### Spirotetramat Human Health Risk Assessment



## UNITED STATES ENVIRONM E NTAL PROTECTION AGENCY WASHINGTON, D.C. 20460 0413575

OFFICE OF C H EMICA L S AFETY AN D PO LL UTION PR E VENTI O N



## MEMORANDUM



## Date: 16

- 0CT-2014



## Subject: Spirotetramat: H uman Health Risk Assessment for the Propo sed Use on T urfgrass (Sod Farms and Golf Course Turf only).

DP Barcode: 04 1 3575 Registration No.: 432-xxxx PC Code: 392201
Decision No.: 478963
Petit i on N o .: NA
Ri s k Assessment Type: Single Chemica V

Aggregate

T XRNo.: NA Regulatory Action: Section 3 Registrat i on Case No

. : 7452

MRIDNo .: NA CAS No.: 

### 203313-25-1

40 

### CFR: §180.641



## F rom:

Through:

To : Kelly M. Lowe, Environmental Scientist Monique M. Perron, Sc.D., Toxicolo gis t
Risk Assessme nt Branch 1 (RAB 1)

Health Effects Di vis ion (HED ; 7 509P)

Char l es W. 

## Smith II I

, 

## Branch

# Chief**C-~** George F. Kramer, Ph.D.

, Senior Chemist .-:- / 6-



# - -?"' (._.-;;...) .:;;::- RABl!HED (7509 P)**~** L

- -

### ~



## Rita Kumar, RM07 Registration Di v i s ion (RD ; 

## 7505P) 

# -----

The HED ofthe Office of Pesticide Programs (OPP) is charg ed with estimating the risk to human

health from exposure to pesticides

. The RD 

## of O P P has requested t h at HED evaluate hazard and

exposure data and conduct dietary

, occupational

, residential

, and aggregate exposure

assessmen ts, as needed

, to est imate the risk to human health that will result from the prop osed
Sectio n 3 re g i str a t ion use of the active in gredient (ai) spir otetram at (cis-3-(2,5-dimethlyphenyl)-

8-methox y

- 2

- oxo-1-azaspiro[ 4.5]dec

- 3-en-4-yl-ethyl carbonate]) on sod farms and golf course turf.

T h e most recent Section 3 human health risk assessment for s pirot etramat was fo r use on ornamentals 

## in r esidentia l settings

, and was comp l eted on 29 -0CT- 2013 (J. Va n Alstine et al.,

29

- 0CT -201 3 ; D405870). However, subsequent to com pl etion ofthis risk assessment, the use

on orname n tals was wit hdr awn by the Registrant. The risk assessment prior to the ornamental

use was fo r use on taro

, waterc re ss, pomegranates

, bananas, bulb vegetables

, low-growing
berries (except strawberries), bushberries

, globe artichokes, fruiting vegetables

, p om e fruit, citrus

f ruit

, pineapple s, and coffee, and was completed o n 17-APR

- 2013 ( J

. 

## Van A l st in e 

## et al.

, 17-

P age 1 of 14

### Spirotetramat Human Hea l th rusk Assessment 0413575



## APR

- 2013; 0398509)

.

It has been determined that a quantitative risk assessment i s not required at this time for the

current action since the proposed uses will not impact the aggregate assessment. The aggregate

assessment considers exposures from three major sources: food, drinking water, and res i dential

exposures

. A revised dietary assessment (food+ drinking water) is not needed for this registration request since to l erances are not required based on the proposed turfgrass use and the
estima t ed drinking water concentrations (EOWCs) used in 

## the previous dietary risk assessment

st ill represent the highest estimated drinking wa t er exposure scenarios (J

. 

## Melendez, 7

- MA Y

20 13; 0410121 ). In terms of residential exposures, there are no res i dent i al uses of sp i rotetramat
that requi r e a quantitative r i sk assessment. The proposed golf course use could result in potential
post

- application dermal exposure; however, since the l ast risk assessment for spirotetramat (J.



## Van Alstine et al., 29-0CT

- 2013; D405870), a reevaluation of the toxico l ogical database has

been conducted and yielded no dermal hazard

. Therefore, quantification of dermal risk is no t

necessary. Furthermore, the ornamental resident ia l use assessed in t h e last risk assessment was subsequent l y withdrawn by the Registrant; t herefore, there is no residential component for the
aggregate assessment of spirotetramat.

