normochromatic erythrocyte ratios (PCE:NCE) were observed in both doses
at all time points, indicating that the test material was toxic to the
bone marrow.  There was no significant increase in the frequency of
micronucleated polychromatic erythrocytes in bone marrow after any
treatment time.

870.5450	Dominant Lethal Assay - Mice

	47090406 (1982)

Acceptable/guideline

0, 25, 50, or 100 mg/kg/day (total doses of 0, 125, 250, or 500 mg/kg)
Mortality (3/15 males) was noted at 100 mg/kg/day during dosing.  Slight
decreases (p<0.05) in body weight were observed at 50 mg/kg/day and
above during dosing.  There were no treatment-related effects on
fertility, mean number of implantations, or the number of early or late
deaths.  There was no time-related positive response of increased pre-
or post-implantation loss compared to controls.

870.5550	Unscheduled DNA Synthesis in Primary Rat Hepatocytes/Mammalian
Cell	47090407 (2003)

Acceptable/guideline

0, 250, 500, or 1000 mg/kg	The net nuclear grain (NNG) counts in the
treated animals (–3.42 to –2.64) were well below the threshold of
≥5 NNG needed for a positive response, and no increase in the mean
percent of cells in repair was observed.  There was no evidence that
unscheduled DNA synthesis, as determined by radioactive tracer
procedures [nuclear silver grain counts] was induced.

870.6200a

	Acute neurotoxicity screening battery	47090408 (2005)

Acceptable/guideline

0, 125, 250, or 750 mg/kg	NOAEL = 250 mg/kg

LOAEL = 750 mg/kg, based on decreased body weight, body-weight gain,
absolute and relative food consumption, and clinical signs of toxicity,
indicative of a moribund condition, in both sexes: dehydration,
urine-stained abdominal fur, ungroomed coat, ptosis, decreased motor
activity, prostration, limp muscle tone, muscle flaccidity, hypothermia,
hunched posture, impaired or lost righting reflex, scant feces; in
males: red or tan perioral substance, chromodacryorrhea,
chromorhinorrhea and labored breathing, and in females:  piloerection
and bradypnea, and signs of neurotoxicity:  hunched posture in females
and ataxia in males.

870.6200b	Subchronic Neurotoxicity – Feeding Study in Rats	47090410
(2007)

Acceptable/guideline

0, 500, 1500, or 3000 ppm (0/0, 28.9/32.6, 84.3/97.6, and 172.1/185.0
mg/kg/day [M/F])	NOAEL = 1500 ppm (84.3/97.6 mg/kg/day [M/F]).

LOAEL = 3000 ppm (172.1/185.0 mg/kg/day [M/F]) based on decreased
body-weight gain, absolute and relative food consumption; and decreased
hindlimb grip strength in males.

870.7485

	Metabolism and pharmacokinetics

(rat)	47090412 (2006)

Acceptable/guideline

5 or 250 mg/kg	More than 78% of the dose was recovered in the bile and
urine.  Absorption was similar between sexes and between single and
multiple dose regimes.  Absorption is extensive.  The dose was mostly
eliminated within 48 hours. Only 0.04-0.05% of the dose was found in the
expired carbon dioxide.  Most of the radioactivity was excreted in the
bile (47-79% of the dose).  The excretion profile was similar between
sexes.  In the blood, radioactivity partitioned into the red blood
cells.  In both sexes and all groups, concentrations of radioactivity
were relatively high in whole blood, liver and kidneys.  Other organs
with high concentrations included the adrenal glands, spleen, and
pituitary.  The distribution profiles were similar between species, dose
level, and single vs multiple dose regime.  In the whole blood, the
concentrations were proportional to the dose.  The total amount of
radioactivity isolated in the tissues and carcass was <1-3%.
Bioaccumulation was considered unlikely.  The parent was isolated in
only trace amounts in the urine and feces and more than 19 metabolites
were isolated, indicating extensive metabolism.  Metabolism profiles
were similar between sexes.  The metabolic profiles were similar
regardless of the matrix (feces, urine, or bile), the dose, and the sex.
 The primary site for metabolism was the 2-fluorophenyl ring.  The
initial metabolic step was epoxidation followed by either rearrangement
to form the dihydrodiol isomers or to form hydroxy or dihydroxy
metabolites.  The hydroxyl groups on these primary metabolites may then
be either conjugated with glucuronic acid or methylated.  A second,
minor route for metabolism of flutriafol was via the removal of the
triazole ring to form 1-(2 fluorophenyl)-1-(4-fluorophenyl)-ethandiol,
which is then conjugated with glucuronic acid.

870.7600	In vivo dermal penetration

(rat)	47090415 (2006)

Acceptable/guideline

0.02, 0.2 or 2 mg/cm2 skin were tested (10 µl/cm2 skin), and actual
doses were 0.0208, 0.201, and 2.154 mg/cm2 skin	Dermal ranged up to
15.8% of the applied dose.  Absorption was minimal with only 4 h of
exposure.  Absorbable radioactivity (radioactivity in the skin at the
application site and the adjacent skin) was minimal in groups that were
exposed for 10 h and evaluated for an additional 158 h post-exposure.
Thus, almost all of the dose isolated in the skin will be absorbed.
Considering the sum of absorbable and absorbed doses, 4-37% of the
applied dose was recovered in the treatment groups (11%).  Absorption
rate constants were calculated as 0.236, 0.190, and 0.072 h-1 for the 2,
20, and 200 µg/cm2 dose groups.  Absorption mechanisms were saturated
at the high dose.  The elimination half-lives were calculated to be 31,
30, and 37 h for the 2, 20, and 200 µg/cm2 dose groups.  A maximum of
36.56% of the applied dose was noted as absorbed/absorbable (observed
after 24 h exposure to 2 µg/cm2). The dose that is absorbed/absorbable
following a 10 h exposure is 16.54%, 21.31% and 11.39%, respectively, at
2, 20 and 200 µg/cm2.

A.3.	Executive Summaries

A.3.1	Subchronic Toxicity

	870.3100	90-Day Oral Toxicity – Rat

In a subchronic oral toxicity study (MRID 47090345), PP450 (93% a.i.;
Batch No. P10) was administered to 20 Wistar rats/sex/dose in the diet
at dose levels of 0, 20, 200, or 2000 ppm (calculated to be 0, 1.5, 14
and 158 mg/kg bw/day in males, and, 0, 1.6, 22 and 145 mg/kg/day in
females) for 90 days.

No treatment-related effects were noted on mortality, clinical signs of
toxicity, ophthalmoscopic examinations, urinalysis, or gross pathology
at any dose in either sex.

At 2000 ppm, body-weight gains were decreased (p<0.01) throughout the
study by 15-62% in both sexes.  Food consumption was decreased (p<0.05)
by 7-21% in the males (Weeks 1, 3, 5, 8, 10, and 12) and 9-35% in the
females (throughout the study).  Total (Weeks 1-13) food consumption was
decreased (p<0.01) by 7-19% in both sexes.  At 200 ppm, sporadic
decreases (p<0.05) of 5-12% were noted in food consumption and overall
food consumption was decreased by 6-7% in both sexes. At 20 ppm,
sporadic decreases (p<0.05) in food consumption of 4-12% in was observed
in both sexes.

Slight anemia was noted at 2000 ppm as indicated by decreases (p<0.01)
in the following parameters: (i) hemoglobin (↓4-7%) at Weeks 4 and 13;
(ii) hematocrit (↓5%) at Week 13; (iii) mean corpuscular volume
(↓3%) at Week 13; (iv) mean corpuscular hemoglobin (↓3-4%) at Weeks
4 and 13 and (v) mean corpuscular hemoglobin concentration at Weeks 4
and 13 (↓1-3%). The kaolin-cephalin time was decreased (↓13%) at
terminal sacrifice.  APDM activity was increased (p<0.05) by 22-27% in
both sexes, triglycerides were decreased (p<0.01), and cholesterol was
increased (p<0.01) at Weeks 4 and 13 in both sexes.

The target organ was the liver.  At 200 ppm, the absolute and adjusted
liver weights were increased (p<0.05) in females by 5-8% at this dose.
At 2000 ppm, increases (p<0.01) in absolute and adjusted for body weight
liver weights were observed in both sexes.  Increased incidence (#
affected/40) of hepatocyte vacuolation (fatty change) was noted in 25
treated animals vs. 5 controls. Centrilobular hypertrophy (25 treated
vs. 0 controls) with associated proliferation of smooth endoplasmic
reticulum and elevated aminopyridine-N-demethylase (APDM) activity was
also observed in both sexes at this dose.   Smooth endoplasmic reticulum
proliferation in the liver was increased (p<0.01) in the males.

The LOAEL is 2000 ppm (158/145 mg/kg bw/day in males/females) based on
decreased body-weight gain; decreased food consumption and liver
toxicity (increased absolute and adjusted liver weights, increased
endoplasmic reticulum proliferation in the males, and increased APDM
activity).  The NOAEL is 200 ppm (14/22 mg/kg bw/day in males/females).

This study is classified as acceptable/guideline and satisfies the
guideline requirements (OPPTS 870.3100a; OECD 408) for a subchronic oral
toxicity study in the rat.

870.3150	90-Day Oral Toxicity - Dog

In a subchronic toxicity study in dogs (MRID 47090346), Flutriafol
technical (PP450; 93.0% a.i.; batch # P10) was administered to four
beagle dogs/sex/dose group daily by capsule for 90 days at doses of 0,
1, 5, or 15 mg/kg/day.

No adverse, treatment-related effects were observed on mortality,
clinical signs of toxicity, food consumption, ophthalmoscopic
examinations, hematology, urinalysis, or gross pathology.

Mild focal alveolitis/bronchiolitis of the lungs was observed in the
females at 1 (3/4 dogs), 5 (1/4 dogs), and 15 (3/4 dogs) mg/kg/day
compared to (0/4) controls.  However, the Sponsor stated that this
finding was common in Alderley Park beagles, and was probably partly
associated with migration of Ascarid larvae and partly with respiratory
viruses.  Additionally, there was no strong dose relationship, and this
finding was observed in (2/4) control males.  Therefore, this finding
was considered equivocal.

≤0.05) in both sexes during Weeks 4, 8, and 13 by 42-82%, and the
increases became greater in magnitude with time of exposure.
Triglycerides were increased (p≤0.01) by 65% in the 15 mg/kg/day
females during Week 13.  Hepatic aminopyrine N-demethylase activity was
increased (p≤0.01) in both sexes by 149-156%.

Additionally at 15 mg/kg/day, cumulative body-weight gains in the
females were decreased (p≤0.05; except NS during Weeks 2, 5, and 6)
throughout treatment by 39-75%, with body weight losses of 0.3-0.5 kg
occurring during Weeks 1 and 2.  Additionally, males were also noted to
have spleens with hemosiderin content slightly higher than controls in
3/4 dogs compared to 0 controls.

The LOAEL is 15 mg/kg/day, based on adverse liver findings (increases in
organ weight, alkaline phosphatase, aminopyrine N-demethylase activity,
and incidence of hemosiderin-laden Kupffer cells) in both sexes, spleens
with hemosiderin content slightly higher than controls in the males, and
decreased cumulative body-weight gains and increased triglycerides in
the females.  The NOAEL is 5 mg/kg/day.

This study is classified as acceptable/guideline and satisfies the
guideline requirements (OPPTS 870.3150) for a subchronic oral toxicity
study in dogs.

	870.3200	21/28-Day Dermal Toxicity – Rat

In a 28-day dermal toxicity study (MRIDs 47090347 and 47090348),
Flutriafol Technical (Batch # UPL Bx 1; purity 95.1% a.i.) was applied
to the clipped skin of ten Sprague Dawley (Crl:CD[SD]) rats/sex at dose
levels of 0, 250, 500, or 1000 mg/kg bw/day (corrected for purity) in a
dose volume of 1.0 mL/kg for six hours/day on at least 28 consecutive
days.  Rats were sacrificed on Day 29.

No effects of treatment were observed on mortality, clinical signs of
toxicity, neurobehavioral examinations, body weights, body-weight gains,
food consumption, ophthalmology, hematology, clinical chemistry, or
gross or microscopic pathology.

In the 500 mg/kg/day females, increased incidence (total #
observations/# of rats with observation) of erythema grade 1 (24/5; not
significant [NS]) and flaking grade 1 (12/5; p≤0.05) were observed at
the treatment site.  Additionally at 1000 mg/kg/day, increased
(p≤0.01) incidences of erythema grade 1 (52/7 males; 73/10 females),
flaking grade 1 (11/6 males; 49/7 females), and scab(s) (35/5 males
[NS]; 17/4 females) were noted at the treatment site.

The systemic LOAEL was not observed.  The NOAEL is 1000 mg/kg bw/day
(limit dose).

This 28-day dermal toxicity study is classified acceptable/guideline and
satisfies the guideline requirement (OPPTS 870.3200) for a dermal
toxicity study in the rat.

A.3.2	Prenatal Developmental Toxicity

	870.3700a Prenatal Developmental Toxicity Study – Rat

In a developmental toxicity study (MRID 47090349), Flutriafol (PP450;
93%; Batch # P10) in corn oil was administered via daily oral gavage in
a dose volume of 10 mL/kg to 24 presumed pregnant Wistar rats at doses
of 0, 10, 50, or 125 mg/kg/day from gestation days (GD) 6-15.  On GD 21,
all dams were euthanized; each dam’s uterus was removed via cesarean
section and its contents examined.  Fetuses were examined for external,
visceral, and skeletal malformations and variations.

All dams survived until scheduled termination.  There were no
treatment-related macroscopic findings.

Increased incidence of staining of the genital/ventral fur was observed
primarily during the dosing period in 16 dams at 125 mg/kg/day compared
to 7 dams in the control group.  Additionally at 125 mg/kg/day, maternal
body-weight gains were decreased (p≤0.01) during the treatment (decr.
26%) and post-treatment (decr. 33%) intervals, and for the overall study
(decr. 23%).  Overall net weight gain, corrected for gravid uterine
weight, was decreased by 19% compared to controls.  Food consumption was
decreased by 14-17% at this dose compared to controls during the
treatment and post-treatment intervals.

The maternal LOAEL is 125 mg/kg bw/day based on increased incidence of
ventral/genital staining of the fur and decreased maternal body-weight
gains and food consumption.  The maternal NOAEL is 50 mg/kg bw/day.

The number and percent of early intrauterine deaths were increased at
125 mg/kg/day (40 deaths; 14.8%) compared to controls (15 deaths; 6.3%),
with a significantly higher (p≤0.05) proportion of dams affected at
125 mg/kg/day (14/21 dams) compared to controls (6/20 dams).  Similarly,
the number and percent of late intrauterine deaths were increased at 125
mg/kg/day (46 deaths; 18.7%) compared to controls (0 deaths; 0%), with a
significantly higher (p≤0.05) proportion of dams affected at 125
mg/kg/day (14/21 dams) compared to controls (0/20 dams).  The increases
in early and late intrauterine deaths were reflected by an increased
post-implantation loss at this dose (33.5% affecting 17/21 dams)
compared to controls (6.3% affecting 6/20 dams).

