PINE
OIL
TOXICOLOGY
CHAPTER
September
8,
2004
TABLE
OF
CONTENTS
1.0
HAZARD
CHARACTERIZATION
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
3
2.0
REQUIREMENTS
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
5
3.0
DATA
GAPS
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
6
4.0
HAZARD
ASSESSMENT
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
6
4.1
Acute
Toxicity
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
6
4.2
Subchronic
Toxicity
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
7
4.3
Prenatal
Developmental
Toxicity
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
8
4.4
Reproductive
Toxicity
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
9
4.5
Mutagenicity
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
9
4.6
Neurotoxicity
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
10
4.7
Metabolism.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
10
5.0
TOXICITY
ENDPOINT
SELECTION
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
11
5.1
See
Section
9.2
for
Endpoint
Selection
Table.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
11
5.2
Dermal
Absorption
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
11
5.3
Classification
of
Carcinogenic
Potential
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
11
6.0
FQPA
CONSIDERATIONS
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
11
6.1
Special
Sensitivity
to
Infants
and
Children
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
11
6.2
Recommendation
for
a
Developmental
Neurotoxicity
Study
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
11
7.0
ENDOCRINE
DISRUPTION
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
12
8.0
OTHER
ISSUES
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
12
9.0
REFERENCES
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
13
10.0
APPENDICES
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
14
10.1
Toxicity
Profile
Summary
Tables
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
14
10.1.1
Acute
Toxicity
Table
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
14
10.1.2
Subchronic,
Chronic
and
Other
Toxicity
Tables
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
14
10.2
Summary
of
Toxicological
Dose
and
Endpoints
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
16
Page
3
of
17
1.0
HAZARD
CHARACTERIZATION
Pine
Oil
is
a
complex
combination
of
terpenes
produced
by
high
temperature
distillation
of
oil
of
turpentine
or
by
catalytic
hydration
of
terpenes.
Pine
oil
contains
a
minimum
of
60%
isomeric
secondary
and
tertiary
cyclic
terpene
alcohols
and
a
maximum
of
40%
non­
terpene
alcohol
components
consisting
of
terpene
hydrocarbons,
terpene
ether,
and
ketones.
This
complex
blend
forms
a
sole
active
ingrediet
containing
no
impurities.
The
CSPA
Pine
Oil
joint
venture
has
submitted
toxicology
data
to
the
Office
of
Pesticide
Programs
on
different
`
blends'
of
pine
oil,
namely,
Pine
Oil
Blend
1687,
Pine
Oil
Blend
110990,
and
Pine
Oil
Blend
012494.

Pine
Oil
is
registered
for
use
as
a
disinfectant,
sanitizer,
microbiocide/
microbiostat,
virucide,
and
insecticide
for
indoor
food
use,
indoor
non­
food
use,
indoor
residential
use,
indoor
medical
use,
and
aquatic
non­
food
industrial
use.
There
are
at
least
23
registered
labels
containing
pine
oil
or
pine
oil
mixed
with
a
quaternary
ammonium
compound.
Some
of
these
formulations
are
allowed
for
use
as
hard
surface
disinfectants
in
eating
establishments
where
there
may
be
the
potential
for
indirect
transfer
to
food.

The
acute
toxicity
of
Pine
Oil
is
relatively
low.
Acute
toxicity
studies
submitted
to
the
Office
of
Pesticide
Programs
show
low
toxicity
by
the
oral
route
(
Toxicity
Category
III,
oral
LD
50
=
2.7
g/
kg
),
dermal
route
(
Toxicity
category
III,
dermal
LD50
>
2000
mg/
kg),
and
inhalation
route
(
LC50
>
3.67
mg/
L,
Toxicity
category
IV).
Pine
oil
is
a
moderate
skin
irritant
(
category
III)
but
shows
a
stronger
response
as
an
eye
irritant
(
category
II).

Non­
acute
toxicity
testing
with
Pine
oil
is
limited
to
one
14­
day
dermal
toxicity
test,
one
90­
day
dermal
toxicity
test,
and
one
developmental
toxicity
test
in
the
rat.
In
the
14­
day
dermal
toxicity
test
(
submitted
under
MRID
40515401),
no
significant
adverse
effects
were
observed
up
to
a
dose
of
940
mg/
kg.
In
the
90­
day
test,
the
highest
dose
that
could
be
tested
without
significant
dermal
irritation
was
226
mg/
kg/
day,
and
no
significant
systemic
or
dermal
effects
were
observed
up
to
and
including
this
dose
level.

