ECOLOGICAL
RISK
ASSESSMENT
Re­
registration
"
Acid
Blue
9"(
Erioglaucine)
and
"
Acid
Yellow
23"
(
Tartrazine
)
Dyes
Used
together
in
the
End­
use
Products
Aquashade,
Aquashade
OA,
Admiral
Liquid,
Admiral
WSP
and
Pond
Care
AlgaBlocker
for
Control
of
Algal
Growth
and
Other
Undesirable
aquatic
Plants
Acid
Yellow
23:
PC
Code
Number:
110302
CAS
Registry
No.:
1934­
21­
0
Acid
Blue
9
(
trisodium
salt)
PC
Code
Number:
110301
CAS
Registry
No.:
Trisodium
salt:
2650­
18­
2
Triammonium
salt:
3844­
45­
9
FD&
C
Blue
1
In
theTriammonium
salt.
the
counter
cation
is
ammonium
instead
of
sodium.
The
nature
of
the
counter
cation
is
not
relevant
because
it
is
the
anion
what
is
responsible
for
color
of
the
dyes
Reviewers:
Ecological
Effects:
James
Goodyear,
Biologist
Environmental
Fate:
Silvia
C.
Termes,
Chemist
Secondary
Reviewers:
Mark
Corbin
Stephanie
Syslo
Brian
Anderson
Branch
Chief:
Daniel
Rieder
Ecological
Risk
Branch
III
Environmental
Fate
and
Effects
Division
Table
of
Contents
I.
Executive
Summary                  ..
3
A.
Nature
of
the
Chemical
Stressor
B.
Potential
Risks
to
Non­
target
Organisms
C.
Conclusions
to
the
Exposure
Characterization
D.
Conclusions
to
the
Effects
Characterization
E.
Uncertainties
and
Data
Gaps
II.
Problem
Formulation                  
8
A.
Stressor
Source
and
Identity
B.
Receptors
C.
Assessment
End­
points
D.
Conceptual
Model
E.
Measures
to
Evaluate
Risk
Hypothesis
F.
Measures
of
Ecosystems
and
Receptor
Characteristics
G.
Analysis
Plan
III.
Analysis                       
19
A.
Use
Characterization
B.
Exposure
Characterization
C.
Ecological
Effects
Characterization
IV.
Risk
Characterization                  
32
V.
Literature
Cited                    .
39
Appendices
Appendix
A
Data
Gaps
Appendix
B
Environmental
Fate
Appendix
C
Aquatic
Exposure
and
Modeling
Appendix
D
Ecological
Toxicity
and
Uncertainty
in
Ecological
Toxicity
Data
Appendix
E
Status
of
Data
Requirements
Appendix
F
Environmental
Fate
Bibliography
Appendix
G
Ecological
Toxicity
Bibliography
Appendix
H
Endangered
and
Threatened
Species
Appendix
K
Risk
Quotient
Calculation
and
Interpretation
Appendix
L
Papers
accepted
for
ECOTOX,
but
not
used
in
science
chapter
I.
Executive
Summary
A.
Nature
of
the
Chemical
Stressor
The
chemical
stressors
are
the
dyes
Acid
Yellow
23
and
Acid
Blue
9,
which
are
used
together
in
several
end­
use
products
intended
to
control
algae
and
other
undesirable
aquatic
plants
in
artificial,
confined
water
bodies
(
ornamental
ponds,
golf
course
lakes,
fountains;
hatcheries;
swimming
pools).
The
dyes
block
sunlight
energy
necessary
for
algal
growth
and,
at
the
same
time,
impart
a
greenish
blue
coloration
to
the
water.
The
two
dyes
are
also
regulated
under
FIFRA
as
"
inerts"
used
as
colorants
in
some
pesticide
formulations.
In
addition,
they
are
regulated
under
the
Food,
Drug,
and
Cosmetic
Act
as
colorants
for
food
(
food
additives),
drug
formulations,
and
cosmetics1.

The
specific
end­
use
products
containing
Acid
Blue
9
and
Acid
Yellow
23
are:
(
a)
Pond
Care
Algae
Blocker
(
2.36%
Acid
Blue
9
and
0.24%
Acid
Yellow
23;
EPA
Reg.
No.
8709­
6);
(
b)
Aquashade
(
23.6%
Acid
Blue
9
and
2.39%
Acid
Yellow
23,
EPA
Reg.
No.
3306­
1);
(
c)
Aquashade
OA
(
2.36%
Acid
Blue
9
and
0.24%
Acid
Yellow
23;
EPA
Reg.
No.
33068­
2);
(
d)
Admiral
Liquid
(
15.31%
Acid
Blue
9
and
1.00
%
Acid
Yellow
23;
EPA
Reg.
No.
67064­
2);
(
e)
Admiral
WSP
(
49.72%
Acid
Blue
9
and
3.27%
Acid
Yellow
23;
EPA
Reg.
No.
67064­
1).
Except
for
Admiral
WSP,
all
the
products
are
liquid
formulations.

The
Agency
had
assigned
the
PC
110301
to
Acid
Blue
9,
110302
to
Acid
Yellow
23,
and
110303
to
Aquashade,
the
mixture
of
23.6%
Acid
Blue
9
and
2.39%
Acid
Yellow
23
(
an
end­
use
product).
In
this
document,
the
names
Acid
Blue
9
and
Acid
Yellow
23
were
used
when
the
percentage
of
each
dye
was
not
specified
or
when
individual
dyes
were
described
separately.
"
Aquashade"
was
used
when
referring
to
the
test
substance
used
in
the
ecological
toxicity
studies.

B.
Potential
Risks
to
Non­
target
Organisms
Based
on
available
toxicity
data
and
the
exposures
expected
when
used
according
to
the
label,
the
Environmental
Fate
and
Effects
Division
(
EFED)
does
not
believe
that
"
Aquashade"
and
other
products
containing
Acid
Blue
9
and
Acid
Yellow
23
2
1
The
specific
names
used
for
"
Acid
Blue
9"
under
the
FDCA
are
"
FD&
C
Blue
No.
1"
for
the
trisodium
salt
and
"
D&
C"
"
Blue
No.
4"
for
the
triammonium
salt
(
also
known
as
erioglaucine).
Both
are
salts
of
the
same
anion.
Acid
Yellow
23
is
FD&
C
"
Yellows",
also
known
as
Tartrazine.

2
The
ecological
toxicity
studies
were
conducted
with
an
Aquashade
end­
use
product
containing
23.6%
Acid
Blue
9
and
2.365
Acid
Yellow
23.
The
PC
Number
assigned
to
the
product
is
110303,
which
has
created
some
confusion
because
this
product
is
a
combination
of
two
dyes,
each
with
and
assigned
PC
Number,
4
pose
a
direct
acute
risk
to
freshwater
fish
and
invertebrates,
birds,
amphibians,
reptiles
or
mammals,
including
endangered
species.
These
dyes
are
not
expected
to
harm
terrestrial
animals
that
drink
treated
water.
Note
that
even
though
we
do
not
have
toxicity
data
on
amphibians
and
reptiles,
aquatic
phase
amphibians
are
represented
by
fish,
and
reptiles
and
terrestrial
phase
amphibians
are
represented
by
birds.
It
is
possible
that
endangered
terrestrial
animals
may
be
affected
indirectly
by
loss
of
food
in
water
bodies
where
treatment
occurs.
The
extremely
limited
area
in
which
this
may
occur,
given
the
label­
specified
water
bodies
and
the
method
of
application
(
i.
e.
by
hand)
may
allow
a
further
analysis
to
reduce
this
apparent
possibility
to
be
almost
nonexistent.
However,
this
analysis
has
not
been
completed.

Aquashade
is
an
aquatic
herbicide
and
will
kill
algae
and
nontarget
submerged
aquatic
plants.
If
any
treated
habitats
contained
endangered
aquatic
plants
at
the
time
of
application,
effects
are
possible.
Exposure
to
emerged
plants
and
terrestrial
plants
is
assumed
to
be
low,
resulting
in
little
or
no
risk.

The
Environmental
Fate
and
Effects
Division
is
unable
to
assess
the
potential
chronic
risk
to
animals
due
to
lack
of
data
and
therefore
cannot
conclude
that
potential
chronic
risk
to
these
taxa
(
and
the
taxa
for
which
they
are
surrogates)
does
not
exist.
However,
because
of
its
low
acute
toxicity,
mode
of
action
which
is
not
toxicological,
and
because
Acid
Yellow
23
dye
is
shown
not
to
be
chronically
toxic
to
mammals,
EFED
does
not
believe
that
chronic
effects
are
likely
and
therefore,
chronic
studies
are
not
needed.

C.
Conclusions­
Exposure
Characterization
Status
of
Data
Requirements
All
of
the
environmental
fate
data
requirements
were
placed
in
"
Reserved"
in
1993,
depending
on
the
results
of
the
required
ecological
toxicity
studies.
Because
the
risk
assessment
did
not
identify
risks
to
fish,
aquatic
invertebrates,
or
mammals,
the
environmental
fate
studies
may
be
"
Waived".
The
environmental
fate
in
this
assessment
is
qualitative,
based
mostly
on
data
from
the
open
literature
on
structurally
related
dyes.
Although
structure­
activity
relationship
(
SAR)
estimates
using
EPI
Suite/
EPIWIN3
were
performed,
these
programs
do
not
adequately
estimate
physical
and
chemical
properties
of
salts,
particularly
those
of
a
large
anion
such
as
in
Acid
Blue
9
and
Acid
Yellow
23.
For
this
reason,
the
qualitative
assessment
relies
on
open
literature
information.

No
environmental
fate
data
from
Subdivision
N
guideline
studies
are
required
for
the
present
uses.
If
new
uses
and/
or
changes
in
rate
or
method
of
application
or
products
are
proposed,
the
need
for
new
data
will
be
reevaluated.

3
EPI
(
Estimation
Programs
Interface)
SuiteTM
(
formerly
known
as
EPIWIN)
5
Exposure
in
the
Environment
Unlike
the
uses
on
food,
drugs
and
cosmetics,
the
dyes
are
exposed
to
an
open
(
but
contained)
aquatic
environment
when
used
as
herbicides.
Because
the
concentrated
products
are
added
directly
to
a
water
body,
the
dyes
(
which
do
not
react
chemically
with
each
other)
become
diluted
in
the
treated
water
body.
Neither
runoff
nor
spray
drift
are
routes
of
exposure
because
a
specified
amount
of
product
is
directly
applied
to
the
water
body
to
attain
recommended
target
concentrations
of
the
product
of
either
"
1
ppm
or
2
ppm",
depending
on
the
weed
to
be
controlled.
To
attain
these
target
concentrations,
the
labels
specify
the
volume
of
product
to
be
added
per
volume
of
water
to
be
treated.
These
maximum,
target
concentrations
were
assumed
to
be
maintained
after
treatment.
That
is,
no
degradation
was
assumed.

The
major
route
of
dissipation
of
the
dyes
in
an
aquatic
environment
is
likely
indirect
photolysis,
which
depends
of
on
the
nature
and
concentration
of
natural
photosensitizers
as
well
as
on
the
geographical
location
where
and
season
when
the
products
are
used.
However,
biotransformation
under
anaerobic
conditions
may
also
contribute
to
the
dissipation
of
each
dye.
The
specific
chemical
nature
of
photoproducts
and
metabolites
is
not
known.

The
dyes
are
predominantly
associated
with
the
water
column
and
have
no
potential
to
volatilize
from
water.
Although
the
dyes
are
not
applied
to
soils,
they
are
unlikely
to
volatilize
from
soils.
Acid
Blue
9
and
Acid
Yellow
23
do
not
have
the
potential
to
bioaccumulate
in
fish.

D.
Conclusions­
Effects
Characterization
Aquashade
is
a
formulation
that
contains,
as
its
active
ingredients,
a
yellow
and
a
blue
dye.
These
dyes,
Acid
Yellow
23
and
Acid
Blue
9
are
also
used
as
food
drug
and
cosmetic
colorants
for
human
consumption.
Their
mode
of
herbicidal
action
is
not
toxicological,
therefore
little
toxicity
is
expected.
The
toxicity
information
available
to
characterize
the
toxicity
to
wildlife
includes
testing
with
formulations
such
as
Aquashade
and
Admiral
WSP
and
also
testing
with
the
individual
dyes.
Data
used
to
characterize
effects
include
studies
conducted
by
a
registrant
(
testing
with
Aquashade
and
Admiral
WSP)
and
studies
reported
in
published
literature.
Published
literature
were
obtained
by
the
Office
of
Research
and
Development's
(
ORD)
Middle
Ecological
Division
in
Duluth
through
their
literature
searched
conducted
as
part
of
the
ECOTOX
program.
All
testing
indicates
that
these
dyes,
whether
alone,
or
in
formulations
cause
no
toxicity
to
fish,
aquatic
invertebrates,
birds
or
mammals
at
doses
and
concentrations
far
above
those
expected
in
the
environment.
No
studies
were
required
or
found
that
tested
the
dyes
or
formulations
on
aquatic
or
terrestrial
plants.
In
as
much
as
this
is
a
control
agent
for
submerged
aquatic
vegetation
and
algae,
it
is
assumed
to
have
an
adverse
effect
on
aquatic
plants.
The
label
also
warns
that
if
it
gets
on
emergent
vegetation,
some
burning
will
occur.
This
indicates
that
these
dyes
and
formulations
may
be
toxic
to
terrestrial
plants.
However,
the
method
of
application
would
preclude
exposure
to
terrestrial
plants,
so
toxicity
to
this
group
of
organisms
is
not
considered
necessary.
If
treated
waters
are
used
for
irrigation,
exposure
to
terrestrial
plants
may
be
possible
and
toxicity
testing
for
terrestrial
plants
would
be
needed
to
assess
risk.
6
E.
Uncertainties
and
Data
Gaps
1.
Exposure
Exposure
to
the
dyes
were
based
on
the
target
dilution
concentration
of
a
directly
applied
volume
of
product
to
a
volume
of
water
body
to
attain
the
concentrations
specified
in
the
labels.
(
1
or
2
ppm;
1
or
2
mg/
L).
Thus,
the
exposure
concentrations
in
water
are
the
same
as
the
target
concentrations
in
the
labels.
It
is
assumed
that
the
dyes
are
stable
in
the
water
and
that
the
target
concentration
remains
constant.
That
is,
routes
and
rates
of
dissipation
were
not
taken
into
account,
as
data
are
not
available.

Indirect
photolysis
has
been
identified
as
the
major
route
of
dissipation
of
the
dyes
in
aquatic
systems.
However,
the
specific
nature
of
transformation
products
for
Acid
Blue
9
and
Acid
Yellow
23
is
not
known.
Although
no
kinetics
data
are
available
to
assess
how
fast
each
dye
photodegrades,
the
geographical
location,
season,
and
nature/
concentration
of
natural
photosensitizers
would
control
the
rate
of
photolysis.
Therefore,
an
uncertainty
exists
on
the
chemical
nature
and
concentration
of
photoproducts.
In
addition,
anaerobic
biotransformation
may
also
be
a
route
of
dissipation,
but
the
chemical
identity
of
the
metabolites
specific
to
each
dye
is
not
known.

Purity
of
the
Dyes
Three
major
issues
have
been
identified
that
are
associated
with
the
purity
of
the
dyes
and/
or
test
substance
used
in
the
ecological
toxicity
studies.

a.
The
experimental
characterization
of
the
physical
and
chemical
properties
required
under
FIFRA
was
not
apparently
done
with
a
100%
pure
dye.
The
Acid
Blue
9
is
reported
to
be
50%
pure
(
43503401)
and
Acid
Yellow
23
as
being
28%
pure
(
43503402).
Although
the
presence
of
chemical
impurities
affect
the
physical
and
chemical
properties
of
a
chemical
substance,
the
extent
by
which
they
affect
those
of
the
dyes
is
unknown.
Some
of
the
physical
and
chemical
properties
reported
in
these
studies
are
not
consistent
with
those
expected
for
dyes
and
these
differences
may
be
related
to
the
unspecified
impurities.
The
high
vapor
pressure
reported
in
the
studies
may
be
that
of
a
volatile
impurity.

b.
The
above
studies
were
submitted
in
support
of
Aquashade
(
23.6%
Acid
Blue
9
and
2.39
Acid
Yellow
2.39%
registration
It
is
unclear
if
this
percent
composition
take
into
account
the
purity
of
each
dye.
In
addition,
it
is
not
known
if
the
impurities
in
the
test
substances
used
in
the
characterization
of
physical
and
chemical
properties
are
the
same
as
those
declared
as
impurities
in
the
technical
and/
or
end­
use
product
(
Confidential
Business
Information).
It
is
unclear
if
the
purity
of
the
dyes
in
other
products
is
the
same
as
in
Aquashade.

c.
The
Aquashade
product
that
was
used
in
the
"
basic
six"
studies
was
labeled
as
containing
23.63
%
Acid
Blue
9
and
2.39%
Acid
Yellow
23.
The
study
authors
stated
that
the
test
substance
characterization
provided
by
the
sponsor
indicated
a
purity
of
13.9%
"
azure
blue
dye".
The
use
of
the
term
"
azure"
is
unclear,
as
it
could
describe
a
color
or
a
series
of
structurally
7
related
"
azure
dyes".
These
azure
dyes
are
not
structurally
related
to
Acid
Blue
9
or
Acid
Yellow
23.
Therefore
the
exact
composition
and
purity
of
the
toxicant
that
was
used
in
the
studies
is
not
certain.
The
registrant
must
clarify
what
is
meant
by
"
azure
blue
dye".

2.
Effects
EFED
has
toxicity
data
for
both
formulations
containing
the
yellow
and
blue
dye,
plus
testing
with
the
dyes
separately.
Tests
with
the
Aquashade
product
containing
23.6%
Acid
Blue
9
and
2.36%
Acid
Yellow
23
as
the
test
substance
were
conducted
on
birds,
fish
and
aquatic
invertebrates.
The
Health
Effects
Division
(
HED)
has
a
rat
study
conducted
with
Admiral
WSP,
which
showed
that
the
LD50
was
equal
or
higher
than
5,000
mg/
kg.
There
are
also
mammal
acute
toxicity
data
with
both
the
blue
and
yellow
dyes
also
indicating
low
toxicity.

No
chronic
studies
using
aquashade
or
other
formulations
with
animals
have
been
submitted,
so
it
is
not
known
for
certain
what
would
happen
over
an
extended
exposure
period.
However,
there
is
mammal
chronic
testing
with
the
Acid
Yellow
23
dye
which
indicates
it
does
not
have
chronic
effects
on
mammals.
Furthermore,
these
dyes
are
used
as
colorants
in
human
foods
drugs
and
cosmetics,
so
long
term
effects
from
either
the
yellow
or
blue
dye
to
mammals
are
not
expected.
While
directly
correlating
lack
of
long­
term
mammal
toxicity
to
birds
is
uncertain,
given
the
mode
of
herbicidal
action
and
low
level
of
exposure
it
is
considered
unlikely
that
long­
term
exposures
would
have
adverse
effects
on
birds.
The
effects
of
chemical
impurities
in
the
dyes
is
not
known.

No
studies
on
aquatic
plants
have
been
required,
because
they
are
the
target
of
Aquashade
and
other
end­
use
products
and
it
is
assumed
that
all
submerged
aquatic
plants
in
the
treated
pond
will
be
killed.
No
terrestrial
plant
studies
have
been
required,
because
there
is
no
expected
exposure
unless
treated
water
are
used
for
irrigation.

Because
of
the
mode
of
action
(
blocking
sunlight
energy
that
enters
bodies
of
water)
and
the
low
acute
toxicity
of
this
pesticide,
it
is
believed
that
additional
ecological
toxicity
studies
are
not
needed
unless
new
uses,
methods
of
applications
or
new
products
are
proposed.
8
II.
Problem
Formulation
A.
Stressor
Source
and
Distribution
1.
Source
and
Identity
of
the
Stressor
Pond
Care
Algae
Blocker,
Aquashade,
and
Admiral
are
trade
names
for
products
containing
the
two
dyes4
Acid
Blue
9
and
Acid
Yellow
23
which
do
not
react
chemically
with
each
other.
Each
product
has
a
different
ratio
of
the
dyes
(
see
below),
but
in
all
of
the
product
formulations
the
percent
of
Acid
Blue
9
is
higher
than
that
of
Acid
Yellow
23.
The
use
pattern
of
the
products
are
essentially
the
same
in
the
sense
that
all
of
the
products
are
applied
directly
(
as
a
volume
of
product
or
number
of
packets)
to
artificial
(
or
natural),
confined
water
bodies,
including
swimming
pools
(
provided
that
chlorine
disinfectants
are
not
used).
Minimal
or
no
outflow
are
specified
in
the
labels
because
the
mode
of
action
is
such
that
allowing
treated
water
to
outflow
would
eliminate
the
effectiveness
of
the
products.
These
dyes
must
remain
in
the
water
column
and
block
the
sunlight
energy
for
a
sufficient
time
to
inhibit
photosynthesis
and
cause
the
plants
to
die.
They
are
not
intended
for
use
in
non­
confined
natural
water
bodies,
drinking
water
sources,
or
artificial
water
bodies
with
outflow.
Use
of
the
treated
water
for
irrigation
is
not
precluded
on
the
label,
and
this
is
an
uncertainty
because
irrigation
could
result
in
exposure
to
nontarget
terrestrial
plants,
and
there
are
no
tests
with
terrestrial
plants.

