
systemic
effects
up
to
and
including
a
limit
dose.

4.4
Reproductive
Toxicity
Adequacy
of
database
for
Reproductive
Toxicity:
There
are
no
data
on
the
reproductive
toxicity
of
azadioxabicyclooctane.
For
the
indirect
food
use
of
this
chemical,
this
study
is
being
held
in
reserve.

4.5
Chronic
Toxicity
Adequacy
of
database
for
Chronic
Toxicity:
There
are
no
data
on
chronic
toxicity
of
azadioxabicyclooctane.
Metalworking
cutting
fluid
use
triggers
the
requirement
for
chronic
toxicity
studies
as
described
on
the
1987
Antimicrobial
Data
Call
In
Notice.

4.6
Carcinogenicity
Adequacy
of
database
for
Carcinogenicity:
A
determination
of
carcinogenic
potential
cannot
be
made
based
on
the
available
data.
Metalworking
cutting
fluid
use
triggers
the
requirement
for
carcinogenicity
studies
as
described
on
the
1987
Antimicrobial
Data
Call­
In
Notice.

4.7
Mutagenicity
Azadioxabicyclooctane
has
been
tested
for
mutagenic
activity
in
several
assays,
including
a
bacterial
DNA
damage/
repair
assay
(
MRID
93050019;
93050020),
dominant
lethal
assay
(
MRID
00088953),
unscheduled
DNA
synthesis
assay
(
MRID
93050021;
42711001;
41642401),
micronucleus
assay
(
41728601),
in
vitro
cell
transformation
assay
(
MRID
93050022),
and
the
Ames
assay
(
MRID
93050017;
93050018).
In
the
Ames
Salmonella
test,
azadioxabicyclooctane
was
negative
in
all
strains
tested,
but
deficiencies
in
the
studies
were
identified
and
these
deficiencies
must
be
addressed
to
upgrade
the
studies.
Azadioxabicyclooctane
was
also
negative
in
the
micronucleus
assay,
but
was
positive
in
one
unscheduled
DNA
synthesis
assay
(
93050021)
and
the
bacterial
DNA
damage/
repair
assay.
Several
tests
(
MRID
numbers
93050022
through
93050025)
were
conducted
with
C3H/
10T1/
2
cell
cultures
examining
cell
transformation
in
vitro.
Although
results
were
largely
negative,
these
studies
collectively
did
not
properly
identify
the
test
material
being
examined.
In
fact,
only
one
study
(
MRID
93050018)
properly
characterized
the
test
material.
Thus,
although
the
data
suggest
largely
negative
responses,
the
lack
of
test
article
characterization
(
especially
in
light
of
positive
responses
observed
in
two
studies)
points
to
the
need
for
proper
test
article
characterization
before
an
adequate
conclusion
can
be
made
about
the
mutagenicity
of
azadioxabicyclooctane.

Salmonella/
Mammalian
Activation
Gene
Mutation;
OPPTS
870.5265
[
§
84­
2]
In
a
reverse
gene
10
mutation
assay
in
bacteria
(
MRID
93050017
reformat
of
00088956),
strains
TA98,
TA100,
TA1535,
TA1537
and
TA1538
of
S.
typhimurium
were
exposed
to
Nuosept
®
95
(
Lot
No.
005­
267)
in
water
at
concentrations
of
0,
0.01,
0.05,
0.23,
0.45
and
0.9
µ
L/
plate
in
the
presence
and
absence
of
mammalian
metabolic
activation
(
S9­
mix).
The
S9­
fraction
was
obtained
from
Aroclor
1254
induced
male
Sprague­
Dawley
rat
liver.

Nuosept
95
was
tested
up
to
cytotoxic
concentrations.
A
preliminary
assay
with
TA100
showed
no
cytotoxicity
at
concentrations
up
to
0.29
µ
L/
plate
and
virtually
complete
cell
killing
at
the
0.94
µ
L/
plate
and
higher
concentrations.
The
highest
concentration
tested
in
the
mutagenicity
assay,
0.9
µ
L/
plate
was
cytotoxic
as
evidenced
by
a
reduction
in
the
number
of
revertants
per
plate
in
all
strains
compared
to
the
solvent
controls.
At
no
Nuosept
95
concentration,
with
or
without
S9­
mix,
was
there
at
least
a
doubling
of
the
mutant
frequency
in
any
tester
strain
over
the
solvent
control
values.
The
greatest
increase
was
seen
in
strain
TA100
where
the
mutant
frequency
at
0.45
µ
L/
plate,
both
with
and
without
S9­
mix,
was
approximately
1.9
times
the
solvent
control
value.
The
positive
and
solvent
controls
induced
the
appropriate
responses
in
the
strains
in
which
they
were
tested.
There
was
no
evidence
of
induced
mutant
colonies
over
background.

This
study
is
classified
as
unacceptable/
guideline
due
to
incomplete
positive
control
data.
In
the
absence
of
S9­
mix,
only
strains
TA1535
and
TA1537
were
treated
with
a
positive
control
and
in
the
presence
of
S9­
mix,
only
strains
TA98,
TA100
and
TA1538
were
treated
with
the
positive
control.

Salmonella/
Mammalian
Activation
Gene
Mutation;
OPPTS
870.5265
[
§
84­
2]
In
a
reverse
gene
mutation
assay
in
bacteria
(
MRID
93050018
reformat
of
00088974),
strains
TA98,
TA100,
TA1535,
TA1537
and
TA1538
of
S.
typhimurium
were
exposed
to
Nuosept
®
95
(
Lot
No.
005­
267)
in
water
at
concentrations
of
0,
5,
10,
50,
100
and
250
µ
g/
plate
in
an
initial
assay
and
to
concentrations
of
0,
375,
500,
625
and
750
µ
g/
plate
in
a
second
assay.
Testing
was
in
the
presence
and
absence
of
mammalian
metabolic
activation
(
S9­
mix)
from
Aroclor
1254
induced
rat
liver.

Nuosept
95
was
tested
up
to
cytotoxic
concentrations.
Toxicity,
as
evidenced
by
changes
in
the
background
lawn
of
bacteria,
was
seen
in
all
tester
strains
at
750
µ
g/
plate
in
the
absence
of
S9­
mix
and
in
TA1535
in
the
presence
of
S9­
mix.
Toxicity
was
seen
in
the
absence
of
S9­
mix
at
500
µ
g/
plate
in
TA98
and
TA1538
and
at
625
µ
g/
plate
in
all
strains
except
TA1535.
The
mitotic
index
(
number
of
revertants
on
Nuosept
95
treated
plates/
number
of
revertants
on
solvent
control
plates)
remained
below
two
in
all
tester
strains
at
all
concentrations,
with
or
without
S9­
mix.
In
almost
all
cases
the
mitotic
index
was
1.0
or
lower.
The
positive
controls
induced
the
appropriate
responses
in
the
presence
of
S9­
mix.
There
was
no
evidence
of
induced
mutant
colonies
over
background.