≤0.01) by 16-27%.

Incidences of the following skeletal variations, indicating skeletal
retardation, were increased (p≤0.05) over concurrent controls and/or
the provided historical control data:  (i) in all treated groups -
incompletely ossified unilateral and/or bilateral calcanea, partially
ossified occipital, and not ossified odontoid; (ii) in the 50 and 125
mg/kg/day fetuses - unilateral and/or bilateral cervical rib and
unilateral and/or bilateral extra (14) ribs; (iii) at 125 mg/kg/day -
partially ossified parietals, increased fontanelle, partially ossified
cervical arches between and including #3 and #6, partially ossified 1st
sternebra, partially ossified 2nd sternebra, not ossified 5th sternebra,
partially ossified, not ossified 6th sternebra, and partially ossified
frontals.  Mean scores for ossification of the manus were increased in
all treated groups (2.66-3.13) compared to concurrent (2.42) and
historical (1.88-2.59) controls.  Similarly, mean scores for
ossification of the pes were increased in all treated groups (3.06-3.63)
compared to concurrent (2.72) and historical (2.53-3.05) controls.
Aside from the variations listed above indicating skeletal retardation,
there were no treatment-related external, visceral, or skeletal
variations.

There were no treatment-related external, visceral, or skeletal
malformations.

The developmental LOAEL is 50 mg/kg bw/day based on delayed ossification
or non-ossification of the skeleton in the fetuses.  The developmental
NOAEL is 10 mg/kg bw/day.

This study is classified unacceptable/guideline and does not satisfy the
guideline requirement for a developmental toxicity study (OPPTS
870.3700; OECD 414) in rats.

	870.3700a Prenatal Developmental Toxicity Study – Rat

In a developmental toxicity study (MRID 47521303), Flutriafol (95.1%;
Batch # UPL Bx 1 (2001)) in corn oil was administered via daily oral
gavage in a dose volume of 4 mL/kg to 22 presumed pregnant Wistar rats
per dose group at doses of 0, 2, 5, 10, or 75 mg/kg bw/day from
gestation days (GD) 6-20.  On GD 21, all dams were euthanized; each
dam’s uterus was removed via cesarean section and its contents
examined.  Fetuses were examined for external, visceral, and skeletal
malformations and variations.

All dams survived until scheduled termination.  There were no clinical
signs of toxicity throughout the study, and no gross abnormalities were
observed at necropsy.  At 75 mg/kg/day, absolute maternal body-weight
gains were decreased by 33% compared to controls during GD 6-9 and by
20% during GD 9-12.  Body-weight gains for the overall (GD 6-21)
treatment period were decreased by 8% compared to controls; when
corrected for gravid uterine weight, body-weight gains were 28% lower
than controls.  When expressed as a percent of body weight on GD 6,
body-weight gains were significantly decreased (p≤0.01) beginning the
first day after dosing (GD 6-7) and continuing through GD 17.  The
decrease was most pronounced for GD 6-7 (decr 100%) and diminished to
13% lower than controls on GD 14.  Although not statistically
significant, relative body-weight gains at 75 mg/kg/day were also
decreased (decr 11-12%) on GD 18 and 19.  Additionally at this dose,
mean body-weight gain for the overall (GD 6-20) treatment period was
decreased by 9% (p≤0.05) compared to controls.  Food consumption at
this dose was decreased by 11-15% compared to controls throughout the
treatment interval.

The maternal LOAEL is 75 mg/kg bw/day based on decreased body-weight
gains and food consumption.  The maternal NOAEL is 10 mg/kg bw/day.

There were no abortions, premature deliveries, dead fetuses, or complete
litter resorptions and no effects of treatment on the number of litters
or sex ratio.  The number of late resorptions at 75 mg/kg/day was higher
than controls (21 treated vs 1 control), with the number of late
resorptions per dam significantly increased (p≤0.05) at this dose
(1.0/dam treated vs 0.0/dam controls).  The number of early resorptions
was also increased at 75 mg/kg/day (17 treated vs 12 controls), with the
number of early resorptions per dam increased at this dose (0.8/dam
treated vs 0.5/dam controls), although this increase was not
significant.  The increases in early and late resorptions, particularly
late resorptions, resulted in a significantly decreased (p≤0.05)
number of live fetuses/dam (11.7 treated vs 13.1 controls).  Although
numerically minor, there was a reduction in fetal weights at 75
mg/kg/day that was statistically significant and would correspond to the
delay in development (non-ossified, incompletely ossified bones) at this
dose level.

Treatment-related malformations were observed in the hyoid at 75
mg/kg/day compared to 0 concurrent and historical controls, including
incidences of:  misshapen arch (1% fetuses; 5% litters); absent body (1%
fetuses; 5% litters); interrupted body (7% fetuses; 18% litters); and
bent body (2% fetuses; 9% litters).  Short intestine was noted in a
single 75 mg/kg/day fetus and was not observed in the historical
controls.  Cleft palate was noted in a single fetus at 75 mg/kg/day.
This uncommon malformation was also observed in a single fetus at 100
mg/kg/day in the supplementary range-finding study (MRID 47521302).  It
should also be noted that cleft palate occurred in a single historical
control fetus.  There were no other treatment-related external,
visceral, or skeletal malformations.

Treatment-related visceral variations included misshapen nasopharynx
lumen and displaced common carotid artery origin, which were observed at
75 mg/kg/day, but were not found in any concurrent or historical
controls.

The following skeletal variations at 75 mg/kg/day were considered to be
due to the test material because the fetal and litter incidences were
dose-related and exceeded concurrent and historical controls:  (i)
additional ossification of the squamosal or zygomatic process of the
maxilla; (ii) zygomatic arch fusion; (iii) blue-stained focus on the
maxilla or mandible; (iv) accentuated curvature of the hyoid body; (v)
long cervical rib; (vi) rudimentary cervical rib; (vii) caudal
displacement of the pelvic girdle; (viii) bilateral radius and ulna
bent; and (ix) cervical rib cartilage fused with thoracic rib 1
cartilage.

≤0.05) at 75 mg/kg/day over concurrent controls and exceeded the range
of historical controls:  (i) incompletely ossified sternebra 6; (ii)
unilateral left supernumerary rib; (iii) unilateral left rudimentary
rib; (iv) unilateral right supernumerary rib;

(v) unilateral right rudimentary rib; (vi) supernumerary unilateral left
costal cartilage;

≤0.05) at 75 mg/kg/day (0%) compared to concurrent controls and fell
below the range of historical controls.

≤0.05); however, both the fetal and litter incidences fell within the
range of historical controls.  The incidence of cervical vertebral body
2 was lower at this dose compared to concurrent and historical controls,
with the litter incidence attaining statistical significance (p≤0.05);
however, incidences of this finding fell within the historical control
range.  Although the incidences of several of these findings were not
significantly increased and/or fell within the range of historical
controls, they were considered treatment-related because of their
increase over concurrent controls and their corroboration of the
generalized skeletal retardation.

≤0.05) at 75 mg/kg/day compared to concurrent controls.  Although the
incidences of these findings fell within the range of historical
controls, they were considered to be due to treatment due to the
substantial and statistically significant increases over concurrent
controls and the fact that this developmental delay is consistent with
the other indications of skeletal retardation.  Furthermore, the fact
that these incidences fall within the range of historical controls is
attributed to a single study (No. 857932).

In summarizing, administration of flutriafol to dams at 75 mg/kg/day
results in teratogenicity (external, visceral and skeletal
malformations), embryo-lethality, skeletal variations, a generalized
delay in fetal development and fewer live fetuses.

The developmental LOAEL is 75 mg/kg bw/day based on:  increased late
resorptions; malformations (cleft palate and multiple hyoid
malformations) and variations in the hyoid; variations in the
maxilla/mandible, rudimentary and long cervical ribs, pelvic girdle, and
radius/ulna; numerous skeletal retardations detailed above and
corresponding decrease in fetal weights. The developmental NOAEL is 10
mg/kg bw/day.

This study is classified acceptable/guideline and satisfies the
guideline requirement for a developmental toxicity study (OPPTS
870.3700a; OECD 414) in rats.

870.3700b Prenatal Developmental Toxicity Study - Rabbit

In a developmental toxicity study (MRID 47090350), Flutriafol (PP450;
93%; Batch # P10) was administered daily in gelatin capsules to 24
presumed pregnant Dutch rabbits at doses of 0, 2.5, 7.5, or 15 mg/kg/day
from gestation days (GD) 6-18.  On GD 29, each surviving female was
euthanized, and the uterus was removed via cesarean section and its
contents examined.  Fetuses were examined for external, visceral, and
skeletal malformations and variations.

At 7.5 mg/kg/day, one doe (#52) aborted part of its litter on GD 20.  At
15 mg/kg/day, one doe (#70) was killed in extremis after observations
that the animal had not been eating or drinking and that it had lost
weight and was in poor condition.  No other maternal deaths could be
attributed to treatment.  Loose feces on the cage floor and/or fur of
the animals was observed in 2/15 rabbits at 7.5 mg/kg/day and 4/15
rabbits at 15 mg/kg/day.  These findings were observed only once per
female, except for one doe at 7.5 mg/kg/day for which the observation
was made on 2 days and one doe at 15 mg/kg/day for which loose feces was
noted on three days.

At 15 mg/kg/day, maternal body weight gains were decreased during the
treatment interval (-79 g treated vs 48 g controls) and for the overall
(GD 0-29) study, both when uncorrected for (149 g treated vs 230 g
controls) and when corrected for (-158 g treated vs -55 g controls)
gravid uterine weights.  Additionally at this dose, maternal food
consumption was increased by 24% (p≤0.01) over controls during the
pre-treatment interval, but was decreased by 22% (not significant)
during treatment.

In two of the females examined at 15 mg/kg/day, the stomach was found to
contain a fur ball and was otherwise empty or contained little food.
Only a single female at 7.5 mg/kg/day had little to no food in the
stomach.  Additionally at 15 mg/kg/day, one of the aforementioned does
had dark pitted areas on the mucosal surface of the glandular portion of
the stomach.

The maternal LOAEL is 15 mg/kg/day based on decreased corrected and
uncorrected maternal body weight gains and food consumption.

The number of early intrauterine deaths was higher at 15 mg/kg/day than
controls (36 deaths; 31.0%) compared to controls (11 deaths; 10.4%).
Similarly, the number of late intrauterine deaths was increased at this
dose (19 deaths; 16.4%) compared to controls (1 death; 1.0%).  Complete
litter resorptions were significantly higher (p<=0.05) at 15 mg/kg/day,
occurring in 5/14 does compared to 0/15 controls.  These findings
resulted in a significantly increased (p<=0.01) post-implantation loss
at 15 mg/kg/day (45.5% vs 13.1% controls); a decreased number of litters
(9 vs 15); and a decreased total (61 vs 94) and mean (4.0 vs 6.5;
p<=0.05) number of live fetuses.

There were no treatment-related effects on growth or development of the
fetuses.  Fetal body weights and litter weights of the treated groups
were comparable to controls.  Reduced/delayed ossification was observed
in several bones in the skeleton (skull, vertebrae, and sternebrae) at
an increased incidence over controls.  However, these findings were
minor in incidence and were not significantly different from the
controls.  Furthermore, mean scores for ossification of the manus and
pes in all treated groups were comparable to controls.

There were no treatment-related external, visceral, or skeletal
malformations or variations.  Two fetuses, one at 7.5 mg/kg/day and
another at 15 mg/kg/day, had multiple abnormalities; however, historical
control data showed that similar findings were previously noted in
individual fetuses (e.g., cleft palate, gastroschisis, malformed eyes,
and shortened/flexed limbs with reduced number of digits).  Furthermore,
the findings in the fetus at 7.5 mg/kg/day were more severe than those
in the 15 mg/kg/day fetus.  All other findings were unrelated to dose,
minor in incidence, and/or not significantly different from the
controls.

The developmental LOAEL is 15 mg/kg/day based on decreased number of
live fetuses, complete litter resorptions and increased
post-implantation loss.   The developmental NOAEL is 7.5 mg/kg/day.

This study is classified acceptable/guideline and satisfies the
guideline requirement for a developmental toxicity study (OPPTS
870.3700; OECD 414) in rabbits.

A.3.3	Reproductive Toxicity

	870.3800 Reproduction and Fertility Effects – Rat

In a two-generation reproduction toxicity study (MRID 47090351),
Flutriafol (93%; Batch # P10) was administered in the diet to 15 male
and 30 female Wistar rats/sex/dose group at dietary levels of 0, 60,
240, or 1000 ppm (0, 4.8, 20.6 and 88.7 mg/kg bw/day in P  males; and,
0, 5.5, 21.9 and 103 mg/kg bw/day in  P females)  The P generation
animals were fed the test diets for 12 weeks prior to mating to produce
the F1a litters. After weaning of the F1a litter, the females were mated
with a different male to produce the F1b litters.  On post-natal day
(PND) 36, offspring from the F1b litters were selected to be parents and
were fed the same test diet concentration as their dam for 11 weeks
prior to mating to produce the F2a litters.  This procedure continued
through weaning of the F2b litters which was produced by mating of the
F1b.

There were no treatment-related deaths or clinical signs of toxicity.

Treatment-related effects on body-weight gain, food consumption, and
food utilization during the pre-mating period were observed at 1000 ppm.

Body-weight gains were decreased (p≤0.05, unless otherwise noted) at
1000 ppm compared to controls:  throughout the pre-mating period in the
P males (decr. 6-8%); beginning at Week 7 in the P females (decr. 4-6%,
not significant [NS] at Week 8); and throughout pre-mating in the F1
females (decr. 6-18%; NS at Week 6).

≤0.05) at 1000 ppm compared to controls:  generally throughout
pre-mating in the P males (decr. 4-7%), resulting in a decrease in total
food consumption (Weeks 1-12) of 2% (p≤0.01) compared to controls;
beginning at Week 6 in the P females (decr. 6-8%), resulting in a
decrease of 5% (p≤0.01) in total food consumption; and at Week 8 in
the F1 females (decr. 6%; p≤0.01).

Food utilization was increased by 3% (p≤0.01) compared to controls in
the 1000 ppm P males for Weeks 1-4.  Food utilization was increased by
10% (p≤0.05) over controls in the F1 females at this dose for Weeks
1-4, resulting in an increase of 8% (p≤0.01) for the overall (Weeks
1-11) pre-mating period.