The
specific
names
of
the
end­
use
products
containing
Acid
Blue
9
and
Acid
Yellow
23
as
the
active
ingredients
are
AlgaeBlocker
and
Aquashed
OA
(
both
2.36%
Acid
Blue
9
and
0.24%
Acid
Yellow
23),
Aquashade
(
23.63%
Acid
Blue
9
and
2.39%
Acid
Yellow
23),
Admiral
Liquid
(
15.31%
Acid
Blue
9
and
1.00%
Acid
Yellow23),
and
Admiral
WSP
(
49.72%
Acid
Blue
9
and
3.27%
Acid
Yellow
23).
Additional
information
on
these
end­
use
products
is
presented
within
the
"
Use
Characterization"
section.

The
dyes
Acid
Blue
9
and
Acid
Yellow
23
are
known
by
several
different
names,
which
may
cause
confusion
about
the
chemical
identity
of
the
dye.
For
this
reason,
each
one
is
also
identified
by
the
chemical
name
used
by
the
Office
of
Pesticide
Programs,
the
Chemical
Abstracts
Registry
Number,
and
their
synonyms
(
Table
1).
"
Acid
Blue
9"
could
refer
to
either
the
trisodium
and
triammonium
salts,
each
one
having
a
different
Chemical
Abstracts
Registry
Number.
In
most
of
the
product
labels,
the
blue
dye
is
identified
as
"
Acid
Blue
9"
alone,
without
indicating
which
salt
is
used
in
the
formulation.
Throughout
the
document,
the
names
"
Acid
Blue
9"
and
"
Acid
Yellow
23"
are
used,
as
they
are
identified
as
such
in
the
product
labels.
In
the
Admiral
products,
they
are
identified
as
"
tartrazine"
for
the
yellow
dye
and
as
"
erioglaucine"
for
the
blue
dye.
The
nature
of
the
counter
cation
for
Acid
Blue
9
in
the
products
is
unclear,
although
the
Color
Index
International
identifies
it
as
the
triammonium
salt.
It
is
likely
that
the
blue
dye
used
in
the
products
is
the
triammonium
salt.
Further
information
on
the
chemical
identity
of
the
dyes
are
included
in
Table
2.
However,
because
it
is
the
anion
of
the
blue
dye
what
exerts
herbicidal
activity
by
blocking
sunlight
energy,
the
identity
of
the
counter
cation
is
not
relevant
to
the
exposure
and
risk
assessment
when
used
as
herbicides.

4
A
dye
is
defined
as
a
chemical
compound
that
is
capable
of
imparting
color
and
that
is
soluble
in
the
vehicle
in
which
it
is
applied.
9
In
addition
to
being
regulated
under
the
Federal
Insecticide,
Fungicide,
and
Rodenticide
Act
(
FIFRA),
the
two
dyes
are
regulated
by
the
Food
and
Drugs
Administration
(
FDA)
under
the
Food,
Drugs,
and
Cosmetics
Act
(
FDCA)
as
color
additives
(
colorants)
in
food,
drugs
and
cosmetics
.
Acid
Yellow
23
(
FD&
C
Yellow
5)
is
a
food,
drug,
and
cosmetics
colorant.
The
trisodium
salt
of
Acid
Blue
9
is
a
FD&
C
colorant
under
the
name
"
Blue
1",
whereas
the
triammonium
salt
(
Blue
4;
erioglaucine)
is
a
colorant
in
drugs
and
cosmetics
(
D&
C).
Use
on
foods,
drugs,
and
cosmetics
require
batch5
certification
prior
to
use.
While
the
nature
of
the
counter
cation
is
important
for
regulation
under
FDCA,
under
FIFRA
the
pesticide
active
species
is
the
anion.
It
is
the
anion
what
gives
color
to
the
treated
water.
Therefore,
for
the
use
of
these
blue
dyes
as
an
algicide
and
aquatic
herbicide
for
submerged
plants,
the
nature
of
the
counter
cation
is
irrelevant.
While
the
use
of
the
dyes
in
food,
drug
formulations
and
cosmetics
require
a
certification
of
their
purity,
the
purity
of
the
dyes
used
as
an
algicide/
aquatic
herbicide
is
unclear.

Acid
Blue
9
and
Acid
Yellow
23
are
also
regulated
under
FIFRA
as
"
Inerts"
used
to
impart
color
to
pesticide
formulations.
On
December
21,
2004
the
Agency
completed
the
"
Reassessment
of
the
Exemptions
from
the
Requirement
of
a
Tolerance
for
the
FDA­
Certified
FD&
C
Additives
Blue
No.
1
[
Acid
Blue
9];
erioglaucine]
and
Yellow
5
[
Acid
Yellow
23;
tartrazine]"
6.
This
reassessment
was
based
on
structure­
activity
relationships
(
EPISuite)
and
does
not
include
a
discussion
of
transformation
products..
However,
physical
and
chemical
properties
and
environmental
fate
estimates
using
EPIWIN
are
uncertain
because
salts
of
large
anions
are
not
adequately
handled
by
EPIWIN.

The
Society
of
Dyers
and
Colourists
and
the
American
Association
of
Textile
Chemists
and
Colorists
have
developed
a
Color
Index
International7
standard
for
identification
of
pigments
and
dyes.
The
Color
Index
classifies
dyes
and
pigments
by
their
composition.
Individual
pigment
or
dye
are
identified
by
a
unique
Color
Index
Generic
Name
(
C.
I.
Name)
and
a
Color
Index
Constitution
Number
(
C.
I.
Number),
although
the
C.
I.
Generic
Name
is
most
commonly
used..

Expanded
listings
of
other
names
used
for
Acid
Yellow
23
and
Acid
Blue
9
can
be
found
from
the
Unilever
Center
for
Molecular
Informatics,
Cambridge
University,

5
"
Batch"
is
defined
as
"
an
homogeneous
lot
of
color
additive
or
color
additive
mixture
produced
by
an
identified
production
operation,
which
is
set
apart
and
held
as
a
unit
for
the
purpose
of
obtaining
certification
of
such
quantity".
The
"
Batch
Number"
is
the
number
assigned
to
a
batch
of
colorant
by
the
person
who
requests
certification
of
the
batch.
A
"
Lot
Number"
is
the
identifying
number
or
symbol
assigned
by
the
FDA
to
a
batch
of
color
additive
after
certification.
Note
than
only
the
trisodium
salt
(
Blue
1)
can
be
used
in
food.
6
"
Reassessment
of
the
Exemption
from
the
Requirement
of
a
Tolerance
for
the
FDA­
Certified
Color
Additives
FD&
C
Blue
No.
1,
FD&
C
Red
No.
40,
and
FD&
C
Yellow
No.
5
(
Tartrazine)".
U.
S.
Environmental
Protection
Agency,
Office
of
Pesticides
Programs.
December
21,
2004.

7
http://
www.
colour­
index.
org/
10
Acid
Blue
9:
http://
www.
dspace.
cam.
ac.
uk/
handle/
1810/
5713
Acid
Yellow
23:
http://
www.
dspace.
cam.
ac.
uk/
handle/
1810/
6131
Table
1
Chemical
Nomenclature
and
Structure
of
the
Dyes
Acid
Yellow
23
Acid
Blue
9
Common
Name
as
Appearing
in
the
labels
of
the
Admiral
Products
Tartrazine
Erioglaucine
(
diammonium
salt)

Chemical
Name(
s)
4,5­
Dihydro­
5­
oxo­
1­(
4­
sulfophenyl)­
4­
((
4­
sulfophenyl)
azo)­
1H­
pyrazole­
3­
carboxylic
acid
trisodium
salt
3­
carboxy­
5­
hydroxy­
1­
p­
sulfophenyl­
4­
p­
sulfophenylazopyrazole
trisodium
salt
5­
hydroxy­
1­(
p­
sulfophenyl)­
4­((
psulfophenyl
azo)
pyrazole­
3­
carboxylic
acid
trisodium
salt
N­
Ethyl­
N­(
4­[(
4­{
ethyl[(
3­
sulfophenyl)
methyl]
amino}
phenyl)(
2­
sulfophenyl)
methylene]­
2,5­
cyclohexadien­
l­
ylidene)­
3­
sulfobenzenemethanaminium
hydroxide
inner
salt,
trisodium
salt
N­
Ethyl­
N­(
4­[(
4­{
ethyl[(
3­
sulfophenyl)
methyl]
amino}
phenyl)(
2­
sulfophenyl)
methylene]­
2,5­
cyclohexadien­
l­
ylidene)­
3­
sulfobenzenemethanaminium
hydroxide
inner
salt,
triammonium
salt
Molecular
Structure
Only
the
structure
of
the
disodium
salt
is
represented.
The
diammonium
salt
has
ammonium
instead
of
sodium
as
the
counter
cation.
It
is
the
organic
anion
what
is
responsible
for
the
color
of
the
dye.
11
Synonyms
FD
&
C
Yellow
No.
5:

21CFR74.705
(
Foods)
21CFR74.1705
(
Drugs)
21CFR74.2705
(
Cosmetics)
These
FD&
C
dyes
are
of
high
purity
and
require
batch
certification
analysis
when
used
in
food,
drugs,
or
cosmetics.

CI
Food
Yellow
4
CI
Number
15985
CAS
Registry
No.
1934­
21­
0
Sodium
Salt:
Brilliant
Blue
CI
Food
Blue
2
Erioglaucine
trisodium
salt
CI
Food
Blue
2.
CI
Number
42090
CAS
Registry
No.
2650­
18­
2
Triammonium
salt
Erioglaucine
diammonium
salt
Alphazurine
CI
Blue
8
CI
42090
CAS
Reg.
No.
3844­
54­
9
FD&
C
Blue
1
21CFR74.101
(
Foods)
21CFR74.1101
(
Drugs)
21CFR74.2101
(
Cosmetics)

D&
C
Blue
4
21CFR74.1104
(
Drugs)
21CFR.
74.2104
(
Cosmetics)
Chemical
Family
Azo
(
monoazo)
dye;
Contains
one
azo
group,
­
N=
N­)
Aminotriphenylmethane
dye
Other
uses
of
the
dyes
include;
Acid
Yellow
23­
Dye
for
wool
and
silk;
As
indicator
for
chlorine
estimations
in
biochemistry
Acid
Blue
9­
Biological
stain;
Textile
dye;
Wood
stain;
Indicator
2.
Physical
and
Chemical
Properties
and
Environmental
Fate
of
the
Stressor
Physical
and
Chemical
Properties
The
most
distinct
characteristic
of
dyes
is
that
they
absorb
energy8
strongly
within
the
wavelength
range
of
the
visible
spectrum
(
360
to
750
nm)
and
that
they
are
highly
soluble
in
the
vehicle
in
which
they
are
used,
in
this
case
water.
Dyes
are
highly
conjugated
systems
(
i.
e.,
multiple
double
bonds),
for
which
multiple
resonance
structures
can
be
written.
These
resonance
structures
cause
shifting
or
appearance
of
absorption
bands
into
the
visible
spectrum
of
electromagnetic
radiation.
Multiple
resonance
structures
increase
the
stability
of
a
molecule.
The
color
of
a
dye
or
other
colored
materials
is
determined
by
the
energy
(
wavelength)
of
maximum
absorption
(
electronic
absorption)
of
incident
light
by
the
molecule.
The
observed
color
is
determined
by
the
wavelength
region
in
which
the
substance
does
not
absorb
light
(
i.
e.,
the
reflected
color).
In
an
"
acid
dye",
the
chromophore
is
part
of
a
negative
ion
(
anion)
9
.
Therefore,
the
color
is
independent
of
the
counter
cation.
Table
2
summarizes
information
on
the
light
absorption
characteristics
of
each
dye.

8
Light
is
energy.
The
color
observed
in
dyes
(
or
in
gems)
is
caused
by
electronic
transitions
between
energy
levels
of
a
molecule.
The
color­
giving
absorption
band
is
known
as
a
chromophore
("
color
bearer")

9
Although
Acid
Blue
9
and
Acid
Yellow
23
belong
to
different
chemical
families,
both
dyes
contain
sulfonate
groups.
12
Table
2­
Electronic
Absorption
Spectra
of
Acid
Yellow
23
and
Acid
Blue
9
in
Water
Electronic
Absorption
Spectra
Acid
Yellow
23
Acid
Blue
9
Wavelength
of
Absorption
Maximum
(
8max,
in
nm)
in
water
420­
430
range,
8max,
425
620­
639
range;
8max,
630
Reflected
Color
Yellow
Blue
The
physical
and
chemical
properties
of
Acid
Yellow
23
and
Acid
Blue
9
are
summarized
in
Table
3.

Table
3­
Physical
and
Chemical
Properties
of
Acid
Yellow
23
and
Acid
Blue
9
(
Estimated);
Intrinsic
properties
Information
Acid
Yellow
23
Acid
Blue
9
Empirical
Formula
C16H9N4Na3O9S2
C37H34N2Na2
O9
S3
(
disodium
salt)

C37H34N2(
NH4)
2
O9
S3
(
diammonium
salt)

Molecular
Weight
534.37
(
Marmion)
792.84
(
disodium
salt)
(
Marmion)
782.96
(
diammonium
salt)
Physical
State,
as
pure
chemicals
Bright
orange­
yellow
powder
(
Marmion)
Reddish­
violet
powder
(
Marmion)
Vapor
pressure,
at
25E
C,
in
mmHg
7.43
x
10­
22
there
is
some
uncertainty
in
these
estimated
values,
however
they
do
not
significantly
impact
the
assessment
as
the
vapor
pressure
are
very
low
EPI
Suite,
2004
25.0
(
Reported
in
43503402)
2.97
x
10­
42
there
is
some
uncertainty
in
these
estimated
values,
however
they
do
not
significantly
impact
the
assessment
as
the
vapor
pressure
are
very
low
EPI
Suite,
2004
28.6
(
Reported
in
43503401)
Solubility
in
Water,
25EC
in
mg/
L
Log
n­
Octanol/
Water
Partition
Coefficient
1
x
106;
EPI
Suite,
2004
"
Completely
soluble"
(
Reported
in
43503402)

­
10.7
EPI
Suite,
2004
and
SRC
1.45
mg/
L;
EPI
Suite,
2004
"
Completely
soluble"
(
Reported
in
43503401)

­
1.50
EPI
Suite,
2004
and
SRC
Data
Source
Marmion=
Handbook
of
U.
S.
Colorants­
Food,
Drugs,
and
Medical
Devices,
Third
Edition.,
1991.

EPI
Suite,
2004=
Tolerance
Reassessment
Document
Submitted
studies=
40503401
(
Acid
Blue
9)
and
40503402
(
Acid
Yellow
23);
According
to
the
studies,
the
purity
of
Acid
Blue
9
was
50%
(
43503401)
and
that
of
Acid
Yellow
23
was
28%.
Thus,
the
reported
physical
chemical
properties
do
not
reflect
those
of
the
pure
materials..
The
high
vapor
pressure
reported
for
the
two
dyes
are
likely
to
be
associated
with
volatile
impurities.

SRC=
2004.
Syracuse
Research
Corporation.
Interactive
Physical
Properties
(
PHYSPROP)
Database
Demo.
Search
terms:
FD&
C
Blue.
No.
1,
FD&
C
Red
No.
40,
FD&
C
Yellow
No.
5.
(
November
28,
2004);
http://
www.
syrres.
com/
esc/
physdemo.
htm
13
Dissociation
Constant
(
pKa):
Acid
dyes
are
weak
acids.
The
pKa
values
for
Acid
Blue
9
and
Acid
Yellow
23
are
below
4.
Therefore,
they
are
completely
dissociated
in
the
environmentally
significant
pH
range
of
5
to
9.

Environmental
Fate
Parameters
(
Extrinsic
Properties)

Dyes
such
as
Acid
Yellow
23
and
Acid
Blue
9
are
not
readily
biodegradable
under
aerobic
conditions
nor
degrade
via
abiotic
hydrolysis
(
no
hydrolyzable
groups)
10,
Although
they
absorb
energy
strongly
in
the
visible
range
of
sunlight,
they
appear
to
undergo
slow
direct
photolysis11
Direct
photolysis
would
be
only
significant
in
clear,
shallow
water.
But
dyes
have
varying
susceptibility
to
fading
when
exposed
to
sunlight.
Light
fastness12
is
the
degree
to
which
a
dye
resists
fading,
which
varies
from
dye
to
dye.
Indirect
photolysis
is
likely
to
be
the
cause
of
fading
in
water
and
one
of
the
major
routes
of
transformation
in
the
environment.
Unlike
uses
in
food,
drugs,
and
cosmetics,
the
dyes
when
used
as
algicides,
are
exposed
to
sunlight
in
an
open
aquatic
environment.
Given
that
sunlight
energy
varies
according
to
latitude
and
time
of
the
year,
their
rate
of
fading
are
expected
to
vary
depending
on
geographical
location
and
season.
Biotransformation
under
anaerobic
conditions
has
as
also
been
documented
for
other
structurally
related
dyes.
Acid
Blue
9
and
Acid
Yellow
23
do
not
react
chemically
with
each
other.

Although
environmental
fate
properties
for
Acid
Blue
9
and
Acid
Yellow
23
were
estimated
with
Structure­
Activity
Relationships
using
the
EPI
(
Estimation
Programs
Interface)
SuiteTM
(
formerly
known
as
EPIWIN)
13
,
it
was
considered
that
the
estimates
are
not
reliable
because
EPIWIN
is
not
suitable
for
handling
salts,
particularly
of
those
with
large
anions.
Therefore,
EPIWIN
introduces
a
high
degree
of
uncertainty
for
dyes.
In
addition,
EPIWIN
does
not
provide
any
information
on
the
products
that
may
form
as
the
result
of
photoreactions
or
anaerobic
biotransformation.

The
vapor
pressure
and
the
Henry's
Law
Constants
of
dyes
are
very
low
and
it
is
unlikely
that
they
volatilize
from
soil
or
water.
Dyes
such
as
Acid
Blue
9
and
Acid
Yellow
23
are
very
hydrophilic
(
Log
Kow
<<
1)
and,
therefore
,
they
are
not
likely
to
bioaccumulate
in
fish.

10
Lynch,
D.
G.
"
Estimating
the
Properties
of
Synthetic
Organic
Dyes",
in
Handbook
of
Property
Estimation
Methods
for
Chemicals­
Environmental
Health
Sciences,
Edited
by
Robert
S.
Boethling
and
Donald
Mackay.
Published
by
Lewis
Publishers,
Boca
Raton,
FL,
2000;
Pages
447­
467.
And
pertinent
references
therein.

11
Even
though
it
has
the
necessary
condition
(
i.
e.,
absorb
energy
within
the
spectrum
of
sunlight)
to
undergo
direct
photolysis
that
process
is
slow.
However,
they
can
degrade
via
indirect
photolysis.
For
further
information
on
indirect
photolysis
refer
to
Footnote
17.

12
In
artist's
color
language,
a
dye
or
pigment
that
fades
is
said
to
be
"
fugitive"
and
those
that
do
not
fade
are
called
"
permanent
colors".

13
http://
www.
epa.
gov/
opptintr/
exposure/
docs/
episuite.
htm
14
3.
Pesticide
Type,
Class,
and
Mode
of
Action
There
are
five
products
that
contain
both
Acid
Blue
9
and
Acid
Yellow
23
in
their
formulation:
(
1)
Care
Pond
AlgaeBlocker
(
one
label);
(
2)
Aquashade
(
two
labels)
and
(
3)
Admiral
(
two
labels).
The
products
are
used
to
control
the
growth
of
algae
in
ornamental
ponds,
recreational
man­
made
(
closed­
system
ponds,
such
as
golf
course
lakes),
rearing
lakes
for
nonedible
fish.,
and
fountains
containing
fish.
The
dyes
do
not
directly
kill
the
algae,
but
prevent
growth
by
blocking
sunlight
energy
necessary
for
photosynthesis
and
survival
of
the
algal
and
other
undesirable
aquatic
plants.
Thus,
these
products
serve
as
algicides
or
as
herbicides
for
undesirable
aquatic
plants.
In
addition,
the
dyes
confer
a
bluish
green
coloring
to
the
water
body.

4.
Overview
of
Pesticide
Usage
These
products
are
to
be
applied
directly
to
the
water
body
by
adding
a
recommended
amount
of
the
product
to
a
specified
volume
of
water
to
attain
1
or
2
ppm
according
to
the
target
weed.
The
extent
of
use,
timing
of
application,
and
where
it
is
used
is
not
known,
but
geographical
location
and
season
will
affect
the
rate
of
fading
and
frequency
of
application.
The
dose
to
be
apply
varies
with
the
products
and
is
specified
in
the
labels
and
takes
into
account
the
volume
and
depth
of
the
water
to
be
treated.
The
specific
times
of
application
are
not
included
in
the
labels.
However,
some
of
the
labels
recommend
that
the
product
be
applied
"
early
in
the
season"
(
presumably
Spring)
and
also
indicate
that
the
product
can
be
added
to
an
ice
surface
of
the
water
body
(
i.
e.,
before
melting).