This
study
is
classified
as
unacceptable/
guideline.
Few
experimental
details
were
provided
and
positive
controls
were
not
used
in
non­
activated
tests.
In
addition,
the
composition
of
the
S9­
mix
was
not
provided.
11
In
Vitro
Mammalian
Cytogenics
­
Micronucleus
Assay
in
Mouse
Bone
Marrow
Polychromatic
Erythrocytes
OPPTS
870.5395
[
§
84­
2]
In
a
Swiss­
Webster
mouse
bone
marrow
micronucleus
assay
(
MRID
41728601),
five
mice/
sex/
dose
were
treated
by
oral
gavage
(
two
treatments
separated
by
24
hours)
with
Nuosept
®
95
(
Lot
No.
129593,
purity
not
reported)
at
doses
of
0,
300,
600
and
1200
mg/
kg
body
weight
in
males
and
0,
600,
1200
and
2400
mg/
kg
body
weight
in
females.
Bone
marrow
cells
were
harvested
at
24
and
48
hours
post­
treatment.
The
vehicle
was
sterile
deionized
water.

There
were
signs
of
toxicity
during
the
study.
A
preliminary
toxicity
assay
was
used
to
determine
doses
used
in
the
micronucleus
assay.
In
the
micronucleus
assay,
all
mice
in
the
solvent
control
and
the
300
or
600
mg/
kg
Nuosept
95
dose
groups
appeared
normal.
One
male
in
the
1200
mg/
kg
group
had
rough
fur,
a
humped
back
and
labored
breathing
on
day
2
and
was
found
dead
on
day
3.
Nine
of
a
total
of
17
females
dosed
at
2400
mg/
kg
had
rough
fur
and
humped
backs
with
four
also
showing
labored
breathing.
Four
females
from
the
2400
mg/
kg
dose
groups
were
found
dead
on
day
3
and
three
others
had
rough
fur
and
humped
backs.
Other
clinical
signs
seen
in
the
2400
mg/
kg
groups
were
lacrimation
from
both
eyes
and
closed
eyes.
The
solvent
controls
showed
a
frequency
of
micronucleated
PCEs
at
24
and
48
hours
of
0.14%
and
0.16%,
respectively,
in
males
and
0.14%
and
0.12%,
respectively,
in
females.
The
frequency
of
micronucleated
PCEs
in
Nuosept
95
treated
mice
did
not
exceed
0.20%
in
males
or
0.16%
in
females
at
any
concentration
at
either
harvest
time.
No
increase
was
statistically
significant.
The
urethane
positive
control
produced
a
frequency
of
micronucleated
PCEs
in
males
at
24
and
48
hours
of
2.59%
and
1.16%,
respectively.
These
increases
were
statistically
significant
(
p<
0.05).
No
positive
control
was
used
in
females.
Positive
and
solvent
control
values
were
acceptable.
There
was
no
significant
increase
in
the
frequency
of
micronucleated
polychromatic
erythrocytes
in
bone
marrow
PCEs
in
either
sex
at
any
tested
dose
at
either
harvest
time.

This
study
is
classified
as
unacceptable/
guideline
and
upgradable.
The
test
substance
purity
and
composition
is
required.

Rat
Dominant
Lethal
Mutagenicity
Assay;
OPPTS
870.5450
[
§
84­
2]
In
a
CD1
Sprague­
Dawley
rat
dominant
lethal
assay
(
MRIDs
240259303
and
93050026,
reformat
of
00088953),
male
rats
were
treated
for
10
consecutive
weeks
with
Nuosept
®
95
(
Lot
No.
005­
267)
in
their
drinking
water
at
doses
of
0,
125,
625
or
3125
pom
of
active
ingredient
(
estimated
daily
average
for
the
10
week
period
of
0,
14,
66
and
268
mg/
kg
body
weight/
day
of
Nuosept
95
active
ingredients).
After
the
10
week
dosing
period,
each
male
was
mated
with
two
untreated
females
per
week
for
two
weeks.

Nuosept
95
was
tested
to
a
dose
limited
by
taste
aversion.
Average
daily
water
consumption
was
comparable
in
the
solvent
control
and
two
lower
dose
Nuosept
95
groups;
however,
the
overall
average
daily
water
consumption
for
the
3125
pom
group
was
approximately
69%
of
the
solvent
control
group.
The
average
weight
gain
of
the
rats
over
the
10
week
dosing
period
was
280
g
for
the
solvent
control,
255
g
for
the
positive
control
and
251,
284
and
244
g
for
the
125,
625
and
3125
pom
Nuosept
95
dosed
groups,
respectively.
The
lower
weight
gain
was
considered
a
result
of
reduced
water
intake
rather
than
a
toxic
effect
of
the
test
material.
No
other
clinical
signs
were
12
reported.
All
treated
mice
survived
until
the
end
of
the
study.
The
females
were
sacrificed
16
days
after
the
first
day
of
cohabitation
and
examined
for
pregnancy,
living
fetuses,
early
and
late
fetal
death
and
corpora
lutea
and
for
intercurrent
infections.
The
following
parameters
were
determined:
total
implants
per
pregnant
female
(
live
fetuses
plus
early
and
late
fetal
deaths),
dead
implants
per
pregnant
female
(
early
and
late
fetal
deaths),
total
number
of
dead
implants
per
total
implants,
corpora
lutea
per
pregnant
female,
live
implants
per
pregnant
female
and
preimplantation
loss
(
the
difference
between
total
corpora
lutea
and
total
implants),
the
number
of
Corpora
lutea,
implantations,
living
embryos
and
fetuses,
and
dead
implantations
(
sum
of
the
deciduomata,
resorptions
and
dead
embryos
and
fetuses).
Pre­
and
post­
implantation
losses
were
determined.
With
one
exception,
there
were
no
other
statistically
significant
differences
between
Nuosept
95
treated
groups
and
solvent
controls
in
any
of
the
measured
parameters
at
any
dose
in
either
the
first
or
second
week
of
mating.
The
one
exception
was
a
statistically
significant
reduction
in
the
average
number
of
corpora
lutea
per
female
in
the
625
pom
group
in
the
second
week
of
mating
compared
to
the
solvent
control
(
15.40
vs
17.53).
This
difference
was
not
considered
biologically
significant
by
the
testing
laboratory
because
the
count
was
within
normal
limits
(
which
were
not
given)
and
comparable
to
that
of
all
groups
in
the
first
mating
week.
Positive
and
solvent
control
values
were
appropriate.
There
was
no
significant
difference
between
the
control
group
and
Nuosept
95­
treated
groups
with
respect
to
fertilization
rate,
dead
implantations,
living
implantations,
preimplantation
loss
or
total
implantations.

This
study
is
classified
as
unacceptable/
guideline.
The
route
of
administration
of
the
test
substance
is
insufficient.
The
only
statistically
and
biologically
significant
effect
observed
in
the
high­
dose
group
was
reduced
water
consumption
and
resultant
reduced
body
weight.
Because
Nuosept
95
was
administered
in
the
drinking
water,
this
effect
may
be
due
to
taste
aversion
of
the
test
substance.
If
the
test
substance
had
been
administered
by
another
method,
such
as
oral
gavage,
higher
doses
may
have
been
evaluated.