Developmental
toxicity
testing
of
pine
oil
in
the
rat
(
submitted
under
MRID
40515201)
showed
maternal
toxicity
in
the
form
of
excess
salivation,
alopecia,
and
ungroomed
coat
at
doses
of
600
and
1200
mg/
kg/
day
as
well
as
increased
adrenal
weight
at
1200
mg/
kg/
day.
Maternal
animals
who
died
at
the
1200
mg/
kg/
day
dose
level
exhibited
ataxia,
decreased
motor
activity,
and
impaired
righting
reflex
prior
to
death,
raising
the
possibility
of
a
neurotoxic
effect.
External
examination
of
fetuses
showed
an
increased
fetal
and
litter
incidence
of
depressed
eye
bulge
at
the
1200
mg/
kg/
day
dose.
Visceral
examination
showed
dilation
of
the
lateral
ventricles
at
1200
mg/
kg/
day,
and
increased
incidence
of
micropthalmia.
Skeletal
examination
showed
increased
fetal
and
litter
incidence
of
non­
ossified
or
incompletely
ossified
skull,
vertebrae,
manubrium,
sternebrae,
and
pelvis.
The
maternal
and
developmental
toxicity
NOAEL
were
determined
to
be
50
mg/
kg/
day
in
this
study
and
were
considered
somewhat
conservative
based
on
the
lack
of
any
other
studies
available
for
non­
acute
toxicity
of
pine
oil.
Page
4
of
17
Mutagenicity
testing
of
Pine
oil
in
various
assays
(
Ames
Salmonella
assay,
micronucleus
assay,
and
mammalian
cells
in
culture
assay)
showed
the
chemical
blend
to
be
negative,
but
the
cell
culture
assay
was
considered
unacceptable
and
must
be
repeated.

On
February
24,
2004
,
the
Antimicrobials
Division's
Toxicology
Endpoint
Selection
Committee
(
ADTC)
reviewed
the
available
Toxicology
data
for
Pine
Oil
and
discussed
endpoint
selection
for
use
as
appropriate
in
occupational/
residential
exposure
risk
assessments.
The
potential
for
increased
susceptibility
of
infants
and
children
from
exposure
to
pine
oil
was
also
evaluated
by
the
committee
in
order
to
meet
the
statutory
requirements
of
the
Food
Quality
Protection
Act
(
FQPA)
of
1996.
Page
5
of
17
2.0
TOXICOLOGY
DATA
REQUIREMENTS
The
requirements
(
CFR
158.340)
for
the
registered
uses
for
Pine
Oil
are
in
Table
1.
Use
of
the
new
guideline
numbers
does
not
imply
that
the
new
(
1998)
guideline
protocols
were
used.

Table
1.
Toxicity
Data
Requirements
for
Pine
Oil
Test
Technical
Required
Satisfied
870.1100Acute
Oral
Toxicity
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.1200Acute
Dermal
Toxicity
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.1300Acute
Inhalation
Toxicity
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.2400
Primary
Eye
Irritation
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.2500
Primary
Dermal
Irritation
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.2600
Dermal
Sensitization
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
no
870.3100
Oral
Subchronic
(
rodent)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.3150
Oral
Subchronic
(
nonrodent)
.
.
.
.
.
.
.
.
.
.
.
.
870.3250
90­
Day
Dermal
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.3465
90­
Day
Inhalation
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
yes
no
yes
no
no
yes1
870.3700
Developmental
Toxicity
(
rodent)
.
.
.
.
.
.
.
.
.
870.3700
Developmental
Toxicity
(
nonrodent)
.
.
.
.
.
.
.
870.3800Reproduction
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
yes
no
yes
yes
no
870.4100Chronic
Toxicity
(
rodent)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.4100Chronic
Toxicity
(
nonrodent)
.
.
.
.
.
.
.
.
.
.
.
.
870.4200
Oncogenicity
(
rat)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
870.4200
Oncogenicity
(
mouse)
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
no
no
no
no
870.5100Mutagenicity
C
Gene
Mutation
­
bacterial
.
.
.
870.5375Mutagenicity
C
CHO
Assay
.
.
.
.
.
.
.
.
.
.
.
.
.
870.5550Mutagenicity
C
Other
Genotoxic
Effects
.
.
.
.
yes
yes
yes
yes
no
yes
870.6100
Acute
Delayed
Neurotox.
(
hen)
.
.
.
.
.
.
.
.
.
.
870.6300
Developmental
Neurotoxicity
.
.
.
.
.
.
.
.
.
.
.
.
no
no
­­