B.
Receptors
1.
Aquatic
Effects
For
aquatic
ecosystems,
ecological
receptors
include
all
aquatic
life
(
fish,
amphibians,
invertebrates,
plants)
and
those
terrestrial
animals
(
e.
g.,
birds
and
mammals)
that
consume
water
and/
or
aquatic
organisms.
Since
these
products
are
directly
applied
to
contained
water
bodies,
any
effects
on
aquatic
life
and
terrestrial
animals
would
come
from
consumption
of
water
or
fish
in
rearing
lakes
or
fountains.
Immersion
in
the
treated
water
may
also
have
effects,
such
as
irritation
or
changes
in
color
caused
by
the
dyes.
Terrestrial
animals
may
also
be
affected
indirectly
if
they
depend
on
the
aquatic
plants
in
the
treated
water
bodies
for
food.

The
labels
for
these
products
state
that
non­
target
aquatic
plants
(
water
lilies,
hyacinths,
cattails)
may
suffer
contact
burns
if
the
chemicals
are
accidentally
poured
directly
into
them
and
that
desirable
submerged
plants
may
also
be
affected
due
to
reduced
levels
of
sunlight.

Aquatic
organisms
that
depend
on
sunlight
for
survival
may
be
affected
by
the
reduction
of
sunlight.
Examples
include
non­
target
plants,
animals
that
consume
non­
target
or
target
plants
or
prey
on
the
animals
that
consume
aquatic
plants.

Risk
to
aquatic
animals
is
based
on
registrant
submitted
acute,
dietary
laboratory
studies
with
aquatic
vertebrates
(
rainbow
trout
and
bluegill
sunfish)
and
invertebrates
(
water
fleas).
15
The
taxa
evaluated
for
ecological
effects
are
presented
in
Table
4.

Table
4.
Taxa
evaluated
for
ecological
effects
in
screening
level
risk
assessments.
Taxon
Surrogate
Species
Used
in
Risk
Assessment
Birds
a
Mallard
duck
(
Anas
playtrhynchos)
Bobwhite
quail
(
Colinus
virginianus)
Mammals
Laboratory
rat
(
Rattus
norvegicus)

Freshwater
fish
b
Bluegill
sunfish
(
Leopomis
macrochirus)
Rainbow
trout
(
Oncorhynchus
mykiss))
Freshwater
invertebrates
Water
flea
(
Daphnia
magna)

Estuarine/
marine
fish
Not
Required
because
estuarine
exposure
is
considered
unlikely
Estuarine/
marine
invertebrates
Not
Required
because
estuarine
exposure
is
considered
unlikely
Terrestrial
plants
Not
Required
because
exposure
to
terrestrial
plants
is
not
expected
providing
treated
water
is
not
used
for
irrigation
Aquatic
plants
and
algae
Not
Required
because
aquatic
plants
are
the
target
and
assumed
to
be
controlled
by
use
of
aquashade
a
Birds
are
used
as
surrogates
for
amphibians
(
terrestrial
phase)
and
reptiles.

b
Freshwater
fish
may
be
surrogates
for
amphibians
(
aquatic
phase).

Because
of
the
very
low
exposure
potential
and
mode
of
action
and
very
restricted
use
pattern,
only
the
avian
oral,
avian
dietary,
and
acute
freshwater
studies
were
required.
An
acute
rat
study
was
submitted
to
OPP's
Health
Effects
Division
(
HED)
and
EFED
used
their
review
as
a
surrogate
for
wild
mammals.

2.
Terrestrial
Effects
Terrestrial
plants
are
not
receptors
for
the
direct
application
of
Aquashade
to
contained
water.
Terrestrial
animals
(
e.
g.
birds
and
mammals)
may
become
receptors
if
they
consume
or
immerse
in
the
water
treated
with
these
dyes.

Risk
to
terrestrial
animals
will
be
based
on
registrant
submitted
acute
laboratory
tests
with
birds
(
bobwhite
quail
and
mallard
duck)
and
acute
and
chronic
tests
with
mammals
to
represent
all
terrestrial
vertebrates.
In
addition,
effects
data
from
open
literature
will
also
be
considered.

3.
Ecosystems
at
Risk
All
of
the
products
are
directly
applied
to
relatively
small,
ornamental
or
decorative
water
bodies
without
outlets
and
are
not
to
be
used
in
or
discharged
to
streams
or
rivers.
The
ecosystems
at
risk
are
those
aquatic
habitats
in
the
treated
water
bodies
and
the
areas
occupied
by
terrestrial
animals
that
might
consume,
or
immerse
themselves
in,
the
treated
water.
The
labels
allow
treatment
of
ponds
in
golf
courses,
which
suggests
that
"
natural
looking"
ponds
may
be
treated.
However,
it
is
assumed
that
these
water
bodies
are
managed
regularly
for
esthetic
16
reasons
(
i.
e.
to
control
weed
population
and
to
add
color
to
the
water
body).
The
terminology
"
relatively
small"
is
intended
to
differentiate
the
potential
use
areas
from
extremely
large
water
bodies
(
e.
g.
large
lakes
and
rivers).
These
products
are
used
to
curtail
photosynthesis
in
weeds
and
algae
by
blocking
sunlight
energy
and
therefore,
use
of
these
products
to
control
aquatic
plants
and
algae
in
extremely
large
or
flowing
water
bodies
is
impractical.

C.
Assessment
Endpoints
Ecological
Effects
Assessment
endpoints
are
the
"
explicit
expressions
of
the
environmental
value
that
is
to
be
protected."
Defining
an
assessment
endpoint
involves
two
steps:
1)
identifying
the
valued
attributes
of
the
environment
that
are
considered
to
be
at
risk,
and
2)
operationally
defining
the
assessment
endpoint
in
terms
of
an
ecological
entity
(
i.
e.,
a
community
of
fish
and
aquatic
invertebrates)
and
its
attributes
(
i.
e.,
survival
and
reproduction).
Therefore,
selection
of
the
assessment
endpoints
is
based
on
valued
entities
(
i.
e.,
ecological
receptors),
the
ecosystems
potentially
at
risk,
the
migration
pathways
of
pesticides,
and
the
routes
by
which
ecological
receptors
are
exposed
to
pesticide­
related
contamination.
The
selection
of
clearly
defined
assessment
endpoints
is
important
because
they
provide
direction
and
boundaries
in
the
risk
assessment
for
addressing
risk
management
issues
of
concern.

Table
5.
Summary
of
assessment
and
measurement
endpoints.
Assessment
Endpoint
Measurement
Endpoint
1.
Abundance
(
i.
e.,
survival
and
growth
and
reproduction)
of
individuals
and
populations
of
birds.
1a.
Bobwhite
quail
and
mallard
duck
acute
oral
LD50.
1b.
Bobwhite
quail
and
mallard
duck
subacute
dietary
LC50.
1c.
Avian
reproductive
NOAEL
(
in
this
case,
no
avian
chronic
data
are
available,
low
chronic
toxicity
is
inferred
based
on
low
chronic
toxicity
to
mammals)
2.
Survival
of
individual
mammals
and
reproductive
effects.
2a.
Laboratory
rat
acute
oral
LD50
and
reproduction
test
NOAEL.
3.
Survival
and
reproduction
of
freshwater
fish
and
invertebrates
3a.
Rainbow
trout
and
bluegill
sunfish
acute
LC50.
3c.
Water
flea
acute
LC50.
3d.
Fish
and
invertebrate
chronic
toxicity
NOAEL.
4.
Survival
and
Growth
of
Aquatic
Plants
4.
Algae
and
vascular
plant
EC50
and
NOAEL
5.
Survival
and
Growth
of
Terrestrial
Plants
5.
Terrestrial
monocot
and
dicot
vegetative
vigor
and
seedling
emergent
EC25
and
NOAEL
LD50
=
Lethal
dose
to
50%
of
the
test
population.
LC50
(
EC50)
=
Lethal
(
effective)
concentration
to
50%
of
the
test
population.
EC25
=
Estimated
concentration
at
which
25%
response
in
tested
population
would
be
expected
NOAEL
=
Highest
dose
or
test
level
at
which
no
statistically
significant
effects
were
observed.

D.
Conceptual
Model
1.
Risk
Hypothesis
Risk
hypotheses
are
specific
assumptions
about
potential
adverse
effects
(
i.
e.,
changes
in
assessment
endpoints)
and
may
be
based
on
theory
and
logic,
empirical
data,
mathematical
17
models,
or
probability
models
(
EPA,
1998).
For
this
assessment,
the
risk
is
stressor­
linked,
where
the
stressors
are
the
release
of
Acid
Yellow
23
and
Acid
Blue
9
by
direct
application
into
artificial
and
confined
water
bodies
(
i.
e.
no
outlets)
and
in
accordance
.
The
following
hypotheses
are
presumed
for
this
screening
level
assessment
when
the
products
are
used
in
accordance
with
instructions
and
the
precautions
stated
in
the
labels.

Aquashade
represents
a
group
of
end­
use
products
containing
the
two
dyes
Acid
Blue
9
and
Acid
Yellow
23.
They
are
used
to
control
weeds
in
contained
water
bodies
(
no
outflow)
by
blocking
sunlight
to
aquatic
weeds
and
thereby
inhibiting
photosynthesis.
They
are
applied
directly
to
these
contained
water
bodies.
The
risk
hypothesis
is
that
when
used
as
directed,
Aquashade
and
the
other
end­
use
products
may
represent
a
risk
to
aquatic
and
terrestrial
organisms."

2.
Conceptual
Model
Water
is
treated
directly
either
by
pouring
the
product
directly,
or
by
dropping
packets.

Aquatic
plants
and
animals
are
exposed
directly.

Terrestrial
animals
may
be
exposed
by
drinking
the
treated
water
or
immersing
in
it.

Terrestrial
plants
are
unlikely
to
be
exposed
unless
treated
water
is
used
for
spray
irrigation.

E.
Analysis
Plan
1.
Preliminary
Identification
of
Data
Gaps
and
Methods
Environmental
Fate
As
a
first
step,
the
current
status
of
FIFRA's
Subdivision
N
Data
Requirements
was
identified
for
Acid
Blue
9
and
Acid
Yellow
23.
The
161­
1
(
Abiotic
Hydrolysis),
161­
2
(
Direct
Photolysis
in
Water),
162­
3
(
Anaerobic
Aquatic
Metabolism),
and
162­
4
(
Aerobic
Aquatic
Metabolism)
guideline
studies
were
"
Reserved"
based
on
low­
volume
of
use
of
these
chemicals
(
when
use
as
pesticides)
14,
but
pending
the
results
of
the
ecological
toxicity
studies
(
Guidelines
71
through
72).
Physical
and
chemical
properties
were
submitted
in
1994
for
the
individual
dyes
(
MRID
Numbers
43503401
and
43503402),
reviewed,
and
deemed
acceptable
However,
these
two
studies
were
not
conducted
with
a
high
purity
dye
(
50%
for
Acid
Blue
9
and
28%
for
Acid
Yellow
23)).
These
two
studies
were
submitted
in
support
of
the
Aquashade
product
of
23.6%
Acid
Blue
9
and
2.39%
Acid
Yellow
23.

In
December
2004,
the
Agency
completed
a
tolerance
reassessment
for
these
two
chemicals
when
used
as
inert
components
in
pesticide
formulations.
Most
of
the
environ
mental
14
Memorandum:
"
Review
of
Phase
4
List
D
Package
for
Aquashade"
(
EFGWB
#
93­
0119,
93­
0120;
Chemical
#
110301
and
110302;
Case
#
819437
and
819438;
DP
Barcode
D184289
and
D184278).
Dated
February
4,
1993.
18
fate
information
in
the
document
come
from
structure­
activity
relationships
using
computational
tools
(
EPIWIN).
In
addition,
a
"
Robust
Summary
for
Acid
Yellow
23
(
tartrazine)
15
was
submitted
to
the
Agency
under
the
High
Production
Volume
(
HPV)
Challenge
Program",
which
also
relies
on
EPIWIN
estimates.
As
already
indicated,
EPIWIN
is
not
a
suitable
estimation
program
for
salts,
particularly
for
salts
of
a
large
anion.
Physical
and
chemical
properties
estimated
by
EPI
Suite/
EPIWIN
have
been
included
in
Table
3
and
come
from
the
2004
tolerance
reassessment
document.

Information
from
the
open
literature
was
included
to
present
a
more
comprehensive,
but
qualitative,
assessment
of
the
environmental
fate
of
the
two
dyes
beyond
structure­
activity
estimates
(
Refer
to
the
Exposure
Characterization­
Environmental
Fate
and
Transport
section
of
this
document).

Given
that
the
ecological
risk
assessment
concluded
that
products
containing
Acid
Blue
9
and
Acid
Yellow
23
did
not
pose
risk
to
non­
target
organisms,
no
further
environmental
fate
data
are
required
unless
new
uses
or
products
are
proposed.

Ecological
toxicity
Measures
of
effects
are
obtained
from
a
suite
of
registrant­
submitted
guideline
studies
conducted
with
a
limited
number
of
surrogate
species
and
from
open
literature.
The
test
species
are
not
necessarily
intended
to
be
representative
of
the
most
sensitive
species
but
rather
were
selected
based
on
their
ability
to
thrive
under
laboratory
conditions.
Acute
measures
of
effect
are
the
concentrations
that
produce
50%
mortality
or
growth
reduction
in
the
test
organisms
(
LC50s
and
EC50s,
respectively).
Chronic
measures
are
the
no
observed
adverse
effect
level
or
NOAEL
which
is
the
highest
test
level
at
which
significant
effects
were
not
observed.

2.
Measures
to
Evaluate
Risk
Hypotheses
and
Conceptual
Model
a.
Measures
of
Exposure
from
Contaminated
Water
For
these
products,
which
are
directly
applied
to
the
water
body
in
a
specified
volume
of
the
liquid
product
or
number
of
packets,
(
no
spraying),
the
concentration
of
each
dye
depends
on
the
amount
added
and
the
ratio
of
each
dye
in
the
products.
The
amount
to
be
added
depends
on
where
it
is
used
and
the
size
and
depth
of
the
water
body.
Neither
runoff
nor
spray
drift
are
routes
of
exposure.
Therefore,
simulation
models
(
GENEEC;
PRZM
and
EXAMS)
are
not
applicable
to
estimate
environmental
exposure
concentrations
in
the
treated
water.

To
estimate
exposure
to
terrestrial
animals,
estimates
of
water
consumption
were
used
to
derive
daily
doses
of
the
formulation
aquashade
and
compared
to
acute
toxicity
endpoints.

b.
Measures
of
Effects
15
http://
www.
epa.
gov/
chemrtk/
ciacdylo/
c15133tc.
htm
"
Robust
Summaries
&
Test
Plans:
C.
I.
Acid
Yellow
23
(
FD&
C
Yellow
5)",
Submitted
ti
the
Agency
by
the
International
Association
of
Color
Manufacturers
on
March
10,
2004.
19
Since
Aquashade
(
23.6%
Acid
Blue
9
and
2.39%
Acid
Yellow
23)
is
composed
of
two
dyes
that
are
never
used
individually
as
algicide/
aquatic
herbicide,
all
of
the
studies
submitted
were
conducted
with
the
end­
use
product
(
Refer
to
the
Memorandum
cited
in
Footnote
13).
Studies
submitted
to
fulfill
the
"
basic
six"
requirements
(
71­
1b
avian
oral,
71­
2b
avian
dietary,
72­
1b
and
72­
1d
Freshwater
fish
acute,
and
72­
2b
Aquatic
invertebrate
acute
toxicity).
Based
on
low
toxicity
and
no
effects
observed
at
the
highest
test
level
in
the
acute
toxicity
test,
the
mode
of
action,
the
low
chronic
toxicity
of
the
yellow
dye
demonstrated
to
mammals
in
chronic
testing,
and
the
fact
that
both
of
the
dyes
are
used
as
food
coloring
for
human
foods
and
often
used
in
other
animal
foods
with
no
long­
term
effects
detected
there
is
no
reason
to
believe
that
other
effects
(
reproductive,
estuarine,
marine,
etc.)
are
likely.
Chronic
studies
are
not
required.

For
the
present
ecological
risk
assessment,
EFED
has
used
the
ecological
toxicity
data
from
the
studies
conducted
with
together
with
an
acute
laboratory
rat
toxicity
conducted
with
Admiral
WSP
submitted
to
HED.

In
addition
EFED
has
made
the
assumption
that
the
dyes
do
not
represent
chronic
risks
based
on
a
mammalian
chronic
study
with
the
Acid
Yellow
dye
23.

c.
Measures
of
Ecosystem
and
Receptor
Characteristics
Aquashade's
and
the
labels
of
the
other
end­
use
products
specifically
state
that
treated
water
bodies
are
not
to
be
drained
into
streams,
rivers
or
other
water
systems,
it
is
assumed
that
exposure
to
these
other
water
bodies
will
not
occur.
The
aquatic
organisms
that
occur
in
the
treated
water
bodies
are
generally
assumed
to
have
been
placed
there
by
people
that
manage
these
decorative
ponds
and
containers.
The
submerged
plants
in
the
ponds
are
assumed
to
be
the
target
organisms,
so
the
risk
to
them
is
not
assessed.

Since
it
is
never
applied
to
terrestrial
environments,
exposure
to
terrestrial
animals
assessment
estimating
residues
on
terrestrial
vegetation
and
invertebrates
was
inappropriate.
Instead
calculations
of
the
amount
in
water
consumed
by
birds
or
mammals
drinking
from
the
treated
body
of
water
were
done
to
derive
a
dose
of
Aquashade
to
birds
and
mammals
in
three
size
classes.

The
receptors
addressed
by
the
aquatic
and
terrestrial
risk
assessments
for
Aquashade
are
summarized
in
Table
4.
For
aquatic
assessments,
freshwater
fish
and
invertebrates
were
studied.
Estuarine/
marine
animals
are
not
represented,
because
Aquashade
is
not
applied
to
these
environments
and
is
not
expected
to
drain
into
them.

III.
Analysis
A.
Use
Characterization
The
use
characterization
of
the
products
containing
Acid
Blue
9
and
Acid
Yellow
23
is
based
on
the
current
labels
for
these
products.
Table
7
identifies
these
end­
use
products
with
20
their
EPA
Registration
Numbers,
and
label
date.
Except
for
Admiral
WSP,
all
of
the
products
are
liquid
formulations.
The
products
are
directly
poured
or
droppen
into
water
in
a
specified
volume
of
the
liquid
formulation
or
number
of
packets.
No
spraying
is
involved
which
could
lead
to
spray
drift
or
inadvertent
exposure
to
terrestrial
habitats.

Table
7
Identification
of
the
End­
use
Products
Containing
Acid
Blue
9
and
Acid
Yellow
23
as
the
Active
Ingredients
Product
EPA
Reg.
No
Registrant
Label
Date
Composition
(
by
weight)
Type
of
Formulation
Target
pests
Pond
Care
Algae
Blocker
8709­
6
Aquarium
Pharmaceuticals,
Inc.
February
6,
2004
2.36
%
Acid
Blue
9
0.24
%
Acid
Yellow
23
Liquid
formulation
Control
of
growth
of
many
algae
and
underwater
aquatic
weeds
Aquashade
OA
Aquashade
33068­
2
Aquashade
,
Inc.
February
23,
1993
3306­
1
Aquashade,
Inc.
1981
2.36
%
Acid
Blue
9
0.24
%
Acid
Yellow
23
Liquid
formulation
23.63%
Acid
Blue
9
2.39%
Acid
Yellow
23
Liquid
formulation
Control
of
growth
of
many
algae
and
underwater
aquatic
weeds
Control
of
growth
of
many
algae
and
underwater
aquatic
weeds
Admiral
Liquid
67064­
2
Becker
Underwood
March
14,
2000
15.31%
Acid
Blue
8
1.00%
Acid
Yellow
23
Liquid
formulation
Control
algae
and
aquatic
vegetation
and
color
the
water
Admiral
WSP
67064­
1
Becker
Underwood
August
17,
2001
49.72%
Acid
Blue
9
3.27%
Acid
Yellow
23
Concentrated
blend
in
pre­
measured
water
soluble
packages
Control
algae
and
aquatic
vegetation
and
color
the
water
The
use
sites
and
timing
of
application
are
summarized
in
Table
8.
Note
that
the
uses
are
limited
to
confined
water
bodies
with
minimal
or
no
outflow,
but
that
the
Admiral
products
has
a
wider
range
of
use
sites
than
AlgaeBlocker
and
Aquashade.
Only
the
Admiral
products
have
uses
on
swimming
ponds,
but
treated
swimming
ponds
must
not
be
treated
with
a
chlorine
disinfectant
to
avoid
fading
of
the
dyes.
Likewise,
no
carbon
filters
should
be
used
to
prevent
removal
of
the
dyes
from
water.

Table
8­
Use
Sites
and
Timing
of
Application
for
the
End­
use
Products
Containing
Acid
Blue
9
and
Acid
Yellow
23
as
the
Active
Ingredient
Product
Use
Sites
Time
of
Application
Pond
Care
Algae
Blocker
Ornamental
Ponds
and
recreational
man­
made,
closed
system
ponds
(
Golf
course
lakes);
Rearing
lakes
for
non­
edible
fish;
Fountains
containing
fish.
Not
specified
"
Reapply
as
needed"
21
Aquashade
OA
Aquashade
Fountains
Aquarium
Ornamental
Ponds
Re­
circulated
or
artificial
waterscapes
Ideal
Settings:
Shopping
malls
Executive
office
parks
Botanical
gardens
Recreational
and
amusement
parks.