Because of the reasons above, a quantitative risk assessment has not been completed for spirotetramat for the current action

. A summary of the revised hazard assessment is prov i ded
herein since a reevaluation of the toxico l ogical database was conducted since the last risk

assessment. I n addition, the proposed sod farm and golf cou r se uses will result in occupational

exposure; therefore, an occupational/residential exposure assessment was conducted and can be
found in the following memo: K. 

## Lowe

, 16

- 0CT-2014; 0414958

.

Att achments:

Attac h ment A . R evised Hazard Charac t erizat i on and Dose-Response Assessment for Spirotetramat.
Attac h ment B. Toxico l ogy Profile and Executive Summa r ies

Page 2 of 14

### Spirotetramat Human Health Risk Assessment D 413575

Page 3 of 14 

## Attachment A. Re vised Hazard Characterization and Dose

- Response Assessment for Spirotetramat.

4.0 Hazard Characterization and Dose

- Response Assessment

Spirotetramat belongs to the class of cyclic ketoenol insecticides. The proposed insecticidal
mode of action for these agents is based on inhibition of acetyl

- CoA carboxylase, a key enzyme in fatty acid biosynthesis in eukaryotes and prokaryotes. Inhibition of lipogenesis, resulting in decreased lipid content (notably triglycerides and free fatty acids) in treated insect s, hinders the ability of younger insects to develop and ultimately their ability to reproduce as adults. The mammalian toxicity database for spirotetramat does not indicate changes in plasma lipid
parameters

, such as plasma levels of triglycerides and ch olestero l; therefore, it is unclear if this mode of action also operates in mammals.

4.1 Toxicology Studies Available for Analysis

The toxicology database for spirotetramat is adequate for characterization of its hazard, toxicity endpoint selection, and FQPA SF consideration. As part of the revised 40 CFR Toxicology Data Requirements, subchronic inhalation and subchronic neurotoxicity (SCN) studies are necessary for registered uses; however

, t he Hazard and Science Policy Council (HASPOC) concluded using a weight

- of

- evidence approach that these studies are not required at this time (TXR# 0056480)

. Although the SCN study is not required according to HASPOC, a SCN study has
been submitted

, but not reviewed (note that there are no neurotoxic effects seen in the SCN study after preliminary review)

.

4.2 Absorption, Distribution, Metabolism, and Excretion (ADME)

Absorption of orally

- administered spirotetramat in rats was rapid and nearly complete. In females, the time to maximum plasma levels ( T max 

## ) at the low dose tested of 2 mg/kg bw was estimated to be ~10 minutes, while at the high dose of 100 mg/kg bw, it was prolonged to 42 minutes. In males, T max 

## was between 0.7 and 1 hour at both dose levels. The extent of absorption was 89

- 98% by 48 hours following single low (2 mg/kg bw) and high (100 mg/kg bw) gavage doses, as well as repeated low doses (2 mg/kg bw). Systemic exposure estimated by the area under the plasma concentration

- time curve (AUC) was higher for male than female rats. At 2 mg/kg bw (single or repeated dosing), the AUC for males was about 60% higher than females and much greater at the high dose. Less than 0.2% of the administered dose was detected in the body at 48 hours after sacrifice and the highest concentrations were detecte d at similar levels in the liver and kidney. Spirotetramat was completely metabolized in the rat since no parent compound was detected in the excreta. The main metabolic reaction was cleavage of the ester group, which resulted in formation of the major m etabolite BYI 08330

- enol (53

- 87% of administered dose). All other identified metabolites could be derived from the primary enol metabolite. Male rats exhibited much higher rates of demethylation of BYI 08330

- enol to BYI
08330

- desmethyl

- enol (25

- 37%) whe n compared to female rats (5

- 10 %). Excretion of radioactivity was relatively rapid with 88

- 95% of the administered dose eliminated via urine and

2

- 11% via feces by 48 hours of exposure.

4.2.1 Dermal Absorption

### Spirotetramat Human Health Risk Assessment D 413575

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## A dermal

- absorption factor was not needed fo r this risk assessment because no dermal endpoint was chosen ( Section 4.5.1). An in vivo dermal penetration study in rats exists in the database
that can be used to estimate a dermal absorption factor in the future if needed

.