870.7485
General
Metabolism
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
no
1although
the
90­
day
study
is
classified
as
unacceptable/
guideline,
the
data
from
the
90­
day
dermal
toxicity
study
in
conjunction
with
the
14­
day
dermal
toxicity
study
(
contained
within
the
90­
day
study)
provide
enough
data
for
regulatory
purposes.
Page
6
of
17
3.0
DATA
GAPS
Data
gaps
exist
for
a
two­
generation
reproduction
toxicity
test
and
a
90­
day
oral
toxicity
test
with
neurotoxic
endpoint
evaluation.
These
aspects
of
Pine
Oil
are
not
adequately
addressed
by
the
current
toxicology
database,
and
uncertainty
factors
have
been
used
in
the
absence
of
these
data.

In
addition,
data
gaps
exist
for
the
dermal
sensitization
assay
and
the
in
vitro
mammalian
cells
in
culture
assay.
These
studies
must
be
repeated.
The
sensitization
assay
used
an
inadequate
number
of
animals
to
make
a
determination
of
sensitization,
and
the
CHO
assay
had
significant
problems
with
the
integrity
of
the
cell
cultures
that
compromised
the
results
of
the
study.

4.0
HAZARD
ASSESSMENT
4.1
Acute
Toxicity
Adequacy
of
data
base
for
acute
toxicity:
The
data
base
for
acute
toxicity
is
considered
incomplete.
The
dermal
sensitization
assay
must
be
repeated.

The
acute
toxicity
data
on
technical
Pine
Oil
is
summarized
below
in
Table
2.

Table
2.
Acute
Toxicity
Profile
for
Pine
Oil
Guideline
No./
Study
Type
MRID
No.
Results
Toxicity
Category
870.1100
Acute
oral
toxicity­
Rat
40253502
LD50(
combined)
=
2.7
g/
kg
III
870.1200
Acute
Dermal
toxicity­
Rat
40253503
LD50
>
2000
mg/
kg
III
870.1300
Acute
inhalation
toxicity­
Rat
43375208
LC50
>
3.67
mg/
L
IV
870.2500
Dermal
irritation­
Rabbit
43375210
erythema/
edema
up
to
7
days
post
dose
III
870.2400
Primary
Eye
Irritation
43375209
irritation
lasting
up
to
16
days
II
870.2600
Skin
sensitization­
Guinea
pig
not
a
sensitizer
study
unacceptable
Page
7
of
17
4.2
Subchronic
Toxicity
Adequacy
of
data
base
for
subchronic
toxicity:
The
data
base
for
subchronic
toxicity
of
pine
oil
is
incomplete.
A
subchronic
oral
toxicity
with
neurotoxicity
evaluation
is
considered
necessary
for
adequate
evaluation
of
the
hazard
from
repeated
oral
exposure.
Only
a
90­
day
dermal
toxicity
study
exists
which
had
deficiencies.

870.3250
90­
Day
Dermal
Toxicity
B
Rat
Executive
Summary:
In
a
90­
day
dermal
toxicity
study
(
MRID
40515401),
Pine
Oil
Blend
CSMA
1687
(%
a.
i.
and
batch/
lot
number
not
reported)
was
applied
to
the
shaved
skin
of
10
Crl:
CD
®
BR
rats/
sex/
dose
at
dose
levels
of
0,
50,
113,
and
226
mg/
kg
bw/
day,
6
hours/
day
for
5
days/
week
during
a
90­
day
period.
The
application
method
involved
applying
the
test
article
dosage
to
one
quadrant
of
the
shaved
area
on
the
back
of
each
rat
(
reportedly
25%
of
the
surface
area
was
shaved)
with
each
quadrant
being
exposed
once
every
four
days
throughout
the
study
period.