Natural
and
manmade
contained
ponds
and
lakes
including
ornamental,
recreational,
fish
rearing
and
fish
farming
ponds,
golf
course
ponds
Do
not
apply
directly
to
streams,
other
natural
bodies,
or
any
body
of
water
not
under
control
of
the
user.
Do
not
apply
to
water
that
will
be
used
for
human
consumption.
Not
specified
Recommends
application
before
weed
growing
season.

May
be
applied
while
ice
is
still
covering
the
water
body
Admiral
Liquid
Natural
or
manmade
ponds,
lakes,
fountains,
fish
farms,
fish
hatcheries,
golf
courses
and
swimming
ponds
Prior
to
or
early
in
the
weed
growing
season
for
optimum
results
May
be
applied
while
ice
is
still
covering
the
water
body
Admiral
WSP
Natural
or
manmade
ponds,
lakes,
fountains,
fish
farms,
fish
hatcheries,
golf
courses
and
swimming
ponds
Prior
to
or
early
in
the
weed
growing
season
for
optimum
results
Reapply
"
as
needed"

The
liquid
concentrates
AlgaeBlocker,
Aquashade,
Aquashade
OA,
and
Admiral
liquid
are
added
directly
to
water
in
a
specified
volume
of
product
according
to
the
volume
and
depth
of
the
water
body
to
be
treated.
to
reach
a
target
concentration.
Admiral
WSP
is
also
directly
applied
to
water
in
a
prescribed
number
of
packets
for
specific
volume
of
the
water
bodies.
The
resulting
concentration
of
the
products
in
water
is
fixed
to
"
1
or
2
ppm",
depending
on
the
target
weed..
The
dosing
specified
in
the
labels
are
summarized
in
Table
9
.

Table
9.­
Dose
Recommended
for
the
End­
Use
Products
Containing
Acid
Blue
9
and
Acid
Yellow
23
as
the
Active
Ingredients
Product
Dose
Pond
Care
Algae
Blocker
Ornamental
ponds
and
fountains
less
than
2
feet
deep:
8
mL
of
product
per
every
100
gallons
of
pond
water
(
378
L);
Fish
rearing
ponds
and
large
ornamental
ponds
over
2
feet
deep:
4
mL
for
every
100
gallons
of
water
Maximum
application
rate,
1
ppm
22
Aquashade
OA
Aquashade
"
Small
water
bodies"
(
unspecified)
One
drop
for
each
gallon
of
water
`
Larger
water
bodies"
1
oz
per
1,000
gallons
of
water
For
controlling
submerged
weed,
the
depth
of
water
must
be
2
feet
Maximum
application
rate,
1
ppm
One
gallon
of
product
(
1
ppm)
per
acre
with
4
foot
average
depth
when
submerged
weeds
and
algae
are
growing
at
depths
>
2
feet.
target
pests
are
leafy
pondweed,
Chara,
slender
naiad,
filamentous
green
and
blue
algae,
spyrogyra­
Water
milfoil.

2
ppm
to
prevent
production
of
Hydrilla's
tubers.
Admiral
Liquid
The
amount
of
the
product
added
depends
on
the
volume
of
the
water
body.
Amount
to
be
added
(
volume)
per
volume
of
water
body
is
specified
in
the
label.

Regardless
of
the
amount
added
and
volume
of
the
water
body,
the
concentration
regime
in
the
water
is
:

1
ppm
for
filamentous
blue
and
blue
algae,
Chara,
Leafy
Pond
weed,
spirogyra,
slender
naiad,
and
water
milfoil
2
ppm
is
used
to
prevent
Hydrilla
tuber
production
after
a
herbicide
treatment
Admiral
WSP
Same
as
Admiral
Liquid
Most
of
the
labels
contain
precautionary
language
and/
or
environmental
hazard
statements,
but
the
language
is
not
consistent
across
the
labels.
Table
10
summarizes
the
"
Precautionary
Language",
whereas
Table
11
summarizes
the
"
Environmental
Hazard
Statements"
as
they
appear
in
each
label.

Table
10
Precautionary
Language
Included
in
the
End­
use
Products
Containing
Acid
Blue
9
and
Acid
Yellow
23
as
the
Active
Ingredients
Products
Precautionary
Language
Included
in
the
Labels
Pond
Care
Algae
Blocker
Non­
target
plants
(
water
lilies,
hyacinths,
cattails)
may
suffer
contact
burns
if
material
is
accidentally
poured
directly
into
them
Desirable
submerged
plants
may
also
be
affected
due
to
reduced
levels
of
sunlight
Aquashade
OA
Aquashade
None
None
Admiral
Liquid
Water
must
be
under
the
complete
control
of
the
user
and
have
little
or
no
outflow
(
dye
concentration
must
be
maintained).
Do
not
apply
directly
to
streams,
or
any
body
of
water
that
may
be
used
for
human
consumption
Admiral
WSP
Same
as
for
Admiral
Liquid
23
Table
11
Environmental
Hazard
Statements
Included
in
the
Labels
of
the
End­
use
Products
Containing
Acid
Blue
9
and
Acid
Yellow
23
as
the
Active
Ingredients
Products
Environmental
Hazard
Statements
Included
in
the
Labels
Pond
Care
Algae
Blocker
Do
not
contaminate
domestic
livestock,
irrigation
water
and
streams
with
outflow.
Use
only
in
impounded
water
with
no
outlet
and
under
the
total
control
of
the
applicator.
Do
not
use
in
recreational
waters
intended
for
swimming
or
in
waters
intended
for
edible
fish
Aquashade
OA
Aquashade
Desirable
plants
such
as
water
lilies
may
suffer
contact
burn
if
material
is
accidentally
poured
on
them
directly
Shoreline
non­
target
plants
(
cattails,
water
lilies)
may
suffer
contact
burn
if
material
is
accidentally
poured
on
them.
Do
not
contaminate
water
by
cleaning
of
equipment
or
disposal
of
wastes.
Admiral
Liquid
Shoreline
non­
target
plants
(
cattails;
water
lilies)
may
suffer
contact
burn
if
the
material
is
accidentally
poured
on
them.
Apply
this
product
only
as
specified
in
the
label.
Do
not
make
applications
when
weather
conditions
favor
drift
from
nontarget
areas.
Do
not
apply
where
runoff
is
likely
to
occur.
Admiral
WSP
None
specified
B.
Exposure
Characterization
1.
Environmental
Fate
and
Transport
No
experimental
environmental
fate,
Subdivision
N
Guideline
studies
were
conducted
with
each
individual
dye
as
the
test
substance
or
an
end­
use
product.
These
studies
were
placed
in
"
Reserved"
pending
the
results
of
the
required
ecological
toxicity
studies,
as
indicated
in
the
"
Analysis
Plan"
section.
For
consistency
with
the
tolerance
reassessment
for
inerts,
the
environmental
fate
assessment
originally
based
findings
on
estimates
from
Structure­
Activity
Relationship
(
SAR)
using
EPISUITE
(
EPIWIN).
However,
these
estimates
are
considered
uncertain
because
EPIWIN
is
not
suitable
for
salts,
particularly
those
of
large
anions,
such
as
in
dyes.
A
literature
search
for
environmental
fate
studies
conducted
with
Acid
Yellow
23
and
Acid
Blue
9
as
the
test
substances
did
not
produced
sufficient
experimental
data
specific
to
these
two
dyes.
Therefore,
most
of
the
environmental
fate
information
comes
from
dyes
structurally
related
to
Acid
Yellow
23
(
azo
dyes)
and
Acid
Blue
9
(
triphenylmethane
dyes).
Further
information
was
obtained
from
the
"
Handbook
of
U.
S.
Colorants­
Food,
Drugs,
and
Medical
Devices"
(
Marmion,
1991).
Thus,
most
of
the
present
environmental
fate
assessment
uses
published
information
about
the
properties
of
synthetic
dyes
and
other
relevant,
open
literature
sources
(
Lynch,
2000;
Lynch,
D.,
OPPTS.
private
communication,
August
2005).
The
environmental
fate
assessment
is
only
qualitative
in
nature.

Acid
Blue
9
and
Acid
Yellow
23
belong
to
different
chemical
families
but
both
contain
sulfonate
groups.
Because
none
of
these
two
dyes
contain
hydrolyzable
groups,
abiotic
hydrolysis
is
not
a
degradation
pathway
for
these
two
dyes.
24
As
chemical
substances
absorbing
sunlight
energy,
photodegradation
in
water
can
occur
via
direct
or
indirect
photolysis.
The
direct
photolysis
of
most
dyes
appears
to
be
slow
and
would
be
only
significant
in
clear,
shallow
water
(
which
could
be
the
case
for
ornamental
uses,
such
as
in
fountains
exposed
to
sunlight).
Because
the
presence
of
natural
photosensitizers
in
the
environment16,
indirect
photolysis
has
been
identified
as
a
major
transformation
pathway
for
dyes
in
environmental
water.
Besides
the
concentration
of
photosensitizers
in
water,
the
rate
of
photolysis
would
also
be
a
function
of
the
solar
photon
flux,
which
depends
on
latitude
and
season.
For
example,
photolysis
would
be
faster
in
the
Summer
than
in
the
Spring
and
faster
in
the
Southern
than
in
the
Northern
parts
of
the
country.
Acid
Yellow
23
appears
to
be
more
photolytically
stable
than
Acid
Blue
9,
as
the
latter
fades
faster
(
Marmion,
1991).
Photodegradation
of
dyes
can
also
occur
on
soil
surfaces.

There
are
no
experimental
data
that
identifies
the
photoproducts
of
Acid
Blue
9
or
Acid
Yellow
23.
However,
for
azo
dyes
(
Acid
Yellow
23
is
an
azo
dye),
azo
bond
cleavage
and
photoredox
reactions
occur
in
most
cases.
Potential
products
are
aminobenzyl
sulphonic
acids,
but
their
specific
chemical
identity
and
toxicity
are
not
known.

In
general,
most
azo
dyes
appear
to
resist
biodegradation
under
aerobic
conditions.
However,
microbes
living
in
an
anaerobic
environment
can
reduce
azo
bonds
resulting
in
significant
loss
of
color
(
Brown
and
Laboureur,
1983;
Weber
and
Adams,
1995;
Jank,
et
al.,
1998;
Baughman,
1995).
Some
of
the
products
of
the
anaerobic
transformation
of
azo
dyes
are
aromatic
amines.
Therefore,
in
the
environment
azo
dyes
can
lose
their
color
via
photoredox
reactions
and
by
anaerobic
biodegradation.
Anaerobic
biodegradation
is
most
likely
to
occur
in
sediments.

Fading
can
also
occur
by
reaction
with
trace
metals,
such
as
zinc,
tin,
aluminum,
iron,
and
copper,
mostly
via
redox
reactions.
However,
fading
is
primarily
a
photochemical
process.

Dyes
such
as
Acid
Blue
9
and
Acid
Yellow
23
are
completely
dissociated
in
the
environmentally
significant
pH
range
of
5
to
9.
Because
the
chromophoric
species
is
the
anion,
the
dyes
do
not
adsorb
strongly
to
soils
or
sediments
and
are
predominantly
associated
with
the
water
column.
Acid
Blue
9
has
been
long
used
as
a
hydrological
tracer
because
it
does
not
adsorb
strongly
on
soil
particulates
(
Mon
et
al.,
2005;
Tsai,
et
al.,
2004)
.
The
very
low
vapor
pressure
and
Henry's
Law
Constant
of
the
days
indicate
that
volatilization
would
not
be
a
significant
process.
As
highly
hydrophilic
chemicals
with
Log
Kow
<<
1,
they
are
not
likely
to
bioaccumulate
in
fish.

2.
Measures
of
Aquatic
Exposure
a.
Aquatic
Exposure
Modeling
16
Natural
photosensitizers
in
environmental
waters
include
dissolved
organic
matter
(
DOM),
singlet
oxygen
(
1O2),
and
hydroxy
radicals
(@
OH).
These
species
are
photooxidants.
In
additions,
surfaces
of
semiconducting
metal
oxides
(
such
as
titanium
dioxide
or
zinc
oxide)
may
serve
as
heterogeneous
photooxidants
and
it
is
a
potential
technology
to
treat
wastewater
containing
.
dye
residues.
(
Baran,
et
al.,
2003).
In
addition,
photoreactions
at
the
surface
of
iron
oxides/
hydroxides
may
also
be
involved.
25
All
of
these
products
are
to
be
applied
directly
to
a
confined
or
mostly
confined
water
body.
Therefore,
the
concentration
of
each
dye
in
water
is
only
the
result
of
direct
application
to
water
and
depends
on
the
relative
ratio
of
the
two
dyes
for
each
product
and
the
recommended
dose.
Neither
spray
drift
nor
runoff
are
routes
of
exposure
for
Acid
Yellow
23
and
Acid
Blue
9
in
the
water.
A
Drinking
Water
Assessment
was
performed.
and
it
was
concluded
that
there
was
no
exposure
from
drinking
water.
This
assessment
was
based
on
the
label
restrictions,
which
indicate
that
treated
water
is
not
to
be
used
as
a
drinking
water
source
for
humans.

For
the
aquatic
assessment,
the
concentrations
of
Acid
Yellow
23
and
Acid
Blue
9
were
those
target
concentrations
specified
in
the
labels
(
1
ppm
or
2
ppm,
depending
on
the
target
weed).
These
concentrations
are
attained
by
directly
adding
a
specified
volume
of
product
per
specified
volume
of
water
to
be
treated
(
i.
e.,
by
dilution
of
the
product
into
a
larger
volume
of
water).
It
should
be
noted
that
the
dosing
language
in
some
labels
(
e.
g.,
Aquashade
OA
)
are
vague
("
one
drop")
in
indicating
the
volume
of
product
to
be
added
to
the
water
body.
It
is
unclear
if
the
specified
concentration
is
in
terms
of
percent
of
product
(
i.
e.,
the
two
dyes
and
inerts)
nor
the
purity
of
the
dyes
are
taken
into
account..

The
concentrations
resulting
from
direct
application
are
summarized
in
Table
12.
Because
there
are
no
kinetics
data
for
indirect
photolysis
or
anaerobic
biodegradation,
it
is
assumed
that
these
concentrations
remain
constant.
For
Acid
Blue
9,
the
estimated
concentrations
do
not
take
into
account
the
type
of
counter
cation
(
i.
e.,
no
molar
fraction
correction,
given
the
uncertainty
in
the
identity
of
the
salt).
Even
though
the
environmental
concentrations
were
estimated
for
each
individual
dye,
there
are
no
ecological
toxicity
for
the
individual
dyes
because
the
test
substance
in
all
of
the
ecological
toxicity
studies
was
an
end­
use
Aquashade
product
containing
23.6%
Acid
Blue
9
and
2.39%
Acid
Yellow
23
of
uncertain
purity.
Therefore,
an
assessment
based
on
each
individual
dye
was
not
possible.
The
ecological
toxicity
of
products
with
a
higher
percentage
of
each
individual
dye
and/
or
different
ratio
of
the
dyes
than
Aquashade
may
potentially
be
under
represented.
These
products
are
Admiral
Liquid
and
Admiral
WSP.

Table
12
Maximum
Concentrations
of
Acid
Blue
9
and
Acid
Yellow
23
Expected
from
Direct
Application
of
Each
Product
at
the
Label
at
Rates
Set
in
the
Labels
and
assuming
100%
purity
of
each
dye
(
Source:
Product
labels)
Product
Application
Rate
of
Product
as
target
concentration,
ppm;
mg/
L
Concentration
of
Acid
Blue
9,
in
mg/
L
Concentration
of
Acid
Yellow
23,
in
mg/
L
Aquashade
OA
Aquashade
1
2
1
2
0.024
0.048
0.24
0.48
0.0024
0.0048
0.024
0.048
Pond
Care
AlgaeBlocker
1
0.024
0.0023
Admiral
Liquid
1
2
0.15
0.3
0.01
0.02
26
Admiral
WSP
1
2
0.497
(
0.5)

0.99
0.033
0.066
b.
Aquatic
Exposure
Monitoring
and
Field
Data
The
use
of
monitoring
data
and/
or
field
data
is
not
applicable
to
these
dyes
when
used
to
control
algal
growth
or
other
undesirable
aquatic
plants
in
confined,
with
no
or
minimal
outlet
water
bodies.
However,
these
dyes
also
have
industrial
applications
and
have
been
detected
in
waste
water,
sewage
systems,
and
water
bodies17.
Ecological
risk
from
those
sources
of
potential
exposure
are
not
addressed
in
this
document.

c.
Measures
of
Terrestrial
Exposure
3.
Terrestrial
Exposure
Modeling
Routinely,
exposure
to
birds
and
mammals
feeding
on
a
treated
field
is
estimated
by
modeling
residues
on
terrestrial
vegetation
and
invertebrates
using
the
Terrestrial
Residue
EXposure
(
TREX)
simulation
model.
Water
treated
with
Aquashade
or
any
of
the
other
products
is
not
expected
to
reach
terrestrial
vegetation
or
invertebrates.
Because
exposure
to
the
dyes
from
residues
on
terrestrial
food
items
is
not
expected,
the
use
of
terrestrial
exposure
models
is
not
applicable.
An
exception
could
be
if
treated
water
was
used
for
irrigation.
If
this
happened,
it
is
possible
the
dyes
in
these
formulations
would
get
on
terrestrial
food
items.
However,
given
the
low
toxicity,
it
is
unlikely
to
result
in
direct
acute
risk
to
terrestrial
animals.

a.
Terrestrial
Exposure
via
Consumption
of
Contaminated
Water
Modeling
To
determine
the
exposure
of
mammals
and
birds
to
"
Aquashade"
via
consumption
of
contaminated
water,
a
single
daily
dose
of
Aquashade
was
estimated
using
the
calculated
volume
of
water
that
birds
and
mammals
are
expected
to
consume
per
day
and
the
concentration
of
Aquashade
in
water
as
follows:

Daily
Exposure
(
mg)
=
Daily
Water
Consumption
(
L)
×
Water
Concentration
(
mg/
L).

To
estimate
the
volume
of
water
that
mammals
and
birds
are
expected
to
consume
per
day,
allometric
equations
from
the
EPA
Wildlife
Exposure
Factors
Handbook
(
US
EPA
1993)
were
used.
For
birds,
the
daily
water
consumption
(
L)
was
calculated
using
the
equation
(
US
EPA
1993,
Equation
3­
15,
p.
3­
8,
for
all
birds):

L
=
"
(
body
weight
in
kg)
$,
where
"
=
0.059
and
$
=
0.67
17
See,
for
example,
"
Background
Document
for
Identification
and
Listing
of
Deferred
Dye
and
Pigment
Wastes,
U.
S.
Environmental
Protection
Agency,
Office
of
Solid
Waste,
Washington,
D.
C.,
June
1999.
27
For
mammals,
the
daily
water
consumption
(
L)
was
calculated
as
using
the
equation
(
US
EPA
1993,
Equation
3­
7,
p.
3­
10,
for
all
mammals):

L
=
"
(
body
weight
in
kg)
$,
where
"
=
0.099
and
$
=
0.9
The
maximum
concentration
(
weight
to
volume)
of
Aquashade
in
water
after
application
at
the
labeled
rate
is
2
mg/
L.
As
summarized
in
Table
13,
a
single
daily
exposure
via
drinking
water
containing
2
mg/
L
Aquashade
was
calculated
for
three
weight
classes
for
birds
(
20,
100,
and
1000g)
and
mammals
(
15,
35,
and
1000g).

Table
13.
Exposure
estimates
for
birds
and
mammals
via
consumption
of
water
contaminated
with
Aquashade
at
a
maximum
calculated
concentration
of
2
mg/
L.
Taxon
Body
Weight
(
g)
Estimated
Water
Consumption
(
L)
Estimated
Exposurea
(
mg)
Birds
20
0.0043
0.008
100
0.013
0.026
1,000
0.059
0.119
Mammals
15
0.0023
0.004
35
0.0048
0.01
1,000
0.099
0.1
a
Estimated
Daily
Exposure
(
mg)
=
Daily
water
consumption
(
L)
×
Water
concentration
2
mg/
L).

b.
Residue
Studies
There
is
no
terrestrial
exposure
from
this
use
pattern.
Therefore,
residue
data
are
not
necessary.

C.
Ecological
Effects
Characterization.

This
document
is
a
screening
level
evaluation
of
the
potential
ecological
risk
from
the
end­
use
products
that
contain
Acid
Yellow
23
and
Acid
Blue
9
dyes,
and
are
represented
by
a
formulation
referred
to
as
Aquashade.
All
of
the
products
are
composed
of
two
dyes
that
are
never
used
individually
as
an
algicide/
aquatic
herbicide.
These
products
are
applied
directly
to
confined
water
bodies
and
never
to
terrestrial
or
aquatic
ecosystems.

Available
data
are
from
tests
conducted
with
either
a
formulation
containing
Acid
Yellow
23
and
Acid
Blue
9,
or
with
the
dyes
separately.