Bacterial
Damage/
Repair
in
Salmonella
typhimurium
Assay;
OPPTS
870.5500
[
§
84­
2]
In
a
differential
killing/
growth
inhibition
assay
in
bacteria
(
MRID
93050019
reformat
of
00088957),
strains
WP2
(
uvr
A+
rec
A+)
and
WP100
(
uvr
A­
rec
A­)
of
Escherichia
coli
and
strains
TA
1978
(
uvr
B+)
and
TA
1538
(
uvr
B­)
of
Salmonella
typhimurium
were
exposed
to
Nuosept
®
95
(
Lot
No.
005­
267)
in
water
in
a
suspension
assay
at
concentrations
of
0.03,
0.3,
0.6
and
0.9
µ
L/
mL
in
the
presence
and
absence
of
mammalian
metabolic
activation
(
S9­
mix).
The
S9­
fraction
was
obtained
from
Aroclor
1254
induced
male
Sprague­
Dawley
rat
liver.

Nuosept
95
was
tested
up
to
cytotoxic
concentrations.
In
the
absence
of
S9­
mix,
no
cell
killing
was
seen
in
either
the
repair
proficient
or
deficient
E.
coli
strains
at
the
lowest
concentration
tested,
0.03
µ
L/
mL;
however,
survival
of
the
repair
deficient
strains
was
virtually
zero
at
all
higher
concentrations
compared
to
average
survivals
ranging
from
54%
to
42%
in
the
repair
proficient
strain.
The
S.
typhimurium
strains
were
somewhat
more
sensitive
with
survivals
at
the
lowest
concentration
of
81%
and
72%
for
the
repair
proficient
and
deficient
strains,
respectively.
Survival
of
the
repair
proficient
strain
ranged
from
31%
to
18%
at
the
higher
concentrations
and
virtually
complete
killing
of
the
repair
deficient
strain
at
all
higher
concentrations.
Decreasing
survival
indices
(
defined
as
the
percent
survival
of
the
repair
deficient
strain
by
the
percent
survival
of
the
repair
proficient
strain)
with
increasing
concentrations
of
test
chemicals
are
13
considered
indicative
of
genotoxicity
in
this
assay.
In
the
presence
of
S9­
mix,
the
results
for
the
E.
coli
strains
were
similar
to
those
in
the
absence
of
S9­
mix.
Survivals
of
the
S.
typhimurium
strains
at
the
two
lowest
concentrations
of
Nuosept
95
were
actually
higher
for
the
repair
deficient
strain
(
72%
and
92%
for
the
repair
proficient
and
deficient
strains
at
0.03
µ
L/
mL,
respectively,
and
20%
and
36%
for
the
repair
proficient
and
deficient
strains
at
0.3
µ
L/
mL,
respectively).
At
0.6
µ
L/
mL,
survival
of
the
repair
deficient
strain
was
15%
compared
to
28%
for
the
repair
proficient
strain
with
comparable
values
at
0.9
µ
L/
mL
of
11%
and
23%.
Only
the
0.6
and
0.9
µ
L/
mL
concentrations
presented
positive
results
in
the
suspension
assay
(<
0.8).
The
negative,
solvent
and
positive
controls
induced
the
appropriate
responses
in
the
corresponding
strains.
There
was
evidence
of
greater
growth
inhibition
or
cell
killing
in
repair­
defective
strains
compared
to
repair
competent
strains.

This
study
is
classified
as
acceptable/
guideline.

Bacterial
Damage/
Repair
in
Escherichia
coli
and
Salmonella
typhimurium
Mutagenicity
Assay
PPTS
870.5500
[
§
84­
2]
In
a
differential
killing/
growth
inhibition
assay
in
bacteria
(
MRID
93050020
reformat
of
00088958),
strains
W3110
(
pol
A+)
and
p3478
(
pol
A­)
of
Escherichia.
coli
and
Salmonella
typhimurium
strains
TA
1978
(
uvr
B+)
and
TA
1538
(
uvr
B­)
were
exposed
to
Nuosept
®
95
(
Lot
No.
005­
267)
in
sterile
distilled
water
at
concentrations
of
50,
100,
500,
1000
and
5000
µ
g/
mL
in
suspension
culture
in
the
presence
and
absence
of
mammalian
metabolic
activation
(
S9­
mix).
The
two
E.
coli
strains
were
also
exposed
to
Nuosept
95
in
a
disk
assay
(
10
µ
L
of
Nuosept
95
on
a
paper
disk
in
the
center
of
the
plate)
but
the
activation
status
was
not
given.
The
S9­
fraction
was
obtained
from
Aroclor
1254
induced
rat
liver.

Nuosept
95
was
tested
up
to
cytotoxic
concentrations.
In
the
disk
assay,
the
diameter
of
the
zone
of
inhibition
(
scored
as
a
ratio
of
the
zone
of
inhibition
and
of
the
pol
A­
to
the
zone
of
inhibition
of
the
pol
A+)
of
Nuosept
95
treated
strains
was
36
mm
for
the
pol
A+
strain
and
43
mm
for
the
pol
A­
strain
compared
to
0.0
mm
for
the
solvent
control
in
both
strains.
A
chemical
is
considered
to
have
a
positive
response
if
the
difference
in
the
zones
of
inhibition
is
greater
than
or
equal
to
3
mm.
The
difference
of
7
mm
in
Nuosept
®
95
treated
bacteria
was
equal
to
the
MNNG
positive
control
difference
(
28
and
35
mm
for
the
pol
A+
and
pol
A­
strains,
respectively).
In
the
suspension
assay,
the
testing
laboratory's
criterion
for
a
positive
response,
a
survival
index
(
ratio
of
survival
of
repair
deficient/
repair
proficient
strains)
of
less
than
0.8,
was
met
at
100,
500,
1000,
and
5000
µ
g/
mL
in
both
tester
pairs
with
and
without
S9­
mix
with
the
exception
of
a
negative
response
at
100
µ
g/
mL
in
the
presence
of
activation.
The
assay
was
negative
for
all
strains
with
and
without
metabolic
activation
at
the
50
µ
g/
mL
concentration.
The
survival
of
all
strains
(
except
E.
coli
(
pol
A+)
only
reduces
to
37.3%
at
5000
µ
g/
mL
with
S9­
mix)
was
reduced
to
zero
at
5000
µ
g/
mL.
Positive
and
solvent
control
values
were
acceptable.
Few
experimental
details
were
provided
with
this
study.
The
number
of
replicate
plates
and/
or
cultures
was
not
given,
individual
plate
counts
were
not
provided
(
just
summary
data),
the
S9­
mix
composition
was
not
given,
and
the
positive
control
for
the
S.
typhimurium
strains
without
S9­
mix
was
not
specified.
There
was
evidence
of
greater
growth
inhibition
or
cell
killing
in
repairdefective
strains
compared
to
repair
competent
strains.

This
study
is
classified
as
unacceptable/
guideline
and
upgradable.
Information
on
individual
14
plate
counts
and
the
S9­
mix
composition
are
required.