There
were
no
compound
related
effects
on
mortality,
clinical
signs,
body
weight,
food
consumption,
hematology,
clinical
chemistry,
organ
weights,
or
gross
pathology.
No
significant
adverse
dermal
reactions
were
observed
due
to
treatment;
very
slight
to
slight
erythema
was
observed
in
the
female
treated
rats,
desquamation
and
scabbing
occurred
sporadically
in
male
and
female
treated
rats,
and
microscopic
changes
included
minimal
to
mild
acanthosis
and
minimal
hyperkeratosis
with
similar
incidences
in
both
the
controls
and
226­
mg/
kg/
day
rats.
The
skin
in
the
low­
dose
animals
did
exhibit
slight
injury
with
evidence
of
desquamation,
scabbing,
and
very
slight
to
slight
erythema,
however.
Again,
these
dermal
irritation
responses
were
sporadic
and
of
a
minimal
severity
but
if
the
skin
had
been
treated
in
accordance
with
guidelines
with
the
entire
shaved
application
area
being
treated
at
least
5
days/
week
for
6
hours
then
more
substantial
injury
to
the
dermis
might
have
occurred.

A
slight
increase
was
observed
in
the
incidence
of
tingable
bodies
in
macrophages
in
the
thymus
of
treated
rats.
There
was
no
dose­
response
in
incidence
or
severity
of
this
finding.
Therefore,
the
systemic
toxicity
no­
observable­
adverse­
effect­
level
(
NOAEL)
is

226
mg/
kg/
day
due
to
a
lack
of
a
treatment­
related
response
in
any
of
the
dose
groups.
A
lowest­
observable­
adverse­
effect­
level
(
LOAEL)
could
not
be
determined
due
to
a
lack
of
treatment­
related
effects.

A
dermal
toxicity
LOAEL
could
not
be
determined;
the
dermal
NOAEL
is

226
mg/
kg/
day
based
on
the
lack
of
consistent
dose­
related
effects
on
the
skin.
It
is
noted,
however,
that
in
the
14­
day
range­
finding
study,
a
dermal
NOAEL
of
940
mg/
kg/
day
could
be
identified.
An
accurate
determination
of
a
dermal
NOAEL
is
complicated
by
the
dosing
methodology
used
in
both
the
14­
day
and
90­
day
studies,
in
which
the
same
area
of
shaved
skin
was
not
dosed
each
day.
Page
8
of
17
This
90­
day
dermal
toxicity
study
in
the
rat
is
unacceptable­
guideline.
Although
the
study
does
not
satisfy
the
guideline
requirement
for
a
90­
day
dermal
toxicity
study
(
OPPTS
870.3250;
OECD
411)
in
the
rat,
the
data
from
the
14­
day
study
provides
enough
additional
information
in
conjunction
with
the
90­
day
study
to
determine
a
NOAEL
value
for
regulatory
purposes.

870.3465
90­
Day
Inhalation
B
Rat
A
90­
day
inhalation
toxicity
study
is
not
available
for
Pine
Oil.
An
oral
endpoint
was
selected
for
use
in
risk
assessment
if
necessary.

4.3
Prenatal
Developmental
Toxicity
Adequacy
of
data
base
for
Prenatal
Developmental
Toxicity:
The
data
base
for
prenatal
developmental
toxicity
is
considered
complete.
No
additional
studies
are
required
at
this
time.

870.3700
Prenatal
Developmental
Toxicity
Study
­
Rat
Executive
Summary:
Pine
Oil
blend
1687
was
administered
to
25
mated
female
Sprague­
Dawley
rats
by
gavage
at
doses
of
0,
50,
600,
or
1200
mg/
kg/
day
from
days
6­
15
of
gestation.
Abnormal
clinical
signs
consisting
of
excess
salivation,
alopecia,
and
ungroomed
coat
were
observed
in
increased
incidence
at
the
600
and
1200
mg/
kg/
day
dose
level
in
maternal
animals.
Body
weight
gain
at
the
1200
mg/
kg/
day
dose
level
was
decreased
60%
during
the
dosing
period,
and
37%
when
corrected
for
gravid
uterine
weight.
Overall
corrected
weight
gain
was
decreased
17%
for
days
0­
20
of
the
study.
Food
consumption
by
contrast
was
decreased
27%
during
the
treatment
phase
and
5%
overall
(
days
0­
20).
Therefore
it
is
possible
that
the
effect
on
body
weight
was
treatment
related
at
the
1200
mg/
kg/
day
dose
level.
Adrenal
gland
weight
was
significantly
increased
in
maternal
animals
at
the
1200
mg/
kg/
day
dose
level.
Maternal
deaths
were
increased
at
the
1200
mg/
kg/
day
dose
level,
where
6
animals
died
after
three
to
seven
doses
had
been
administered.
These
animals
demonstrated
ataxia,
decreased
motor
activity,
and
impaired
righting
reflex
prior
to
death.
Mean
fetal
weight
was
significantly
decreased
at
the
1200
mg/
kg/
day
dose
level
vs.
control,
but
there
were
no
other
abnormal
cesarean
section
observations.