4.3 Toxicological Effects

The thyroid and thymus glands were target organs in oral subchronic and chronic toxicity studies in the dog , which was the most sensitive species . In su b chronic dog studies , lower circulating levels of the thyroid hormones T3 and T4 were seen in both sexe s . Males exhibited the effect at a lower dose than females. It was noted in ADME studies that males achieve much higher systemic exposures than their female counterparts, which helps explain the ir higher sensitivity to spirotetramat exposure

. Decreases in T4 were also observed in a one

- year study in dogs at a lower dose as compared to subchronic exposures. The only histopathological change in the thyroid was a reduction in follicle size in the high

- dose group, a possible indication of reduced
amount of colloid. There were no correlative changes in thyroid weight, thyroid histopathology, or thyroid

- stimulating hormone (TSH) in either sex. In all dog stud ies , there were decreases in thymus weights described microscopically as involution. In the one

- year chronic study, the thymus involution was only observed in male dogs. Although the toxicology database for spirotetramat shows effects in the thymus gland in dogs, the results of a recently submitted and reviewed immunotoxicity study in rats do es not indi cate any functional deficits in immune function. Thymus involution has been demonstrated to occur when hypothyroidism is induced in animals, so it is reasonable to conclude that the thymus involution in dogs was secondary to the thyroid effects rather tha n a direct effect on the immune system.

In rats, t he testes were the target organ following subchronic and chronic exposure to spirotetramat

. In a subchronic study, a bnormal spermatozoa and hypospermia in the epididymis, decreased testicular weight, and testicular degeneration and vacuolation were observed in males

at a relatively high dose (616 mg/kg bw/day). These effects proved to be reversible in most
animals after cessation of treatment. Following 12 or 24 months of oral exposure, a bnormal

sperm or spermatid degeneration in the testes and germ cell exfo liated debris in the epididymis were observed in male rats at the high dose ( ~400 mg/kg bw/day ) . In an investigative study with the enol metabolite of spirotetramat, spermatotoxicity, abnormal sperm, and Sertoli cell
vacuolation were observed in the testes

- epididymides of treated animals. In conjunction with the ADME data, which indicates that the parent chemical is completely metabolized to the enol metabolite, these data support the conclusion that male reproductive toxicity in rats is likely due to the enol metabolite of spirotetramat.

Other effects reported in rats were associated with the kidney. Kidney toxicity in the carcinogenicity study consisted of decreased organ weight and tubular dilation in both sexes . Similar kidney toxicity (decreased organ weight and histopathology) was also observed in parental animals of the F1 generation in the two

- generation rat reproduction toxicity study.

No adverse findings were observed in mice up t o the limit dose in subchronic and chronic studies with spirotetramat.

There was no evidence of increased quantitative susceptibility of offspring following pre

- or postnatal exposure to spirotetramat. In the rat developmental study, there is evidence of qualitative susceptibility with reduced fetal weight and increased incidences of malformations
and skeletal deviations observed in offspring along with decreased maternal body weight and
foo d consumption at the limit dose . In the rabbit developmental stu dy

, no developmental effects

### Spirotetramat Human Health Risk Assessment D 413575

Page 5 of 14 

## were seen. M aternal toxicity was evidenced by sacrifice of animals in moribund condition and clinical signs , such as alopecia, cold ears, altered fecal amount/appearance, and reddish excretions at the highest dose tested (160 mg/kg bw/day)

. Animals sacrificed in moribund
condition di splayed severe body

- weight loss, reduced food consumption, clinical signs, and gross pathological findings at necropsy. Late abortions were noted in only one female at the mid

- dose
and two females at the high

- dose . These females displayed body weight decrements and clinical signs before abortion that were similar to those seen in animals sacrificed in moribund condition indicating the abortions were secondary to maternal toxicity . In the reproduc tion toxicity study, offspring effects (decreased pup body weights) were observed at the same dose as parental effects (body

- weight decrements, decreased kidney weights, and kidney histopathology). Evidence of male reproductive toxicity consistent with th e subchronic and chronic studies in
adult rats was also observed in this study

.

In a 28

- day dermal toxicity study in rats, there were no effects (local or systemic) reported up to the limit dose.