Unscheduled
DNA
Synthesis
in
Primary
Rat
Hepatocytes/
Mammalian
Cell
Cultures
Mutagenicity
Assay
OPPTS
870.5550
[
§
84­
2]
In
an
unscheduled
DNA
synthesis
assay
(
MRID
93050021
reformat
of
00088959),
primary
rat
hepatocyte
cultures
from
male
F­
344
rats
were
exposed
to
Nuosept
®
95
(
lot
number
and
purity
not
provided)
in
culture
medium
at
concentrations
of
0,
0.0156,
0.0316,
0.0625,
0.125,
0.25,
1.0
and
2.0
µ
L/
mL
for
one
hour.

Nuosept
95
was
tested
up
to
cytotoxic
concentrations.
Concentrations,
ranging
from
0.0058
to
47.6
µ
L/
mL,
were
tested
in
a
preliminary
cytotoxicity
assay
(
trypan
blue
exclusion)
with
no
viable
cells
remaining
in
cultures
exposed
to
1.49
µ
L/
mL
and
higher
concentrations.
A
concentration
of
0.74
µ
L/
mL
reduced
viability
to
approximately
45%
of
the
negative
control
at
two
hours
posttreatment
and
to
zero
when
incubation
was
allowed
to
continue
for
24
hours
post­
treatment.
Two
UDS
assays
were
performed.
Results
from
the
first
assay
were
considered
unusable
due
to
poor
cell
morphology
and
distribution
on
the
coverslips
and
an
abnormally
low
positive
control
value.
The
data
from
the
first
assay
were
not
reported.
In
the
second
UDS
assay,
Nuosept
95
concentrations
of
0.5
µ
L/
mL
and
higher
were
quite
toxic
with
no
surviving
cells
24
hours
after
treatment.
Concentrations
of
0.0156
to
0.25
µ
L/
mL
gave
a
range
of
survivals
at
24
hours
posttreatment
from
100%
to
29.1%.
The
average
percentage
of
nuclei
with
six
or
more
grains
was
32.3%
and
25.9%
at
Nuosept
95
concentrations
of
0.0625
and
0.125
µ
L/
mL,
respectively,
thus
exceeding
the
criterion
of
21%
required
for
a
positive
response
as
established
for
this
particular
assay.
The
average
percentage
of
nuclei
with
six
or
more
grains
at
0.25
µ
L/
mL
was
18.0%,
not
quite
reaching
the
level
for
a
positive
response;
however,
cytotoxicity
was
high
at
this
concentration
(
29.1%
survival
at
24
hours)
and
the
variation
between
the
three
cultures
at
this
dose
was
said
to
range
from
8%
to
36%
(
individual
culture
data
were
not
presented).
Cytotoxicity
may
have
inhibited
UDS
activity
at
this
dose.
The
average
percentage
of
nuclei
with
 
6
grains
in
the
negative
(
culture
medium)
and
positive
(
2­
AAF)
controls
were
11.0%
and
84.7%,
respectively.
The
controls
induced
the
appropriate
response.
There
was
evidence
of
unscheduled
DNA
synthesis,
as
determined
by
radioactive
tracer
procedures
[
nuclear
silver
grain
counts].

This
study
is
classified
as
unacceptable/
guideline
and
upgradable.
The
test
substance
purity
and
composition
is
required.

Unscheduled
DNA
Synthesis
in
Primary
Rat
Hepatocytes/
Mammalian
Cell
Cultures;
OPPTS
870.5550
[
§
84­
2].
In
an
ex
vivo
unscheduled
DNA
synthesis
(
UDS)
assay
(
MRID
42711001),
three
male
Fischer­
344
rats
per
dose
group
were
given
a
single
oral
dose
(
gavage)
each
of
Nuosept
®
95
(
lot
No.
05249587,
purity
not
provided)
in
water
at
concentrations
of
0,
875,
1750
or
3500
mg/
kg
body
weight.
Rats
were
sacrificed
two
or
16
hours
after
treatment
and
hepatocytes
isolated
and
analyzed
in
culture
for
induced
UDS
activity.
The
percentage
of
hepatocytes
in
repair
was
also
determined.

Nuosept
95
was
tested
to
toxic
levels
(
approximately
20,
40
and
80%
of
the
estimated
LD50
of
4401
mg/
kg
calculated
from
the
results
of
two
preliminary
toxicity
assays).
Rats
in
the
16­
hour
1750
and
3500
mg/
kg
dose
groups
had
rough
fur
on
the
morning
after
dosing
and
one
rat
in
the
15
16­
hour
3500
mg/
kg
dose
group
died
before
the
scheduled
sacrifice
time.
Viability
of
the
isolated
hepatocytes
averaged
approximately
80%
in
all
Nuosept
95
treated
groups
and
in
the
solvent
control
group
and
approximately
70%
in
the
positive
control
group.
No
increase
in
the
mean
net
nuclear
grain
count
over
the
solvent
control
value
of
­
7.4
was
seen
at
any
tested
Nuosept
95
concentration
at
either
the
two­
hour
or
16­
hour
sacrifice
time.
The
mean
percentage
of
cells
in
repair
did
not
exceed
1%
(
the
solvent
control
value)
at
any
tested
Nuosept
95
concentration
at
either
sacrifice
time.
The
positive
control
(
dimethylnitrosamine)
induced
a
mean
net
nuclear
grain
count
of
18.3
and
a
percentage
of
cells
in
repair
of
25.6%.
The
positive
and
solvent
controls
induced
the
appropriate
responses.
There
was
no
evidence
that
unscheduled
DNA
synthesis,
as
determined
by
radioactive
tracer
procedures
(
nuclear
silver
grain
counts)
was
induced
by
Nuosept
95
as
tested
in
this
study.

This
study
is
classified
as
unacceptable/
guideline
and
upgradable.
The
test
substance
purity
and
composition
is
required.

In
Vitro
Cell
Transformation
Assay
in
C3H/
10T
½
Cells
Other
Genotoxicity
[
§
84­
4]
In
an
in
vitro
cell
transformation
assay
(
MRID
93050022
reformat
of
00088960),
C3H/
10T
½
cell
cultures
were
exposed
without
metabolic
activation
to
Nuosept
®
95
(
Lot
No.
005­
267)
at
concentrations
of
0,
0.0005,
0.001,
0.002,
and
0.004
µ
l/
ml
for
18
hours.
After
replacement
of
the
medium
and
further
incubation
for
5
weeks
with
weekly
refeeding,
cultures
were
fixed,
stained,
and
scored
for
morphological
transformation.
Foci
of
transformed
cells
were
observed
in
5
of
12
cultures.
No
dose
response
was
observed
but
that
is
not
required
for
this
assay.
Cytotoxicity
was
demonstrated
by
reduction
of
relative
cloning
efficiency
below
50%
at
the
two
higher
doses.
The
positive
and
solvent
control
values
were
appropriate.

This
study
is
classified
as
acceptable/
non­
guideline.