All
of
the
toxicity
studies
on
birds,
fish
and
aquatic
invertebrates
submitted
by
the
registrant
were
conducted
with
the
end­
use
product
Aquashade
(
23.6%
Acid
Blue
9
and
2.39%
Acid
Yellow
23)
and
not
with
the
individual
dyes.
The
studies
were
submitted
to
support
the
registration
of
this
"
Aquashade"
end­
use
product
(
EPA
Reg.
No.
33068­
1).
Through
searches
conducted
by
the
ORD
Middle
Ecological
Division
at
Duluth
under
their
ECOTOX
literature
28
search
program,
ecological
toxicity
studies
conducted
with
the
individual
dyes
were
located.
The
results
of
these
published
literature
data
supported
the
results
of,
and
did
not
indicate
greater
toxicity
than,
or
effects
other
than
the
registrant
submitted
tests,
therefore,
they
were
not
used
to
estimate
risk
in
this
assessment.
A
laboratory
rat
study
was
conducted
with
the
Admiral
WSP
product
as
the
test
substance
(
49.62%
Acid
Blue
9
and
3.05%
Acid
Yellow
23).

There
is
an
uncertainty
in
the
purity
of
the
test
substance
used
in
the
ecological
toxicity
studies.
However,
considering
the
low
exposure
level
relative
to
even
the
lowest
assumed
purity,
this
uncertainty
does
not
cause
doubt
in
the
conclusions.

The
studies
submitted
to
fulfill
the
"
basic
six"
requirements
(
an
avian
oral,
two
avian
dietary,
two
fish
acute,
and
one
aquatic
invertebrate
acute
toxicity)
found
very
low
toxicity
(
practically
nontoxic
and
slightly
toxic).
A
study
on
the
laboratory
rat
found
an
LD50
equal
to
or
higher
than
5,000
ppm.
Table
14
summarizes
the
acute
data
for
the
tested
species
used
to
assess
risk.
29
Table
14.
Summary
of
most
sensitive
acute
data.
No
chronic
data
were
required
for
birds,
fish
or
aquatic
invertebrates.
Species
Study
Type1
Source
Results2
Test
Material
Aquashade
Bobwhite
quail
Acute
Avian
Oral
433367­
01
LD50
&
acute
NOAEC
$
2250
mg/
kg
Mallard
duck
Acute
Avian
Oral
433367­
02
LD50
&
acute
NOAEC
$
2250
mg/
kg
Bobwhite
quail
Avian
Dietary
435034­
03
LC50
&
acute
NOAEC
$
5,
620ppm
Mallard
duck
Avian
Dietary
435034­
04
LC50
&
acute
NOAEC
$
5,620ppm
Bluegill
sunfish
Fish
Toxicity
Bluegill
432975­
02
LC50
&
acute
NOAEC
$
96
mg/
L
Rainbow
trout
Fish
Toxicity
Rainbow
Trout
432975­
01
LC50
&
acute
NOAEC
$
96
mg/
L
Daphnia
magna
Invertebrate
Toxicity
432975­
03
LC50
&
acute
NOAEC
$
97
mg/
L
Test
Material
Admiral
WSP
Laboratory
rat
Wild
Mammal
Acute
452811­
01
LD50
%
&
&
$
5,000
mg/
kg
1
All
were
performed
with
the
Typical
End­
use
Product,
Aquashade
or
Admiral
WSP
2
The
results
refer
to
the
concentration
of
the
end­
use
product.

Birds
and
mammals
are
expected
to
drink
from
ponds
that
have
been
treated
with
the
end­
use
products.
EFED
calculated
an
estimated
dose
that
they
would
receive
from
that
water
(
see
Table
10).

1.
Terrestrial
Effects
Avian
effects
The
avian
oral
acute
toxicity
studies
for
both
the
bobwhite
quail
and
the
mallard
duck
found
LD50'
s
and
NOAECs
equal
to
or
greater
than
2,250
mg/
kg.
The
avian
dietary
toxicity
studies
found
LC50s
and
NOAELs
equal
to
or
greater
than
5,620
ppm.
Aquashade
is
considered
to
be
practically
nontoxic
to
birds.
Higher
tiered
studies
are
not
required.

Mammalian
effects
An
acute
toxicity
study
that
used
Admiral
WSP
with
the
laboratory
rat
was
reviewed
by
OPP's
Health
Effects
Division
found
an
LD50
equal
to
or
greater
than
5,000
mg/
kg.
EFED
30
considers
this
to
be
practically
nontoxic.
The
Health
Effects
Division
wrote
that,
".
.
.
this
formulation
contained
49.62
%
Acid
Blue
dye
and
3.05%
Acid
Yellow
dye
(
presumably
Acid
Blue
9
[
Erioglaucine],
and
Acid
Yellow
23
[
Tartrazine],
respectively).
A
chronic
mammal
toxicity
test
with
Acid
Yellow
23
indicated
low
likelihood
of
adverse
chronic
effects
at
environmentally
relevant
exposure
levels.

Calculating
Daily
Dose
Terrestrial
animals
drink
water
from
ponds
and
other
bodies
and
may
be
affected
by
the
Aquashade
or
any
of
the
other
end­
use
products
in
the
water.
Exposure
estimates
for
terrestrial
animals
via
drinking
contaminated
water
were
expressed
in
terms
of
a
single
daily
dose
(
mg)
for
three
different
body
weight
classes.
All
of
the
exposures
were
less
than
0.2
mg
a.
i./
day.

Birds
The
weight
classes
for
birds
were
20,
100,
and
1,000
g.
To
obtain
the
toxicity
value
for
a
daily
dose
(
mg
of
Aquashade)
for
each
weight
class
for
birds
termed
the
"
derived
median
dose,"
the
acute
oral
LD50
($
2,250
mg/
kg­
bw)
was
multiplied
by
body
weight
(
kg).
The
calculated
daily
doses
for
birds
for
each
weight
class
are
summarized
in
Table
13.

Mammals
Exposure
estimates
for
terrestrial
animals
via
drinking
of
contaminated
water
were
expressed
in
terms
of
a
single
daily
dose
(
mg)
for
three
different
body
weight
classes
(
15,
25,
and
1,000
g).
Therefore,
to
obtain
the
lethal
dose
(
mg
of
Aquashade)
for
each
weight
class
for
mammals,
the
acute
rat
oral
LD50
value
of
$
5,000
mg/
kg­
bw
(
MRID
452811­
01)
was
multiplied
by
body
weight
(
kg).
The
calculated
daily
doses
for
mammals
for
each
weight
class
are
summarized
in
Table
15.

Table
15.
Acute
toxicity
values
for
birds
and
mammals
drinking
from
contaminated
water,
expressed
in
terms
of
a
single
Aquashade
dose
(
mg).
Animals
Body
Weight
(
kg)
LD50
(
mg/
kg­
bw)
Derived
Median
Lethal
Dose
(
mg)
a
Birds
0.020
$
2,250
$
45
0.100
$
2,250
$
225
1.000
$
2,250
$
2,250
Mammals
0.015
$
5,000
$
75
0.035
$
5,000
$
175
1.000
$
5,000
$
5,000
a
Derived
Median
Lethal
Dose
(
mg)
=
body
weight
(
kg)
×
LD50
(
mg/
kg
body
weight).
31
Terrestrial
Plant
effects
The
method
of
application,
i.
e.
pouring
and
dropping
specific
volumes
or
packets
into
the
target
water
body,
is
not
expected
to
result
in
drift
or
runoff
resulting
in
exposure
to
terrestrial
plants.
Since
it
will
not
be
applied
to
terrestrial
plants
no
plant
studies
were
required.
One
uncertainty
exists,
however,
in
that
not
all
the
labels
preclude
use
of
treated
water
for
irrigation.
If
it
was
used
for
irrigation,
the
potential
routes
of
exposure
would
be
drift
and/
or
runoff.
To
assess
risk
to
terrestrial
plants,
tier
1
or
tier
2
terrestrial
plant
testing
would
be
used.
If
the
labels
stated
that
treated
water
must
not
be
used
for
irrigation,
such
testing
would
be
unnecessary.
Some
of
the
labeling
indicates
that
emergent
plants
may
experience
burning
if
exposed
to
the
dye
formulations.
Based
on
this,
it
is
assumed
that
any
inadvertent
exposure
to
terrestrial
plants
might
cause
adverse
effects.

2.
Aquatic
Effects
The
registrant
submitted
studies
that
determined
the
toxicity
of
Aquashade
to
aquatic
animals.
The
bluegill
sunfish
and
rainbow
trout
studies
found
LC50s
and
NOAECs
equal
to
or
greater
than
96
mg/
L.
This
level
is
considered
by
EFED
to
be
slightly
toxic.
The
level
for
being
considered
practically
nontoxic
is
100
mg/
L
for
aquatic
animals.

The
aquatic
invertebrate
(
Daphnia
magna)
study
found
that
the
LC50
and
the
NOAEC
were
equal
to
or
greater
than
97
mg/
L.
This
level
is
considered
to
be
slightly
toxic.
No
further
studies
are
required.

Aquashade,
Admiral,
and
AlgaBlocker
are
algicides
or
herbicides
for
undesirable
aquatic
plants.
They
are
not
applied
to
terrestrial
environments
nor
if
treated
water
used
for
irrigation.
Because
submerged
aquatic
plants
are
the
target
species
and
it
is
assumed
that
all
submerged
plants
will
be
killed,
no
aquatic
plant
studies
were
required.

3.
ECOTOX
and
other
Published
information
The
Agency
also
obtained
summary
toxicity
information
from
published
literature
found
as
part
of
the
ECOTOX
program
maintained
by
the
EPA
Office
of
Research
and
Development
Middle
Ecological
Division
(
MED)
at
Duluth.
This
program
regularly
searches
open
literature
for
toxicity
information.
OPP
requested
of
MED
Duluth
to
search
its
holdings
for
toxicity
information
on
aquashade
and
the
dye
components
of
the
formulations
being
assessed
for
reregistration.
Toxicity
information
was
located
on
mammals
and
aquatic
animals
from
tests
with
either
of
the
dyes.
In
all
cases,
the
test
results
located
through
ECOTOX
corroborated
the
findings
of
the
registrant
submitted
data,
and
did
not
indicate
effects
unforeseen
from
the
registrant
studies.
Neither
did
these
published
literature
studies
indicate
adverse
effects
to
organisms
other
than
those
tested
by
the
registrant.
None
of
the
data
located
through
ECOTOX
were
used
in
the
assessment.
See
Appendix
F
for
a
summary
table
of
data
located
from
ECOTOX.
32
IV.
Risk
Characterization
Acid
Yellow
23
and
Acid
Blue
9
are
applied
together
via
the
end­
use
products
Aquashade
Admiral
Liquid.
Admiral
WSP,
and
AlgaBlocker.
These
products
are
applied
to
confined
water
bodies
or
water
bodies
with
minimal
outflow.
None
of
the
products
have
terrestrial
uses
and
would
not
directly
contact
terrestrial
food
items
for
birds
and
mammals.
Therefore,
the
RQs
based
on
dietary
exposure
from
Aquashade
residues
on
food
items
were
not
calculated.
However,
it
could
affect
them
via
drinking
water.

Thus,
all
of
the
potential
risks
of
Acid
Yellow
23
and
Acid
Blue
9
are
associated
with
direct
exposure
of
terrestrial
animals
through
drinking
water
or
immersion
in
the
treated
water.
In
addition,
aquatic
organisms
dwelling
in
treated
water
are
also
exposed
to
the
dyes
(
refer
to
"
Conceptual
Model")
Risks
were
estimated
based
on
"
environmental"
concentrations
in
a
water
body
resulting
from
direct
application
of
the
product(
s)
that
is,
a
dilution
concentration
based
on
the
application
rates
indicated
in
the
labels
(
see
Table
10).
It
must
be
kept
in
mind
that
the
characterization
of
effects
was
based
in
terms
of
the
combined
two
dyes
for
the
Aquashade
enduse
product
(
23.6%
Acid
Blue
9
and
2.36%
Acid
Yellow
23)
used
in
the
ecological
toxicity
studies
and
not
for
the
individual
dyes.

A.
Risk
Estimation
­
Integration
of
Exposure
and
Effects
Data
1.
Birds
and
Mammals
Exposure
to
birds
and
mammals
was
expected
primarily
through
consumption
of
drinking
water
treated
with
the
dyes.
Birds
are
surrogates
for
reptiles
and
terrestrial
phase
amphibians
because
it
is
assumed
for
screening
level
assessment
purposes
that
reptiles
and
terrestrial
phase
amphibians
are
not
more
sensitive
to
oral
toxicity
than
birds.

Acute
Risk
Quotients
(
RQs)
via
Consumption
of
Contaminated
Water
Acute
RQs
for
birds
and
mammals
exposed
to
Aquashade
via
consumption
of
contaminated
water
were
calculated
for
each
of
three
body
weight
classes
using
the
daily
exposure
value
expressed
as
milligrams
of
Aquashade
(
Table
13)
and
the
toxicity
value
expressed
in
terms
of
milligrams
of
Aquashade
(
Table
14).
The
RQs
are
summarized
in
Table
16.
33
Table
16.
Acute
Risk
Quotients
(
RQ)
for
birds
and
mammals
exposed
to
Aquashade
through
consumption
of
contaminated
water.
Taxa
Body
weight
(
g)
Estimated
Environmental
Exposurea
(
mg)
Derived
Medium
Lethal
Dose
(
DMDL)
b
Acute
RQ
EEC/
DMDL
Birds
20
0.01
$
45
<
0.01
100
0.02
$
225
<
0.01
1,000
0.12
$
2,250
<
0.01
Mammals
15
<
0.01
$
75
<
0.01
35
<
0.01
$
175
<
0.01
1,000
<
0.10
$
5,000
<
0.01
a
Estimated
Environmental
Exposure
(
EEC)
=
Daily
water
consumption
(
L)
×
Water
concentration
(
mg/
L).
Rounded
to
two
decimal
places.
b
Derived
Median
Lethal
Dose
(
mg)
=
body
weight
(
kg)
×
LD50
(
mg/
kg
body
weight).
c
Acute
RQ
=
Estimated
Environmental
Exposure
/
Concentration
Derived
Median
Lethal
Dose
RQs
are
below
the
Levels
of
Concern
(
LOCs)
for
acute
risk
(
LOC
0.5),
acute
restricted
use
(
LOC
0.2),
and
acute
endangered
species
(
LOC
0.1).

Acute
RQs
are
below
the
Levels
of
Concern
(
LOCs)
for
endangered
species
(
0.1).

Use
of
Treated
Water
for
Irrigation
While
it
is
considered
highly
unlikely
that
treated
water
would
be
used
for
irrigation,
given
the
kinds
of
water
bodies
treated,
if
this
happened,
exposure
to
birds
and
mammals
might
occur
through
ingesting
terrestrial
food
items
that
had
been
exposed
to
the
water.
Given
the
low
toxicity
of
the
dyes,
and
the
relative
low
concentration
in
the
treated
water
(
no
more
than
2
ppm),
it
is
unlikely
that
this
route
of
exposure
would
cause
direct
toxicity
to
birds
or
mammals.
If
2
inches
of
irrigation
water
containing
2
ppm
were
sprayed
onto
terrestrial
vegetation,
the
resulting
"
application
rate"
of
the
dyes
would
be
approximately
1
lb
ai/
acre.
Applying
1
lb
ai/
acre
to
short
grass
would
result
in
residues
no
greater
than
240
ppm.
For
a
small
(
15g)
mammal
that
eats
close
to
its
body
weight
this
would
still
result
in
a
dose
per
animal
much
less
than
the
lowest
toxicity
value
for
mammals.
See
appendix
E
for
calculations.

2.
Non­
target
Aquatic
Animals
and
Plants
Aquashade
and
any
of
the
other
end­
use
product
containing
Acid
Blue
9
and
Acid
Yellow
23
are
only
applied
directly
to
ponds
and
other
aquatic
systems
without
an
outlet.
It
is
not
expected
to
come
in
contact
with
non­
target
aquatic
animals
outside
of
those
systems.

This
is
a
screening
level
assessment
of
the
acute
risk
of
using
Aquashade,
therefore,
the
highest
EECs
(
2
mg/
L;
for
Admiral
Liquid
and
Admiral
WSP)
and
the
lowest
toxicity
values
were
used.
The
concentration
in
the
treated
systems
will
be
higher
than
any
body
of
water
that
34
accidentally
receives
(
by
flooding,
etc.)
a
dose
of
Aquashade.
A
summary
of
aquatic
acute
RQs
is
presented
in
Table
17.

Table
17.
Estimated
acute
risk
quotients
(
RQ)
for
aquatic
animals
exposed
to
Aquashade.
In
this
screening
analysis
only
the
highest
Estimated
Environmental
Concentration
(
EEC
=
2
mg/
L)
and
lowest
NOAECs
were
used.
Animal
EEC
(
mg/
L)
NOAEC
RQ
(
EEC
/
NOAEC)
LOCs
Exceeded
Bluegill
sunfish
2
96
mg/
L
<
0.01
none
Rainbow
trout
2
96
mg/
L
<
0.01
none
Daphnia
magna
2
97
mg/
L
<
0.01
none
Fish
are
used
as
surrogated
for
aquatic
phase
amphibians,
so
lack
of
risk
to
fish
results
in
a
conclusion
of
low
risk
to
amphibians.

Aquatic
Plants:
This
is
a
screening
level
assessment
based
on
an
Aquashade
product.
Aquashade,
AlgaeBlocker,
and
Admiral
products
are
aquatic
algicides
or
herbicides
for
undesirable
aquatic
plants
and
are
expected
to
kill
all
submerged
aquatic
plants.
However,
since
it
is
only
applied
to
ponds
with
little
or
no
outlet,
it
is
not
expected
to
come
into
contact
with
nontarget
aquatic
out
side
of
the
target
pond.
All
of
the
submerged
plants
in
the
pond
are
considered
to
be
targets.
Therefore,
the
RQs
were
not
calculated
for
aquatic
plants.
If
the
water
treated
by
these
products
were
used
for
irrigation,
it
is
unlikely
the
concentration
of
dyes
in
the
runoff
from
such
irrigation
would
be
sufficient
to
cause
effects
to
aquatic
plants
in
receiving
water
bodies.

3.
Non­
target
Terrestrial
Plants
in
Dry­
land
and
Semi­
aquatic
Environments
Terrestrial
plants
growing
in
dry­
land
and
semi­
aquatic
environments
are
not
expected
to
be
exposed
to
the
end­
use
products,
since
they
products
are
poured,
or
applied
with
an
eyedropper
into
confined
water
bodies
or
water
bodies
with
minimal
outflow.
Because
the
labels
do
not
allow
application
to
terrestrial
ecosystems,
a
terrestrial
plant
risk
characterization
was
not
performed.
An
uncertainty
exists
in
that
the
labels
do
not
preclude
use
of
treated
water
for
irrigation.
If
this
happened,
there
is
a
potential
for
adverse
effects
to
terrestrial
plants
because
some
labels
indicate
that
if
emerged
plants
are
exposed
to
the
formulations,
burning
would
be
expected.

B.
Risk
Description
­
Interpretation
of
Direct
Effects
Aquashade
does
not
exceed
the
level
of
concern
for
animals
with
which
it
is
expected
to
come
in
contact.
Because
of
the
method
of
application,
no
drift
or
runoff
onto
terrestrial
habitats
is
expected
precluding
exposure
to
terrestrial
plants,
and
to
food
items
consumed
by
terrestrial
animals.
Thus,
the
risk
hypothesis
is
proven
wrong
for
these
organisms.

The
mode
of
herbicidal
action
and
intended
use
will
result
in
adverse
effects
to
target
aquatic
plants,
therefore
the
risk
hypothesis
that
these
formulations
adversely
affect
aquatic
plants
is
correct.
However,
in
all
cases,
the
treated
water
body
is
treated
for
the
express
reason
of
35
controlling
(
eliminating)
submerged
aquatic
plants
therefore,
this
effect
is
not
interpreted
as
an
adverse
ecological
effects
unless,
as
discussed
below,
there
are
endangered
aquatic
plants
in
these
treated
water
bodies.

C.
Endangered
and
Threatened
Species
The
use
of
these
dyes
may
affect
endangered
aquatic
plants
possibly
occurring
in
ponds
where
this
might
be
used.
It
may
also
have
the
potential
to
affect
endangered
animal
species
which
depend
on
aquatic
plants
in
these
managed
water
bodies
for
food
and/
or
habitat.
The
potential
for
this
direct
effects
to
listed
aquatic
plants,
and
indirect
effects
to
listed
animals
is
very
limited
because
of
the
method
of
application
(
i.
e.
by
hand),
and
the
kinds
of
water
bodies
treated
(
golf
course
ponds
and
other
managed
ponds)
where
keeping
the
water
free
of
all
vegetation
is
aesthetically
desirable.
These
water
bodies
do
not
typically
have
outflow
to
streams
or
rivers,
which
would
preclude
endangered
fish
and
invertebrates
from
entering.
However,
endangered
amphibians
may
enter
such
water
bodies,
and
the
loss
of
plants
that
serve
as
food
and
shelter
for
these
amphibians
have
the
potential
indirectly
affect
these
organisms.