External
examination
of
fetuses
showed
an
increased
fetal
and
litter
incidence
of
depressed
eye
bulge
at
the
1200
mg/
kg/
day
dose.
Visceral
examination
showed
dilation
of
the
lateral
ventricles
at
1200
mg/
kg/
day,
and
increased
incidence
of
micropthalmia.
Skeletal
examination
showed
increased
fetal
and
litter
incidence
of
non­
ossified
or
incompletely
ossified
skull,
vertebrae,
manubrium,
sternebrae,
and
pelvis.
Page
9
of
17
The
dose
spacing
in
this
study
(
i.
e.
50,
600,
1200
mg/
kg/
day)
along
with
the
appearance
of
effects
mainly
at
1200
but
also
at
600
mg/
kg/
day
suggests
that
effects
could
have
arisen
at
doses
lower
than
1200
mg/
kg/
day
but
greater
than
600
mg/
kg/
day.
This,
in
conjunction
with
the
fact
that
there
is
only
one
developmental
toxicity
study
for
Pine
Oil
submitted
to
the
Agency,
leads
to
a
conservative
effect
level
(
LOAEL)
of
600
mg/
kg/
day
with
a
NOAEL
of
50
mg/
kg/
day
for
both
maternal
and
developmental
toxicity.

4.4
Reproductive
Toxicity
Adequacy
of
data
base
for
Reproductive
Toxicity:
The
data
base
for
reproductive
toxicity
is
considered
incomplete.
Reproductive
toxicity
is
not
represented
in
any
of
the
available
toxcity
data
for
Pine
oil.
As
there
are
indirect
food
uses
for
this
chemical,
and
in
accordance
with
the
interim
working
policy
of
AD
regarding
indirect
food
use
data
requirements,
a
reproductive
toxicity
study
must
be
submitted
in
order
to
properly
characterize
these
risks.

4.5
Mutagenicity
Gene
Mutation
870.5100,
Ames
Salomella
Assay
Accession
#
(
MRID
43375212)
This
study
is
classified
as
acceptableguideline
In
an
Ames
Salmonella
assay,
Pine
oil
blend
012494
was
used
in
strains
TA98,
TA100,
TA1535,
TA1537
at
concentrations
of
0.1,
1.0,
100,
1000,
and
10000
ug/
plate.
No
evidence
of
an
increase
in
induced
mutant
colonies
over
background
in
the
absence
or
presence
of
metabolic
activation.

Cytogenetics
870.5375,
In
vitro
mammalian
cells
in
culture
This
study
is
unacceptable
In
an
in
vitro
cytogenetics
assay
using
Chinese
hamster
ovary
(
CHO)
cells,
CHO
cells
were
exposed
to
Pine
Oil
blend
1687
at
100,
130,
170,
200,
250
nl/
ml
(
no
metabolic
activation)
or
50,
100,
200,
300,
400
nl/
ml
(
S9
activation).
No
evidence
of
induction
of
mutant
colonies
over
background.
Study
not
acceptable
and
must
be
repeated.
Cells
not
checked
for
mycoplasma
contamination,
and
several
cultures
were
lost
to
contamination
during
conduct
of
the
assay.

Other
Genotoxicity
870.5550,
Micronucleus
assay
MRID
40341403
This
study
is
acceptable
Single
i.
p
injection
with
Pine
Oil
blend
1687
at
doses
of
0,
116,
578,
1155
mg/
kg.
Compound
toxic
at
the
1155
mg/
kg
dose.
No
significant
increase
in
frequency
of
micronucleated
polychromatic
erythrocytes
in
bone
marrow
at
any
test
concentration
or
harvest
time.
Page
10
of
17
4.6
Neurotoxicity
Adequacy
of
data
base
for
Neurotoxicity:
The
neurotoxicity
of
pine
oil
can
be
addressed
through
the
conduct
of
neurotoxicity
screening
as
specified
in
the
90­
day
oral
toxicity
guideline.

870.6100
Delayed
Neurotoxicity
Study
­
Hen
Study
not
required.

870.6300
Developmental
Neurotoxicity
Study
Study
not
required
4.7
Metabolism
Study
is
not
required.
Page
11
of
17
5.0
TOXICITY
ENDPOINT
SELECTION
5.1
See
Section
9.2
for
Endpoint
Selection
Table.