Throughout gestation, cumulative body-weight gains were decreased by
3-25% at 1000 ppm in the P dams during both litters and in the F1 dams
during the F2b litter.  With the exception of the P females on GD 8 and
22 during the F1a litter and the F1 females on GD 22 during the F2b
litter, these decreases were significantly (p≤0.05) different from
controls.

Absolute and adjusted (for body weight) liver weights were increased
(p≤0.01) by 11-29% over controls in both sexes in both generations,
with the exception of the absolute liver weight in the P females, which
was increased by 6% over controls (NS).

Treatment-related microscopic findings were found in the liver.
Centrilobular hypertrophy was observed in 1000 ppm males in the P
generation (2/15 treated vs. 0/15 controls) and F1 generation (4/15
treated vs. 0/15 controls).  Increased incidences of fatty change in the
liver were observed at this dose in the P generation males (8/15 treated
vs. 0/15 controls) and females (5/30 treated vs. 1/30 controls) and in
the F1 generation males (13/15 treated vs. 0/15 controls) and females
(3/30 treated vs. 0/30 controls).  Fatty change was also observed in the
240 ppm F1 males (5/15 treated vs. 0/15 controls).

The LOAEL for parental toxicity is 1000 ppm (88.7/103 mg/kg bw/day in
males/females) based on decreased body-weight gains and food consumption
and on effects on the liver (increased liver weights, centrilobular
hypertrophy, and fatty change). The NOAEL is 240 ppm (20.6 mg/kg bw/day
in P males and 21.9 mg/kg bw/day in P females).

Although no data were provided, it was stated that the offspring
generally remained in good clinical condition and that there were no
clinical abnormalities which could be related to treatment.  There were
no effects of treatment on the offspring survival indices (percent pups
surviving to PND 22 and proportion of litters with all pups surviving to
PND 22) or cumulative pup body-weight gains throughout the post-natal
period.  At necropsy, no macroscopic findings could be attributed to
treatment.

≤0.05) throughout the post-natal period in the F1b litter (decr.
17-18%) and in the F2a litter (decr. 22-23%).  The percent of pups born
alive was decreased (p≤0.05) in the F2a litter (94.9% treated vs. 100%
controls) and F2b litter (90.1% treated vs. 99.3% controls).  The
proportion of litters with all pups born alive was decreased at this
dose in the F2a litter (15/20 treated vs. 18/18 controls) and F2b litter
(19/29 treated vs. 24/26 controls).  In the liver, fatty change was
observed at 1000 ppm in the F1b male pups (1/7 treated vs. 0/6 controls)
and F1b female pups (1/6 treated vs. 0/8 controls).  Fine vacuolar
hepatocyte vacuolation/fatty change was observed in the F2b males (5/10
treated vs. 0/10 controls) and F2b females (1/10 treated vs. 0/10
controls).

The LOAEL for offspring toxicity is 1000 ppm (88.7/103 mg/kg bw/day in
males/females) based on decreased live birth index and litter size and
on effects on the liver (fatty change/vacuolation).  The NOAEL is 240
ppm (approximately equivalent to 20.6 mg/kg bw/day in males and 21.9
mg/kg b w/day in females).

There was no apparent effect of treatment on estrous cycle duration or
periodicity in the P generation.  There were no effects of treatment on
precoital interval, gestation duration, or fertility in either litter in
either generation.

The LOAEL for reproductive toxicity was not observed.  The NOAEL is 1000
ppm (approximately equivalent to 88.7/103 mg/kg bw/day in
males/females).

This study is classified as acceptable/guideline and satisfies the
guideline requirements (OPPTS 870.3800; OECD 416) for a two-generation
reproduction study in the rat.

A.3.4	Chronic Toxicity

	870.4100a (870.4300) Chronic Toxicity – Rat

In a combined chronic toxicity/carcinogenicity study (MRID 47090352), 52
Alpk:AP rats/sex/dose were exposed to flutriafol (93% a.i.; Batch No.:
P10) for up to 24 months in the diet at concentrations of 0, 20, 200, or
2000 ppm (calculated to be, 0, 1.02, 10.0, and 102 mg/kg bw/day in
males; and 0, 1.27, 12.2 and 122 mg/kg bw/day in females).
Additionally, 12 rats/sex/dose were treated similarly for up to 12
months.

No treatment-related effects were observed on mortality, ophthalmology,
clinical chemistry, or urinalysis.

Grossly,  small discolored foci were commonly observed after 2 years of
treatment.  After 1 year of treatment, an increased incidence of fatty
change in the liver was observed in the 200 and 2000 ppm males (21-93%
of treated rats vs 7% controls).  The severity was minimal in the
controls and 200 ppm males, but was minimal to marked in the 2000 ppm
males.  After 2 years of treatment of the 200 and 2000 ppm males,
increased incidences of minimal to severe hepatic fatty change (54-96%
treated vs 24% controls) and clear cell foci of hepatocytes (40-50%
treated vs 18% controls) were observed.

At 2000 ppm, systemic toxicity was noted in both sexes as follows.  More
rats appeared thin and fewer rats had distended abdomens.  Final body
weights were decreased by 12-22%, and cumulative body-weight gains were
decreased by 12-48% throughout the study.  Weekly food consumption was
frequently decreased by 4-24% throughout treatment, and total food
consumption was decreased by 8-12% for the Weeks 1-13 interval.  Food
utilization (g food/g growth) was increased by 8-11% for the Weeks 1-4
interval, and by 7% (each sex) for the Weeks 1-12 interval.

A slight treatment-related anemia was noted in the 2000 ppm group as
indicated by the following decreases (p≤0.05) in hematological
parameters: (i) hemoglobin in males (↓4-7%) during Weeks 4-65 and
females (↓4-9%) during Weeks 13-52, 78, and 92; (ii) hematocrit in
males (↓3-8%) during Weeks 26-65 and females (↓5-11%) during Weeks
13-52, 78, and 104; (iii) mean cell volume in males (↓3-8%) during
Weeks 4-104 and females (↓2-10%) during Weeks 4-104; and (iv) mean
cell hemoglobin in males (↓4-7%) during Weeks 4, 26, 39, and 78-104
and females (↓4-10%) during Weeks 4-52 and 78-104.  The total iron
binding capacity of the 2000 ppm females was increased (p≤0.01) by
40%.  Increased (p≤0.05) lymphocytes were observed in the 2000 ppm
females (↑22-61%) during Weeks 26-78 and 104, and increased (p≤0.05)
total leukocytes were noted at Weeks 26, 39, and 78 (↑20-38%).  The
hematological changes were not considered to be an adverse effect due to
the minor decreases in magnitude without corroborating clinical signs.

At 2000 ppm, the following toxicologically significant differences
(p≤0.05) were observed: (i) increased plasma cholesterol in the
females throughout the study (↑24-49%; NS at Week 91); (ii) decreased
plasma triglycerides in the males during Weeks 4-65 (↓40-68%); (iii)
decreased alkaline phosphatase in the males during Weeks 13-91
(↓12-33%); (iv) increased plasma total protein in the females
throughout treatment (↑4-9%); and (v) increased plasma alanine
transaminase during Weeks 4 and 13 (↑54-82%).

At 2000 ppm, hepatoxicity was noted in both sexes.  In both sexes,
increased liver weights, both absolute and adjusted for body weight,
were observed after 1 year of treatment (incr 11-37%) and after 2 years
(incr 27-34%, except similar to control for absolute liver weight of the
females).  There was hepatic enlargement, often coupled with the
presence of numerous discolored foci, commonly observed in both sexes.
These liver findings were observed after 2 years of treatment, but not
after 1 year of treatment.  After 2 years of treatment, the following
histological hepatic lesions were increased in incidence in the females:
 (i) minimal to severe fatty change (65% treated vs 23% controls); (ii)
bile duct proliferation/ cholangiolarfibrosis (67% treated vs 44%
controls); (iii) hemosiderin accumulation in Kupffer cells (55% treated
vs 0% controls); and (iv) centrilobular hypertrophy (8% treated vs 0%
controls).  Hepatic centrilobular hypertrophy was increased in incidence
at the interim sacrifice in males (71%) and females (31%), but only
minor increases were noted at terminal sacrifice in both sexes (6-8%)
with 0% in the controls.  An increased incidence of foci of cortical
macrophages in adrenal glands was observed in the 2000 ppm females (80%
treated vs 25% controls); however, there was no corroborating evidence
of toxicity in the adrenal gland, and this lesion alone was not
considered adverse.

The LOAEL is 2000 ppm (102/122mg/kg bw/day in males/females), based on
adverse liver effects (increased liver weights, fatty change, bile duct
proliferation/cholangiolarfibrosis, hemosiderin accumulation in Kupffer
cells and centrilobular hypertrophy), and clinical chemistry findings.
The NOAEL is 200 ppm (10.0/12.2 mg/kg bw/day in males/females).

At the doses tested, there was not a treatment related increase in tumor
incidence when compared to controls.  Dosing was considered adequate
based on decreased body-weight gain and food consumption, increased food
utilization, and hepatotoxicity observed in both sexes.

This study is classified as Acceptable/guideline and satisfies the
guideline requirements (OPPTS 870.4300; OECD 453) for a combined chronic
toxicity/carcinogenicity study in rats.

	870.4100b Chronic Toxicity – Dog

In a chronic toxicity study in dogs (MRID 47090353), flutriafol (PP450;
93.0% a.i.; batch # P10) was administered to four beagle dogs/sex/dose
group daily by capsule for at least 52 weeks at doses of 0, 1, 5, or 20
mg/kg bw/day.

No adverse, treatment-related effects were observed on mortality,
clinical signs of toxicity, food consumption, ophthalmoscopic
examinations, or gross pathology.

One 20 mg/kg/day female was observed to be subdued and not eating during
Week 15.  This dog was dehydrated and thin, had pale mucus membranes,
loud intestinal sounds and a tense abdomen, with marked reduction in
muscle mass.  Mucus containing blood was present in the feces, and vomit
was present on the pen floor on three days.  This animal was killed for
humane reasons during Week 16.  In view of the limited toxicity noted in
the other dogs receiving 20 mg/kg/day, it was unclear if the poor
condition of this dog was due to administration of the test compound.

The liver was a target organ.  At 20 mg/kg/day, liver weights were
increased (p≤0.01) by 27-39% in both sexes.  The following
treatment-related alterations in clinical chemistry parameters were
observed:  (i) alkaline phosphatase was increased (p≤0.01, except not
significant [NS] for Week 13 males) by 60-268% in both sexes during
Weeks 4, 13, 26, and 52, and the increases became greater in magnitude
with time of exposure; (ii) albumin was decreased (p≤0.05) by 15-19%
during Weeks 4, 13, 26, and 52 in the males, and by 9-10% during Weeks
13 and 26 in the females; and (iii) triglycerides were increased
(p≤0.05) by 31% during Week 52 in the males, and by 62-71% during
Weeks 4 and 13 in the females.  Additionally, minimal to slight
increased centrilobular hepatocyte lipid in the liver was noted in 3/4
females vs. 0/4 controls.

Body weight losses (p≤0.05) of 0.02-0.17 kg were observed in the
females during Weeks 1 and 2.  These animals did not recover over the
course of the study, and demonstrated decreased (not significant [NS])
cumulative body-weight gains for the study period (Weeks 0-52; decr.
28%).  Adrenal weights were increased (p≤0.01) by 38% in the females,
and increased cortical vacuolation of the zona fasciculata was observed
in the adrenal in 4/4 males (slight severity) and 4/4 females (minimal
to slight severity) vs. 0/4 controls of both sexes.  Additionally in the
males, during Weeks 26 and 52, hemoglobin was decreased (p≤0.05) by
7-9%, hematocrit was decreased (p≤0.05 for Week 52; NS for Week 26) by
8-11%, and red blood cell counts were decreased (NS). Marked liver
sinusoidal cell hemosiderin pigmentation was observed in 4/4 males and
4/4 females vs. minimal to moderate in 4/4 controls of both sexes, and
marked hemosiderin pigmentation in the spleen was noted in 4/4 males and
4/4 females vs. minimal to moderate in 4/4 controls of both sexes.

At 5 mg/kg/day, a minimal severity of cortical vacuolation of the zona
fasciculata was present in one female and one male out of a total of 8
animals.  RAB1 toxicologists concluded that this minimal effect in the
adrenal gland was not an adverse effect.

The LOAEL is 20 mg/kg/day, based on:  adverse liver findings (increased
liver weights, increased centrilobular hepatocyte lipid in the liver,
and increases in alkaline phosphatase, albumin and triglycerides),
increased adrenal cortical vacuolation of the zona fasciculata, and
marked hemosiderin pigmentation in the liver and spleen in both sexes;
mild anemia (characterized by decreased hemoglobin, hematocrit, and red
blood cell count) in the males; and initial body weight losses,
decreased cumulative body-weight gains, and increased adrenal weights in
the females.  The NOAEL is 5 mg/kg/day.

This study is classified as acceptable/guideline and satisfies the
guideline requirements (OPPTS 870.4100) for a chronic oral toxicity
study in dogs.

A.3.5	Carcinogenicity

	870.4200a Carcinogenicity Study – Rat

(see A.3.4 870.4100a Chronic Toxicity- rat)

	870.4200b Carcinogenicity (feeding) - Mouse

In a carcinogenicity study (MRID 47090354), Flutriafol (93% a.i.; Batch
No. P10) was administered in the diet to C57BL/10JfCD-1/Alpk mice
(50/sex/dose) for up to 2 years at doses of 0 (two control groups), 10,
50, or 200 ppm (calculated to be 0, 1.1, 5.9, and 24 mg/kg bw/day in
males; and 0, 1.4, 7.4 and 31 mg/kg bw/day in females).

No adverse treatment-related effects were observed on mortality or food
consumption.

≤0.05) generally throughout the study in both sexes (decr. 2-8%).
Overall (Weeks 1-104) body-weight gains were decreased in the males
(decr. 18%; p≤0.01) and females (decr. 8%; not statistically
significant [NS]); and decreased (p≤0.01) food efficiency was observed
in the males during Weeks 1-4 (decr. 38%) and 1-12 (decr. 21%).
Additionally, increased (p≤0.05) platelet (incr 42%), white blood cell
(incr 62%), neutrophil (incr. 81%), and lymphocyte (incr 58%) counts
were noted in the males.  Hepatotoxicity was also noted.  Increased
(p<0.01) liver weights (absolute and adjusted for body weight) were
observed in males (incr. 32-37%) and females (incr. 17-26%).
Furthermore, increased incidences (# affected/50 in treated vs controls)
of minimal to marked hepatic centrilobular fatty change were noted in
the males (23 vs. 1) and females (17 vs. 0); and minimal to moderate
hepatic centrilobular hypertrophy were noted in the males (14 vs 0-1)
and females (3 vs 0).