Spirotetramat is classified as having low acute toxicity via the oral, dermal and inhalation routes (Toxicity Category III or IV). It was found to be an eye irritant and dermal sensitizer, but not a
dermal irritant.

In an acute neurotoxicity study in rats, the only evidence of neurotoxicity was d ecreased motor and locomotor activity which occurred only at relatively high

- dose levels (200 mg/kg bw) with no confirmatory findings of neurological pathology

. A preliminary review of a recently submitted SCN study in rats does not display any neurotoxic effects up to the highest dose tested.

There is no evidence of carcinogenicity in rats or mice

. Spirotetramat was also negative for mutagenicity and clastogenicity in sever al standard in vivo and in vitro assays.

4.4 Safety Factor for Infants and Children (FQPA SF)

The RAB1 risk assessment team recommends that the FQPA SF be reduced to 1X for all exposure scenarios based on the following considerations outlined in more det ail in the sections below: (1) the toxicology database for spirotetramat is adequate for FQPA SF consideration; (2) spirotetramat is not considered neurotoxic in mammals; (3) evidence of qualitative susceptibility to offspring is protected for by selected endpoints; and (4) the exposure databases are sufficient and unlikely to underestimate exposure.

4.4.1 Completeness of the Toxicology Database

The existing toxicology database for spirotetramat is complete for FQPA SF evaluation. The following acceptab le studies are available: developmental toxicity studies in ra ts and rabbits,

one

- and two

- generation reproduction toxicity studies in rats, and neurotoxicity studies in rats.

4.4.2 Evidence of Neurotoxicity

Although there is evidence of neurotoxicity i n the acute neurotoxicity study (see section 4.3),
concern is low since the effect s are well

- characterized with clearly established no

- observed
adverse

- effect level (NOAEL)/lowest

- observed adverse

- effect level (LOAEL) values

, the selected endpoints are pro tective of the observed neurotoxic effect

, there are no neurotoxic

### Spirotetramat Human Health Risk Assessment D 413575

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## effects seen in the SCN study after preliminary review, and the existing toxicological database indicates that spirotetramat is not a neurotoxic chemical.

4.4.3 Evidence of Sensitivity/Sus ceptibility in the Developing or Young Animal

There was no evidence of quantitative susceptibility of offspring following pre

- or postnatal exposure. There is evidence of qualitative susceptibility in the rat developmental study (see Section 4.3); however

, concern is low since offspring effects were only seen at the limit dose and selected endpoints are protective of the offspring effects.

4.4.4 Residual Uncertainty in the Exposure Database

The acute dietary food and drinking water exposure asse ssment utilizes existing and proposed
tolerance

- level residues, DEEM 7.81 default processing factors (where provided), and 100% CT information for all commodities. By using these screening

- level assessments, actual exposures/risks will not be underestimat ed. The chronic dietary food and drinking water exposure assessment utilizes average field trial residues for some commodities, tolerance

- level residues for the remaining commodities, and 100% CT. Some empirical processing factors were
used in the chroni c assessment along with DEEM 7.81 default processing factors (where provided). The chronic assessment is somewhat refined; however, since it is based on reliable data, it will not underestimate exposure/risk. The drinking water assessments utilize water
concentration values from the watercress application scenario generated by the Tier 1 Rice Model v.1.0. The resulting screening

- level EDWCs provide conservative, health

- protective, high

- end estimates of water concentrations that will not likely be exceede d. There are no residential exposures that have been quantitatively assessed for spirotetramat.

4.5 Toxicity Endpoint and Point of Departure Selections

4.5.1 Dose

- Response Assessment

Tables 4.5.4.1 and 4.5.4.2 summarize the toxicological doses and endpoints selected for human
health risk assessment. S ince the last risk assessment, dermal and inhalation exposure endpoints have been updated.

Acute Dietary Endpoint (General Population): The acute dietary endpoint was based on clinical signs and de creased motor activity observed in the acute neurotoxicity study at the LOAEL of 200 mg/kg bw (NOAEL = 100 mg/kg bw). The acute population

- adjusted do se (aPAD) of 1.0 mg/kg bw/day is derived from the NOAEL of 100 mg/kg bw and a 100

- fold uncertainty factor (10X for interspecies extrapola tion , 10X for intraspecies variation, and 1X for FQPA SF).