In
Vitro
Cell
Transformation
Assay
in
C3H/
10T
½
Cells;
Other
Genotoxicity
[
§
84­
4].
In
an
in
vitro
cell
transformation
assay
(
MRID
93050023
reformat
of
00088961),
C3H/
10T
½
cell
cultures
were
exposed
without
metabolic
activation
to
Nuosept
®
95
(
Lot
No.
005­
267)
at
concentrations
of
0,
0.0003,
0.001,
and
0.003
µ
l/
ml
for
24
hours.
After
replacement
of
the
medium
and
further
incubation
for
4­
6
week
with
twice
weekly
refeeding,
cultures
were
fixed,
stained,
and
scored
for
morphological
transformation.
No
Type
II
or
Type
III
foci
of
transformed
cells
were
observed
in
any
treatment
group.
Cytotoxicity
was
evident
at
the
high
dose,
demonstrated
by
reduction
of
relative
cloning
efficiency
below
50%.
Positive
and
solvent
control
values
were
appropriate.

This
study
is
classified
as
unacceptable/
non­
guideline
and
upgradable
because
it
cannot
be
ascertained
whether
T1597
could
have
been
tested
at
a
considerably
higher
dose
and
because
the
duration
of
incubation
of
the
cell
cultures
following
treatment
was
not
precisely
specified.
If
data
can
be
presented
to
show
that
a
significantly
higher
dose
would
be
too
toxic
(>
90%
reduction
of
cloning
efficiency)
and
to
specify
the
duration
of
post­
treatment
incubation
of
the
cultures,
the
study
could
be
upgraded
to
acceptable.

In
Vitro
Cell
Transformation
Assay
in
C3H/
10T
½
Cells;
Other
Genotoxicity
[
§
84­
4].
In
an
in
vitro
cell
transformation
assay
(
MRID
93050024
reformat
of
00088962),
C3H/
10T
½
cell
cultures
16
were
exposed
to
R­
1162
(
Lot
number
and
purity
not
reported)
without
metabolic
activation
to
R­
1162
at
concentrations
of
0,
0.0098,
0.0195,
0.039,
and
0.078
µ
l/
ml
for
18
hours.
After
replacement
of
the
medium
and
further
incubation
for
5
weeks
with
weekly
refeeding,
cultures
were
fixed,
stained,
and
scored
for
morphological
transformation.
Foci
of
transformed
cells
were
not
observed
at
any
dose
level.
The
number
of
transformed
foci
in
the
positive
and
solvent
controls
was
appropriate.
Cytotoxicity
was
demonstrated
by
reduction
of
relative
cloning
efficiency
below
50%
at
the
highest
dose,
although
unusually
low
survival
in
the
solvent
control
cultures
caused
relative
cloning
efficiencies
to
be
higher
than
expected
in
the
cell
transformation
assay.

This
study
is
classified
as
unacceptable/
non­
guideline
and
upgradable.
Information
on
the
test
substance
lot
number,
purity,
and
composition
was
not
provided.

In
Vitro
Cell
Transformation
Assay
in
C3H/
10T
½
Cells;
Other
Genotoxicity
[
§
84­
4].
In
an
in
vitro
cell
transformation
assay
(
93050025
reformat
of
00088963),
C3H/
10T
½
cell
cultures
were
exposed
without
metabolic
activation
to
R­
1143
(
Lot
#
11­
20­
1979)
at
concentrations
of
0,
0.0006,
0.0012,
0.0024,
and
0.0048
µ
l/
ml
for
18
hours.
After
replacement
of
the
medium
and
further
incubation
for
5
weeks
with
weekly
refeeding,
cultures
were
fixed,
stained,
and
scored
for
morphological
transformation.
Foci
of
transformed
cells
were
not
observed
at
any
dose
level.
The
number
of
transformed
foci
for
the
positive
and
solvent
controls
was
appropriate.
Cytotoxicity
was
demonstrated
by
reduction
of
relative
cloning
efficiency
below
50%
at
the
highest
dose
tested.

This
study
is
classified
as
unacceptable/
non­
guideline
and
upgradable.
Information
on
the
test
substance
lot
number,
purity,
and
composition
was
not
provided.

4.8
Neurotoxicity
Adequacy
of
database
for
Neurotoxicity:
From
the
available
repeat
dose
toxicity
studies,
there
was
no
evidence
of
neurotoxicity
of
azadioxabicyclooctane.

4.9
Metabolism
and
Pharmacokinetics
Adequacy
of
database
for
Metabolism
and
Pharmacokinetics:
The
database
does
not
provide
information
on
the
metabolism
and
pharmacokinetics
of
azadioxabicyclooctane.

5.0
TOXICITY
ENDPOINT
SELECTION
5.1
See
Section
7.1,
Summary
of
Toxicological
Doses
and
Endpoint
Selection,
Table
3.

5.2
Dermal
Absorption
Dermal
Absorption
Factor:
Since
a
dermal
endpoint
was
selected
from
a
dermal
toxicity
study,
a
dermal
absorption
factor
is
not
needed
for
azadioxabicyclooctane.
17
5.3
Classification
of
Carcinogenic
Potential
There
are
no
carcinogenicity
data
for
azadioxabicyclooctane.
A
determination
of
carcinogenic
potential
cannot
be
made
based
on
the
available
data.
Metalworking
cutting
fluid
use
triggers
the
requirement
for
carcinogenicity
studies
and
chronic
toxicity
studies
as
described
on
the
1987
Antimicrobial
Data
Call
In
Notice.

6.0
FQPA
CONSIDERATIONS
6.1
Reproductive
Toxicity
Study
Conclusions
There
are
no
data
on
the
reproductive
toxicity
of
azadioxabicyclooctane.
For
the
indirect
food
use
of
this
chemical,
this
study
is
being
held
in
reserve.

6.2
Pre­
and/
or
Postnatal
Toxicity
A.
Determination
of
Susceptibility
The
ADTC
concluded
that
there
is
no
evidence
for
susceptibility
to
azadioxabicyclooctane
from
the
available
data,
but
the
data
are
very
limited
to
one
developmental
toxicity
study.
For
this
reason,
extra
uncertainty
factors
will
be
employed
to
account
for
the
lack
of
a
complete
database
until
more
data
are
made
available.

B.
Degree
of
Concern
Analysis
and
Residual
Uncertainties
Uncertainties
exist
with
respect
to
determination
of
both
developmental
and
reproductive
effects
of
azadioxabicyclooctane,
as
the
data
are
limited
to
the
one
study
mentioned
above.