At 50 ppm, a slight effect was observed on body weights and body-weight
gains in males.  Body weights were decreased by 5% (p≤0.05) on Week
104, and overall (Weeks 1-104) body-weight gains were decreased by 8%
(NS).  Furthermore, a treatment-related increased incidence of hepatic
centrilobular fatty change was noted in 6/50 males (1 minimal, 4 slight,
and 1 marked severity).

The LOAEL is 200 ppm (24/31 mg/kg bw/day in males/females), based on
hepatotoxicity (increased fatty change) in both sexes.  The NOAEL is 50
ppm (5.9/7.4 mg/kg bw/day in males/females).

At the doses tested, there was not a treatment related increase in tumor
incidence when compared to controls.  There was an apparent increase in
the incidence of generalized composite lymphomas in the 200 ppm female
decedents (100% treated vs 62% controls).  Although this finding was
statistically significant (p≤0.05), the difference was no longer
evident when all animals were considered (92% treated vs 81-91%
controls).  Furthermore, the effect was not clearly dose-dependent.
Dosing was considered adequate based on decreases in body weights and
body-weight gain in both sexes, decreased food efficiency in males,
hematological findings in males, and hepatoxicity in both sexes.

This study is classified as acceptable/guideline and satisfies the
guideline requirement for a carcinogenicity study [OPPTS 870.4200; OECD
451] in mice.

A.3.6	Mutagenicity

Gene Mutation

870.5100, In vitro Bacterial Gene Mutation (Salmonella typhimurium)/
mammalian activation gene mutation assay

MRID 47090401 Acceptable/guideline	0, 1.6, 8, 40, 200, 1000, or 5000
µg/plate ( Trial 1) or 0, 8, 40, 200, 1000, 2500, or 5000 µg/plate
(Trial 2); Both trials were performed w/wo S9-activation. There were no
marked increases in the mean number of revertants/plate in any strain.
There was no evidence of induced mutant colonies over background.

870.5300, In Vitro Gene Mutation assay in mouse lymphoma cells

MRID 47090402

Acceptable/guideline	0, 10, 33, 100, 333, or 1000 µg/mL (+S9, Trial 1);
0, 150, 300, 450, 600, or 750 µg/mL (-S9, Trial 1); 0, 150, 300, 450,
600, 750, 900, 1050, or 1200 µg/mL (+S9, Trial 2); or 0, 200, 300, 400,
500, 600, 700, or 800 µg/mL (-S9, Trial 2).  There was a dose-related
increase in mutant frequency (7.0-9.0x10-5 treated vs. 3.0x10-5
controls) and absolute mutant numbers (70-148 colonies/plate vs. 63
controls) at 100 µg/mL and above in Trial 1 and a marked increase in
mutant frequency at 750 µg/mL (6.5x10-5 treated vs. 1.2x10-5 controls)
in Trial 2 attributable to severe cytoxicity (2% relative survival).
However, the increases in mutant frequency did not achieve the threshold
value for a positive response (>10x10-5) in either trial and there was
no marked increase in absolute mutant numbers at 750 µg/mL in Trial 2.
In the absence of S9, there were no marked increases in mutant frequency
or absolute mutant numbers compared to controls in either trial.  There
was no convincing evidence of induced mutant colonies over background in
the presence or absence of S9-activation.

Cytogenetics

870.5375, In vitro Mammalian Cytogenetics (Chromosomal Aberration Assay
in Human Peripheral Blood Lymphocytes)

MRID 47090403

Acceptable/guideline	No significant increases in the numbers of cells
with aberrations (excluding gaps) were observed in either donor in the
presence or absence of S9. There was no evidence of chromosome
aberrations induced over background in the presence or absence of
S9-activation.

870.5385, In vivo Mammalian Cytogenetics – [Bone Marrow Chromosomal
Aberration Test

MRID 47090404 Acceptable/guideline	0, 15, 70, or 150 mg/kg. There was no
evidence of chromosome aberration induced over background.

870.5395, In Vivo Mammalian Cytogenetics - Erythrocyte Micronucleus
Assay in Mice

MRID47090405

Acceptable/guideline	0, 93.8, or 150 mg/kg. Decreased (p<0.01)
polychromatic erythrocyte to normochromatic erythrocyte ratios (PCE:NCE)
were observed in both doses at all time points, indicating that the test
material was toxic to the bone marrow. There was no significant increase
in the frequency of micronucleated polychromatic erythrocytes in bone
marrow after any treatment time.

	Other Genotoxicity

870.5450, Dominant Lethal Assay – Mice

MRID 47090406

Acceptable/guideline	0, 25, 50, or 100 mg/kg/day (total doses of 0, 125,
250, or 500 mg/kg). Mortality (3/15 males) was noted at 100 mg/kg/day
during dosing.  Slight decreases (p<0.05) in body weight were observed
at 50 mg/kg/day and above during dosing.  There were no
treatment-related effects on fertility, mean number of implantations, or
the number of early or late deaths. There was no time-related positive
response of increased pre- or post-implantation loss compared to
controls.

870.5550, Unscheduled DNA Synthesis in Primary Rat Hepatocytes/Mammalian
Cell

MRID 47090407 (2003)

≥ 5 NNG needed for a positive response, and no increase in the mean
percent of cells in repair was observed. There was no evidence that
unscheduled DNA synthesis, as determined by radioactive tracer
procedures [nuclear silver grain counts] was induced.

A.3.7	Neurotoxicity

		870.6200 Acute Neurotoxicity Screening Battery - Rat

In an acute neurotoxicity study (MRID 47090408), Flutriafol (95.1% a.i.;
Lot # UPL Bx 1 [2001]) was administered once via gavage (10 mL/kg) to 10
Sprague-Dawley rats/sex/group at dose levels of 0, 125, 250, or 750
mg/kg.  Neurobehavioral assessment (functional observational battery
[FOB] and motor activity testing) was performed on 10 rats/sex/group at
pre-dosing and Days 1 (approximately 8 hours post-dosing; estimated time
of peak effect), 8, and 15.  At study termination, 5 rats/sex/group were
anesthetized and perfused in situ for neuropathological examination.
The tissues from the perfused animals in the control and 750 mg/kg
groups were subjected to histopathological evaluation of brain and
peripheral nervous system tissues.  Acceptable positive control data
were provided.

No compound-related effects were observed in brain weights or gross or
neuropathology.

At 125 mg/kg, males exhibited a dose-dependent body weight loss of -4.2%
on Days 1-2.  At 250 mg/kg and above, dose-dependent losses (-4.2-28.5
g) (p<0.01) in body weight were observed in both sexes on Days 1-2.  On
Days 2-3, body-weight gains were increased (52-74%) (p<0.05) in both
sexes at 250 mg/kg but remained decreased in both sexes at 750 mg/kg.
The increase in the 250 mg/kg females was sufficient to compensate for
the initial body weight loss and allowed the body weights in this group
to remain similar to controls for the remainder of the study.  Despite
the increase at 250 mg/kg, the body-weight gain remained decreased in
the 250 mg/kg males during the interval, 1-3 days (-76%).  At 750 mg/kg,
overall (Days 1-16) body-weight gain was decreased in the males (-20%)
(p<0.01) and females (-17%) (NS).  In the males, dose-dependent
decreases (p<0.01) were noted in both absolute and relative food
consumption on Days 1-2 in the 250 mg/kg group (64% and 63%,
respectively) and the 750 mg/kg group (89% and 88%, respectively).  This
initial decrease resulted in a continued reduction in both absolute and
relative food consumption in males on Days 1-3.  In the 750 mg/kg males,
overall (Days 1-16) absolute food consumption was decreased (-15%)
(p<0.01) compared to controls while no statistically significant
decrease in relative food consumption was observed.  In the 750 mg/kg
females, no statistically significant decreases in absolute or relative
food consumption were observed during the remainder of the study or in
the overall (Days 1-16) values.

Additionally at 750 mg/kg, 4/10 males and 2/10 females were sacrificed
in moribund condition on Days 2 or 3.  Increases (p<0.01) were noted in
the incidence of the following clinical signs in the males (unless
otherwise stated):  dehydration (both sexes), chromorhinorrhea,
urine-stained abdominal fur (both sexes), ungroomed coat (both sexes),
decreased motor activity, chromodacryorrhea, ptosis, lost righting
reflex, scant feces, and red or tan perioral substance.  The following
additional clinical signs of toxicity were noted in the animals
sacrificed in a moribund condition:  males (prostration, limp muscle
tone, muscle flaccidity, hypothermia, hunched posture, and labored
breathing) and females (ptosis, prostration, piloerection, bradypnea,
decreased motor activity, impaired righting reflex, lost righting
reflex, limp muscle tone, scant feces, and hypothermia).

At 125 mg/kg, effects were limited to minor decreases (p<0.01) in
body-weight gain and absolute and relative food consumption in the
males.

At 750 mg/kg, increased (p<0.01) incidence (# affected/10 vs. 0
controls) of the following neurological effects were noted at 8 hours
post-dosing during the FOB: (i) hunched posture in 6 males and 4 females
and (ii) slight ataxia in 3 males.  All findings were resolved by Day 8.
 No statistically significant differences were observed on either the
interval or total session motor activity (number of movements or time
spent in movement).  However, at 8 hours post-dosing, total session
number of movements and time spent in movement were slightly decreased
in both sexes.  These parameters remained decreased in the males on Day
8.  These changes in motor activity were observed at a dose that
resulted in moribundity and are considered related indirectly to the
overall toxicity of the test material.  No treatment-related microscopic
lesions were observed.

The LOAEL is 750 mg/kg, based on decreased body weight, body-weight
gain, absolute and relative food consumption, and clinical signs of
toxicity, indicative of a moribund condition, in both sexes:
dehydration, urine-stained abdominal fur, ungroomed coat, ptosis,
decreased motor activity, prostration, limp muscle tone, muscle
flaccidity, hypothermia, hunched posture, impaired or lost righting
reflex, scant feces; in males: red or tan perioral substance,
chromodacryorrhea, chromorhinorrhea and labored breathing, and in
females: piloerection and bradypnea, and signs of neurotoxicity: hunched
posture in females and ataxia in males.  The NOAEL is 250 mg/kg.

The study is classified as acceptable/guideline and satisfies the
guideline requirement (OPPTS 870.6200a) for a neurotoxicity screening
battery in rats.

870.6200 Subchronic Neurotoxicity - Rat

In a subchronic neurotoxicity study (MRID 47090410), Flutriafol (95.1%
a.i.; Lot # UPL Bx 1 [2001]) was administered in the diet to 10
Sprague-Dawley rats/sex/group at dose levels of 0, 500, 1500, or 3000
ppm (equivalent to 0/0, 28.9/32.6, 84.3/97.6, and 172.1/185.0 mg/kg/day
[M/F], respectively) for 92 days.  Neurobehavioral assessment
(functional observational battery [FOB] and motor activity testing) were
performed in 10 rats/sex/group at pre-dosing and Weeks 2, 4, 8, and 13.
At study termination, 5 rats/sex/group were anesthetized and perfused in
situ for neuropathological examination.  The tissues from the perfused
animals in the control and 3000 ppm groups were subjected to
histopathological evaluation of brain and peripheral nervous system
tissues.  Acceptable positive control data were provided.

No compound-related effects were observed in mortality, clinical signs
of toxicity, ocular effects, motor activity, brain weights, or gross or
neuropathology.

At 1500 ppm, body-weight gains were decreased (p≤0.05) in the males by
28% compared to controls during the first week of dosing and overall
(Days 1-92) body-weight gain was decreased (p≤0.05) by 19% in the
females.  Likewise, absolute and relative food consumption were
decreased (p≤0.01) by 15-16% in both sexes during Week 1.

At 3000 ppm, body weights were decreased throughout the study in the
males (decr. 5-14%) and females (decr 5-10%) and attained statistical
significance (p≤0.05) at Days 8 and 57 through 92 in the males and
Days 50, 64, and 85 through 92 in the females.  During the first week of
dosing, body-weight gains were decreased (p≤0.05) by 108% in both
sexes at this dose compared to controls.  During Week 2, body-weight
gain was increased (p≤0.01) by 36% in the males.  Overall (Days 1-92)
body-weight gain was decreased (p≤0.05) by 23-34% in the males and
females compared to controls.  Similarly, absolute and relative food
consumption were decreased (p≤0.01) by 35-40% in both sexes during the
first week of dosing. Additionally in the females, absolute food
consumption was decreased (p≤0.05) by 9-14% at most intervals
throughout the exposure period.  Overall absolute food consumption was
decreased (p≤0.01) by 10-13% in both sexes.  During Week 2, relative
food consumption was increased (p≤0.01) by 10% in the males.  In the
females, relative food consumption was slightly lower throughout the
rest of the exposure period, but only attained statistical significance
(decr 8%; p≤0.01) on Days 29-36.  Overall relative food consumption
was only slightly decreased (decr 5%; not statistically significant) in
the females.

Additionally at 3000 ppm, hindlimb grip strength was decreased
(p≤0.05) in the males by 17% compared to controls during Week 2.  The
decreased hindlimb grip strength was considered to be a
treatment-related neurotoxic effect.

No treatment-related effects were observed at 500 ppm in either sex.

The LOAEL was 3000 ppm (equivalent to 172.1/185.0 mg/kg/day [M/F]) based
on decreased body-weight gain, and absolute and relative food
consumption and decreased hindlimb grip strength.  The NOAEL is 1500 ppm
(equivalent to 84.3/97.6 mg/kg/day [M/F]).

The study is classified as acceptable/guideline and satisfies the
guideline requirement (OPPTS 870.6200b) for a subchronic neurotoxicity
study in rats.

A.3.8	Special

	870.7485	Metabolism – Rat

In rat metabolism studies (MRIDs 47090412, 47090413, and 47090414),
14C-flutriafol (>97% radiochemical purity) in polyethylene glycol 600
was administered to rats as a single oral gavage dose at 5 or 250 mg/kg
body weight.  Group sizes were 1 rat/sex in a preliminary study at 5
mg/kg, 2 rats/sex/dose in bile duct-cannulation studies, one group of 6
females at 250 mg/kg, and 4-5 rats/sex/dose in other dose groups.  One
group of 4 rats/sex received 14 consecutive daily doses at 5 mg/kg/day.
14C-carbinol-flutriafol was administered to all groups, except one group
of 2 rats/sex was treated with 5 mg/kg 14C-triazole-flutriafol.  Excreta
(urine, feces, and bile [in some groups]) were collected, and analyzed
for radioactivity concentration.  Additionally, pools of selected
excreta were analyzed to identify and quantify metabolites.  Animals
were sacrificed at 48 hours in the preliminary experiment and at 72 or
168 hours post-dose or post final dose in the other studies.  Tissues
were collected and analyzed for radioactivity concentration.

More than 78% of the administered dose was recovered in the bile and
urine of the single 5 mg/kg (both radiolabels) and 250 mg/kg dose
groups.  Absorption was generally similar between sexes, radiolabels,
and between single and multiple dose regimes.  Comparing absorption in 5
mg/kg groups to the 250 mg/kg groups, absorption remains extensive;
however, a longer time is required for absorption to complete.

