Document ID: EPA-HQ-OPP-2011-0276-0002
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2011-04-26T04:00Z

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<EPA BIOPESTICIDES AND POLLUTION PREVENTION DIVISION COMPANY NOTICE OF
FILING FOR PESTICIDE PETITIONS PUBLISHED IN THE FEDERAL REGISTER  >

<EPA Biopesticides and Pollution Prevention Division contact: Linda
Hollis #17 (703-308-8367)>

<INSTRUCTIONS:  Please utilize this outline in preparing the pesticide
petition.  In cases where the outline element does not apply, please
insert “NA-Remove” and maintain the outline. Please do not change
the margins, font, or format in your pesticide petition. Simply replace
the instructions that appear in green, i.e., “[insert company
name],” with the information specific to your action.>

<SUBMISSION: E-mail the completed template to: hollis.linda@epa.gov.>

<TEMPLATE:>

Arkion Life Sciences

<[Insert petition number]>

<	EPA has received a pesticide petition ([insert petition number]) from
Interregional Research Project Number 4(IR-4) 500 College Road East,
Suite 201W Princeton, New Jersey on behalf of Arkion Life Sciences, 551
Mews Drive Suite J, New Castle DE 19720 proposing, pursuant to section
408(d) of the Federal Food, Drug, and Cosmetic Act (FFDCA), 21 U.S.C.
346a(d), to amend 40 CFR part 180.>

<(Options (pick one)>

<	1. by establishing a tolerance for residues of>

<	2. to establish an exemption from the requirement of a tolerance for>

	

<	3. to establish an amendment/expansion of an existing tolerance
exemption for the>

<(Options (pick one)>

<	1. microbial pesticide  [insert name of active ingredient]>

<	2. biochemical pesticide 9,10 anthraquinone>

	

<	3. plant-pesticide [insert name of active ingredient] in or on [insert
commodity].>

<	Pursuant to section 408(d)(2)(A)(i) of  FFDCA, as amended,
Interregional Research Project Number 4(IR-4) on behalf of Arkion Life
Sciences has submitted the following summary of information, data, and
arguments in support of their pesticide petition. This summary was
prepared by Interregional Research Project Number 4(IR-4) on behalf of
Arkion Life Sciences and EPA has not fully evaluated the merits of the
pesticide petition. The summary may have been edited by EPA if the
terminology used was unclear, the summary contained extraneous material,
or the summary unintentionally made the reader conclude that the
findings reflected EPA’s position and not the position of the
petitioner.>

<I. Arkion Life Sciences  Petition Summary>

<	[Insert petition number]>

<A. Product Name and Proposed Use Practices>

<	Avipel liquid corn seed treatment.  Avipel will be used for the
protection of planted field and sweet corn seed against consumption by
blackbirds, cranes, grackles and pheasants. Application will be
performed to corn seed either by commercial seed treatment businesses or
on farm as a planter box seed treatment.>

<B. Product Identity/Chemistry>

<	1. Identity of the pesticide and corresponding residues.>> 

	

	The active ingredient in Avipel is 9,10 anthraquinone (CAS 84-65-1). 
Residues of 9,10 anthraquinone, 1-hydroxy anthraquinone and 2-hydroxy
anthraquinone have been quantitatively and qualitatively measured using
a GLP Magnitude of Residue study conducted under the auspices of IR-4
and included in this filing.

<	2. Magnitude of residues at the time of harvest and method used to
determine the residue.> 

	