C.
Proposed
Hazard­
based
Special
FQPA
Safety
Factor(
s):

The
ADTC
recommended
that
the
special
10x
hazard­
based
safety
factor
under
the
FQPA
be
retained
(
10x)
for
azadioxabicyclooctane.
There
is
only
one
prenatal
dermal
developmental
toxicity
study
available
for
azadioxabicyclooctane,
and
there
is
no
reproductive
toxicity
information
or
studies.
While
the
developmental
study
showed
no
evidence
of
susceptibility
of
offspring
to
this
chemical,
the
route
of
administration
is
not
a
good
indicator
of
potential
ffects
from
oral
exposures.
In
addition,
as
noted
above,
information
on
the
test
article
characterization
is
not
resolved
at
this
time.
The
retention
of
10x
FQPA
hazard­
based
safety
factor
is
consistent
with
the
Agency's
published
guidance.
(
see
http://
www.
epa.
gov/
oppfead1/
trac/
science/
determ.
pdf)

6.3
Recommendation
for
a
Developmental
Neurotoxicity
Study
The
ADTC
reserved
its
recommendation
for
a
developmental
neurotoxicity
study,
pending
fulfillment
of
the
data
gaps
for
developmental
and
reproductive
toxicity.
18
7.0
SUMMARY
OF
TOXICOLOGICAL
DOSES
AND
ENDPOINTS
FOR
AZADIOXABICYCLOOCTANE
FOR
USE
IN
HUMAN
RISK
7.1
Summary
Table
of
Toxicological
Dose
and
Endpoint
Selection
(
Table
2)

Exposure
Scenario
Dose
Used
in
Risk
Assessment,
UF
Special
FQPA
SF*
and
Level
of
Concern
for
Risk
Assessment
Study
and
Toxicological
Effects
Acute
Dietary
(
gen.
pop.)
NOAEL
=
10.6
mg/
kg/
day
UF
=
100
FQPA
SF
=
10x
aPAD
=
acute
RfD
FQPA
SF
=
0.01
mg/
kg/
day
90
day
oral
toxicity
in
rats
LOAEL
=
10.6
mg/
kg/
day
based
on
decreased
water
consumption
at
56.5
mg/
kg/
day
in
males.

Acute
Dietary
(
females
13+)
NOAEL
=
10.6
mg/
kg/
day
UF
=
100
FQPA
SF
=
10x
aPAD
=
acute
RfD
DB
UF
=
0.01
mg/
kg/
day
90
day
oral
toxicity
in
rats
LOAEL
=
10.6
mg/
kg/
day
based
on
decreased
water
consumption
at
56.5
mg/
kg/
day
in
males.

Chronic
Dietary
(
All
populations)
NOAEL
=
10.6
mg/
kg/
day
UF
=
300
FQPA
SF
=
10x
cPAD
=
chronic
RfD
FQPA
SF
=
0.003
mg/
kg/
day
90
day
oral
toxicity
in
rats
LOAEL
=
10.6
mg/
kg/
day
based
on
decreased
water
consumption
at
56.5
mg/
kg/
day
in
males.

Short­
Term/

Intermediateterm
Incidental
Oral
(
1­
30
days;
30
days­
6
months)
No
endpoint
required
19
Exposure
Scenario
Dose
Used
in
Risk
Assessment,
UF
Special
FQPA
SF*
and
Level
of
Concern
for
Risk
Assessment
Study
and
Toxicological
Effects
Dermal
(
all
durations)
dermal
LOAEL=
100
mg/
kg/
day
Occupational
MOE
=
300
(
ST
and
IT)
=
1000
(
LT)
Co­
critical
studies:

21­
day
dermal
toxicity
in
rats
LOAEL
=
100
mg/
kg/
day
(
severe
dermal
effects)

developmental
toxicity
in
rats
LOAEL
=
100
mg/
kg/
day
(
severe
dermal
effects)

Inhalation
(
all
durations
)
90­
day
oral
study
NOAEL=
10.6
mg/
kg/
day
(
inhalation
absorption
rate
=
100%)
Occupational
MOE
=
3000
90
day
oral
toxicity
in
rats
LOAEL
=
10.6
mg/
kg/
day
based
on
decreased
water
consumption
at
56.5
mg/
kg/
day
in
males.

Cancer
(
oral,
dermal,
inhalation)
no
cancer
data
available
UF
=
uncertainty
factor,
FQPA
SF
=
Special
FQPA
safety
factor,
NOAEL
=
no
observed
adverse
effect
level,
LOAEL
=
lowest
observed
adverse
effect
level,
PAD
=
population
adjusted
dose
(
a
=
acute,
c
=
chronic)
RfD
=
reference
dose,
MOE
=
margin
of
exposure,
LOC
=
level
of
concern,
NA
=
Not
Applicable
NOTE:
The
Special
FQPA
Safety
Factor
recommended
by
the
ADTC
assumes
that
the
exposure
databases
(
dietary
food,
drinking
water,
and
residential)
are
complete
and
that
the
risk
assessment
for
each
potential
exposure
scenario
includes
all
metabolites
and/
or
degradates
of
concern
and
does
not
underestimate
the
potential
risk
for
infants
and
children.

8.0
TOXICITY
PROFILE
TABLES
8.1
Acute
Toxicity
Profile
Table
­
(
See
Section
4.1,
Acute
Toxicity,
Table
1).

8.2
Subchronic,
Chronic
and
Other
Toxicity
Profiles
Table
(
Table
3)
20
Guideline
Number/
Study
Type/
Test
Substance
(%
a.
i.)
MRID
Number
(
Year)/
Citation/
Classification/
Doses
Results
870.3100
(
§
82­
1)
90­
day
Oral
­
Rat
Azadioxabicyclooctane
purity
50%
41641606,
reformat
of
00109245,
supplemental
410801010
Acceptable
­
Guideline
0,
10.6,
56.5,
or
228.9
mg/
kg/
day
(
males)
0,
12.2,
64.3,
or
255.9
mg/
kg/
day
(
females)
1
Oral
Toxicity
(
males)
NOAEL
=
10.6
mg/
kg/
day
(
males)
=
12.2
mg/
kg/
day
(
females)
LOAEL
=
56.5
mg/
kg/
day
(
males)
=
64.3
mg/
kg/
day
(
females)
based
on
reduced
water
consumption.

870.3200
(
§
82­
2)
21­
day
Dermal
­
Rabbit
Azadioxabicyclooctane
purity
not
reported
41641605
Acceptable
­
Non­
guideline
0,
100,
300,
or
1000
mg/
kg/
day
Dermal
Toxicity
NOAEL
not
established
LOAEL
=
100
mg/
kg/
day
(
severe
dermal
effects)
Systemic
Toxicity
NOAEL
 
1000
mg/
kg/
day
(
highest
dose
tested)
LOAEL
>
1000
mg/
kg/
day
(
not
established)

870.3700a
(
§
83­
3)
Developmental
­
Rat
Azadioxabicyclooctane
purity
50.38%
41537501,
reformat
of
41699001
Acceptable
­
Non­
guideline
0,
100,
300,
or
1000
mg/
kg/
day
Dermal
Maternal
Toxicity
NOAEL
not
established
LOAEL
=
100
mg/
kg/
day
(
severe
dermal
irritation)
Systemic
Maternal
Toxicity
NOAEL
=
300
mg/
kg/
day
LOAEL
=
1000
mg/
kg/
day
(
decreased
body
weight
gain)
Developmental
Toxicity
NOAEL
 
1000
mg/
kg/
day
(
highest
dose
tested)
LOAEL
>
1000
mg/
kg/
day
(
not
established)