1.
Action
Area
For
listed
species
assessment
purposes,
the
"
action
area"
is
considered
to
be
the
area
affected
directly
or
indirectly
by
the
Federal
action
and
not
merely
the
immediate
area
involved
in
the
action.
At
the
initial
Level
I
screening
assessment,
broadly
described
taxa
are
considered
and
thus
the
screening
conservatively
assumes
that
listed
species
within
those
broad
groups
are
colocated
with
the
pesticide
treatment
area.
This
means
that
terrestrial
plants
and
wildlife
are
assumed
to
be
located
on
or
adjacent
to
the
treated
site,
and
aquatic
organisms
are
assumed
to
be
located
in
a
surface
water
body
adjacent
to
the
treated
site.
The
assessment
also
assumes
that
the
listed
species
are
located
within
an
assumed
area
that
has
the
relatively
highest
potential
exposure
to
the
pesticide,
and
that
exposures
are
likely
to
decrease
with
distance
from
the
treatment
area.

Aquashade,
Admiral,
and
AlgaBlocker
are
used
in
ornamental
ponds,
recreational
manmade
ponds,
closed
system
ponds,
golf
course
lakes,
rearing
lakes
for
fish,
fountains
containing
fish,
and
recirculated
or
artificial
waterscapes.
Therefore,
these
products
are
only
applied
to
confined
water
bodies.

2.
Taxa
Potentially
at
Risk
In
this
assessment,
it
is
concluded
that
the
only
species
potentially
at
risk
are
aquatic
plants
from
direct
effects
if
they
occur
in
treated
ponds
and
lakes,
and
aquatic
or
terrestrial
animals
that
depend
on
the
vegetation
in
the
treated
water
bodies.

3.
Discussion
of
Risk
Quotients
No
direct
effects
on
listed
animal
species
are
predicted.
No
exposure
to
terrestrial
plants
was
expected,
so
no
terrestrial
plant
RQs
were
calculated.
Even
though
no
RQs
were
calculated
36
for
aquatic
plants,
it
is
assumed
they
will
be
affected,
since
they
are
the
target
organisms
to
be
controlled.
4.
Probit
Dose
Response
Relationship
Since
no
mortality
was
observed
in
the
standard
studies,
no
probit
analysis
was
performed.

5.
Data
Related
to
Under­
represented
Taxa
The
screening
level
assessment
relies
on
RQ
calculations
that
use
toxicity
endpoints
selected
from
the
most
sensitive
species
tested
within
broad
taxonomic
groups.
There
may
be
situations
in
which
additional
effects
data
from
one
or
more
sources
may
suggest
that
a
given
suite
of
listed
taxa
may
be
more
or
less
sensitive
than
suggested
by
the
effects
data
used
for
RQ
calculations.
No
such
relationships
were
found
in
this
analysis.

6.
Implications
of
Sublethal
Effects
The
Level
I
screening
assessment
normally
relies
on
the
acute
mortality
endpoint
as
well
as
a
suite
of
sublethal
responses
to
the
chemical
stressor.
EFED
decided
that,
based
on
its
mode
of
action,
the
low
chronic
toxicity
of
at
least
one
of
the
dyes
to
mammals,
low
potential
for
exposure
based
on
use
pattern,
and
its
history
as
a
human
food
dye,
Aquashade
did
not
need
chronic
toxicity
studies.
Sublethal
effects
were
not
analyzed.

7.
Indirect
Effects
Analysis
This
assessment
indicates
that
Aquashade
may
impact
listed
invertebrates,
fish
and
amphibians
that
depend
upon
submerged
aquatic
plants
and
algae.
There
is
also
a
potential
for
indirect
effects
to
listed
terrestrial
birds
and
mammals
that
occur
near
treated
water
bodies,
and
that
depend
on
submerged
aquatic
vegetation
or
algae
for
food
or
other
survival
or
reproductive
processes.
The
likelihood
of
potential
indirect
effects
has
not
been
quantified
or
refined.
Such
refinement
could
incorporate
information
on
specific
use
locations
relative
to
specific
locations
of
listed
species.
Subsequent
refinement
may
also
include
additional
analyses
on
potential
indirect
effects
to
listed
species.

8.
Critical
Habitat
Aquashade
may
impact
invertebrates
and
fish
that
depend
upon
the
target
species
(
submerged
aquatic
plants
and
algae).
The
likelihood
of
potential
indirect
effects
has
not
been
quantified.
Subsequent
refinement
may
include
additional
analyses
on
potential
indirect
effects
to
listed
species.

9.
Co­
occurrence
Analysis
The
goal
of
the
analysis
for
co­
location
is
to
determine
whether
sites
of
pesticide
use
are
geographically
associated
with
known
locations
of
listed
species.
Listed
species
potentially
at
risk
include
aquatic
plants,
and
animals
that
depend
on
aquatic
effects
from
indirect
effects.
Aquashade's
use
pattern
makes
it
(
potentially)
possible
that
it
will
be
used
in
every
county
in
the
37
United
States.
Therefore,
EFED's
database
to
find
which
counties
harbor
which
listed
species
("
LOCATES")
was
not
used.
However,
there
are
possible
effects
to
D.
Description
of
Assumptions,
Limitations,
Uncertainties,
Strength,
and
Data
Gaps
1.
Environmental
fate
Data
Source
The
environmental
fate
assessment
is
only
at
the
qualitative
level.
Structure­
activity
estimations
(
EPIWIN)
of
physical
and
chemical
properties/
characteristics
are
uncertain.
EPIWIN
is
not
suitable
for
estimating
properties
of
salts
of
large
anions,
such
as
in
acid
dyes.
Therefore,
this
qualitative
environmental
fate
assessment
was
primarily
based
on
the
general,
known
behavior
of
dyes
in
the
environment
and
not
on
the
specific
behavior
of
Acid
Blue
9
and
Acid
Yellow
23.

Reaction
kinetics
and
transformation
products
Although
indirect
photolysis
was
identified
as
a
major
route
of
dissipation
of
the
dyes
in
aquatic
media,
the
identity
of
the
photoproducts
(
and
hence,
their
toxicity)
is
not
known.
The
rate
of
phototransformation
is
expected
to
be
a
function
of
geographical
location
and
season.
Biotransformation
under
anaerobic
conditions
may
also
contribute
to
the
dissipation
of
the
dyes
and
can
occur
predominately
in
sediment,
but
the
chemical
identity
of
the
metabolites
is
not
known.

Impurities
The
characterization
of
the
physical
and
chemical
properties
required
under
FIFRA
were
not
done
with
a
100%
pure
dye.
The
Acid
Blue
9
is
reported
to
be
50%
pure
and
Acid
Yellow
23
as
being
28%.
Although
the
presence
of
chemical
impurities
affect
the
physical
and
chemical
properties
of
a
chemical
substance,
the
extent
by
which
they
affect
those
of
the
dyes
is
not
known.
The
physical
and
chemical
properties
were
submitted
in
support
of
Aquashade
(
23.6%
Acid
Blue
9
and
2.39
Acid
Yellow
2.39%
registration
In
is
unclear
if
these
percentages
take
into
account
the
purity
of
each
dye.
Furthermore,
the
percent
purity
of
each
dye
in
the
other
products
is
unknown.

Given
the
low
exposure
of
these
dyes
in
the
environment,
when
used
as
pesticides,
no
Subdivision
N
guideline
studies
are
needed
for
the
intended
uses.
However,
if
new
uses
are
petitioned,
the
EFED
will
reevaluate
the
status
of
data
requirements.

2
Ecological
toxicity
The
assessment
assumes
that
Aquashade
and
related
products
will
be
used
strictly
in
accordance
with
label
instructions.
It
assumes
that
there
will
be
no
large
spills
of
the
product
The
ecological
toxicity
studies
were
not
conducted
with
the
individual
dyes,
but
with
the
mixture
of
the
two
dyes
as
the
test
substance
(
23.6%
Acid
Blue
9
and
2.36
%
Acid
Yellow
23,
An
38
Aquashade
end­
use
product).
There
are
two
other
products
that
contain
a
higher
percent
of
the
dyes
than
the
tested
substance,
namely
Admiral
Liquid
(
15.31%
Acid
Blue
9
and
1.00%
Acid
Yellow
23)
and
Admiral
WSP
(
49.
%
Acid
Blue
9
and
%
Acid
Yellow
23).
Therefore,
the
ecological
toxicity
of
these
products
is
not
known.
However,
the
maximum
estimated
environmental
concentration
resulting
from
direct
application
to
a
water
body
is
below
the
Level
of
Concern.

The
chronic
toxicity
RQs
cannot
be
calculated,
because
there
are
no
studies
on
chronic
risk
with
the
pesticide.
The
low
acute
toxicity
values,
along
with
testing
with
the
Acid
Yellow
dye
23
with
mammals,
and
the
mode
of
action
of
Aquashade
and
other
products,
suggest
low
chronic
toxicity
and
EFED
has
not
required
any
chronic
studies.

The
Aquashade
product
that
was
used
in
the
"
basic
six"
studies
was
labeled
as
containing
23.63
%
Acid
Blue
9
and
2.39%
Acid
Yellow
23.
The
study
authors
stated,
"
The
test
substance
characterization
provided
by
the
sponsor
indicated
a
purity
of
13.9%
azure
blue
dye."
Therefore
the
exact
composition
of
the
toxicant
that
was
used
in
the
studies
is
not
certain.
The
use
of
the
word
"
azure"
could
describe
color,
but
also
the
word
"
azure"
could
be
associated
with
a
series
of
structurally
related
dyes
that
are
totally
different
from
Acid
Blue
9
or
Acid
Yellow
23.
39
V.
Literature
Cited
E.
Open
Literature
Books
Lynch,
D.
G.
"
Estimating
the
Properties
of
Synthetic
Organic
Dyes",
in
Handbook
of
Property
Estimation
Methods
for
Chemicals­
Environmental
Health
Sciences,
Edited
by
Robert
S.
Boethling
and
Donald
Mackay.
Published
by
Lewis
Publishers,
Boca
Raton,
FL,
2000;
Pages
447­
467.
And
pertinent
references
therein.

Marmion,
D,
M.
Handbook
of
U.
S.
Colorants­
Food,
Drugs,
and
Medical
Devices,
Third
Edition.,
1991.
Published
by
John
Wiley
and
Sons,
New
York
Helz,
G.
R.,
Zepp,
R.
G.,
and
Crosby,
D.
G,
Editors.
1994.
Aquatic
and
surface
photochemistry.
Lewis
Publishers,
Boca
Raton,
Florida.

Journal
Articles
Jank.
M.,
Köser,
H.,
Lücking,
F.,
Martienssen,
M.,
and
Wittchen,
S.
1998,
"
Decolorization
and
Degradation
of
Erioglaucine
(
Acid
Blue
9)
Dye
in
Wastewater",
Environmental
Technology,
v.
19(
7),
pp.
741­
747.

Weber,
E.
J.
and
Adams,
R.
L.
1995.
"
Chemical
and
Sediment­
Mediated
Reduction
of
the
Azo
Dye
Disperse
Blue
79",
Environ.
Sci.
Technol.
v.
29,
pp.
1163­
1170.

Baughman,
G.
L.
1995.
"
Fate
of
azo
dyes
in
aquatic
systems.
Part
3:
The
role
of
suspended
sediments
in
adsorption
and
reaction
of
acid
and
direct
dyes",
Dyes
and
Pigments,
v.
27,
pp.
197­
210.

Brown,
D.
and
Laboureur,
P.
1983,
"
The
Degradation
of
Dyestuffs:
Part
I­
Primary
biodegradation
under
anaerobic
conditions",
Chemosphere,
v.
122,
pp.
397­
404.

Baran,
W.,
Makowski,
A.,
Wardas,
W.
2003.
"
The
influence
of
FeCl3
on
the
photocatalytic
degradation
of
dissolved
azo
dyes
in
aqueous
TiO2
suspensions",
Chemosphere,
v.
53,
pp
82­
95.

Tai,
W.
T.,
Chang,
C.
Y,
Ing,
C.
H.,
and
Chang,
C.
F.
2004.
"
Adsorption
of
acid
dyes
from
aqueous
solutions
on
activated
bleaching
earth",
J.
Colloid
and
Interface
Science,
vol.
275,
pp
72­
78.

Mon,
J.,
Flury,
M.,
and
Harsh,
J.
B.
2005
."
Sorption
of
four
triarylmethane
dyes
in
a
sandy
soil
determined
by
batch
and
column
experiments",
Geoderma.
in
press.

Mon,
J.,
Flury.
M.,
and
Harsh.
2005
"
A
quantitative
structure­
activity
relationship
(
OSAR)
analysis
of
triarylmethane
dye
tracers",
Journal
of
Hydrology,
in
press.
40
Characterization
of
Physical
and
Chemical
Properties
Submitted
to
the
Agency
in
Support
of
Aquashade
(
23.6%
and
2.39%
Acid
Yellow
23:

Ellison.
F.
E.
1994.
"
Aqua
Shade
Blue
9'­
Physical
and
Chemical
Characteristics
of
Aqua
Shade
Blue
9:
Color,
Physical
State,
Odor,
Boiling
Point,
Specific
Gravity,
Solubility,
Vapor
Pressure,
pH,
and
Stability".
Performed
by
Case
Consulting
Laboratories,
Inc.
Whippany,
NJ
and
sponsored
by
Applied
Biochemistry,
Inc.,
Milwaukee,
WI.
Report
Dated
September
12,
1994.
MRID
43503401
Ellison.
F.
E.
1994.
"
Aqua
Shade
Yellow
23"­
Physical
and
Chemical
Characteristics
of
Aqua
Shade
Blue
9:
Color,
Physical
State,
Odor,
Boiling
Point,
Specific
Gravity,
Solubility,
Vapor
Pressure,
pH,
and
Stability".
Performed
by
Case
Consulting
Laboratories,
Inc.
Whippany,
NJ
and
sponsored
by
Applied
Biochemistry,
Inc.,
Milwaukee,
WI.
Report
Dated
September
12,
1994.
MRID
43503402
5.
Data
Generation
Structure­
activity
Estimates:
EPI
(
Estimation
Programs
Interface)
SuiteTM
(
formerly
known
as
EPIWIN):
http://
www.
epa.
gov/
opptintr/
exposure/
docs/
episuite.
htm
SRC.
2004.
Syracuse
Research
Corporation.
Interactive
Physical
Properties
(
PHYSPROP)
Database
Demo.
Search
terms:
FD&
C
Blue.
No.
1,
FD&
C
Red
No.
40,
FD&
C
Yellow
No.
5.
(
November
28,
2004)
http://
www.
syrres.
com/
esc/
physdemo.
htm
E.
U.
S.
Environmental
Protection
Agency
Documents:

"
Robust
Summaries
&
Test
Plans:
C.
I.
Acid
Yellow
23
(
FD&
C
Yellow
5)",
Submitted
ti
the
Agency
by
the
International
Association
of
Color
Manufacturers
on
March
10,
2004.
http://
www.
epa.
gov/
chemrtk/
ciacdylo/
c15133tc.
htm
"
Background
Document
for
Identification
and
Listing
of
Deferred
Dye
and
Pigment
Wastes,
U.
S.
Environmental
Protection
Agency,
Office
of
Solid
Waste,
Washington,
D.
C.,
June
1999.

US
Environmental
Protection
Agency.
1998.
Guidelines
for
Ecological
Risk
Assessment.
Risk
Assessment
Forum.
EPA/
630/
R­
95/
002F,
April
1998
U.
S.
EPA.
1993.
U.
S.
Environmental
Protection
Agency.
Wildlife
Exposure
Factors
Handbook.
Volume
I
of
II.
EPA/
600/
R­
93/
187a.
Office
of
Research
and
Development,
Washington,
D.
C.
20460.

E.
U.
S.
Environmental
Protection
Agency,
Office
of
Pesticide
Programs
Documents:
41
Memorandum:
"
Review
of
Phase
4
List
D
Package
for
Aquashade"
(
EFGWB
#
93­
0119,
93­
0120;
Chemical
#
110301
and
110302;
Case
#
819437
and
819438;
DP
Barcode
D184289
and
D184278).
Dated
February
4,
1993.

"
Reassessment
of
the
Exemption
from
the
Requirement
of
a
Tolerance
for
the
FDACertified
Color
Additives
FD&
C
Blue
No.
1,
FD&
C
Red
No.
40,
and
FD&
C
Yellow
No.
5
(
Tartrazine)".
U.
S.
Environmental
Protection
Agency,
Office
of
Pesticides
Programs.
December
21,
2004
D.
Internet
Resources
Food
and
Drugs
Administration:
http://
www.
cfsan.
fda.
gov/~
dms/
opa­
col2.
html
Color
Index
International:
http://
www.
colour­
index.
org/

Additional
information
was
also
obtained
from
TOXNET.
http://
toxnet.
nlm.
nih.
gov/
42
Appendix
A­
Data
Gaps
Environmental
Fate
All
of
the
Subdivision
N,
Environmental
Fate
Data
Requirements
maybe
waived
because
of
very
low
risk
potential.
They
were
identified
as
"
Reserved"
in
1993
(
Memorandum:
"
Review
of
Phase
4
List
D
Package
for
Aquashade";
EFGWB
#
93­
0119,
93­
0120;
Chemicals
110301
and
110302;
Case
#
819437
and
819438;
DP
Barcode
D184289
and
D184278(.
Dated
February
4,
1993),
pending
on
the
results
of
required
ecological
toxicity
studies.

There
are
two
major
uncertainties
on
the
environmental
fate
behavior
of
Acid
Blue
9
and
Acid
Yellow
23:

(
1)
Structure­
activity
estimates
of
physical,
chemical,
and
environmental
fate
behavior
are
not
reliable
for
salts
of
large
anions,
such
as
Acid
Yellow
23
and
Acid
Blue
9.

(
2)
The
major
route
of
transformation
of
dyes
such
as
Acid
Blue
9
and
Acid
Yellow
23
is
indirect
photolysis.
However,
the
chemical
identity
of
the
photoproducts
of
these
two
dyes
are
not
known.
Biotransformation
under
anaerobic
conditions
may
also
contribute
to
the
dissipation
of
the
dyes
in
water­
sediments.
The
chemical
identity
of
the
metabolites
is
not
known.

However,
after
the
review
of
the
ecological
toxicity
studies
and
the
risk
assessment,
the
environmental
fate
data
requirements
placed
in
1993
as
"
Reserved'
can
be
waived.
The
Environmental
Fate
and
Effects
Division
is
not
requiring
any
new
environmental
fate
data
for
the
two
dyes
individually
or
for
any
of
the
end­
use
products
(
Aquashade,
Aquashade
OA,
Pond
Care
AlgaeBlocker,
Admiral
Liquid,
and
Admiral
WSP).

Ecological
Toxicity
b.
Effects
The
only
EFED
ecological
toxicity
data
available
is
for
the
Aquashade
product
(
23.6%
Acid
Blue
9
and
2.36%
Acid
Yellow
23)
as
the
test
substance.
There
are
no
EFED
ecological
toxicity
data
for
Admiral
Liquid
or
Admiral
WSP,
but
HED
received
a
study
that
found
a
laboratory
rat
LD50
equal
to
or
more
than
5,000
mg/
kg.
However,
since
organisms
will
be
only
exposed
to
either
1
or
2
ppm
in
the
environment,
this
uncertainty
is
minor.

No
chronic
studies
have
been
required,
so
it
is
not
known
what
would
happen
over
an
extended
exposure
period.
No
studies
on
aquatic
plants
have
been
required,
because
they
are
the
targets
of
Aquashade
and
it
is
assumed
that
all
submerged
aquatic
plants
in
the
treated
pond
will
be
killed.
No
terrestrial
plant
studies
have
been
required,
because
there
is
little
expected
exposure.
An
exception
would
be
if
any
of
these
products
43
might
be
applied
to
water
used
for
irrigation.
If
that
occurred,
exposure
to
terrestrial
plants
may
occur.

It
is
not
known
what
would
happen
to
plants
on
the
margins
of
a
treated
pond
or
growing
as
emergent
vegetation
in
the
shallows
of
the
pond
if
the
technical
product
or
the
end­
use
product
was
applied
to
them.
Some
labels
warn
that
directly
applying
the
products
to
emerged
or
terrestrial
plants
may
cause
burning.

Because
of
the
mode
of
action
(
blocking
light
that
enters
bodies
of
water),
the
low
acute
toxicity
of
this
pesticide,
and
the
relatively
limited
area
involved
in
treatment,
it
is
believed
that
additional
studies
are
not
needed.
44
Appendix
B
Environmental
Fate
No
Subdivision
N
Environmental
Fate
studies
were
submitted,
as
these
data
requirements
were
waived
in
1993.
Therefore,
no
data
from
guideline
studies
are
included
in
this
Appendix.

Although
the
Environmental
Fate
and
Effects
Division
does
not
usually
use
open
literature
data
in
its
assessments,
published
data
on
the
chemistry
and
behavior
of
dyes
in
aquatic
systems
was
used
to
supplement
the
environmental
fate
assessment.
Most
of
the
data
comes
from
structurally
related
dyes.
Acid
Blue
9
and
Acid
Yellow
23
are
also
food,
drugs,
and
cosmetic
colorants
regulated
by
the
Food
and
Drugs
Administration.

The
environmental
fate
behavior
of
dyes
such
as
Acid
Yellow
23
and
Acid
Blue
9
can
be
summarized
as
follows:

The
dyes
are
fully
ionized
in
the
environmentally
significant
pH
range
of
5
to
9.