5.2
Dermal
Absorption
Dermal
absorption
data
are
not
needed
for
pine
oil.
Route
specific
data
are
available
from
the
14­
day
and
90­
day
dermal
toxicity
studies
from
which
an
endpoint
was
selected
for
this
route
of
exposure.

5.3
Classification
of
Carcinogenic
Potential
Pine
oil
has
not
been
classified
as
to
carcingenicity;
however,
these
data
are
not
required
at
this
time.

6.0
FQPA
CONSIDERATIONS
6.1
Special
Sensitivity
to
Infants
and
Children
The
Antimicrobials
Division
Toxicology
Endpoint
Selection
Committee
(
ADTC)
concluded
that
there
is
no
evidence
for
susceptibility
to
exposure
to
pine
oil
from
the
limited
data
available.
The
ADTC
also
concluded
that
the
hazard­
based
special
FQPA
safety
factor
can
be
removed
(
1X)
when
assessing
dietary
risks
resulting
from
the
uses
of
pine
oil,
based
on
the
use
of
conservative
NOAEL
values
from
the
developmental
toxicity
study.
However,
a
database
uncertainty
factor
of
10x
applies
to
pine
oil,
based
on
the
lack
of
specific
neurotoxicity
endpoints
and
a
reproductive
toxicity
study.

6.2
Recommendation
for
a
Developmental
Neurotoxicity
Study
A
developmental
neurotoxicity
study
is
not
required
pine
oil.

7.0
Endocrine
Disruption
The
Agency
is
required
under
the
Federal
Food,
Drug
and
Cosmetic
Act
(
FFDCA),
as
amended
by
FQPA,
to
develop
a
screening
program
to
determine
whether
certain
substances
(
including
all
pesticide
active
and
other
ingredients)
"
may
have
an
effect
in
humans
that
is
similar
to
an
effect
produced
by
a
naturally
occurring
estrogen,
or
other
such
endocrine
effects
as
the
Administrator
may
designate."
Following
the
recommendations
of
its
Endocrine
Disruptor
Screening
and
Testing
Advisory
Committee
(
EDSTAC),
EPA
determined
that
there
was
scientific
bases
for
including,
as
part
of
the
program,
the
androgen
and
thyroid
hormone
systems,
in
addition
to
the
estrogen
hormone
system.
EPA
also
adopted
EDSTAC's
recommendation
that
the
Program
include
evaluations
of
potential
effects
in
wildlife.
For
pesticide
chemicals,
EPA
will
use
FIFRA
and,
to
the
extent
that
effects
in
wildlife
may
help
determine
whether
a
substance
may
have
an
effect
in
humans,
FFDCA
authority
to
require
the
wildlife
evaluations.
As
the
science
develops
Page
12
of
17
and
resources
allow,
screening
of
additional
hormone
systems
may
be
added
to
the
Endocrine
Disruptor
Screening
Program
(
EDSP).

8.0
OTHER
ISSUES
In
the
review
of
the
available
toxicology
database
for
pine
oil,
the
ADTC
included
a
10x
factor
for
the
lack
of
data
on
reproductive
toxicity
(
required
data
for
indirect
food
uses)
and
neurotoxicity
(
from
observations
of
neurotoxicity
in
the
available
data).
Since
the
production
of
this
risk
assessment,
the
registrant
has
made
the
Agency
aware
of
data
submitted
to
EPA
under
the
high
production
volume
program
that
in
their
view
addresses
reproductive
concerns
of
pine
oil.
These
data
have
not
been
formally
reviewed
by
the
Office
of
Pesticide
Programs
for
compliance
with
the
reproductive
or
neurotoxicity
guidelines,
and
only
summary
data
were
available
on
the
Agency's
High
Production
Volume
Challenge
Program.
With
regard
to
the
neurotoxicity
concern,
the
registrant
stated
that
there
was
a
90­
day
oral
toxicity
study
conducted
for
the
Food
and
Drug
Administration
in
1967
that
in
their
view
supports
the
lack
of
neurotoxicity
of
pine
oil.
As
with
the
reproductive
toxicity
data,
the
original
report
is
not
currently
available
to
the
EPA,
and
only
a
summary
was
provided.