Chronic Dietary (All Populations): The chronic dietary endpoint was based on the thymus
involution in males in the chronic dog study at the LOAEL of 20 mg/kg bw /da y (NOAEL = 5 mg/kg bw/day). The chronic population

- adjusted dose (cPAD) of 0.05 mg/kg bw/day is derived from the NOAEL of 5 mg/kg bw/day and a 100

- fold uncertainty factor (10X for interspecies extrapolation , 10X for intraspecies variation, and 1X for FQPA SF).

Dermal (Short

- and Intermediate

- Term ) : A dermal endpoint was not selected. Dermal endpoints were chosen in previous risk assessments; however

, reevaluation of the toxicological database yielded no dermal hazard. Since there is no concern for enhanced sensitivity/susceptibility to offspring from spirotetramat exposure, the effects of concern in adults and offspring are

### Spirotetramat Human Health Risk Assessment D 413575

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## adequately assessed in the dermal toxicity study where no adverse dermal or systemic effects were observed up to the limit dose.

Inhalation (Short

- and Intermediate

- Term) : The 90

- day oral toxicity study in dogs was selected
to evaluate inhalation exposures (NOAEL = 32 mg/kg bw/da y). The LOAEL of 72 mg/kg bw/day is based on depressed red blood cell parameters, thymus atrophy

, and decreased food consumption. The LOC for inhalation exposure scenarios is for MOEs <100 ( 10X for interspecies extrapolation and 10X for intraspecies vari ation). Since the NOAEL for inhalation risk assessment was chosen from an oral study, it was assumed that toxicity via the inhalation route is equivalent to oral toxicity.

4.5.2 Recommendation for Combining Routes of Exposures for Risk Assessment

Sinc e there are no dermal endpoints for spirotetramat, dermal and inhalation exposures do not need to be combined for occupational assessments.

4.5.3 Cancer Classification and Risk Assessment Recommendation

There is no evidence of carcinogenicity in chronic toxicity/carcinogenicity studies performed in rats and mice. Spirotetramat has been classified as “not likely to be carcinogenic to humans” based on lack of evidence for carcinogenicity in rodent studies.

4.5.4 Summary of Points of Departure and Toxici ty Endpoints Used in Human Risk
Assessment

Table 4.5.4.1. Summary of Toxicological Doses and Endpoints for Spirotetramat for Use in Dietary and Non

Occupational Human Health Risk Assessments.

Exposure Scenario POD Uncertainty/FQPA Safety Factors RfD, PAD, LOC for

Risk Assessment Study and Relevant Toxicological Effects

Acute Dietary (General Population, including Infants and Children) NOAEL =
100 mg/kg/day UF A = 10X

UF H = 10X FQPA SF = 1X aRfD = aPAD = 1.0

mg/kg/day Acute neurotoxicity (rat)
LOAEL = 200 mg/kg based on clinical signs and decreased motor activity in males.

Chronic Dietary (All Populations) NOAEL = 5 mg/kg/day UF A = 10X

UF H = 10X FQPA SF = 1X cRfD = cPAD = 0.05

mg/kg/day Chronic toxicity (dog ) LOAEL = 20 mg/kg/day based on thymus involution in males.

Dermal
Short

- and
Intermediate

- Term (1

- 30 days and 1

- 6 months) In a dermal toxicity study in rats, no adverse effects were observed up to the limit dose. Since there
is no evidence of increased susceptibility, a dermal assessment is not needed

.

Inhalation
Short

- and
Intermediate

- Term (1

- 30 days and 1

- 6 months) NOAEL = 32 mg/kg/day

Inhalation toxicity is
considered equivalent to oral toxicity. UF A = 10X

UF H = 10X FQPA SF = 1X Residential LOC for

MOE =100 90

- Day oral toxicity (dog) LOAEL = 72 mg/kg/day [F] based on depressed RBC parameters (red blood cell count, hemoglobin level and hematocrit), thymus atrophy and decreased food consumption

. Cancer (Oral, Dermal, Inhalation) Classification: “not likely to be carcinogenic to humans” based on lack of evidence of carcinogenicity in rats and mice. Abbreviations: UF = uncertainty factor, UF A = extrapolation from animal to human (interspecies), UF H = potential variation in sensit ivity among
members of the human population (intraspecies), FQPA SF = FQPA Safety Factor, NOAEL = no

- observed adverse

- effect level, LOAEL =
lowest

- observed adverse

- effect level, RfD = reference dose (a = acute, c = chronic), PAD = population

- adjusted dose, LOC = level of concern.