1.
There
no
hydrolyzable
groups
in
these
two
dyes
and,
therefore,
abiotic
hydrolysis
is
not
an
important
degradation
pathway
in
the
environment.

2.
Direct
photolysis
is
slow,
as
indicated
by
the
fading
of
the
dyes
in
homogeneous
aqueous
media
exposed
to
sunlight.
However,
indirect
photolysis
has
been
identified
as
the
major
route
of
degradation
in
environmental
media.

3.
The
dyes
are
resistant
to
biodegradation
under
aerobic
conditions.
Structural
related
dyes
are
difficult
to
biodegrade
in
activated
sludge.
However,
redox
reactions
under
anaerobic
conditions
appear
to
degrade
the
dyes,
at
least
to
some
extent.

4.
The
two
dyes
are
highly
hydrophilic
and
do
not
have
the
potential
to
bioaccumulate
in
fish.
45
Appendix
C
Aquatic
Exposure
and
Modeling
The
end­
use
products
containing
Acid
Blue
9
and
Acid
Yellow
23
are
directly
applied
to
a
confined
water
body
and
at
the
dose
specified
in
the
labels
to
attain
a
concentration
of
1
ppm
or
2
ppm
depending
on
the
target
pest.
Because
there
are
no
runoff
or
spray
drift
components,
the
use
of
the
simulation
models
GENEEC
or
PRZM
and
EXAMS
is
not
appropriate.

Aquatic
exposure
was
based
on
the
target
concentration
specified
in
the
product
labels,
depending
on
the
weed
to
be
controlled.
The
maximum
target
concentration
is
2
ppm
(
2
mg/
L)
which
is
the
concentration
used
to
estimate
risk
for
both
aquatic
and
terrestrial
animals
46
Appendix
D.
Ecological
Toxicity
and
Uncertainty
in
Ecological
Toxicity
Data
EFED
requires
the
submission
of
six
studies
for
all
chemicals
that
are
submitted
for
registration.
Which
additional
data
that
is
required
depends
upon
the
circumstances
of
the
chemical
and
its
use
pattern.
Birds
are
used
as
surrogates
for
terrestrial
phase
amphibians
and
reptiles.
Freshwater
fish
are
used
as
surrogates
for
aquatic
phase
amphibians.

The
end­
use
products
are
not
applied
to
a
terrestrial
environment.
However,
terrestrial
animals
(
e.
g.,
mammals
and
birds)
are
exposed
to
the
dyes
when
they
use
treated
water
as
the
source
of
drinking
water.

In
addition
to
data
that
was
submitted
by
the
registrants,
EFED
also
reviewed
studies
from
the
open
literature.
These
studies
were
obtained
through
the
ORD
Middle
Ecological
Division's
ECOTOX
literature
search
and
retrieval
program
managed
in
Duluth,
Minnesota
(
http://
www.
epa.
gov/
ecotox).
These
data
are
consistent
with
the
contention
that
the
dyes
found
in
Aquashade
and
the
other
end­
use
products
are
practically
nontoxic
to
the
organisms
tested.

Table
D­
1.
Summary
of
most
sensitive
acute
toxicity
endpoints
multiplied
by
13%
based
on
percent
purity
of
the
dyes
in
aquashade.

Species
Study
Type1
Source
Extrapolated
Endpoint
Bobwhite
quail
Acute
Avian
Oral
433367­
01
LD50
&
NOAEL
$
2250
x
0.13
$
292.5
mg/
kg
Mallard
duck
Acute
Avian
Oral
433367­
02
LD50
&
NOAEL
$
2250
x
0.13
$
292.5
mg/
kg
Bobwhite
quail
Avian
Dietary
435034­
03
LC50
&
NOAEL5
$
620
x
0.13
$
80.6
ppm
Mallard
duck
Avian
Dietary
435034­
04
LC50
&
NOAEL5
$
620
x
0.13
$
80.6
ppm
Bluegill
sunfish
Acute
Toxicity
432975­
02
LC50
&
NOAEC
$
96
x
0.13
$
12.48
mg/
L
Rainbow
trout
Acute
Toxicity
432975­
01
LC50
&
NOAEC
$
96
x
0.13
$
12.48
mg/
L
Daphnia
magna
Acute
Toxicity
432975­
03
LC50
&
NOAEC
$
97
x
0.13
$
12.61
mg/
L
1
All
original
tests
were
performed
with
the
Typical
End­
use
Product,
Aquashade.
No
testing
was
performed
on
birds,
fish
and
invertebrates
with
the
end­
use
products
Admiral
Liquid
and
Admiral
WSP,
which
have
a
higher
percentage
of
the
dyes
than
the
tested
Aquashade
product.

In
addition
to
the
uncertainty
about
the
possible
percent
purity
adjustment,
Admiral
WSP
(
49.27%
acid
blue
9
and
3.27%
acid
yellow
23)
has
about
50%
greater
concentration
of
dyes
than
Aquashade.
To
take
this
higher
concentration
into
account,
each
toxicity
endpoint
could
be
multiplied
by
2
resulting
in
an
extrapolated
avian
acute
NOAEL
$
146
mg/
kg
and
aquatic
organism
NOAEC
$
6.3
mg/
L.
47
Table
D­
2.
EcoEffects
Data
including
Data
from
Open
Literature
Guideline
/
MRID
or
Citation
Discussion
Results
71­
1a
/
433367­
01
Acute
oral
bobwhite
quail
1
71­
1a
/
433367­
02
Acute
oral
mallard
duck
1
The
formulated
product
(
23.6%
acid
blue
and
2.39%
acid
yellow
dyes)
was
used.
The
study
concentration
was
not
adjusted
for
the
purity
of
the
active
ingredients
(
the
two
dyes).
The
results
are
milligrams
of
the
end­
use
product.
Five
experimental
doses
were
used
­
292,
486,
810,
1350,
2250
mg/
L.
There
were
no
mortalities
or
other
adverse
effects.
LD50
$
2250
mg
eup
/
kg
NOAEC
=
2250
mg
eup
/
kg
71­
2b
/
435034­
03
Acute
dietary
bobwhite
quail
1
71­
2a
/
435034­
04
Acute
dietary
waterfowlmallard
duck
1
The
formulated
product
(
23.6%
acid
blue
and
2.39%
acid
yellow
dyes)
was
used.
The
study
concentration
was
not
adjusted
for
the
purity
of
the
active
ingredients
(
the
two
dyes).
The
results
are
parts
per
million
of
the
end­
use
product.
Five
experimental
doses
were
used
­
562,
1000,
1780,
3160,
and
5620
ppm.
There
were
no
mortalities
or
other
adverse
effects.
LC50
eup
$
5,620
ppm
NOAEC
eup
=
5,620
ppm
eup
71­
3
/
452811­
01
Acute
Wild
Mammal
Toxicity
using
the
Laboratory
rat
2
The
study
used
Admiral
WSP,
a
purple
powder
formulation.
The
dyes
were
not
otherwise
identified.
A
group
of
7­
10
week
old
rats
(
5/
sex)
received
a
single
dose
of
5000
mg/
kg
by
gavage
and
were
observed
for
14
days.
There
were
no
mortalities,
therefore
the
chemical
was
categorized
as
"
Toxicity
Category
IV."
Oral
LD50
%
&
&

>
5,000
mg
eup
/
kg
Oral
LD50
%
&
&

.2,500
mg
a.
i.
/
kg
83­
4
/
434109­
01
Reproduction
study
using
the
laboratory
rat
3
Three
generations
of
Long­
Evans
rats
were
given
tartrazine
(
92%
a.
i.)
at
7.5,
75,
225,
and
750
mg/
kg/
day
in
their
diet.
This
is
consistent
with
no
effects
and
addresses
whether
acid
yellow
23
causes
reproductive
toxicity.
There
were
no
adverse
treatment
related
effects.
NOAEL
%
&
&
=
750
mg/
kg/
day
72­
1c
/
432975­
01
Acute
coldwater
fish­
Rainbow
trout
1
72­
1a
/
432975­
02
Acute
warmwater
fish­
bluegill
sunfish
1
The
formulated
product
(
23.6%
acid
blue
and
2.39%
acid
yellow
dyes)
was
placed
in
the
water
of
aquaria.
The
study
concentration
was
not
adjusted
for
the
purity
of
the
active
ingredients
(
the
two
dyes).
The
results
are
milligrams
of
the
end­
use
product.
The
only
dose
used
was
96
mg/
L.
LC50
eup
$
96
mg/
L
NOAEC
eup
=
96
mg/
L
48
Table
D­
2.
EcoEffects
Data
including
Data
from
Open
Literature
Guideline
/
MRID
or
Citation
Discussion
Results
72­
2a
/
432975­
03
Freshwater
invertebrate
Daphnia
magna
1
The
formulated
product
(
23.6%
acid
blue
and
2.39%
acid
yellow
dyes)
was
placed
in
the
water
of
aquaria.
The
study
concentration
was
not
adjusted
for
the
purity
of
the
active
ingredients
(
the
two
dyes).
The
results
are
milligrams
of
the
end­
use
product.
The
only
dose
used
was
97
mg/
L.
LC50
eup
$
97
mg/
L
NOAEC
eup
=
97
mg/
L
72­
1a
Warne
&
Schifko,
1999
FW
Cladoceran
Freshwater
cladocerans
(
Ceriodaphnia
dubia)
were
exposed
to
a
dye
for
two
days.
The
EC50
put
the
chemical
in
the
category
"
practically
nontoxic."
EC50
=
5707
mg/
L
72­
3a
Estuarine/
Marine
Acute
Toxicity
with
Sheepshead
Minnow,
Oyster,
Mysids
Exposure
to
marine
habitats
not
expected
N/
A
72­
4a
Freshwater
Fish
Early
Life
Stage
Fathead
minnow
Exposure
to
marine
habitats
not
expected
N/
A
72­
4b
Freshwater
Invertebrate
Life
Cycle­
Water
flea
Chronic
effects
not
expected
based
on
mode
of
action
and
lack
of
effects
in
acute
tests.
N/
A
123­
1a
Tier
II
Terrestrial
Plant
Seedling
Emergence
(
GF
871)
If
the
label
is
changed
and
the
water
is
not
used
for
irrigation,
exposure
to
terrestrial
plants
would
be
unlikely.
Reserved
123­
1b
Tier
II
Terrestrial
Plant
Vegetative
Vigor
(
GF
871)
If
the
label
is
changed
and
the
water
is
not
used
for
irrigation,
exposure
to
terrestrial
plants
would
be
unlikely.
Reserved
123­
2
Tier
II
Aquatic
Plant
Studies
Aquatic
plants
are
the
targets.
Therefore,
testing
is
not
required.
Release
from
treated
ponds
to
other
water
bodies
considered
a
very
low
probability.
N/
A
49
Table
D­
2.
EcoEffects
Data
including
Data
from
Open
Literature
Guideline
/
MRID
or
Citation
Discussion
Results
141­
1
Honey
Bee
Acute
Contact
Toxicity
Mode
of
action
suggests
low
likelihood
of
toxicity.
The
method
of
application
minimizes
exposure
to
terrestrial
insects.
N/
A
Nonguideline
Spencer,
1984
Crayfish
Crayfish
were
exposed
to
3
levels
of
Aquashade
for
5
days.
The
oxygen
use
was
effected.
The
study
was
categorized
as
"
slightly
toxic."
NOAEL
=
15
mg/
L
Nonguideline
Borzelleca
and
Hallagan,
1988.
Chronic
toxicity/
carcinogenicity
3
Laboratory
rats
were
fed
4
levels
of
yellow
5
for
900
days.
The
study
was
categorized
as
"
practically
nontoxic."
NOAEL
=
984
mg/
kg/
day
LOAEL
=
2641
mg/
kg/
day
Nonguideline
Borzelleca,
et
al.,
1990
Chronic
toxicity/
carcinogenicity
in
mice
3
House
mice
(
Mus
musculus)
were
given
food
with
3
levels
of
dye
in
their
diet
for
720
days.
The
study
was
characterized
as
"
practically
nontoxic."
7354
mg/
kg/
day
1
The
study
was
done
with
Aquashade.
The
doses
were
calculated
with
the
end­
use
product.
The
percentages
of
the
dyes
making
up
liquid
is
not
clear.
2
The
study
was
done
with
Admiral
Lake
Colorant
WSP.
The
LD50
is
based
on
the
dry
weight
of
the
end­
use
product.
The
composition
of
the
powder
was
not
determined,
but
it
is
usually
49.62%
blue
dye
and
3.05%
yellow
dye.
The
LD50
a.
i.
cannot
be
determined,
but
is
probably
50%
of
the
stated
>
5,000
mg/
kg.
3
The
study
was
done
with
FD&
C
yellow
dye
(
acid
yellow
23
or
5),
which
is
not
an
end­
use
products.

Uncertainty
in
toxicity
information
Most
of
the
toxicity
results
used
for
risk
assessment
were
based
on
effects
testing
done
with
Aquashade
(
23.6%
acid
blue
9
and
2.39%
acid
yellow
23)
except
the
acute
mammal
LD50
was
done
with
Admiral
WSP
and
the
reproductive
mammalian
NOAEL
was
done
with
acid
yellow.

There
is
some
uncertainty
in
this
approach
because
1)
the
percent
of
the
dyes
in
this
formulation
is
unclear,
and
2)
some
formulations
have
a
higher
percent
of
each
dye
than
Aquashade,
and
it
is
not
clear
if
the
doses
were
expressed
in
terms
of
end­
use
product
or
active
ingredients.
The
dyes
in
Aquashade
are
reported
as
being
13%
pure.
To
demonstrate
the
degree
of
uncertainty,
the
toxicity
values
may
be
multiplied
by
0.1
to
50
take
these
percentages
into
account.
Note
that
the
risk
assessment
was
conducted
with
actual
toxicity
test
results,
not
with
these
adjusted
toxicity
values.

Table
F­
1.
Summary
of
acute
toxicity
endpoints
multiplied
by
13%
to
composite
for
the
possibility
of
the
tested
materials
being
only
13%
pure.

Species
Study
Type1
Source
Extrapolated
Endpoint
Bobwhite
quail
Acute
Avian
Oral
433367­
01
LD
50
&
NOAEL
$
2250
x
0.13
$
292.5
mg/
kg
Mallard
duck
Acute
Avian
Oral
433367­
02
LD
50
&
NOAEL
$
2250
x
0.13
$
292.5
mg/
kg
Bobwhite
quail
Avian
Dietary
435034­
03
LC
50
&
NOAEL
$
620
x
0.13
$
80.6
ppm
Mallard
duck
Avian
Dietary
435034­
04
LC
50
&
NOAEL
$
620
x
0.13
$
80.6
ppm
Bluegill
sunfish
Acute
Toxicity
432975­
02
LC
50
&
NOAEC
$
96
x
0.13
$
12.48
mg/
L
Rainbow
trout
Acute
Toxicity
432975­
01
LC
50
&
NOAEC
$
96
x
0.13
$
12.48
mg/
L
Daphnia
magna
Acute
Toxicity
432975­
03
LC
50
&
NOAEC
$
97
x
0.13
$
12.61
mg/
L
1
All
original
tests
were
performed
with
the
typical
end­
use
product,
Aquashade.
No
testing
was
performed
on
birds,
fish
and
invertebrates
with
the
end­
use
products
Admiral
Liquid
and
Admiral
WSP,
which
have
a
higher
percentage
of
the
dyes
than
the
tested
Aquashade
product.

In
addition
to
the
uncertainty
about
the
possible
percent
purity
adjustment,
Admiral
WSP
(
49.27%
acid
blue
9
and
3.27%
acid
yellow
23)
has
about
50%
greater
concentration
of
dyes
than
Aquashade.
To
take
this
higher
concentration
into
account,
each
toxicity
endpoint
could
be
multiplied
by
2
resulting
in
an
extrapolated
avian
acute
NOAEL
$
146
mg/
kg
and
aquatic
organism
NOAEC
$
6.3
mg/
L.
51
Appendix
E
Status
of
Data
Requirements
1.
Environmental
Fate
Subdivision
N,
Environmental
Fate
Data
Requirements­
Status
of
Data
Requirements
Data
Requirement
Status
in
1993
Status
after
the
Risk
Assessment
for
Reregistration
Additional
Data
Required
161­
1
[
Abiotic]
Hydrolysis
Reserved
May
be
Waived
No
161­
2
[
Direct]
Photolysis
in
Water
Reserved
May
be
Waived
No
161­
3
Photolysis
on
Soil
Not
Required
Not
Required
No
162­
1
Aerobic
Soil
Metabolism
Not
Required
Not
Required
No
162­
2
Anaerobic
Aquatic
Metabolism
Not
Required
Not
Required
No
162­
3
Anaerobic
Aquatic
Metabolism
Reserved
May
be
Waived
No
162­
4
Aerobic
Aquatic
Metabolism
Reserved
May
be
Waived
No
163­
1
Mobility
in
Soils/
Sediments
Reserved
May
be
Waived
No
164­
1
Terrestrial
Field
Dissipation
Not
Required
Not
Required
No
164­
2
Aquatic
Field
Dissipation
Reserved
May
be
Waived
No
165­
4
Bioaccumulation
in
Fish
Reserved
May
be
Waived
The
Log
Kow
does
not
trigger
this
data
requirement
No
Ecological
effects
data
requirements
for
pesticide
registration
of
Aquashade
PC
Code:
110303
(
a
mixture
of
110301
and
110302),
only
TEPs
were
studied
Guideline
Number
DATA
REQUIREMENTS
(
Not
Reserved)
FULFILLS
REQ'S
(
Y/
N)
MRID
DATE
STATUS
Acute
Avian
Oral
Quail
TEP
Y
433367­
01
9­
7­
94
Acceptable
71­
1(
b)
Acute
Avian
Oral
Duck
TEP
Y
433367­
02
9­
7­
94
Acceptable
71­
2(
b)
Avian
Dietary/
Quail
TEP
Y
435034­
03
4­
26­
95
Acceptable
71­
2(
b)
Avian
Dietary/
Duck
TEP
Y
435034­
04
4­
26­
95
Acceptable
71­
3
Wild
Mammal
Toxicitya
Y
452811­
01
12­
5­
00
Acceptable
71­
4(
a)
Avian
Reproductive/
Quail
Waived
71­
4(
b)
Avian
Reproductive/
Duck
Waived
72­
1(
b)
Acute
Toxicity
Bluegill
TEP
Y
432975­
02
4­
25­
95
Acceptable
72­
1(
d)
Acute
Toxicity
Rainbow
Trout
TEP
Y
432975­
01
4­
25­
95
Acceptable
52
Ecological
effects
data
requirements
for
pesticide
registration
of
Aquashade
PC
Code:
110303
(
a
mixture
of
110301
and
110302),
only
TEPs
were
studied
Guideline
Number
DATA
REQUIREMENTS
(
Not
Reserved)
FULFILLS
REQ'S
(
Y/
N)
MRID
DATE
STATUS
72­
2(
b)
Acute
Invertebrate
Toxicity
TEP
Y
432975­
03
4­
25­
95
Acceptable
a
This
study
was
performed
with
Admiral
WSP
No
additional
effects
data
are
being
required.
53
Appendix
F
Environmental
Fate
Bibliography
Books
Lynch,
D.
G.
"
Estimating
the
Properties
of
Synthetic
Organic
Dyes",
in
Handbook
of
Property
Estimation
Methods
for
Chemicals­
Environmental
Health
Sciences,
Edited
by
Robert
S.
Boethling
and
Donald
Mackay.
Published
by
Lewis
Publishers,
Boca
Raton,
FL;
Pages
447­
467.
And
pertinent
references
therein.

Marmion,
D,
M.
Handbook
of
U.
S.
Colorants­
Food,
Drugs,
and
Medical
Devices,
Third
Edition.,
1991.
Published
by
John
Wiley
and
Sons,
New
York
Helz,
G.
R.,
Zepp,
R.
G.,
and
Crosby,
D.
G,
Editors.
1994.
Aquatic
and
surface
photochemistry.
Lewis
Publishers,
Boca
Raton,
Florida.

Journal
Articles
Jank.
M.,
Köser,
H.,
Lücking,
F.,
Martienssen,
M.,
and
Wittchen,
S.
1998,
"
Decolorization
and
Degradation
of
Erioglaucine
(
Acid
Blue
9)
Dye
in
Wastewater,"
Environmental
Technology,
v.
19(
7),
pp.
741­
747.

Weber,
E.
J.
and
Adams,
R.
L.
1995.
"
Chemical
and
Sediment­
Mediated
Reduction
of
the
Azo
Dye
Disperse
Blue
79,"
Environ.
Sci.
Technol.
v.
29,
pp.
1163­
1170.

Baughman,
G.
L.
1995.
"
Fate
of
azo
dyes
in
aquatic
systems.
Part
3:
The
role
of
suspended
sediments
in
adsorption
and
reaction
of
acid
and
direct
dyes,"
Dyes
and
Pigments,
v.
27,
pp.
197­
210.

Brown,
D.
and
Laboureur,
P.
1983,
"
The
Degradation
of
Dyestuffs:
Part
I­
Primary
biodegradation
under
anaerobic
conditions,"
Chemosphere,
v.
122,
pp.
397­
404.