Anthraquinone, the active ingredient in Avipel( liquid formulation and
Avitec( dry powder formulation, is a pesticide used on field corn for
control of sandhill crane.  To provide the sponsor with residue
chemistry data to support the establishment of a tolerance for
anthraquinone in or on field corn,   HYPERLINK  \l "_Appendix_1_Field" 
15 field trials  were conducted for this study during the 2006 and 2007
growing seasons, two each in Michigan (EPA Region 5) and New York
(Region 1); three in North Dakota (one Region 5, two Region 7); one
each in Ohio (Region 5), South Dakota (Region 5), and Texas (Region 6);
and five in Wisconsin (Region 5).  Six varieties of field corn seed
were treated with Avipel( liquid formulation at targeted rates of
approximately 0.005 lb ai/lb seed (1X), and at 0.025 lb ai/lb seed
(5X)(2006 trials), or 0.05 lb ai/lb seed (10X) (2007 trials).  In
addition, at the Wisconsin 06-WI07 and North Dakota 06-ND04 trials, seed
was treated with Avitec( dry powder formulation at rates of
approximately 0.005 lb ai/lb seed (1X) and 0.025 lb ai/lb seed (5X). 
The seed was planted and samples of kernels plus cob with husks removed
(K+CWHR) and samples of forage were collected at the milk stage. 
Samples of grain and stover were collected at crop maturity.

 HYPERLINK  \l "_Appendix_2_Analytical" Sample analysis  for residues of
anthraquinone, 1-hydroxyanthraquinone, and 2-hydroxyanthraquinone was
conducted by Cornell Analytical Laboratories, Geneva, NY.  The procedure
used was “Residue Analysis of Anthraquinone on Corn (field) including
Kernels plus Cob with Husks Removed (K+CWHR), Forage, Grain, and Stover
by LC/MS Detection”, Version #1 

The  HYPERLINK  \l "TableC3" results  from the 15 trials show that no
residues above the LLMV of 0.10 ppm of anthraquinone,
1-hydroxyanthraquinone, and 2-hydroxyanthraquinone were observed in any
of the samples following seed treatment at rates of approximately 0.025
lb ai/lb seed (5X), or 0.05 lb ai/lb seed (10X) in any grain sample.
Residues of 0.12 ppm 1-hydroxyanthraquinone were observed in one stover
sample treated with Avipel( liquid formulation at the 10X rate from the
Texas trial.  The sample was reanalyzed at a later date and no residues
above the LLMV were observed.  

<	3. A statement of why an analytical method of detecting and measuring
the levels of the pesticide residue are not needed.> 

	An analytical method is available.  HYPERLINK  \l
"_Appendix_2_Analytical" Sample analysis  for residues of anthraquinone,
1-hydroxyanthraquinone, and 2-hydroxyanthraquinone was conducted by
Cornell Analytical Laboratories, Geneva, NY.  The procedure used was
“Residue Analysis of Anthraquinone on Corn (field) including Kernels
plus Cob with Husks Removed (K+CWHR), Forage, Grain, and Stover by LC/MS
Detection”, Version #1. 

<C. Mammalian Toxicological Profile>

<	The information submitted on this formulation as Flight Control for
outdoor nonfood use was previously reviewed by BPPD and states: 
Adequate mammalian toxicology data are available and support
registration of the active ingredient 9,10 – anthraquinone.  The
registrant, (Arkion Life Sciences), submitted acceptable acute toxicity
studies for both the Technical Grade Active Ingredient (TGAI) and the
end-use product.>>  

For the TGAI: (i) acute oral LD50 in rats was >5000 mg/kg (Tox category
IV); (ii) acute dermal LD50 in rabbits was >5000 mg/kg (Tox category
IV); (iii) acute inhalation LC50 in rats was >2.11 mg/L (Tox category
IV); and (iv) the EP caused mild ocular irritation symptoms in rabbits
which cleared in 72 hours post-instillation (Tox category III).  

For the EP: (i) acute oral LD50 in rats was >5000 mg/kg (Tox category
IV); (ii) acute dermal LD50 in rats was >5000 mg/kg (Tox category IV);
(iii) acute inhalation LD50 in rats was > 2.04 mg/L (Tox category IV);
(iv) the EP caused mild ocular irritation symptoms in rabbits which
cleared by 72 hours post instillation (Tox category III); (v) the EP
caused slight dermal irritation symptoms in rabbits which cleared by 24
hours post dosing (Tox category III); and (vi) the EP was shown to be a
contact sensitizer in guinea pigs using the Buehler Method.