If
such
studies
provide
relevant
and
useful
information
to
the
Agency,
the
data
could
be
considered
in
refining
any
concerns
with
respect
to
reproductive
toxicity
or
neurotoxicity.
However,
at
the
present
time,
there
is
not
enough
information
from
these
studies
provided
to
reach
a
different
conclusion
than
previously
stated.
Page
13
of
17
9.0
REFERENCES
in
MRID
order
MRID
#
REFERENCES
40515201
Dearlove
G.
(
1988)
"
Developmental
Toxicity
(
embryo/
fetal
toxicity
potential)
Study
of
Pine
Oil
Blend
1687
administered
orally
via
gavage
to
Crl:
CD
Presumed
Pregnant
Rats."
Argus
Research
Laboratories,
Inc.

43375212
Ferrante
S.
(
1994)
"
Pine
Oil:
Salmonella
typhimurium
Reverse
Mutation
Assay."
Toxikon
Corporation.

40515401
Lavelgia
J,
Nass
D.
(
1988).
"
90­
Day
Dermal
Toxicity
Study
in
Rats
with
Pine
Oil
Blend
CSMA
1687."
WIL
Research
Laboratories,
Inc.

40253503
Naas
D.
(
1987)
"
Acute
Dermal
Toxicity
(
LD50)
study
in
Albino
Rabbits
with
Pine
Oil
Blend
CSMA
1687."
WIL
Research
Laboratories,
Inc.

40253502
Naas
D.
(
1987)
"
Acute
Oral
Toxicity
(
LD50)
study
in
Albino
Rats
with
Pine
Oil
Blend
CSMA
1687."
WIL
Research
Laboratories,
Inc.

40341403
Putman
D.
(
1987)
"
Micronucleus
Cytogenetic
Assay
in
Mice,
Final
Report."
Microbiological
Associates,
Inc.

43375208
Robbins
G.
(
1994).
"
Pine
Oil:
Acute
Inhalation
Toxicity
Study
in
Rats."
Cosmopolitan
Safety
Evaluation,
Inc.

43375211
Robbins
G.
(
1994).
"
Pine
Oil:
Dermal
Sensitization
(
Buehler)
in
Guinea
Pigs."
Cosmopolitan
Safety
Evaluation,
Inc.

43375210
Robbins
G.
(
1994).
"
Pine
Oil:
Primary
Dermal
Irritation
Study
in
Rabbits."
Cosmopolitan
Safety
Evaluation,
Inc.

43375209
Robbins
G.
(
1994).
"
Pine
Oil:
Primary
Eye
Irritation
Study
in
Rabbits."
Cosmopolitan
Safety
Evaluation,
Inc.

40341403
Yang
L.
(
1987)
"
CHO/
HGPRT
Mutation
Assay."
Microbiological
Associated,
Inc.
Page
14
of
17
10.0
APPENDICES
Tables
for
Use
in
Risk
Assessment
10.1
Toxicity
Profile
Summary
Tables
10.1.1
Acute
Toxicity
Table
­
See
Section
4.1
10.1.2
Subchronic,
Chronic
and
Other
Toxicity
Tables
Guideline
No./
Study
Type
MRID
No.
(
year)/
Classification/
Doses
Results
870.3250
90­
day
Dermal
Toxicity
in
Rats
40515401
(
1988)
unacceptable
Doses:
0,
50,
113,
226
mg/
kg/
day
to
shaved
skin,
6
hrs/
day,
5days/
wk,
13
weeks
Systemic
Toxicity
NOAEL
=
>
226
mg/
kg/
day
Systemic
Toxicity
LOAEL
=
>
226
mg/
kg/
day
NOAEL
can
be
estimated
from
a
14­
day
rangefinding
study
that
showed
a
dermal
NOAEL
of
940
mg/
kg/
day.

870.3700
Developmental
Toxicity
in
Rats
40515201
acceptable/
guideline
Doses:
0,
50,
600,
1200
mg/
kg/
day
Maternal/
Developmental
Toxicity
NOAEL
=
50
mg/
kg/
day
Maternal/
Developmental
Toxicity
LOAEL
=
600
mg/
kg/
day
The
dose
spacing
in
this
study
(
i.
e.
50,
600,
1200
mg/
kg/
day)
along
with
the
appearance
of
effects
mainly
at
1200
but
also
at
600
mg/
kg/
day
suggests
that
effects
could
have
arisen
at
doses
lower
than
1200
mg/
kg/
day
but
greater
than
600
mg/
kg/
day.
This
leads
to
a
conservative
assumption
of
50
mg/
kg/
day
as
the
NOAEL
for
maternal
and
developmental
toxicity.