Total recoveries at 168 hours post-dose were 97-99% of the administered
single dose and 115-125% daily dose in the multiple dose study.  The
administered dose was mostly eliminated within 48 hours at 5 mg/kg
(86-97% of the single dose or 104% daily dose of the multiple dose
groups) and at 250 mg/kg (68-85% dose, except bile duct-cannulated
females which was 38% dose).

Only 0.04-0.05% of the dose was found in the expired carbon dioxide in a
preliminary study.  In the bile duct-cannulation study, most of the
radioactivity was excreted in the bile (47-79% of the dose).  In the
single dose 5 mg/kg group (not bile duct-cannulated), similar amounts of
radioactivity were excreted in the feces as in the urine, but only
approximately half as much was excreted in the feces as in the urine at
250 mg/kg.  Slightly more radioactivity was found in the urine of the
multiple dosed animals compared to the single dosed animals.  The
excretion profile was generally similar between the sexes, and was also
similar following 1, 5, 10, and 14 doses.

Tissue distribution was examined in animals sacrificed 168 hours
post-dose.  In the blood, radioactivity partitioned into the red blood
cells.  In animals receiving multiple daily 5 mg/kg doses,
concentrations of radioactivity were higher in the blood cells than
plasma of males (218-fold) and females (129-fold).  Excluding blood
cells and GI tract measurements, the highest concentrations were found
in whole blood in males (190 ng equivalents flutriafol/g tissue in the
single 5 mg/kg dose group, 8040 ng equiv/g in the 250 mg/kg dose group,
and 1450 ng equiv/g in the multiple 5 mg/kg/day dose group) and in
females (140 ng equivalents flutriafol/g tissue in the single 5 mg/kg
dose group, 6740 ng equiv/g in the 250 mg/kg dose group, and 519 ng
equiv/g in the multiple 5 mg/kg/day dose group).  In both sexes and all
groups, concentrations of radioactivity were relatively high in both
liver and kidneys.  Other organs with high concentrations in one or more
groups included the adrenal glands, spleen, and pituitary.  The
distribution profiles were generally similar between species, dose
level, and single vs multiple dose regime.  A 50-fold increase in dose
resulted in an approximately 42-48-fold increase in radioactivity
concentrations in the whole blood; thus, the concentrations were roughly
proportional to the dose.

The total amount of radioactivity isolated in the tissues and carcass
was miniscule: <1% of the administered dose (single dose groups) or 3%
of the daily administered dose (multiple dose group).  Also, the amount
of the dose remaining in the body (GI tract and contents, tissues, and
remaining carcass) after 168 hours was <1.1% of the administered dose
regardless of sex, radiolabel position, or dose.  For these reasons,
bioaccumulation in all dose groups was considered unlikely.

The parent was isolated in only trace amounts in the urine and feces
(<0.5% of the administered dose) and more than 19 metabolites were
isolated, indicating extensive metabolism of flutriafol.  In general,
metabolism profiles were similar between sexes.  The metabolism profile
in urine was similar between the 250 mg/kg dose group and the multiple 5
mg/kg dose group, but the metabolism profiles in feces resulted in the
isolation of greater amounts of identified compounds in the high dose
group.  Summarizing the Sponsor’s stated results in MRID 47090413
(data not provided); the metabolic profiles were similar regardless of
the matrix (feces, urine, or bile), the dose, the sex, or the
radiolabel.

The primary site for metabolism was the 2-fluorophenyl ring.  The
initial metabolic step was probably epoxidation followed by either
rearrangement to form the dihydrodiol isomers or to form hydroxy or
dihydroxy metabolites.  The hydroxyl groups on these primary metabolites
may then be either conjugated with glucuronic acid or methylated.  A
second, minor route for metabolism of flutriafol was via the removal of
the triazole ring to form 1-(2
fluorophenyl)-1-(4-fluorophenyl)-ethandiol, which is then conjugated
with glucuronic acid.

This metabolism study in the rat is classified acceptable/guideline and
satisfies the guideline requirement for a metabolism study [OPPTS
870.7485, OECD 417] in rats.

	870.7600	Dermal Absorption - Rat

In a dermal penetration study (MRID 47090415), 14C-carbinol-flutriafol
(98-99% radiochemical purity as applied; Batch No. Rad164) was applied
to the skin (10 cm2) of Sprague Dawley rats (4 males for each time point
at each dose level).  Nominal doses of 0.02, 0.2 or 2 mg/cm2 skin were
tested (10 µl/cm2 skin), and actual doses were 0.0208, 0.201, and 2.154
mg/cm2 skin.  The exposure durations were 0.5, 1, 2, 4, 10, and 24 h,
and the animals were terminated at the end of the exposure period.  An
additional group was exposed for 10 h and maintained for another 158 h
in the metabolism unit prior to termination.

Recovery of the applied dose (mass balance) was 96-103%.  The majority
of the dose was not absorbed (sum of soap and water wash of the
application site with the dose site appliance wash; generally 75-97% of
the applied dose), with the greatest amount of radioactivity being
recovered from the soap wash of the application site (generally 51-87%
of the applied dose).   Dermal absorption (based on the sum of residues
in urine, feces, cage wash, blood, and carcass) ranging up to 15.8% of
the applied dose was noted.  Absorption was minimal with only 4 h of
exposure (<1.5% of the applied dose), and was saturated in the high dose
(maximum absorption of only 3.7% dose).  Absorbable radioactivity
(radioactivity in the skin at the application site and the adjacent
skin) was minimal (<0.75% of the applied dose) in groups that were
exposed for 10 h and evaluated for an additional 158 h post-exposure.
Thus, the data suggests that almost all of the dose isolated in the skin
will be absorbed.  Considering the sum of absorbable and absorbed doses,
4-37% of the applied dose was recovered in the treatment groups (mean of
11% and median of 9%).  Absorption rate constants were calculated as
0.236, 0.190, and 0.072 h-1 for the 2, 20, and 200 µg/cm2 dose groups,
respectively; thus, absorption mechanisms were saturated at the high
dose.  The elimination half-lives were calculated to be 31, 30, and 37 h
for the 2, 20, and 200 µg/cm2 dose groups, respectively.

As almost all of the absorbable dose (radioactivity in the skin at the
application site and the adjacent skin) will be absorbed, the most
conservative estimation of absorption would consider both the absorbed
dose and the absorbable dose.  In this study, a maximum of 36.56% of the
applied dose was noted as absorbed/absorbable (observed after 24 h
exposure to 2 µg/cm2).  This value is most conservative.  However, it
is likely that any area exposed to the chemical will be washed within 10
h.  The applied dose that is absorbed/absorbable following a 10 h
exposure  is 16.54%, 21.31% and 11.39% ,, respectively, at 2,  20 and
200 µg/cm2.

This study is classified as acceptable/guideline and satisfies the
guideline requirements (OPPTS 870.7600; OECD none) for a dermal
penetration study in rats.

A.4.	References

MRID 47090344 Doe, J.E. (1982) PP450 (Flutriafol technical):  28-Day
feeding study in rats. Imperial Chemical Industries PLC, Central
Toxicology Laboratory, Alderley Park, Cheshire, UK.  Laboratory Study
No.: PR0429, September 23, 1982.  Unpublished.

MRID 47090345 Pigott, G.H. (1982) PP450 (Flutriafol technical):  90-Day
feeding study in rats. Imperial Chemical Industries PLC, Central
Toxicology Laboratory, Alderley Park, Cheshire, UK.  Laboratory Study
No.: PR0432, Laboratory Report No. CTL/P/744, October 21, 1982.
Unpublished.

                         MRID 47090346 Doe, J. E. (1982) PP450
(flutriafol technical):  90-day oral dosing study in dogs. Imperial
Chemical Industries, PLC, Central Toxicology Laboratory, Alderley Park,
Macclesfield, Cheshire, UK.  Laboratory Report No.:  CTL/P/733; Study
No.:  PD0433, November 2, 1982.  Unpublished.

MRID 47090347 Barnett, Jr., J. F. (2007) Percutaneous repeated dose
28-day toxicity study of flutriafol in rats.  Charles River Laboratories
Preclinical Services, Horsham, PA.  Laboratory Project ID.:  TQC00010,
January 31, 2007. Unpublished.

MRID 47090348 Barnett, Jr., J. F. (2006) Percutaneous repeated dose
14-day toxicity dosage range-finding study of flutriafol in rats.
Charles River Laboratories Preclinical Services, Worcester, MA.
Laboratory Project ID.:  TQC00009, June 30, 2006.  .  Unpublished.

			MRID 47090349 PP450 (Flutriafol):  Teratogenicity study in the rat.
Imperial Chemical Industries PLC, Cheshire, UK.  Laboratory Study No.
RR0211, Report No. CTL/P/756, October 27, 1982.  Unpublished.

	MRID 47090350 PP450 (Flutriafol):  Teratogenicity study in the rabbit.
Imperial Chemical Industries PLC, Cheshire, UK.  Laboratory Study No.
RR0211; Report No. CTL/P/756, October 20, 1982.  Unpublished.

MRID 47090351 Flutriafol:  Two generation reproduction study in the rat.
 Imperial Chemical Industries PLC, Cheshire, UK.  Laboratory Study No.
RR0229, Report No. CTL/P/1368, April 17, 1986.  Unpublished.

MRID 47090352 Pigott, G.H. (1986) Flutriafol:  2 year feeding study in
rats.  Imperial Chemical Industries PLC, Central Toxicology Laboratory,
Macclesfield, Cheshire, UK.  Laboratory Study No.: PR0542.  Laboratory
Report No.:  CTL/P/1220, May 22, 1986.  Unpublished.

MRID 47090353 Stonard, M. D. (1988) Flutriafol:  one year oral dosing
study in dogs.  Imperial Chemical Industries, PLC, Central Toxicology
Laboratory, Alderley Park, Macclesfield, Cheshire, UK.  Laboratory Study
No.:  PD0667; Report No.:  CTL/P/2019, May 18, 1988.  Unpublished.

MRID 47090354 Hext, P.M. (1988) Flutriafol:  two year feeding study in
mice.  Imperial Chemical Industries PLC, Cheshire, UK.  Laboratory
Study:  PM0637, Report No. CTL/P/1930, June 3, 1988.  Unpublished.

MRID 47090401 Longstaff, E. (1988) PP450 (Flutriafol) - An evaluation in
the Salmonella/ microsome mutagenicity assay.  Imperial Chemical
Industries PLC, Central Toxicology Laboratory, Alderley Park, Cheshire,
UK.  Laboratory Study No.:  CTL Study Nos. YV0603 and YV0625, February
8, 1988.  Unpublished.

MRID 47090402 McGregor, D.B. (1986) PP450 (Flutriafol) - Assessment of
mutagenic potential in the mouse lymphoma mutation assay.  Inveresk
Research International, Musselburgh, Scotland.  Laboratory Study No.:
730420, May 15, 1986.  Unpublished.

MRID 47090403 Howard, C.A.. (1989) Flutriafol: An evaluation in the In
Vitro cytogenetic assay in human lymphocytes.  Imperial Chemical
Industries PLC, Central Toxicology Laboratory, Alderley Park, Cheshire,
UK.  Laboratory Study No.:  SV0343, June 21, 1989.  Unpublished.

MRID 47090404 Styles, J.A. (1982) PP450 (Flutriafol) – A cytogenetic
study in the rat.  Imperial Chemical Industries PLC, Central Toxicology
Laboratory, Alderley Park, Cheshire, UK.  Laboratory Study No.:
CTL/P/725, September 9, 1982.  Unpublished.

MRID 47090405 Richardson, C.R. (1986) Flutriafol - An evaluation in the
mouse micronucleus test.  Imperial Chemical Industries PLC, Central
Toxicology Laboratory, Alderley Park, Cheshire, UK.  Laboratory Study
No.:  CTL Study No. SM0216, April 2, 1986.  Unpublished.

MRID 47090406 Longstaff, E. (1982) PP450 (Flutriafol) - Dominant lethal
study in the mouse.  Imperial Chemical Industries PLC, Central
Toxicology Laboratory, Alderley Park, Cheshire, UK.  Laboratory Study
No.:  CTL Study No. RM0206, September 28, 1982.  Unpublished.

MRID 47090407 Trueman, R.W. (1987) Flutriafol – Assessment for the
induction of unscheduled DNA synthesis in rat hepatocytes in vivo.
Imperial Chemical Industries PLC, Central Toxicology Laboratory,
Alderley Park, Cheshire, UK.  Laboratory Study No.:  SR0259, June 16,
1987.  Unpublished.

MRID 47090408 Barnett Jr., J.F. (2006) Oral acute neurotoxicity study of
Flutriafol in rats.  Charles River Laboratories Preclinical Services,
Horsham, PA.  Laboratory Project ID: TQC00006, December 1, 2006.
Unpublished.

MRID 47090409 Barnett Jr., J.F. (2006) Acute oral (gavage) dosage
range-finding study of Flutriafol in rats.  Charles River Laboratories
Preclinical Services, Horsham, PA.  Laboratory Project ID: TQC00005,
June 26, 2006.  Unpublished.

MRID 47090410 Barnett Jr., J.F. (2007) Oral (diet) subchronic
neurotoxicity study of Flutriafol in rats.  Charles River Laboratories,
Preclinical Services, Horsham, PA.  Laboratory Project ID: TQC00008,
February 1, 2007.  Unpublished.

MRID 47090411 Barnett Jr., J.F. (2006) Oral (diet) dosage range-finding
subchronic neurotoxicity study of Flutriafol in rats.  Charles River
Laboratories, Preclinical Services, Horsham, PA.  Laboratory Project ID:
TQC00007, Experimental Completion Date: October 6, 2005; Final Pilot
Report Date June 30, 2006.  Unpublished.

MRID 47090412 Jones, B.K. (1982) PP450 (Flutriafol): Excretion and
tissue retention of a single oral dose (5 mg/kg) in the rat.  Imperial
Chemical Industries PLC, Macclesfield, Cheshire, UK.  Laboratory Report
No.:  CTL/P/751, October 7, 1982.  Unpublished.

MRID 47090413 Jones, B.K. (1986) Flutriafol: Biotransformation in the
Rat.  Imperial Chemical Industries PLC, Macclesfield, Cheshire, UK.
Laboratory Study-Report No.:  UR0149-CTL/P/856, June 12, 1986.
Unpublished.

MRID 47090414 Millais, A.J. (2004) [14C]-Flutriafol metabolism in rats
after single and repeated doses.  Huntingdon Life Sciences Ltd.,
Huntingdon, Cambridgeshire, England.  Laboratory Project No.:  CHV
081/024050, August 16, 2004.  Unpublished.