The National Toxicology Program (NTP) has conducted a series of studies
on anthraquinone. All these studies were conducted with anthraquinone
samples produced from a process based on anthracene.  The current
process does not give rise to this impurity.  The sample of
anthraquinone used in the NTP studies were contaminated with up to 0.11%
9-nitroanthracene (9-NA) and 0.6% of mutagenic impurities. The sample
was produced using an anthracene-derived process.  This process has not
been used for decades and is not used in any of the Arkion Life Sciences
material. 

The level of 9-nitroanthracene in the studies were calculated at roughly
4 ppm in the diet at 3750 ppm anthraquinone  The high dose tested in the
rat study was 7500 ppm anthraquinone. The contamination of the
anthraquinone sample with the potent mutagen 9-nitroanthracene was well
documented in both the NTP report and the open literature.  Estimates
based on mutagenic potency indicate it is plausible that the 9-NA and
other contaminants could have been responsible for a portion, if not
all, of the tumor induction observed in the NTP study.

30,000 ppm females. The concentrations of α-2u-globulin in the kidneys
were significantly greater in all exposed groups of males. Thyroid gland
follicular cell hypertrophy was present in all males and females exposed
to 3,750 ppm or greater. Incidences of inflammation and transitional
cell hyperplasia in the urinary bladder of 30,000 ppm females were
greater than those in the controls.  This study was performed with
samples of anthraquinone containing the impurity 9-aminoanthracene, a
known toxicant.  The US EPA Guideline sets the limit dose for 90-day
studies at 1000 mg/kg.  The high dose in this study exceeded the limit
dose.

The NTP has conducted a subchronic oral toxicity study in mice. Groups
of 10 male and 10 female B6C3F1 mice were fed diets containing 0, 1,875,
3,750, 7,500, 15,000, or 30,000 ppm anthraquinone (equivalent to average
daily doses of approximately 250, 500, 1,050, 2,150, or 4,300 mg/kg to
males and 300, 640, 1,260, 2,600, or 5,300 mg/kg to females) for 14
weeks. All mice survived until the end of the study. Mean body weights
and feed consumption were similar among exposed and control groups. A
responsive anemia occurred in exposed mice at week 14 but a clear dose
response was not evident and the response was weak. Liver weights of
exposed groups of mice were significantly greater than those of the
control groups. The incidences of centrilobular hypertrophy in the liver
of mice exposed to 3,750 ppm or greater were significantly greater than
those in the controls, and the severities increased with increasing
exposure concentration. Cytoplasmic alteration of the urinary bladder
was observed in all exposed mice, and the severities increased with
increasing exposure concentration. The incidences of hematopoietic cell
proliferation were increased in all exposed groups of males and females,
and pigmentation was observed in the spleen of all exposed mice (except
one male and one female in the 30,000 ppm groups). This study was
performed with samples of anthraquinone containing the impurity
9-aminoanthracene, a known toxicant. The US EPA Guideline sets the limit
dose for 90-day studies at 1000 mg/kg.  The high dose in this study
exceeded the limit dose.

 and α2u-globulin concentrations at 3 months and for organ weights and
histopathology at 12 months. These dietary anthraquinone concentrations
resulted in average daily doses of approximately 20, 45, 90, and180
mg/kg to males and 25, 50, 100, and 200 mg/kg to females. Additional
groups of 18 males given 469, 938, 1,875, or 3,750 ppm for 8 days and 10
males and 10 females given 469, 938, or 1,875 ppm for 3, 6, 12, or 18
months were designated for toxicokinetic studies. Survival of all groups
of males was similar, and survival of exposed groups of females was
greater than that of the controls. Mean body weights of exposed groups
of males during the latter part of the study and mean body weights of
exposed females throughout most of the study were less than those of the
controls. Feed consumption by exposed groups was similar to that by the
controls.

pathy were increased in females, and severities of nephropathy were
increased in males. At 3 months, the concentration of α2u-globulin in
the kidney of 3,750 ppm males was greater than that in the control
group. 