870.5265
(
§
84­
2)
Salmonella
typhimurium
reverse
mutation
test
Azadioxabicyclooctane
purity
50%
93050017,
reformat
of
00088956
Unacceptable
­
Guideline
0,
0.01,
0.05,
0.23,
0.45,
or
0.9
µ
L/
plate
Negative
There
was
no
evidence
of
induced
mutant
colonies
over
background.
Positive
and
solvent
controls
induced
the
appropriate
responses
in
the
strains
in
which
they
were
tested;
however,
in
the
absence
of
S9­
mix,
only
strains
TA1535
and
TA1537
were
treated
with
a
positive
control
and
in
the
presence
of
the
S9­
mix,
only
strains
TA98,
TA100,
and
TA1538
were
treated
with
a
positive
control.
21
Guideline
Number/
Study
Type/
Test
Substance
(%
a.
i.)
MRID
Number
(
Year)/
Citation/
Classification/
Doses
Results
870.5265
(
§
84­
2)
Salmonella
typhimurium
reverse
mutation
test
Azadioxabicyclooctane
purity
50%
93050018,
reformat
of
00088974
Unacceptable
­
Guideline
0,
5,
10,
50,
100,
or
250
µ
g/
plate
(
initial
assay)
0,
375,
500,
625,
or
750
µ
g/
plate
(
second
assay)
Negative
There
was
no
evidence
of
induced
mutant
colonies
over
background.
The
positive
controls
induced
the
appropriate
responses
in
the
corresponding
strains
in
the
presence
of
S9­
mix;
however,
no
positive
control
was
used
in
the
absence
of
S9­
mix.

870.5395
(
§
84­
2)
Mammalian
erythrocyte
micronucleus
test
­
Mouse
Azadioxabicyclooctane
purity
not
reported
41728601
Unacceptable
­
Guideline
0,
300,
600,
or
1200
mg/
kg
(
males)
0,
600,
1200,
or
2400
mg/
kg
(
females)
Negative
There
was
no
significant
increase
in
the
frequency
of
micronucleated
polychromatic
erythrocytes
in
bone
marrow
PCEs
in
either
sex
at
any
tested
dose
at
either
harvest
time.
No
positive
control
was
used
in
females.
Positive
and
solvent
control
values
were
acceptable.

870.5450
(
§
84­
2)
Dominant
Lethal
­
Rat
Azadioxabicyclooctane
purity
50%
240259303
and
93050026,
reformat
of
00088953
Unacceptable
­
Guideline
0,
125,
625,
or
3125
pom
Negative
There
was
no
significant
difference
between
the
control
group
and
any
Nuosept
95
treated
group
with
respect
to
fertilization
rate,
dead
implantations,
living
implantations,
preimplantation
loss,
or
total
implantations.
The
only
statistically
and
biologically
significant
effect
observed
in
the
high­
dose
group
was
reduced
water
consumption
and
resultant
reduced
body
weight.
Because
Nuosept
95
was
administered
in
the
drinking
water,
this
effect
may
be
due
to
taste
aversion
of
the
test
substance.

870.5500
(
§
84­
2)
Bacterial
DNA
Damage
or
repair
tests
­
Escherichia
coli
and
Salmonella
typhimurium
Azadioxabicyclooctane
purity
50%
93050019,
reformat
of
00088957
Acceptable
­
Guideline
0,
0.03,
0.3,
0.6,
or
0.9
µ
L/
mL
Positive
There
was
evidence
of
greater
growth
inhibition
or
cell
killing
in
repair­
defective
strains
compared
to
repair
competent
strains.
The
negative
solvent
and
positive
controls
induced
the
appropriate
responses
in
the
corresponding
strains.
22
Guideline
Number/
Study
Type/
Test
Substance
(%
a.
i.)
MRID
Number
(
Year)/
Citation/
Classification/
Doses
Results
870.5500
(
§
84­
2)
Bacterial
DNA
Damage
or
repair
tests
­
Escherichia
coli
and
Salmonella
typhimurium
Azadioxabicyclooctane
purity
50%
93050020,
reformat
of
00088958
Unacceptable
­
Guideline
0,
50,
100,
500,
1000,
or
5000
µ
g/
mL
Positive
There
was
evidence
of
greater
growth
inhibition
or
cell
killing
in
repair­
defective
strains
compared
to
repair­
competent
strains.
The
number
of
replicate
plates
and/
or
cultures
was
not
given,
individual
plate
counts
were
not
provided
(
just
summary
data),
the
S9­
mix
composition
was
not
given,
no
information
on
the
test
material
except
the
lot
number
was
provided,
and
the
positive
control
for
the
S.
typhimurium
strains
without
S9­
mix
was
not
specified.

870.5550
(
§
84­
2)
Unscheduled
DNA
Synthesis
in
Mammalian
Cells
in
Culture
­
Rat
Azadioxabicyclooctane
purity
not
reported
93050021,
reformat
of
00088959
Unacceptable
­
Guideline
0,
0.0156,
0.0316,
0.0625,
0.125,
0.25,
1.0,
or
2.0
µ
L/
mL
Positive
There
was
evidence
that
unscheduled
DNA
synthesis,
as
determined
by
radioactive
tracer
procedures
[
nuclear
silver
grain
counts].
The
controls
induced
the
appropriate
responses.

870.5550
(
§
84­
2)
Unscheduled
DNA
Synthesis
in
Mammalian
Cells
in
Culture
­
Rat
Azadioxabicyclooctane
purity
not
reported
42711001
Acceptable
­
Guideline
0,
875,
1750,
or
3500
mg/
kg
Negative
There
was
no
evidence
that
unscheduled
DNA
synthesis,
as
determined
by
radioactive
tracer
procedures
[
nuclear
silver
grain
counts],
was
induced
by
Nuosept
95
as
tested
in
this
study.
The
positive
and
solvent
controls
induced
the
appropriate
responses.

(
§
84­
4)
Other
Genotoxicity
In
Vitro
Cell
Transofrmation
Assay
in
CH3/
10T1/
2
Cells
Azadioxabicyclooctane
purity
50%
93050022
reformat
of
00088960
Acceptable
 
Non­
guideline
0,
0.0005,
0.001,
0.002,
or
0.004
µ
L/
mL
Positive
Nuosept
95
was
tested
from
nontoxic
to
toxic
levels
and
both
positive
and
negative
controls
were
appropriate.
Treatment
with
Nuosept
95
induced
transformation
of
the
CH3/
10T1/
2
cells.
The
transformation
frequency
was
low
compared
to
the
positive
control,
there
was
no
dose
response,
and
no
statistical
methods
were
employed
to
demonstrate
significance;
however,
the
spontaneous
transformation
frequency
of
the
test
system
is
virtually
zero,
so
any
induction
of
transformed
foci
in
an
otherwise
adequate
experiment
is
regarded
as
a
positive
result.
23
Guideline
Number/
Study
Type/
Test
Substance
(%
a.
i.)
MRID
Number
(
Year)/
Citation/
Classification/
Doses
Results
(
§
84­
4)
Other
Genotoxicity
In
Vitro
Cell
Transofrmation
Assay
in
CH3/
10T1/
2
Cells
Azadioxabicyclooctane
purity
not
reported
93050023
reformat
of
00088961
Unacceptable
 
Non­
guideline
0,
0.0003,
0.001,
or
0.003
µ
L/
mL
Positive
No
Type
II
or
Type
III
foci
of
transformed
cells
were
observed
in
any
treatment
group.
Cytotoxicity
was
evident
at
the
high
dose,
demonstrated
by
reduction
of
relative
cloning
efficiency
below
50%.
Positive
and
solvent
control
values
were
appropriate.