MRID 47090415 Sved, D.W. (2006) A dermal-absorption study with
[14C]-labeled flutriafol in the rat. WIL Research Laboratories, LLC,
1407 George Rd, Ashland, OH.  Study No.: WIL-206008.  June 7, 2006.
Unpublished.

Appendix B.	Metabolism Assessment

Plant Metabolism Studies:  The petitioner submitted apple, sugar beet,
rapeseed, and wheat/barley (foliar and seed treatment) metabolism
studies conducted with [triazole-3,5-14C]flutriafol and
[carbinol-14C]flutriafol.  HED notes that the wheat/barley metabolism
studies were deemed unacceptable due to numerous deficiencies but are
presented to supplement the acceptable studies.

Apple (47248901.der.doc):  Apple trees were treated with a single foliar
application of [carbinol-14C]flutriafol or [triazole-3,5-14C]flutriafol
during early fruit development at 0.11 lb ai/acre (1x/0.2x the proposed
single/seasonal application rates).  Samples of apple fruit (both
labels) and foliage (triazole label only) were harvested at maturity, 64
days following application.  Table B.1 is a summary of the total TRRs;
TRRs were slightly higher in the triazole labeled samples.

The majority of the radioactivity (72-84% TRR) in apple fruit and
foliage was solvent extracted with acetonitrile (ACN) and ACN/water.  An
additional ~5% TRR was released from apple fruit via mild acid (MA; 0.1
M HCl) and mild base (MB; 0.1 M NaOH) hydrolysis.  Nonextractable
residues were 18-23% TRR (≤0.012 ppm) in apple fruit and 16% TRR
(0.685 ppm) in triazole-label foliage.

Residues were identified by high-performance liquid chromatography
(HPLC) and thin-layer chromatography (TLC) co-elution with reference
standards (flutriafol, TA, TAA, and T).  Total identified residues
accounted for 50-56% of the TRR in apple fruit (Tables B.2 and B.3).
Unknowns represented <5% TRR in fruit.  Similar metabolic profiles were
observed for the two labels.

Sugar beet (47090439.der.doc):  Sugar beets were treated with a single
foliar application of [carbinol-14C]flutriafol or
[triazole-3,5-14C]flutriafol at ~0.12 lb ai/acre (BBCH 49; harvestable
size root).  Samples of sugar beet root and tops were collected 0, 6,
11, 16, and 21 days after treatment (DAT; 6- and 11-DAT samples were not
analyzed).  Table B.1 is a summary of the TRRs; TRRs did not vary with
radiolabel position in sugar beet tops and TRRs in roots were to low to
draw a conclusion (≤0.009 ppm; not analyzed further)

The majority of the radioactivity in sugar beet tops was solvent
extracted with ACN and/or ACN/water (89-97% TRR).  An additional ~3-4%
TRR was released from 21-DAT tops via mild acid and base hydrolysis.
Nonextractable residues were 5-11% TRR (0.029-0.078 ppm) in
carbinol-label tops and 3-9% TRR (0.030-0.040 ppm) in triazole-label
tops.

Residues were identified by HPLC and TLC co-elution with reference
standards (flutriafol, TA, and TAA).  HED notes that although TA and TAA
were listed as reference standards, the behavior of these compounds
under the employed analytical systems were not provided.  A hexose
conjugate of flutriafol was identified based on comparison of the study
results with those of the rapeseed metabolism study.  Total identified
residues accounted for 73-95% TRR in sugar beet tops (Tables B.2 and
B.3).  Unknowns represented <8% TRR in tops.  Similar metabolite
profiles were observed for the two labels.

Rapeseed (47090438.der.doc):  Rapeseed plants were treated with a single
foliar application of [carbinol-14C]flutriafol or
[triazole-3,5-14C]flutriafol at ~0.10 lb ai/acre (BBCH71; early pod
set).  Forage samples were harvested immediately following application;
samples of pod and foliage were harvested 7 and 14 days after treatment
(DAT); samples of whole plant were collected 21 DAT; and samples of
foliage and seed were collected at maturity, 42 DAT.  Samples harvested
7 and 21 DAT were not analyzed.  Table B.1 is a summary of the TRRs;
TRRs did not vary with radiolabel position.

The majority of the radioactivity (60-98% TRR) in forage/foliage was
extracted by sequential extraction with ACN, ACN/water, and/or water;
40-41% TRR was extracted from 14-DAT pods with ACN/water.  Mature seed
was extracted with hexane (27-32% TRR) followed by ACN/water extraction
(42% TRR).  The following additional radioactivity was released via
hydrolysis with enzyme (E; cellulase and hemicellulase), MA, MB, strong
acid (SA; 6 M HCl), and strong base (SB; 2 M NaOH):  14- and 42-DAT
foliage - combined 7-20% TRR; 14-DAT pods -combined 53% TRR; and 42-DAT
seed - combined 14-22% TRR.  Nonextractable residues were as follows:
14-DAT pods - 6-7% TRR in; 42-DAT seed - 4-7% TRR in; and forage/foliage
- 1-11% TRR (≤0.034 ppm) except in 42-DAT carbinol-label foliage which
had nonextractable residues of 20% TRR (0.071 ppm).

n the hydrolysates of 14-DAT pods (27-28% TRR) and 42-DAT foliage (3-11%
TRR).  Flutriafol was identified in the remaining hydrolysates at ≤7%
TRR and defluorinated flutriafol was also identified in the hydrolysates
at ≤15% TRR.

Barley/Wheat (foliar treatment; 47090440.der.doc):
[Carbinol-14C]flutriafol or [triazole-3,5-14C]flutriafol were applied as
a single foliar broadcast application to spring barley and spring wheat
grown in pots maintained outdoors or in a greenhouse, at a rate of ~0.08
lb ai/acre.  The applications were made 4-26 days prior to ear
emergence, except for field-grown carbinol-label wheat and field-grown
triazole-label barley in which applications were made after ear
emergence.  Samples of mature grain and straw were harvested 56-94 days
after application for those samples treated pre-ear emergence and 44-45
days after application for those samples treated post-ear emergence.
TRRs are summarized in Table B.1; TRRs did not vary significantly with
label position for straw but varied with label position in grain with
the triazole labeled yielding higher residues (chaff not analyzed
further).

The majority of the radioactivity from the grain and straw samples
(60-95% TRR; 47% TRR for field-grown triazole-label barley grain) was
solvent extracted with ACN and ACN/water.  Nonextractable residues
accounted for 5-35% TRR (≤0.035 ppm) in the grain samples (both
labels) and 16-40% TRR (0.150-0.336 ppm) in the straw samples (both
labels).  Nonextractable residues in triazole-label wheat straw (23%
TRR) were subjected to limited sequential extraction procedures with
cold and boiling water and cold 1 M ammonia (released an additional 7%
TRR).

Residues were identified using TLC by co-elution or reference standards
(flutriafol, TA, TAA, and triazole lactic acid).  Total identified
residues represented 36-38 % TRR in carbinol labeled grain and straw
samples and 56-84% TRR in all triazole labeled grain and straw samples
excluding the  triazole-label field-grown barley grain sample treated
post-ear emergence (32% TRR; Table B.4).  Unknowns were ≤4% TRR in the
carbinol labeled grain and straw samples, 5-14% TRR in the triazole
straw samples, and 8-34% TRR in the triazole grain samples (sufficient
characterization of these residues were performed).  The metabolic
profile did not vary significantly with label position in straw.
However, the metabolic profile varied significantly with label position
in grain samples due to the identification of TA and TAA in the
triazole-label samples.

The following deficiencies were identified in the barley/wheat foliar
metabolism study:  (1) residues in forage were not investigated; (2)
insufficient attempts were made to characterize nonextractable residues
of barley and wheat straw; (3) a confirmatory method was not used for
the identification of metabolites; (4) the ACN/water barley straw
extract (carbinol label) was not analyzed (20% TRR; 0.142 ppm), (5) no
information concerning storage conditions or durations was provided, and
(6) the GLP statement indicated that since Cheminova (the petitioner)
did not conduct the study and was not the sponsor, they could not be
certain that the study was conducted in accordance with GLP practices
(40 CFR 160).  In addition, supporting information and data were
extremely limited for this study.

arley straw (both labels) and triazole-label barley/wheat grain
(carbinol grain samples were not analyzed due to low TRR (≤0.005
ppm)).

The majority of the radioactivity (84-88% TRR) in barley straw was
solvent extracted with ACN and ACN/water.  ACN released ≤1% TRR from
triazole-label barley/wheat grain while ACN/water released 89-94% TRR.
Nonextractable residues were 6-10% TRR (0.008-0.016 ppm) for
triazole-label barley/wheat grain and 12-16% TRR (0.030-0.041 ppm) for
triazole- and carbinol-label barley straw.

 and 59% TRR in the carbinol labeled straw sample (Table B.5).  Unknowns
were ≤5% TRR in the triazole labeled grain and straw samples and a
combined 19% TRR (0.047 ppm) in the carbinol labeled straw sample.
Significant differences in the metabolic profile were found in carbinol
and triazole labeled grain and straw samples due to the identification
of TA and TAA in the triazole-label samples.

The following deficiencies were identified in the barley and wheat seed
treatment metabolism study:  (1) residues in forage were not
investigated, (2) a confirmatory method was not used for the
identification of metabolites, (3) no information concerning storage
durations was provided; and (4) the GLP statement indicated that since
Cheminova (the petitioner) did not conduct the study and was not the
sponsor, they could not be certain that the study was conducted in
accordance with GLP practices (40 CFR 160).  In addition, supporting
information and data were extremely limited for this study.

Table B.1:  TRRs

Crop	Timing and Applic. No.	PHI

(days)	Matrix	TRR (ppm parent equivalents)1

Carbinol label	Triazole label

Foliar Plant metabolism Studies

Apple	One foliar application made at early fruit development at 0.105 lb
ai/acre	64	fruit	0.041	0.065

	foliage	not analyzed	4.182

Rapeseed	One foliar application made at early pod set at 0.105 lb
ai/acre (carbinol label) or 0.103 lb ai/acre (triazole label)	0	Forage
1.497	0.782

14	Pod	0.779	0.751

	Foliage	1.601	1.165

42	Seed	0.729	1.316

	Mature Foliage	0.355	0.246

Sugar Beet	One foliar application made at the harvestable size root
stage at 0.115 lb ai/acre (carbinol label) or 0.119 lb ai/acre (triazole
label)	0	Root 	<0.001	0.001

	Tops 	1.273	1.368

16	Root	0.005	0.003

	Tops 	0.381	0.342

21	Root	0.005	0.009

	Tops 	0.596	0.747

Barley

(field grown)	One foliar application made 13 days before ear emergence
at 0.080 lb ai/acre.	62	grain	0.007	--

	straw	0.72	--

	One foliar application made after ear emergence at 0.075 lb ai/acre.	44
grain	--	0.10

	straw	--	0.12

Barley

(greenhouse grown)	One foliar application made 26 days before ear
emergence at ~0.080 lb ai/acre.	94	grain	0.02	0.41

	straw	not analyzed	2.10

Wheat

(field grown)	One foliar application made after ear emergence at 0.079
lb ai/acre. 	45	grain	0.006	--

	straw	0.53	--

	One foliar application made 20 days before ear emergence at 0.094 lb
ai/acre.	74	grain	--	0.05

	straw	--	0.65

Wheat

(greenhouse grown)	One foliar application made 4 days before ear
emergence at ~0.080 lb ai/acre.	56	grain	0.01	0.18

	straw	not analyzed	not analyzed

Seed Treatment Metabolism Study

Barley	Seed treatment at 114 ppm (carbinol label) or 121 ppm (triazole
label).	154	grain	0.005	0.17

	straw	0.25	0.24

	chaff	0.06	0.21

Wheat	Seed treatment at 104 ppm (carbinol label) or 73 ppm (triazole
label).	112	grain	0.003	0.14

	straw	0.17	0.23

	chaff	0.08	0.14

1	Samples in bold were analyzed further.

Table B.2:  Summary of Characterization and Identification of TRRs in
Apple, Rapeseed, and Sugar Beet Following Foliar Application of
[Carbinol-14C]Flutriafol.

Compound	Apple	Rapeseed	Sugar Beet

	64-DAT fruit	0-DAT forage	14-DAT Foliage	42-DAT Foliage	14-DAT Pod
42-DAT Seed	0-DAT Tops	16-DAT Tops	21-DAT Tops

	TRR =

0.041 ppm	TRR =

1.497 ppm	TRR =

1.601 ppm	TRR =

0.355 ppm	TRR =

0.779 ppm	TRR =

0.729 ppm	TRR =

1.273 ppm	TRR =

0.381 ppm	TRR =

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	0.063	16.7	0.131	14.3	0.106	2.4	0.028	16.2	0.061	22.0	0.133

Total extractable	76.9	0.032	97.9	1.466	98.6	1.579	79.9	0.283	92.9	0.723
83.3	0.607	93.9	1.195	88.8	0.338	95.1	0.567

Unextractable	23.0	0.009	2.1	0.031	1.4	0.022	20.1	0.071	7.1	0.055	16.8
0.122	6.1	0.078	11.2	0.043	4.8	0.029

Accountability4	100	100	100	100	100	100	100	100	100

1	Includes flutriafol identified in the E, WA, WB, SA, and/or SB
hydrolysates for 14-DAT rapeseed forage (E-2.8% TRR, SA-0.4% TRR,SB-0.7%
TRR; total of 3.9% TRR; ), 42-DAT rapeseed foliage (E-2.8% TRR and
SA-8.2% TRR; total of 11.0% TRR), 14-DAT rapeseed pod (E-15.3% TRR,
WB-3.3% TRR, SA-3.0% TRR, and SB-6.2% TRR; total of 28% TRR), 42-DAT
rapeseed seed (E-0.4% TRR, WB-0.5% TRR, SA-4.9% TRR, and SB-1.5% TRR;
total of 7% TRR), and 21-DAT sugar beet tops (combined WA and WB-1.9%
TRR).

2	Hydrolysates in bold were not analyzed.

3	Largest unknown of 2.8% TRR.

4	Accountability = (Total extractable + Total unextractable)/(TRR from
combustion analysis) * 100.

Table B.3:  Summary of Characterization and Identification of TRRs in
Apple, Rapeseed, and Sugar Beet Following foliar Application of
[Triazole-14C]Flutriafol.