The incidence of urinary bladder transitional epithelial papilloma was
significantly greater in 1,875 ppm males than in the control group, and
the incidences in groups of males exposed to 938 ppm or greater exceeded
the historical control range. There were positive trends in the
incidences of transitional epithelial hyperplasia and papilloma or
carcinoma (combined) of the urinary bladder in females. The incidences
of hepatocellular adenoma or carcinoma (combined) were slightly
increased in exposed males and females; the incidences in groups of
females exposed to 938 ppm or greater exceeded the historical control
range. The incidences of several nonneoplastic liver lesions of minimal
severity were also increased. The incidences of congestion,
pigmentation, and hematopoietic cell proliferation of the spleen were
greater in exposed males and females than in the controls. The
incidences of bone marrow hyperplasia were increased in most groups of
exposed rats, and the incidences of bone marrow atrophy were increased
in exposed females. The incidences of mononuclear cell leukemia were
significantly less in all exposed groups than in the controls at 2
years, and the incidences were less than the historical control ranges.
The NOEL was 469 ppm (20 mg/kg males, 25 mg/kg females) based on
pathological findings in the liver, kidney and bladder at 938 ppm (45
and 50 mg/kg males and females, respectively).

In an NTP study for oncogenicity in mice, groups of 50 male and 50
female B6C3F1 mice were fed diets containing 0, 833, 2,500, or 7,500 ppm
anthraquinone (equivalent to average daily doses of approximately 90,
265, or 825 mg/kg to males and  80, 235, or 745 mg/kg to females) for
105 weeks. Additional groups of 36 males given 833, 2,500, or 7,500 ppm
for 8 days and 10 males and 10 females given 833, 2,500, or 7,500 ppm
for 12 months were designated for toxicokinetic studies. Survival was
decreased for 7,500 ppm males than for the control group. Mean body
weights of 7,500 ppm males during the last 6 months of the study and
mean body weights of 7,500 ppm females at the end of the study were less
than those of the control groups. Feed consumption was similar in all
groups of males and females.

Incidences of hepatocellular neoplasms (including multiple neoplasms)
increased with a positive trend in male and female mice, and the
incidences were increased in all exposed groups. Incidences of
hepatoblastoma were significantly increased in males exposed to 2,500 or
7,500 ppm. The incidences of several nonneoplastic lesions of the liver
were increased in exposed mice. There was a marginal increase in the
incidences of neoplasms of thyroid gland follicular cells in males and
females. Incidences of intracytoplasmic inclusion body of the urinary
bladder and hematopoietic cell proliferation of the spleen in males and
females and thyroid gland follicular cell hyperplasia and kidney
pigmentation in males were greater in exposed groups than in the
controls.

The NOEL was 833ppm (90 mg/kg and 80 mg/kg males and females,
respectively) based on pathological findings in the liver, kidney,
thyroid and spleen at 2500 ppm (265 and 235 mg/kg males and females,
respectively).

For developmental and reproductive toxicity, no studies have been
conducted for anthraquinone. However, a structural analog, emodin (1, 3,
8-trihydroxy-6-methyl-anthraquinone), has been tested for developmental
toxicity by the NTP in rats and mice.  Emodin is a naturally occurring
and pharmacologically active anthraquinone found in the roots and bark
of certain plant species.

Time mated Sprague Dawley (CD®) rats (25/group) were exposed to emodin
at nominal concentrations of 0, 425, 850, and 1700 ppm in NIH-07 ground
rodent diet from gestational day (gd) 6 to 20. Average daily intake was
0, 31, and 57 mg emodin/kg of body weight/day for the control, low- and
mid-dose groups. Average daily intake for the 1700 ppm group was 80-144
mg emodin/kg of body weight/day. There were no differences among groups
for the number of corpora lutea/dam, number of implantation
sites/litter, or percent preimplantation loss/litter. Postimplantation
loss (resorptions or late fetal deaths), live litter size, percent male
fetuses/litter, and average fetal body weight/litter (males, females or
both) were likewise unaffected. Emodin did not affect the incidence of
fetal malformations or variations. The maternal LOAEL was considered to
be 1700 ppm (80-144 mg/kg/day) based on maternal body weight and weight
gain, and the NOAEL was 850 ppm (57 mg/kg/day). The developmental
toxicity NOAEL was greater than or equal to 1700 ppm (80-144 mg/kg/day).
A LOAEL for developmental toxicity was not established in this study.