This
study
is
unacceptable/
non­
guideline
because
it
cannot
be
ascertained
whether
T1597
could
have
been
tested
at
a
considerably
higher
dose
and
because
the
duration
of
incubation
of
the
cell
cultures
following
treatment
was
not
precisely
specified.

(
§
84­
4)
Other
Genotoxicity
In
Vitro
Cell
Transofrmation
Assay
in
CH3/
10T1/
2
Cells
Azadioxabicyclooctane
purity
not
reported
93050024
reformat
of
00088962
Acceptable
 
Non­
guideline
0,
0.0098,
0.0195,
0.039,
or
0.078
µ
L/
mL
Negative
R­
1162
was
tested
from
nontoxic
to
toxic
levels.
Positive
and
solvent
control
values
were
anomalous
when
expressed
as
relative
cloning
efficiency,
probably
due
to
low
survival
in
the
solvent
control
plates.
Since
survival
in
the
test
and
positive
control
cultures
was
not
reduced,
a
toxic
effect
of
the
solvent
is
not
indicated.
No
transformed
foci
were
observed
in
the
solvent
controls,
and
the
positive
controls
produced
the
expected
number
of
transformed
foci,
so
the
assay
remains
valid.
Treatment
with
R­
1162
did
not
induce
morphologic
transformation
of
CH3/
10T1/
2
cells.

(
§
84­
4)
Other
Genotoxicity
In
Vitro
Cell
Transofrmation
Assay
in
CH3/
10T1/
2
Cells
Azadioxabicyclooctane
purity
not
reported
93050025
reformat
of
00088963
Acceptable
 
Non­
guideline
0,
0.0006,
0.0012,
0.0024,
or
0.0048
µ
L/
mL
Negative
R­
1143
was
tested
from
nontoxic
to
toxic
levels.
Positive
and
solvent
control
values
were
appropriate.
Treatment
with
R­
1143
did
not
induce
morphologic
changes.

1
­
Daily
doses
estimated
based
on
the
analytical
concentration
of
the
test
article
in
water
and
the
water
intake
data.
24
9.0
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MRID
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J.
V.
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95
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rats
(
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(
MRID
#):
Hersham,
R.
J.
(
1984).
Summary
of
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of
a
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eye
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Bioresearch
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41641603
(
MRID
#):
Unwin,
S.
E.
(
1983).
Primary
dermal
irritation
test
of
Nuosept
®
95
in
New
Zealand
White
albino
rabbits.
Midwest
Research
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(
MRID
#):
Elliot,
P.
H.,
et
al.
(
1985).
Twenty­
one
Day
Dermal
Toxicity
Study
in
Rabbits
with
Nuosept
95/
Nuosept
C.
Huntingdon
Research
Centre.

41641606
reformat
of
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(
MRID
#):
Sasmore,
DP.
(
1980).
Final
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Effect
of
Nuosept
95
in
Rats
90­
Day
Toxicity
Study
­
Contains
dose
range
study
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Dominant
Lethal
Study
(
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870.3100).
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International.

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(
MRID
#):
Liggett,
M.
P.
and
L.
A.
McRae.
(
1990).
Acute
dermal
toxicity
to
rabbits
of
Nuosept
95
(
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report).
Huntingdon
Research
Centre
Ltd.

41699001
(
MRID
#):
Smith,
J.
A.,
et
al.
(
1988).
A
Study
of
the
Effect
of
Nuosept
95
on
Pregnancy
of
the
Rat.
Huntingdon
Research
Centre.

41728601
(
MRID
#):
O'Loughlin,
K.
G.
(
1990).
Measurement
of
micronuclei
in
bone
marrow
erythrocytes
of
Swiss­
Webster
mice
following
two
treatments
with
Nuosept
95.
SRI
International.

42650901
(
MRID
#):
Cholakis,
J.
M.
and
H.
Sprinz.
(
1983).
Acute
inhalation
toxicity
of
Nuosept
95
in
Sprague­
Dawley
rats.
Midwest
Research
Institute.

42711001
(
MRID
#):
Hamilton,
C.
M.
(
1983).
Measurement
of
unscheduled
DNA
synthesis
in
male
Fischer­
344
rat
hepatocytes
following
in
vivo
treatment
with
Nuosept
95.
SRI
International.

93050017
reformat
of
00088956
(
MRID
#):
Anonymous
(
1979).
Salmonella/
mammalianmicrosome
plate
incorporation
mutagenesis
assay.
An
Evaluation
of
Carcinogenic
Potential
of
NUOSEPT
®
95
Employing
the
C3H/
10T
½
Cell
Transformation
Assay.
EG&
G
Mason
Research
Institute.

93050018
reformat
of
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(
MRID
#):
Van
Goethen,
D.
(
1979).
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studies
on
Nuosept
95
Salmonella/
microsome
test.
Midwest
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93050019
reformat
of
00088957
(
MRID
#):
Anonymous
(
1979).
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DNA
damage/
repair
suspension
assay.
EG&
G
Mason
Research
Institute.

93050020
reformat
of
00088958
(
MRID
#):
Anonymous
(
1979).
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DNA
repair
assay.
Midwest
Research
Institute.
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93050021
reformat
of
00088959
(
MRID
#):
Anonymous
(
1980).
Evaluation
of
Nuosept
®
95
in
the
primary
rat
hepatocyte
unscheduled
DNA
synthesis
assay.
Litton
Bionetics,
Inc.

93050022
reformat
of
00088960
(
MRID
#):
Thilagar,
A.,
et
al.
(
1979).
An
Evaluation
of
Carcinogenic
Potential
of
NUOSEPT
®
95
Employing
the
C3H/
10T
½
Cell
Transformation
Assay.
EG&
G
Mason
Research
Institute.

93050023
reformat
of
00088961
(
MRID
#):
Schechtman,
M.
(
1980).
Activity
of
T1597
in
an
in
vitro
Mammalian
Cell
transformation
Assay
in
the
Absence
of
Exogenous
Metabolic
Activation.
Microbiological
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93050024
reformat
of
00088962
(
MRID
#):
Thilagar,
A.,
et
al.
(
1980).
An
Evaluation
of
Carcinogenic
Potential
of
R­
1162
(
NUOSEPT
®
95)
Employing
the
C3H/
10T
½
Cell
Transformation
System.
EG&
G
Mason
Research
Institute.

93050025
reformat
of
00088963
(
MRID
#):
Thilagar,
A.
(
1980).
An
Evaluation
of
Carcinogenic
Potential
of
R­
1143
(
NUOSEPT
®
95)
Employing
the
C3H/
10T
½
Cell
Transformation
System.
EG&
G
Mason
Research
Institute.

93050026
reformat
of
00088953
(
MRID
#):
Anonymous
(
1979).
Dominant
lethal
study
of
Nuosept
95.
SRI
International.

McMahon,
Timothy,
et
al.
(
2005).
Azadioxabicyclooctane
­
Report
of
the
Antimicrobial
Division
Toxicology
Endpoint
Selection
Committee.