Compound	Apple1	Rapeseed1	Sugar Beet

	64-DAT Fruit	14-DAT Foliage	42-DAT Foliage	14-DAT Pod	42-DAT Seed	0-DAT
Tops	16-DAT Tops	21-DAT Tops

	TRR =

0.065 ppm	TRR =

1.165 ppm	TRR =

0.246 ppm	TRR =

0.751 ppm	TRR =

1.316 ppm	TRR =

1.368 ppm	TRR =

0.342 ppm	TRR =

0.747 ppm

	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	%
TRR	ppm

Flutriafol2	49.9	0.032	82.7	0.963	55.6	0.137	59.4	0.446	67.6	0.889	95.4
1.304	77.7	0.266	70.8	0.529

FHC (R5a)3	--	--	2.7	0.031	5.7	0.014	1.5	0.011	3.8	0.050	--	--	1.1	0.004
5.0	0.038

DF (C6 )	--	--	1.5	0.018	0.8	0.002	12.1	0.091	3.0	0.039	--	--	--	--	--
--

TA	<2	<0.001	--	--	--	--	--	--	--	--	--	--	--	--	--	--

TAA	--	--	--	--	--	--	--	--	--	--	--	--	--	--	--	--

Unknown R1	--	--	0.3	0.003	4.9	0.012	1.6	0.012	3.5	0.046	0.2	0.003	8.1
0.028	3.1	0.023

Unknown R2	--	--	0.6	0.007	1.6	0.004	1.1	0.008	1.7	0.023	--	--	0.7	0.002
4.6	0.034

Unknown R3	--	--	1.2	0.014	1.2	0.003	0.8	0.006	--	--	--	--	0.9	0.003	2.1
0.016

Unknown R4	--	--	1.0	0.012	2.0	0.005	1.2	0.009	--	--	0.1	0.001	0.5	0.002
4.4	0.033

Unknown R5b	--	--	2.6	0.030	6.1	0.015	1.5	0.011	3.6	0.048	0.9	0.012	2.6
0.009	2.4	0.018

Unknown R6	--	--	0.3	0.003	--	--	--	--	--	--	0.2	0.003	--	--	1.0	0.008

Unknown C1	--	--	1.5	0.018	0.4	0.001	6.3	0.047	3.3	0.044	--	--	--	--	--
--

Unknown C2	--	--	0.1	0.001	--	--	0.5	0.004	--	--	--	--	--	--	--	--

Unknown C3	--	--	--	--	--	--	0.3	0.002	--	--	--	--	--	--	--	--

Unknown C4	--	--	0.1	0.001	--	--	0.5	0.004	--	--	--	--	--	--	--	--

Others (unknown)	21.63	0.013	1.9	0.022	2.4	0.006	6.5	0.048	7.8	0.102	0.4
0.005	--	--	2.4	0.018

Unretained	2.0	0.001	--	--	--	--	--	--	--	--	--	--	--	--	--	--

E4	not performed	5.0	0.058	2.6	0.006	17.1	0.128	1.4	0.018	not performed
not performed	not performed

WA4	2.1	0.001	0.2	0.002	0.4	0.001	0.3	0.002	0.4	0.005

	3.8	0.023

WB4	2.4	0.002	0.6	0.007	1.2	0.003	2.8	0.021	3.0	0.039

SA4	not performed	2.8	0.033	5.5	0.014	16.9	0.127	7.9	0.104

	not performed

SB4

2.8	0.033	3.0	0.007	15.8	0.119	9.1	0.120

	Total identified	49.9	0.032	86.9	1.012	62.1	0.153	73.0	0.548	74.4	0.978
95.4	1.304	78.8	0.270	75.8	0.567

Total characterized	28.1	0.017	10.4	0.120	25.8	0.063	21.0	0.156	21.4
0.282	1.8	0.024	12.8	0.044	20.0	0.150

Total extractable	82.2	0.054	97.0	1.130	88.9	0.218	94.1	0.706	95.8	1.26
97.1	1.328	91.1	0.312	95.8	0.716

Unextractable	17.8	0.012	2.9	0.034	11.1	0.027	5.8	0.044	4.3	0.057	2.9
0.040	8.9	0.030	4.2	0.031

Accountability5	100	100	100	100	100	100	100	100

1	Apple foliage and 0-DAT rapeseed forage were also analyzed but these
data are not presented here (64-DAT apple foliage - 64-DAT; 4.182 ppm;
48% TRR flutrifol; 29% TRR unknowns; 16% TRR unextracted; 0-DAT rapeseed
forage – 0.782 ppm; 97% TRR flutriafol).

2	Includes flutriafol identified in the E, WA, WB, SA, and/or SB
hydrolysates for 14-DAT rapeseed forage (E-4.5% TRR, SA-0.7% TRR, and
SB-1.4% TRR; total of 6.6% TRR; ), 42-DAT rapeseed foliage (SA-3.2% TRR;
total of 3.2% TRR), 14-DAT rapeseed pod (E-15.8% TRR, WB-1.9% TRR,
SA-3.2% TRR, and SB-6.1% TRR; total of 27% TRR), 42-DAT rapeseed seed
(E-0.5% TRR, WB-0.8% TRR, SA-2.0% TRR, and SB-3.0% TRR; total of 6.3%
TRR), and 21-DAT sugar beet tops (combined WA and WB-1.9% TRR).

3	Largest unknown of 4.2% TRR.

4	Hydrolysates in bold were not analyzed.

5	Accountability = (Total extractable + Total unextractable)/(TRR from
combustion analysis) * 100.

Table B.4:  Summary of Characterization and Identification of TRRs in
Barley and Wheat Following Foliar Application of [Carbinol-14C]- or
[Triazole-14C]-Flutriafol.

Compound	[Carbinol-14C]-Flutriafol treated	[Triazole-14C]-Flutriafol
treated

	Barley grain (62-DAT; field grown; treated pre-ear emergence)	Barley
Straw (62-DAT; field grown; treated pre-ear emergence)	Wheat straw
(74-DAT; field grown; treated pre-ear emergence)	Barley straw (94-DAT;
greenhouse grown; treated pre-ear emergence)	Barley grain (44-DAT; field
grown; treated after ear emergence)	Barley grain (94-DAT; greenhouse
grown; treated pre-ear emergence)	Wheat grain (74-DAT; field grown;
treated pre-ear emergence)	Wheat grain (56-DAT; greenhouse grown;
treated pre-ear emergence)

	TRR = 0.007 ppm	TRR = 0.72 ppm	TRR = 0.65 ppm	TRR = 2.10 ppm	TRR = 0.10
TRR = 0.41 ppm	TRR = 0.05 ppm	TRR = 0.18 ppm

	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	%
TRR	ppm

Flutriafol	36.3	0.003	37.5	0.270	56.6	0.368	63	1.323	24.0	0.024	--	--	--
--	--	--

TA	--	--	--	--

	--	--	7.6	0.008	40	0.164	57.9	0.029	48	0.086

TAA	--	--	--	--

	--	--	--	--	26	0.107	26.4	0.013	8	0.014

Others (unknowns)	3.6	<0.001	2.9	0.021	13.6	0.088	5	0.105	7.9	0.008	21
0.086	9.9	0.005	34	0.061

Total identified	36.3	0.003	37.5	0.270	56.6	0.368	63	1.323	31.6	0.032	66
0.271	84.3	0.042	56	0.10

Total characterized	38.6	0.002	22.6	0.163	20.2	0.131	15	0.315	32.7	0.034
23.2	0.095	10.6	0.005	34	0.061

Total extractable	60.26	0.005	60.2	0.434	73.8	0.480	81.0	1.701	47.4
0.048	89.2	0.366	94.9	0.047	90.0	0.162

Unextractable	25.7	0.002	39.8	0.287	23.1	0.150	16.0	0.336	35.1	0.035	6.5
0.027	5.1	0.003	5.0	0.009

Accountability1	86	100	97	97	83	96	100	95

1	Accountability = (Total extractable + Total unextractable)/(TRR from
combustion analysis) * 100.

Table B.5:  Summary of Characterization and Identification of TRRs in
Barley and Wheat Matrices Following Seed Treatment with
[Carbinol-14C]Flutriafol or [Triazole-14C]Flutriafol at ~100 ppm.

Compound	Triazole label	Carbinol label

	Barley grain	Wheat grain	Barley straw	Barley straw

	TRR = 0.17 ppm	TRR = 0.14 ppm	TRR = 0.24 ppm	TRR = 0.25 ppm

	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm

Flutriafol	--	--	--	--	35.8	0.086	59.2	0.148

TA	36.4	0.062	55.2	0.077	--	--	--	--

TAA	35.2	0.060	27.8	0.039	28.0	0.067	--	--

Others (unknowns)	4.6	0.008	4.9	0.007	4.7	0.011	18.8	0.047

Total identified	71.6	0.122	83.0	0.116	63.8	0.153	59.2	0.148

Total characterized	12.3	0.021	5.3	0.008	4.7	0.011	18.8	0.047

Total extractable	90.3	0.154	94.4	0.133	87.7	0.211	83.7	0.209

Unextractable	9.7	0.016	5.7	0.008	12.3	0.030	16.4	0.041

Accountability1	100	100	100	100

1	Accountability = (Total extractable + Total unextractable)/(TRR from
combustion analysis) * 100.

Livestock Metabolism Studies:  The petitioner submitted diary cow
([triazole-3,5-14C]flutriafol) and hen ([triazole-3,5-14C]flutriafol and
[carbinol-14C]flutriafol) metabolism studies.  The following is a
summary of these data.

Dairy Cow (47090443.der.doc):  A single dairy cow was orally
administered [triazole-3,5-14C]flutriafol twice a day for seven
consecutive days at a dietary rate of 2 ppm (10x).  Milk was collected
twice daily throughout the study and muscle, fat (subcutaneous, omental,
and peritoneal), liver, and kidney were collected at sacrifice (4 hours
after the final dose).  Table B.6 is a summary of the TRRs.
Radioactivity was highest in liver, and lowest in muscle and fat.
Residues in milk were low (<0.01 ppm).  The majority of the administered
dose was excreted, with urine and feces accounting for ~78% of the
administered dose.

); muscle and fat were not extracted due to low TRRs (≤0.008 ppm).
Bacterial protease hydrolysis of the unextracted liver residues released
and additional 42% TRR.  This hydrolysate was characterized further by
partitioning with ether (ether – 0% TRR; water - 42% TRR) followed by
hydrolysis of the aqueous phase with β-glucuronidase and 6M HCl with
partitioning of the resulting hydrolysates with ether (ether phases –
0% TRR; aqueous phases - 42% TRR).  The aqueous phase was then
hydrolyzed with 0.1M NaOH and partitioned with ether (ether - 15% TRR
(analyzed); aqueous phase - 18% TRR (not analyzed)).  Nonextractable
residues were 3-11% TRR (<0.03 ppm) in milk, kidney, and liver.

Residues were identified by TLC using co-elution with reference
standards including flutriafol, four metabolites isolated from the rat
metabolism study (M1B, M1D, M2B, and M2C), and a methoxyphenyl
metabolite of flutriafol (Compound X).  Reference standards for T, TA,
and TAA were not included.  Total identified residues accounted for
<5-32% TRR in milk, kidney, and liver (Table B.7).

The following deficiencies were identified in the dairy cow metabolism
study:  (1) a confirmatory method was not used for the identification of
metabolites; (2) no information concerning storage durations was
provided; (3) reference standards for the triazole metabolites, T, TA,
and TAA, were not included; and (4) the GLP statement indicated that
since Cheminova (the petitioner) did not conduct the study and was not
the sponsor, they could not be certain that the study was conducted in
accordance with GLP practices (40 CFR 160).  In addition, supporting
information and data were extremely limited for this study.

Hen (47090442.der.doc):  Laying hens were orally administered
[triazole-3,5-14C]flutriafol or [carbinol-14C]flutriafol once a day for
seven consecutive days at a dietary rate of 13.9 ppm (160x) or 11.6 ppm
(130x), respectively.  Eggs were collected twice daily throughout the
study and muscle (composite of breast and thigh), abdominal fat, and
liver were collected at sacrifice (20-24 hours after the final dose).
Table B.6 is a summary of the TRRs.  TRR were consistently higher in the
triazole-label matrices.  Radioactivity for both labels was highest in
liver and lowest in muscle and fat.  The majority (90-91%) of the
administered dose was excreted.

The majority of the radioactivity was extracted from egg and muscle
using ACN/water (64-98% TRR; both labels) and from fat using
acetone/hexane (94-97% TRR; both labels).  ACN/water extraction released
lower levels of TRR from liver (33-41% TRR; both labels).  Additional
residues were released from liver (both labels) via sequential
hydrolysis with pepsin and pancreatin (21-25% TRR; analyzed), 1 N HCl
(4-6% TRR; not analyzed), 1 M NH4OH (8-9% TRR; not analyzed), and 6 N
HCl at reflux (30% TRR carbinol label; 12% TRR triazole label; not
analyzed).  Nonextractable residues (both labels) accounted for 2.4-6.0%
TRR (0.004-0.008 ppm) in eggs, 6.3-36.4% TRR (0.004 ppm) in muscle,
2.9-6.3% TRR in fat (0.001 ppm), and 1.4-8.5% TRR (0.005-0.035 ppm) in
liver.

d as “other unknowns” accounted for ≤5% TRR in eggs, ≤9% TRR in
muscle, 3% TRR in triazole-label fat, and 10-12% TRR (0.043-0.045 ppm)
in liver.  Significantly different metabolic profiles were observed in
the triazole and carbinol labeled samples due to the identification of T
in all of the triazole samples or M3 in the carbinol muscle sample.

Table B.6:  TRRs in Milk, Tissue and Excreta Following Dosing with
[Triazole-3,5-14C]Flutriafol or [Carbinol-14C]Flutriafol.

Matrix	Collection Timing	[Triazole-3,5-14C]Flutriafol (ppm)
[Carbinol-14C]Flutriafol (ppm)

Diary Cow	Hen1	Dairy Cow	Hen1

Milk/Egg	Day 1 am	--	--	Dosing with [Carbinol-14C]Flutriafol was not
performed.	--

	Day 1 pm	0.002	0.001

No sample

	Day 2 pm	0.004	0.041

0.032

	Day 2 am	0.005	0.089

0.016

	Day 3 pm	0.006	0.088

0.051

	Day 3 am	0.006	0.135

No sample

	Day 4 pm	0.007	0.129

0.079

	Day 4 am	0.007	No sample

0.116

	Day 5 pm	0.007	0.145

0.101

	Day 5 am	0.007	0.184

No sample

	Day 6 pm	0.008	0.167

0.117

	Day 6 am	0.007	0.206 (0.205)

0.160 (0.159)

	Day 7 pm	0.008	0.190

0.126

	Day 7 am	0.007	0.204

0.121

	Day 8 am	--	0.184 (0.204)

0.133 (0.134)

Muscle	At sacrifice	0.008	0.060 (0.064)

0.011 (0.011)

Fat	At sacrifice	--	0.038 (0.035)

0.018 (0.016)

Fat, subcutaneous	At sacrifice	0.002	--

--

Fat, omental	At sacrifice	<0.001	--

--

Fat, perirenal	At sacrifice	0.003	--

--

Kidney	At sacrifice	0.061	--

--

Liver	At sacrifice	0.291	0.360 (0.411)

0.343 (0.359)

Heart	At sacrifice	0.011	--

--

1	TRR reported in parentheses were calculated by summing extractable and
nonextractable radioactivity.

Table B.7:  Summary of Characterization and Identification of
Radioactive Residues in Cow Matrices when Dosed with
[Triazole-3,5-14C]Flutriafol at 2 ppm in the Diet.