In an NTP study, time-mated Swiss albino (CD-1®) mice (25/group) were
exposed to emodin at concentrations of 0, 600, 2500 or 6000 ppm in
NIH-07 rodent diet from gestational day (gd) 6 to 17. Average daily
intake was 0, 94, 391, and 1005 mg emodin/kg/day. Nominal concentrations
of emodin (ppm in feed) and calculated intake of emodin (mg/kg/day) are
expressed as the bulk test article (96.2% purity) unless otherwise
noted. There were no differences among groups for the number of corpora
lutea/dam, number of implantation sites/litter, or percent
preimplantation loss/litter. Postimplantation loss (resorptions or late
fetal deaths), live litter size, and percent male fetuses/litter were
likewise unaffected. At 6000 ppm, average fetal body weight/litter
(males, females or both) was reduced by 10-11%, and this effect was
statistically significant.

Ingested emodin failed to significantly affect the incidences of fetal
malformations or variations. At 2500 ppm, external malformations
included exencephaly (3 fetuses in 3 litters) and cleft palate (2
fetuses in 2 litters). However, this increase in external malformations
was not considered to be treatment related due to the clear absence of a
dose-response relationship. At 6000 ppm, no external malformations were
found among 302 fetuses (24 litters), nor had any been observed among
161 fetuses (13 litters) at 6000 ppm in the screening study (NTP, 2000).

The maternal toxicity LOAEL was considered to be 6000 ppm (1005
mg/kg/day) based on reduction of maternal body weight and weight gain,
and the NOAEL was 2500 ppm (391 mg/kg/day). The developmental toxicity
LOAEL was 6000 ppm (1005 mg/kg/day) based on reduction of fetal body
weight, and the NOAEL was 2500 ppm (391 mg/kg/day). 

Emodin was not a developmental toxin.  It is not anticipated that
anthraquinone will be a developmental toxin.

Several mutagenicity studies were conducted by NTP on anthraquinone
using S. typhimurium, and mouse peripherial blood micronuclei 
Statistically significant increases in the frequencies of micronucleated
normochromatic erythrocytes were observed in peripheral blood samples
from male and female mice exposed to anthraquinone (99.8% pure) in feed
for 14 weeks but the frequency was low. However, results of an acute
exposure mouse bone marrow micronucleus test, with anthraquinone
administered by intraperitoneal injection, was negative.  The NTP
concluded that there was evidence of increased erythropoiesis in the
14-week treated mice and concluded that the increased rate of
erythropoiesis may have contributed to the micronucleus responses seen
in the high exposure concentration groups.  (Increased cell
proliferation can produce increased levels of mitotic errors.)  Overall,
NTP concluded that anthraquinone was negative for genotoxicity.

In EPA Guideline studies, anthraquinone was negative in bacterial
mutation tests using Salmonella and E. coli with and without activation.
 Anthraquinone was also negative in a mouse lymphoma gene mutation test
with and without metabolic activation.   Anthraquinone did not induce
chromosomal aberrations in Chinese hamster ovary (CHO) cells in the
presence or absence of activation.  Additionally, another mouse
micronucleus bone marrow study was also negative following an acute
exposure.

In studies conducted as part of the NTP studies, the metabolism and
disposition of anthraquinone in male F344/N rats were examined. Animals
were administered uniformly labeled 14C-anthraquinone by intravenous
injection at 0.35 mg/kg body weight or by gavage at doses ranging from
0.35 to 350 mg/kg.

Following oral administration, anthraquinone was absorbed from the
gastrointestinal tract and distributed to tissues. No indication of
bioaccumulation was apparent in any tissue. The majority of the
radiolabel was eliminated in the feces and the urine by 24 hours after
dosing at all four concentrations. At 96 hours after dosing, less than
5% of the administered dose remained in major tissues. 