Mutagenicity
Gene
Mutation
870.5100
Ames
Salomella
Assay
43375212
(
1994)
acceptable
concentrations
of
0.1,
1.0,
10,
100,
1000,
10000

g/
plate
No
evidence
of
an
increase
in
induced
mutant
colonies
over
background
in
the
absence
or
presence
of
metabolic
activation.
Page
15
of
17
Guideline
No./
Study
Type
MRID
No.
(
year)/
Classification/
Doses
Results
In
vitro
Cytogenetics
870.5300
40341403
unacceptable
100,
130,
170,
200,
250
ng/
mL
(­
S9);
50,
100,
200,
300,
400
nL/
mL
(+
S9)
No
evidence
of
induction
of
mutant
colonies
over
background.
Study
not
acceptable
and
must
be
repeated.
Cells
not
checked
for
mycoplasma
contamination,
and
several
cultures
were
lost
to
contamination
during
conduct
of
the
assay.

870.5395
Micronucleus
Assay
40341403
acceptable
i.
p.
doses
of
0,
116,
578,
1155
mg/
kg
No
significant
increase
in
frequency
of
micronucleated
polychromatic
erythrocytes
in
bone
marrow
at
any
test
concentration
or
harvest
time.

Neurotoxicity
870.6300
Developmental
neurotoxicity
Not
required
Page
16
of
17
10.2
Summary
of
Toxicological
Dose
and
Endpoints
for
Pine
Oil
for
Use
in
Human
Risk
Assessment1
Exposure
Scenario
Dose
(
mg/
kg/
day)
used
in
risk
assessment
UF
/
MOE
Uncertainty
Factors
Study
and
Toxicological
Effects
Dietary
Risk
Assessments
Acute
Dietary
(
gen
pop)
NOAEL
of
50
mg/
kg/
day
Acute
RfD
=
0.05
mg/
kg/
day
UF
=
1000
Developmental
toxicity
study
in
rats
.
Maternal
LOAEL
of
600
mg/
kg/
day
based
on
clinical
observations
of
toxicity,
decreased
body
weight,
weight
gain,
food
consumption
Acute
Dietary
(
females
13+)
NOAEL
of
50
mg/
kg/
day
Acute
RfD
=
0.05
mg/
kg/
day
UF
=
1000
Developmental
toxicity
study
in
rats
Developmental
toxicity
LOAEL
of
600
mg/
kg/
day
based
on
decreased
fetal
weight,
fetal
malformations,
retardation
of
ossification.

Chronic
Dietary
NOAEL=
50
Chronic
RfD
=
0.016
mg/
kg/
day
UF
=
3000
Developmental
toxicity
study
in
rats
Maternal
LOAEL
of
600
mg/
kg/
day
based
on
clinical
observations
of
toxicity,
decreased
body
weight,
weight
gain,
food
consumption
Incidental
Oral
Short­
Term
(
1
­
30
Days)
NOAEL=
50
mg/
kg/
day
UF=
1000
Developmental
toxicity
study
in
rats
Maternal
LOAEL
of
600
mg/
kg/
day
based
on
clinical
observations
of
toxicity,
decreased
body
weight,
weight
gain,
food
consumption
Incidental
Oral
Intermediate­
Term
(
1
­
6
Months)
NOAEL=
50
mg/
kg/
day
UF
=
3000
Developmental
toxicity
study
in
rats
Maternal
LOAEL
of
600
mg/
kg/
day
based
on
clinical
observations
of
toxicity,
decreased
body
weight,
weight
gain,
food
consumption
Non­
Dietary
Risk
Assessments
Dermal
Short
­
term
Endpoint
not
required
Page
17
of
17
Exposure
Scenario
Dose
(
mg/
kg/
day)
used
in
risk
assessment
UF
/
MOE
Uncertainty
Factors
Study
and
Toxicological
Effects
Dermal
intermediate
and
long­
term
NOAEL
>
226
mg/
kg/
day
UF
=
100
90­
day
dermal
toxicity
study
in
rats
Dermal
NOAEL
of
>
226
mg/
kg/
day
(
highest
dose
tested)

Inhalation
All
durations
LOAEL
=
50
mg/
kg/
day
mg/
L
UF
=
1000
(
ST,
IT)

UF
=
3000
(
LT)
Developmental
toxicity
in
rats
Maternal
LOAEL
of
600
mg/
kg/
day
based
on
clinical
observations
of
toxicity,
decreased
body
weight,
weight
gain,
food
consumption
Cancer
no
data
1
UF
=
uncertainty
factor,
FQPA
SF
=
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