Compound	Milk	Kidney	Liver

	TRR = 0.008 ppm	TRR= 0.061 ppm	TRR= 0.291 ppm

	% TRR	ppm	% TRR	ppm	%TRR	ppm

Flutriafol	1	<0.001	7	0.004	29	0.084

4-Hydroxyflutriafol (M1B)	<4	<0.001	<23	<0.014	1	0.003

4-Hydroxy-5-methoxyflutriafol (M1D)	--	--	--	--	2	0.006

Enzyme solubilzed	--	--	--	--	421	0.122

Total identified	<5	<0.001	<30	<0.018	32	0.093

Total characterized	38	0.003	34	0.021	51	0.148

Total extractable	97	0.008	89	0.054	92	0.268

Unextractable	3	<0.001	11	0.007	9	0.026

Accountability2	100	100	101

1	The hydrolysate was further characterized via sequential hydrolysis
followed with ß-glucuronidase, 6M HCl, and 0.1 M NaOH with ether
partitioning between each hydrolysis.  The TLC of the ether fractions
detected flutriafol, M1D and unknowns (“each at <10% TRR”, but no
quantitative data were reported), the aqueous phase was not
chromatographically analyzed and contained 18% TRR (0.052 ppm).

2	Accountability = (Total extractable + Total unextractable)/(TRR from
combustion analysis) * 100.

Table B.8:  Summary of Characterization/Identification of TRRs Hen
Matrices Dosed with [Triazole-3,5-14C]Flutriafol or
[Carbinol-14C]Flutriafol in the Diet.

Compound	[Carbinol-14C]Flutriafol at 11.6 ppm in the Diet
[Triazole-3,5-14C]Flutriafol at 13.9 ppm in the Diet

	Eggs Day 6	Eggs Day 8	Muscle	Fat	Liver	Eggs Day 6	Eggs Day 8	Muscle	Fat
Liver

	TRR=

0.159 ppm	TRR =

0.134 ppm	TRR=

0.011 ppm	TRR=

0.016 ppm	TRR=

0.359 ppm	TRR= 0.205 ppm	TRR = 0.204 ppm	TRR= 0.064 ppm	TRR= 0.035 ppm
TRR= 0.411 ppm

	% TRR	ppm	%TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	%TRR	ppm	%
TRR	ppm	% TRR	ppm	% TRR	ppm

Flutriafol	74.8	0.119	65.7	0.088	--	--	75.0	0.012	1.9	0.007	48.3	0.099
50.5	0.103	--	--	80.0	0.028	3.2	0.013

T	--	--	--	--	--	--	--	--	--	--	29.3	0.060	27.5	0.056	75.0	0.048	11.4
0.004	16.65	0.068

Hydroxylated flutriafols (M5)	5.7	0.009	7.5	0.010	9.1	0.001	--	--	1.9
0.007	4.4	0.009	4.4	0.009	1.6	0.001	2.9	0.001	1.5	0.006

Unknown M3	13.2	0.021	12.7	0.017	45.5	0.005	6.3	0.001	7.0	0.025	8.8
0.018	11.3	0.023	9.4	0.006	2.9	0.001	6.6	0.027

Unknown M4	3.1	0.005	3.7	0.005	--	--	--	--	7.0	0.025	1.5	0.003	2.9	0.006
--	--	--	--	5.8	0.024

Unknown M2	--	--	--	--	--	--	--	--	2.5	0.009	--	--	--	--	--	--	--	--	--
--

Other unknowns	0.5	0.001	4.5	0.006	9.1	0.001	--	--	12.5	0.045	5.4	0.011
0.5	0.001	7.8	0.005	2.9	0.001	10.5	0.043

Oily phase	--	--	--	--	--	--	5.0	0.001	--	--	--	--	--	--	--	--	3.0	0.001
--	--

Enzyme hydrolysate

–DCM phase	not performed	4.53	0.016	not performed	5.4	0.022

Enzyme hydrolysate

–aqueous phase

16.73	0.060

17.16	0.070

1N HCl hydrolysate

5.84	0.021

4.14	0.017

1M NH4OH hydrolysate

8.14	0.029

8.84	0.036

6N HCl hydrolysate  –DCM phase

5.34	0.019

0.74	0.003

6N HCl hydrolysate  –aqueous phase

25.14	0.090

11.24	0.046

Total identified	80.5	0.128	73.2	0.018	9.1	0.001	75.0	0.012	3.8	0.014
82.0	0.168	82.4	0.168	76.6	0.049	94.3	0.033	21.3	0.087

Total characterized	16.9	0.027	20.9	0.028	54.6	0.006	11.3	0.002	94.5
0.339	15.7	0.032	14.7	0.030	17.2	0.011	8.8	0.003	70.2	0.288

Total extractable	97.5	0.155	94.0	0.126	63.6	0.007	93.8	0.015	98.6	0.354
97.6	0.200	97.1	0.198	93.8	0.060	97.1	0.034	91.5	0.376

Unextractable 1	2.5	0.004	6.0	0.008	36.4	0.004	6.3	0.001	1.4	0.005	2.4
0.005	2.9	0.006	6.3	0.004	2.9	0.001	8.5	0.035

Accountability2	99	101	100	89	105	100	111	107	92	114

1	Residues remaining after exhaustive extractions.

2	Accountability = (Total extractable + Total unextractable)/(TRR from
combustion analysis) * 100.

3	The DCM phase of the enzyme hydrolysate was analyzed by TLC (nothing
identified) and aqueous phase was analyzed by HPLC (nothing identified).

4	These hydrolysates were not analyzed further.

5	2.7% TRR found in the enzyme hydrolysate.

6	The DCM phase of the enzyme hydrolysate was analyzed by TLC (unknowns
≤2.9% TRR; ≤0.012 ppm) and aqueous phase was analyzed by HPLC.
Confined Rotational Crop Study:  [Carbinol-14C]flutriafol or
[triazole-3,5-14C]flutriafol was incorporated into bare loam soil at a
target rate of 0.22 lb ai/acre (1x the proposed rate for soybean;
47090451.der.doc).  Rotational crops of wheat, pea, sugar beet, and
rapeseed were planted 30, 120, and 365 days after soil treatment and
maintained in a greenhouse.  Samples of wheat (grain, straw, and chaff),
pea (seed, pod, and foliage), sugar beet (root and top), and rapeseed
(seed, pod, and foliage) were harvested at maturity.  Table B.9 is a
summary of the TRRs in the harvested samples (30-day plantback interval
(PBI) rapeseed samples were not analyzed).  TRRs were generally higher
in the triazole-label matrices than in the carbinol-label matrices and
were lowest at the 365-day PBI.

≤0.05 ppm in all analyzed matrices excluding the following:  carbinol
label - 120-day wheat straw (18% TRR; 0.196 ppm); triazole label -
120-day wheat grain (13% TRR; 0.150 ppm) and 120-day wheat straw (17%
TRR; 0.419 ppm).

Based on the general extraction flowchart provided in the study, the
extracts were TLC analyzed.  However, no details of the system were
provided and the method of metabolite identification was not described
(no reference standards were listed).  Identified residues accounted for
26-43% TRR in the carbinol labeled samples and 43-67% TRR in the
triazole labeled samples (Table B.10).  The metabolic profile varied
with radiolabel position due to the identification of T, TA, and/or TAA
in the triazole labeled samples.

The following deficiencies were identified in the confined rotational
crop study:  (1) the study did not include a leafy vegetable crop; (2)
residues in wheat forage were not investigated; (3) sandy loam soil was
not used and no data were provided concerning the soil characteristics;
(4) insufficient information was provided concerning analytical
methodology and a confirmatory method was not used for the
identification of  metabolites; (5) insufficient information was
provided to determine whether identification/characterization of
residues met Agency requirements (e.g., five unknowns designated
“others” accounted for up to >50% of TRR in carbinol-label sugar
beet tops and were not further investigated; reference standards used
not identified); (6) insufficient attempts were made to characterize
nonextractable residues of 120-day wheat straw and grain and 365-day
wheat straw samples; (7) insufficient storage stability data/information
are available to support the storage interval of at least 4 years; and
(8) insufficient information/data in general were provided to support
the study, including details of sample handling at the field site and
analytical laboratory; the distribution of radioactivity into sample
extracts and fractions; representative chromatograms, raw data, or
example calculations; and storage conditions and durations.

Table B.9:  TRR in Rotated Crop Matrices.1

Crop	Matrix	Plantback interval (days)	[carbinol-14C]flutriafol
[triazole-3,5-14C]flutriafol

	ppm	ppm

Wheat	Grain	30	0.04	1.04

120	0.02	1.22 (1.18)

365	<0.01	0.3 (0.31)

	Straw	30	10.46	6.47

120	0.93 (1.07)	1.32 (2.45)

365	0.13	0.2 (0.16)

	Chaff	30	2.90	1.82

120	0.88	1.58

365	0.10	0.2

Pea	Seed	30	0.01	0.32

120	<0.01	0.32

365	<0.01	0.2

	Pod	30	0.05	0.14

120	0.03	0.10

365	<0.01	0.1

	Foliage	30	1.25	1.08

120	0.33	0.63

365	<0.01	0.1

Sugar beet	Root	30	0.02	0.08

120	<0.01	0.09 (0.12)

365	<0.01	0.03

	Tops	30	0.20	0.60

120	0.19 (0.31)	0.57 (0.56)

365	0.13	0.35

Rapeseed	Seed	30	Not determined

120	0.03	2.16

365	<0.01	0.6

	Pod	30	Not determined

120	0.97	2.13

365	0.13	0.3

	Foliage	30	Not determined

120	0.28	0.67

365	0.04	0.1

1	TRR were initially determined in 1981/1983; TRR re-determined in 1987
are presented in parentheses; only those samples in bold were analyzed
further.

Table B.10:  Summary of Characterization/Identification of TRRs in
Rotational Crop Samples Following Soil Treatment with
[Triazole-3,5-14C]Flutriafol or [Carbinol-14C]Flutriafol.

Compound	[Carbinol-14C]Flutriafol	[Triazole-3,5-14C]Flutriafol

	Sugar beet top

120-day	Wheat straw

 120-day	Sugar beet root

120-day	Sugar beet top

120-day	Wheat grain

120-day	Wheat grain

365-day	Wheat straw

120-day	Wheat straw

365-day

	TRR = 0.31 ppm	TRR = 1.07 ppm	TRR = 0.12 ppm	TRR = 0.56 ppm	TRR = 1.18
ppm	TRR = 0.31 ppm	TRR = 2.45 ppm	TRR = 0.16 ppm

	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	% TRR	ppm	%
TRR	ppm

Flutriafol	25.7	0.080	43.3	0.463	4.3	0.005	17.0	0.095	--	--	--	--	38.2
0.936	30.7	0.049

4-hydroxy flutriafol	--	--	--	--	--	--	2.5	0.014	--	--	--	--	1.5	0.037
--	--

TA	--	--	--	--	19.6	0.024	2.5	0.014	48.5	0.572	50.5	0.157	--	--	1.1
0.002

TAA	--	--	--	--	2.7	0.003	21.0	0.118	18.8	0.222	14.2	0.044	15.5	0.380
22.2	0.036

T	--	--	--	--	17.3	0.021	--	--	--	--	--	--	--	--	--	--

Others1	51.2	0.159	25.5	0.273	--	--	15.9	0.089	--	--	--	--	16.8	0.412
3.0	0.005

Baseline	--	--	0.7	0.007	4.7	0.006	1.4	0.008	--	--	--	--	--	--	--	--

Remainder	0.5	0.002	1.8	0.019	2.6	0.003	0.5	0.003	4.8	0.057	0.6	0.002
5.5	0.135	3.5	0.006

Total identified	25.7	0.080	43.3	0.463	43.9	0.053	43	0.241	67.3	0.794
64.7	0.201	55.2	1.353	54.0	0.087

Total characterized	51.2	0.161	28.0	0.299	7.3	0.009	15.9	0.100	4.8	0.057
0.6	0.002	22.3	0.547	6.5	0.011

Total extractable2	77.4	0.241	71.3	0.762	51.2	0.062	60.8	0.341	72.1
0.851	65.3	0.203	77.5	1.900	60.5	0.098

Unextractable3	7.0	0.022	18.3	0.196	35.6	0.043	3.6	0.020	12.7	0.150	15.2
0.047	17.1	0.419	31.4	0.050

Accountability4	84.8 (138)	90 (103)	88 (117)	64 (63)	84.8 (82.0)	80.6
(83.3)	94.7 (176)	92.5 (74.0)

1	Consisting of at least 5 compounds in carbinol-label sugar beet tops ,
3 compounds in carbinol-label wheat straw, 2 compounds in 120-day
triazole-label wheat straw, an unspecified number of compounds in
365-day triazole-label wheat straw, and 2 compounds in triazole-label
sugar beet tops.

2	Total identified and characterized residues; actual extraction
distributions were not reported.

3	Residues remaining after extraction.

4	Accountability = (Total extractable + Total unextractable)/(TRR) *
100; values in parentheses are calculated using the initial TRR from
1981/1983.

Appendix C.	Tolerance Summary Table

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萑ː葠ː摧侠ü਀tolerances should be revised to reflect the correct
commodity definition and/or numerical tolerance specified in Table C.1.
A revised Section F is requested

Table C.1:  Tolerance Summary for Flutriafol.

Commodity	Proposed Tolerance (ppm)	HED-Recommended Tolerance (ppm)
Comments

Apple	0.2	0.20	Numerical tolerance should be 0.20.

Soybean	0.3	0.35	Based on the field trial data and the tolerance
calculator, the numerical tolerance should be 0.35 ppm and the correct
commodity definition is "Soybean, seed."

Soybean, aspirated grain fractions	0.5	2.2	Based on the field trial and
processing data, the numerical tolerance should be 2.2 ppm and the
correct commodity definition is "Grain, aspirated fractions."

Liver (cattle, goat, hog, horse, sheep)	0.01	--	Incorrect commodity
definition.

Cattle, liver	--	0.02	--

Goat, liver	--	0.02	--

Hog, liver	--	0.02	--

Horse, liver	--	0.02	--

Sheep, liver	--	0.02	--

Eggs	0.01	--	Tolerance not required.

Appendix D:	Chemical Name and Structure Table

Common name;

or

 Category II, based on no toxicity observed up to 1000 mg/kg/day in the
28-day dermal toxicity study in rat.

Flutriafol	Human-Health Risk Assessment		DP# 372347

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