Elimination of over 50% of the administered radioactivity in the feces
and there was substantial excretion of parent and/or metabolites in the
bile. Analysis of the bile samples indicated that less than 3% of the
radioactivity collected was present as the parent compound, suggesting
extensive hepatic metabolism. Analysis of urine from dosed rats revealed
the presence of as many as 11 metabolites. Two of the metabolites
identified were 1-hydroxyanthraquinone and 2-hydroxyanthraquinone.

The plasma half-life in mice was determined to be 4 hours, and in rats
it was between 10-12 hours. 

<D. Aggregate Exposure>

<	1. Dietary exposure.>>

	No exposure is expected based on the lack of measurable residues in the
crop.  A theoretical estimation of dietary risk was conducted using
LifeLineTM version 5.0 with very conservative assumptions. Values of 0.1
ppm were used based on the LODs of the analytical method for the parent
and three presumptive metabolites.  No residues at a 10x use rate were
detected which represents a 10-fold higher residue. A 100% crop
treatment was also assumed.  All LifeLine default processing values were
used.  The NOEL for the chronic study was assumed to be 7 mg/kg, based
on a 3X reduction in the NOEL of the NTP study.  A 10X FQPA safety
factor was assumed as a conservative default.

Using exaggerated assumptions of residue and hazard, chronic dietary
risks did not exceed 5% of the Reference Dose (RfD) with the most
sensitive subpopulation, children ages 3- 5 with an estimate of 4.5% of
the RfD.  Taking the 95th percentile of the chronic assessment for the
acute dietary risk, the highest subpopulation was 7.9% of the chronic
RfD.

Neither acute or chronic exposures are of dietary concern. 

<i. Food.>> 

No exposure is expected based on the lack of measurable residues in the
crop.

<	ii. Drinking water.>> 

	No exposure is expected in drinking water.

<	2. Non-dietary exposure.>>

	Based on its low toxicity and use practice (repellent of a vertebrate
pest), anthraquinone is not subject to the Worker Protection Standards
(WPS).  Moreover, the possibility for dermal, eye and inhalation
exposure is mitigated as long as the product is used according to label
directions which recommends use of gloves, long sleeved shirts, long
sleeved pants and NIOSH filters to prevent inhalation. It is anticipated
that a majority of the treated corn seed will occur within a commercial
seed treatment facility.

	

<E. Cumulative Effects>

<	Anthraquinone is not toxic and therefore there would be no expected
cumulative effects from common mechanisms of toxicity.>

<F. Safety Determination>

	1. U.S. population. 

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cute toxicity studies previously reviewed by the EPA as part of the
Flight Control (66969-1) registration process. Because the inhalation
toxicity studies for anthraquinone showed no toxicity (Toxicity Category
IV), the risks anticipated for this route of exposure are considered
minimal.  Results of the acute dermal study indicated low toxicity
(Toxicity Category IV), and no significant dermal irritation (Toxicity
Category IV).  Based on these results, the anticipated risks from dermal
and inhalation exposures are also considered minimal.  Therefore, the
risks from aggregate exposure via dermal and inhalation exposure are a
compilation of two low risk exposure scenarios and are considered
negligible.  Exposure is further mitigated by use of worker protection
equipment on the label.

<	2. Infants and children.>

	There is no anticipated residue associated with the proposed use of the
anthraquinone.  Therefore, the acute dietary risks should be negligible
based on the lack of exposure.

<G. Effects on the Immune and Endocrine Systems>

	There are no anticipated residues, therefore there is no exposure to
the population to trigger concern for immune and endocrine effects in
the general population or in the high risk population of infants and
children.<>

<H. Existing Tolerances>

<	There is no established direct food tolerance in food for
anthraquinone.  There is an indirect food tolerance issued by FDA for
anthraquinone in paper products used in contact with food.  The
established tolerance in this case for paper products is 2 ppm.>

<I. International Tolerances>

	There is no established direct food tolerance in food for
anthraquinone.  There is an indirect food tolerance issued by FDA for
anthraquinone in paper products used in contact with food.  The
established tolerance in this case for paper products is 2 ppm.

<>

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