Baran,
W.,
Makowski,
A.,
Wardas,
W.
2003.
"
The
influence
of
FeCl3
on
the
photocatalytic
degradation
of
dissolved
azo
dyes
in
aqueous
TiO2
suspensions,"
Chemosphere,
v.
53,
pp
82­
95.

Tai,
W.
T.,
Chang,
C.
Y,
Ing,
C.
H.,
and
Chang,
C.
F.
2004.
"
Adsorption
of
acid
dyes
from
aqueous
solutions
on
activated
bleaching
earth,"
J.
Colloid
and
Interface
Science,
vol.
275,
pp
72­
78.

Mon,
J.,
Flury,
M.,
and
Harsh,
J.
B.
2005
."
Sorption
of
four
triarylmethane
dyes
in
a
sandy
soil
determined
by
batch
and
column
experiments,"
Geoderma.
in
press.

Mon,
J.,
Flury.
M.,
and
Harsh.
2005
"
A
quantitative
structure­
activity
relationship
(
OSAR)
analysis
of
triarylmethane
dye
tracers,"
Journal
of
Hydrology,
in
press.
54
Appendix
G
Ecological
Toxicity
Bibliography
55
Appendix
H.
Endangered
and
Threatened
Species
EFED
also
assessed
the
potential
for
Aquashade
and
other
end­
use
products
containing
the
Acid
Yellow
23
and
Acid
Blue
9
dyes
to
potentially
cause
harm
to
endangered
species.

Terrestrial
Animals
Listed
endangered
or
threatened
species
of
birds,
mammals
and
reptiles
are
not
expected
to
be
affected
directly
because
exposure
does
not
exceed
the
endangered
species
LOC.
However,
terrestrial
animals
may
be
affected
indirectly
if
aquatic
plants
that
are
necessary
for
their
survival
are
eliminated.
The
degree
to
which
this
might
occur
depends
on
the
extent
the
treated
water
bodies
are
a
critical
component
of
their
food
supply.

Terrestrial
Plants
Terrestrial
plants
have
the
potential
for
exposure
if
water
treated
with
Aquashade
or
the
other
dye
containing
end­
use
products
are
used
for
irrigation.
If
irrigation
is
precluded
from
use
on
the
labels,
EFED
concludes
there
is
virtually
no
potential
for
exposure
to
terrestrial
plants.

Aquatic
Animals
Listed
species
of
fish,
invertebrates
or
amphibians
are
not
expected
to
be
affected
directly
because
the
exposure
does
not
exceed
the
endangered
species
LOC.
However,
aquatic
animals
dwelling
in
treated
ponds
might
have
the
potential
for
harm
if
the
plants
on
which
they
depend
for
food
and
habitat
are
eliminated.
The
likelihood
of
this
occurring
depends
on
if
any
listed
aquatic
animal
species
would
occur
in
these
highly
managed,
often
manmade,
normally
relatively
small
water
bodies
with
usually
no
outflow,
or
if
any,
a
very
small
amount.

Aquatic
Plants
Listed
species
of
aquatic
plants
which
begin
their
life
stages
as
submerged
might
be
harmed
if
water
bodies
in
which
they
are
growing
are
treated
while
they
are
submerged.

EFED
has
not
fully
analyzed
the
potential
for
endangered
species
to
be
harmed
through
use
of
Aquashade
and
other
end­
use
products.
Such
analysis
would
focus
on
the
overlap
of
specific
species
with
potentially
treated
water
bodies,
and
in
the
case
of
indirect
effects,
the
likelihood
that
the
treated
water
body
comprises
enough
of
a
component
of
the
listed
species
food
or
habitat
base
to
actually
impact
the
listed
species,
if
that
food
or
habitat
was
lost.
56
Appendix
K.
Risk
Quotient
Calculation
and
Interpretation
Risk
characterization
integrates
the
results
of
the
exposure
and
ecotoxicity
data
to
evaluate
the
likelihood
of
adverse
ecological
effects.
The
means
of
this
integration
is
called
the
quotient
method.
Risk
quotients
(
RQs)
are
calculated
by
dividing
exposure
estimates
by
acute
and
chronic
ecotoxicity
value
and,
in
this
case,
it
is
a
deterministic
approach.

RQ
=
EXPOSURE/
TOXICITY
RQs
are
then
compared
to
OPP's
levels
of
concern
(
LOCs).
These
LOCs
are
used
by
OPP
to
analyze
potential
risk
to
nontarget
organisms
and
the
need
to
consider
regulatory
action.
The
criteria
indicate
that
a
pesticide
used
as
directed
has
the
potential
to
cause
adverse
effects
on
nontarget
organisms.
LOCs
currently
address
the
following
risk
presumption
categories:
(
1)
acute
­­
potential
for
acute
risk;
regulatory
action
may
be
warranted
in
addition
to
restricted
use
classification,
(
2)
acute
restricted
use
­­
the
potential
for
acute
risk,
but
may
be
mitigated
through
restricted
use
classification,
(
3)
acute
endangered
species
­
endangered
species
may
be
adversely
affected,
and
(
4)
chronic
risk
­
the
potential
for
chronic
risk;
regulatory
action
may
be
warranted.
Currently,
EFED
does
not
perform
assessments
for
chronic
risk
to
plants,
acute
or
chronic
risks
to
nontarget
insects,
or
chronic
risk
from
granular/
bait
formulations
to
birds
or
mammals.

The
ecotoxicity
test
values
(
measurement
endpoints)
used
in
the
acute
and
chronic
risk
quotients
are
derived
from
required
studies.
Examples
of
ecotoxicity
values
derived
from
short­
term
laboratory
studies
that
assess
acute
effects
are:
(
1)
LC50
(
fish
and
birds),
(
2)
LD50
(
birds
and
mammals),
(
3)
EC50
(
aquatic
plants
and
aquatic
invertebrates)
and
(
4)
EC25
(
terrestrial
plants).
Examples
of
toxicity
test
effect
levels
derived
from
the
results
of
long­
term
laboratory
studies
that
assess
chronic
effects
are:
(
1)
LOAEC
(
birds,
fish,
and
aquatic
invertebrates),
and
(
2)
NOAEC
(
birds,
fish
and
aquatic
invertebrates).
For
birds
and
mammals,
the
NOAEC
generally
is
used
as
the
ecotoxicity
test
value
in
assessing
chronic
effects,
although
other
values
may
be
used
when
justified.
However,
the
NOAEC
is
used
if
the
measurement
end
point
is
production
of
offspring
or
survival
57
Risk
presumptions
and
the
corresponding
RQs
and
LOCs,
are
tabulated
below.

Table
K­
1:
Risk
presumptions
for
terrestrial
animals
Risk
Presumption
RQ
LOC
Birds
Acute
High
Risk
EEC1/
LC50
or
LD50/
sqft2
or
LD50/
day3
0.5
Acute
Restricted
Use
EEC/
LC50
or
LD50/
sqft
or
LD50/
day
(
or
LD50
<
50
mg/
kg)
0.2
Acute
Endangered
Species
EEC/
LC50
or
LD50/
sqft
or
LD50/
day
0.1
Chronic
Risk
EEC/
NOAEC
1
Wild
Mammals
Acute
High
Risk
EEC/
LC50
or
LD50/
sqft
or
LD50/
day
0.5
Acute
Restricted
Use
EEC/
LC50
or
LD50/
sqft
or
LD50/
day
(
or
LD50
<
50
mg/
kg)
0.2
Acute
Endangered
Species
EEC/
LC50
or
LD50/
sqft
or
LD50/
day
0.1
Chronic
Risk
EEC/
NOAEC
1
1
abbreviation
for
Estimated
Environmental
Concentration
(
ppm)
on
avian/
mammalian
food
items
2
mg/
ft2
3
mg
of
toxicant
consumed/
day
LD50
*
wt.
of
a
bird
58
Table
K­
2
.
Risk
Presumptions
for
Aquatic
Animals
Risk
Presumption
RQ
LOC
Acute
Risk
EEC1/
LC50
or
EC50
0.5
Acute
Restricted
Use
EEC/
LC50
or
EC50
0.1
Acute
Endangered
Species
EEC/
LC50
or
EC50
0.05
Chronic
Risk
EEC/
MATC
or
NOAEC
1
1
EEC
=
(
ppm
or
ppb)
in
water
Table
K­
3.
Risk
presumptions
for
plants
Risk
Presumption
RQ
LOC
Terrestrial
and
Semi­
Aquatic
Plants
Acute
Risk
EEC1/
EC25
1
Acute
Endangered
Species
EEC/
EC05
or
NOAEC
1
Aquatic
Plants
Acute
High
Risk
EEC2/
EC50
1
Acute
Endangered
Species
EEC/
EC05
or
NOAEC
1
1
EEC
=
lbs
ai/
A
2
EEC
=
(
ppb/
ppm)
in
water
59
Appendix
L.
Papers
that
were
accepted
for
ECOTOX
but
that
were
not
used
in
the
risk
assessment
science
chapter.
These
papers
passed
the
screen
for
consideration
in
ecological
risk
assessment,
but
were
ultimately
not
used
because
they
did
not
add
to
or
change
the
risk
presumption.

Acceptable
for
ECOTOX
and
OPP
Borzelleca,
J.
F.,
Depukat,
K.,
and
Hallagan,
J.
B.
1990.
Lifetime
Toxicity/
Carcinogenicity
Studies
of
FD­
and­
C
Blue
No.
1
(
Brilliant
Blue
FCF)
in
Rats
and
Mice.
Food
Chem.
Toxicol.
28:
221­
234.
EcoReference
No.:
76018
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
GRO,
POCM,
MOR,
BEH,
CEL.

Borzelleca,
J.
F.
and
Hallagan,
J.
B.
1988.
A
Chronic
Toxicity/
Carcinogenicity
Study
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FD
&
C
Yellow
No.
5
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Food
Chem.
Toxicol.
26:
189­
194.
EcoReference
No.:
76019
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
MOR,
GRO,
BEH,
CEL.

Collins,
T.
F.
X.,
Black,
T.
N.,
Brown,
L.
H.,
and
Bulhack,
P.
1990.
Study
of
the
Teratogenic
Potential
of
FD
and
C
Yellow
No.
5
when
Given
by
Gavage
to
Rats.
Food
Chem.
Toxicol.
28:
821­
827.
EcoReference
No.:
76021
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
GRO,
BEH,
PHY,
REP,
MOR.

Collins,
T.
F.
X.,
Black,
T.
N.,
O'Donnell,
M.
W.
Jr.,
and
Bulhack,
P.
1992.
Study
of
the
Teratogenic
Potential
of
FD
&
C
Yellow
No.
5
when
Given
in
Drinking­
Water.
Food
Chem.
Toxicol.
30:
263­
268.
EcoReference
No.:
76022
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
GRO,
BEH,
PHY,
REP,
MOR.

Ershoff,
B.
H.
1977.
Effects
of
Diet
on
Growth
and
Survival
of
Rats
Fed
Toxic
Levels
of
Tartrazine
(
FD
&
C
Yellow
No.
5)
and
Sunset
Yellow
FCF
(
FD
&
C
Yellow
No.
6.
J.
Nutr.
107:
822­
828.
EcoReference
No.:
76051
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
GRO,
MOR.

Giri,
A.
K.,
Das,
S.
K.,
Talukder,
G.,
and
Sharma,
A.
1990.
Sister
Chromatid
Exchange
and
Chromosome
Aberrations
Induced
by
Curcumin
and
Tartrazine
on
Mammalian
Cells
In
Vivo.
Cytobios
62:
111­
117.
EcoReference
No.:
76048
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
CEL.

Maekawa,
A.,
Matsuoka,
C.,
Onodera,
H.,
Tanigawa,
H.,
Furuta,
K.,
Kanno,
J.,
Jang,
J.
J.,
Hayashi,
Y.,
and
Ogiu,
T.
1987.
Lack
of
Carcinogenicity
of
Tartrazine
(
FD
and
C
Yellow
No.
5)
in
the
F344
Rat.
Food
Chem.
Toxicol.
25:
891­
896.
EcoReference
No.:
76017
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
CEL,
GRO,
MOR.

Sasaki,
Y.
F.,
Kawaguchi,
S.,
Kamaya,
A.,
Ohshita,
M.,
Kabasawa,
K.,
Iwama,
K.,
Taniguchi,
K.,
and
Tsuda,
S.
2002.
The
Comet
Assay
with
8
Mouse
Organs:
Results
with
39
Currently
Used
Food
Additives.
Mutat.
Res.
519:
103­
119.
EcoReference
No.:
60
75840
Chemical
of
Concern:
AQS,
TBA,
TRZ,
BZO;
Habitat:
T;
Effect
Codes:
MOR,
CEL.

Sobotka,
T.
J.,
Brodie,
R.
E.,
and
Spaid,
S.
L.
1977.
Tartrazine
and
the
Developing
Nervous
System
of
Rats.
J.
Toxicol.
Environ.
Health
2:
1211­
1220.
EcoReference
No.:
76050
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
PHY,
BEH,
GRO,
BCM.
Stefanidou,
M.,
Alevisopoulos,
G.,
Chatziioannou,
A.,
and
Koutselinis,
A.
2003.
Assessing
Food
Additive
Toxicity
Using
a
Cell
Model.
Vet.
Hum.
Toxicol.
45:
103­
105.
EcoReference
No.:
76016
Chemical
of
Concern:
AQS,
NaNO3;
Habitat:
A;
Effect
Codes:
CEL.

Tripathy,
N.
K.,
Patnaik,
K.
K.,
and
Nabi,
M.
J.
1989.
Genotoxicity
of
Tartrazine
Studied
in
Two
Somatic
Assays
of
Drosophila
melanogaster.
Mutat.
Res.
224:
479­
483.
EcoReference
No.:
76049
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
CEL.

Biggs,
D.
F.,
Peterson,
M.
A.,
and
Aaron,
T.
H.
1981.
Tartrazine
Induced
Airway
Constriction
in
Anesthetized,
Paralyzed
Guinea­
Pigs.
Proc.
West.
Pharmacol.
Soc.
24:
363­
365.
EcoReference
No.:
75992
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
PHY.

D'Souza,
S.
J.
A.
and
Biggs,
D.
F.
1987.
Aspirin,
Indomethacin,
and
Tartrazine
Increase
Carotid­
Sinus­
Nerve
Activity
and
Arterial
Blood
Pressure
in
Guinea
Pigs.
Pharmacology
34:
96­
103.
EcoReference
No.:
76170
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
PHY.

Davis,
K.
J.,
Fitzhugh,
O.
G.,
and
Nelson,
A.
A.
1964.
Chronic
Rat
and
Dog
Toxicity
Studies
on
Tartrazine.
Toxicol.
Appl.
Pharmacol.
6:
621­
626.
EcoReference
No.:
76249
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
MOR,
CEL,
GRO,
PHY.

Kobayashi,
N.,
Taniguchi,
N.,
Sako,
F.,
and
Takakuwa,
E.
1977.
A
Screening
Method
for
the
Toxicity
of
Food
Dyes
Using
Artemia
salina
Larvae.
J.
Toxicol.
Sci.
2:
383­
390.
EcoReference
No.:
7542
Chemical
of
Concern:
AQS;
Habitat:
A;
Effect
Codes:
MOR.

Manker,
D.
C.
and
Martin,
D.
F.
1984.
Investigation
of
Two
Possible
Modes
of
Action
of
the
Inert
Dye
Aquashade
on
Hydrilla.
J.
Environ.
Sci.
Health
Part
A
19:
725­
733.
EcoReference
No.:
11634
Chemical
of
Concern:
AQS;
Habitat:
A;
Effect
Codes:
GRO.

Nicklin,
S.
and
Miller,
K.
1985.
Induction
of
a
Transient
Reaginic
Antibody
to
Tartrazine
in
an
Animal
Model.
Int.
Arch.
Allergy
Appl.
Immunol.
76:
185­
187.
EcoReference
No.:
76218
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
PHY.
61
Office
of
Pesticide
Programs
2000.
Pesticide
Ecotoxicity
Database
(
Formerly:
Environmental
Effects
Database
(
EEDB).
Environmental
Fate
and
Effects
Division,
U.
S.
EPA,
Washington,
D.
C.
EcoReference
No.:
344
Chemical
of
Concern:
24DXY,
ACL,
ACP,
ACR,
AQS,
ATZ,
AZ,
BDF,
BMC,
BML,
BMN,
BS,
BT,
Captan,
CBF,
CBL
,
CFE,
CFE,
CLNB,
CMPH,
CPC,
CPY,
CTN,
CTZ,
Cu,
CuO,
CuS,
CYD,
CYF,
CYP,
CYT,
DBN
,
DCNA,
DFT,
DFZ,
DM,
DMB,
DMM,
DMP,
DMT,
DOD,
DPC,
DPDP,
DS,
DSP,
DU,
DZ,
DZ
M,
EFL,
EFS,
EFV,
EP,
FHX,
FMP,
FO,
Folpet,
FPP,
FVL,
GYP,
HCCH,
HXZ,
IPD,
IZP,
LNR,
M
AL,
MB,
MBZ,
MDT,
MFX,
MFZ,
MGK,
MLN,
MLT,
MOM,
MP,
MTC,
MTL,
MTM,
NAA,
Na
led,
NFZ,
NPP,
NTP,
OXF,
OXT,
OYZ,
PDM,
PEB,
PHMD,
PMR,
PMT,
PNB,
PPB,
PPG,
PPM
H,
PQT,
PRB,
PRT,
PSM,
PYN,
PYZ,
RTN,
SMM,
SMT,
SS,
SXD,
SZ,
TBC,
TDC,
TDZ,
TET,
T
FN,
TFR,
TMT,
TPR,
TRB,
WFN,
ZnP;
Habitat:
AT;
Effect
Codes:
MOR,
POP,
PHY,
GRO,
REP.

Peterson,
M.
A.,
Biggs,
D.
F.,
and
Aaron,
T.
H.
1980.
Comparison
of
the
Effects
of
Aspirin,
Indomethacin
and
Tartrazine
on
Dynamic
Pulmonary
Compliance
and
Flow
Resistance
in
the
Guinea
Pig.
Proc.
West.
Pharmacol.
Soc.
23:
121­
124.
EcoReference
No.:
75980
Chemical
of
Concern:
TRZ,
AQS;
Habitat:
T;
Effect
Codes:
CEL,
PHY.

Procter
&
Gamble
Co.
1992.
Initial
Submission:
Repeated
Dermal
Applications
in
Mice
of
FD&
C
Blue
1,
FD&
C
Red
3,
FD&
C
Yellow
5,
D&
C
Red
19
with
Cover
Letter
Dated
08/
26/
92.
EPA/
OTS
Doc.#
88­
920008654.
EcoReference
No.:
76262
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
PHY,
MOR.

Safford,
R.
J.
and
Goodwin,
B.
F.
J.
1985.
Immunological
Studies
on
Tartrazine
and
Its
Metabolites
1.
Animal
Studies.
Int.
Arch.
Allergy
Appl.
Immunol.
77:
331­
336.
EcoReference
No.:
76217
Chemical
of
Concern:
AQS;
Habitat:
T;
Effect
Codes:
PHY.

Sako,
F.,
Taniguchi,
N.,
Kobayashi,
N.,
and
Takakuwa,
E.
1977.
Effects
of
Food
Dyes
on
Paramecium
caudatum:
Toxicity
and
Inhibitory
Effects
on
Leucine
Aminopeptidase
and
Acid
Phosphatase
Activity.
Toxicol.
Appl.
Pharmacol.
39:
111­
117
(
Author
Communication
Used.
EcoReference
No.:
8261
Chemical
of
Concern:
AQS;
Habitat:
A;
Effect
Codes:
MOR.

Schafer,
E.
W.
and
Bowles,
W.
A.
1985.
Acute
Oral
Toxicity
and
Repellency
of
933
Chemicals
to
House
and
Deer
Mice.
Arch.
Environ.
Contam.
Toxicol.
14:
111­
129.
EcoReference
No.:
35426
Chemical
of
Concern:
ADC,
CST,
MOM,
CPC,
ZnP,
DOD,
MLN,
Cu,
AQS,
CuCO;
Habitat
:
T;
Effect
Codes:
MOR.

Upadhyay,
R.
R.
and
Upadhyay,
L.
1994.
Cirrhosis
and
Dysplasia
Caused
by
Sun
Set
Yellow
and
Brilliant
Blue
in
the
Liver
of
Heteropneustes
fossilis.
62
J.
Ecotoxicol.
Environ.
Monit.
4:
275­
277.
EcoReference
No.:
20309
Chemical
of
Concern:
AQS;
Habitat:
A;
Effect
Codes:
HIS.

Wan,
M.
T.,
Watts,
R.
G.,
and
Moul,
D.
J.
1991.
Acute
Toxicity
to
Juvenile
Pacific
Northwest
Salmonids
of
Basacid
Blue
NB755
and
Its
Mixture
with
Formulated
Products
of
2,4­
D,
Glyphosate,
and
Triclopyr.
Bull.
Environ.
Contam.
Toxicol.
47:
471­
478
(
OECDG
Data
File.
EcoReference
No.:
5132
Chemical
of
Concern:
24DXY,
GYP,
TPR,
AQS;
Habitat:
A;
Effect
Codes:
MOR.