### Spirotetramat Human Health Risk Assessment D 413575

Page 8 of 14 Table 4.5.4.2. Summary of Toxicological Doses and Endpoints for Spirotetramat for Use in Occupational Human
Health Risk Assessments.

Exposure Scenario POD Uncertainty Factors LOC for Risk
Assessment Study and Toxicological Effects

Dermal
Short

- and
Intermediate

- Term (1

- 30 days and 1

- 6 months) In a dermal toxicity study in rats, no adverse effects were observed up to the limit dose. Since there
is no evidence of increased susceptibility, a dermal assessment is not needed

.

Inhalation
Short

- and
Intermediate

- Term (1

- 30 days and 1

- 6 months) NOAEL = 32 mg/kg/day

Inhalation toxicity is
considered equivalent to oral toxicity. UF A = 10X

UF H = 10X Occupational LOC for

MOE =100 90

- Day oral toxicity (dog) LOAEL = 72 mg/kg/day [F] based on depressed RBC parameters (red blood cell count, hemoglobin level and hematocrit), thymus atrophy and decreased food consumption

. Cancer (oral, dermal, inhalation) Classification: “not likely to be carcinogenic to humans” based on lack of evidence of carcinogenicity in rats and mice. Abbreviations: UF = uncertainty factor, UF A = extrapolation from animal to human (interspecies), UF H = potential variation in sensitivity among members of the human population (intraspecies), NOAEL = no

- observed adverse

- effect level, LOAEL = lowest

- observed adverse

- effect level, MOE = margin of exposure, LOC = level of concern.

### Spirotetramat Human Health Risk Assessment D 413575

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## Attachment B . Toxicol ogy Profile and Executive Summaries

A.1 Toxicology Data Requirements

The requirements (40 CFR 158.340) for spirotetramat are presented below.

Test Technical Required Satisfied

870.1100 Acute Oral Toxicity
870.1200 Acute Dermal Toxicity
870.1300 Acute Inhalation Toxicity
870.2400 Primary Eye Irritation
870.2500 Primary Dermal Irritation
870.2600 Dermal Sensitization yes

yes
yes
yes
yes

yes yes

yes
yes
yes
yes

yes 870.3100 Oral Subchronic (rodent)
870.3150 Oral Subchronic (nonrodent)
870.3200 21 /28

- Day Dermal
870.3250 90

- Day Dermal
870.3465 90

- Day Inhalation yes
yes
yes

no

no 1 yes
yes
yes

-

- 870.3700a Developmental Toxicity (rodent)
870.3700b Developmental Toxicity (nonrodent)
870.3800 Reproduction and Fertility Effects yes

yes
yes yes

yes
yes 870.4100a Chronic Toxicity (rodent)
870.4100b Chronic Toxicity (nonrodent)
870.4200a Oncogenicity (rat)
870.4200b Oncogenicity (mouse)
870.4300 Chronic/Oncogenicity yes
yes

yes
yes
yes yes
yes

yes
yes
yes 870.5100 Mutagenicity

— Gene Mutation

- bacterial
870.5300 Mutagenicity

— Gene Mutation

- mammalian
870.5400 Mutagenicity

— Structural Chromosomal Aberrations
870.5500 Mutagenicity

— Other Genotoxic Effects yes

yes
yes

no yes

yes
yes

- 870.6100a Acute Delayed Neurotox. (hen)
870.6100b 90

- Day Neurotoxicity (hen)
870.6200a Acute Neurotoxicity Screening Battery (rat)
870.6200b 90

- Day Neuro xicity . Screening Battery (rat)
870.6300 Develop. Neurotoxicity no

no
yes

no

no -

- yes
yes

- 870.7485 General Metabolism
870.7600 Dermal Penetration yes
yes yes
yes

1 The Hazard and Science Policy Council (HASPOC) met on 04

- OCT

- 2012 (TXR# 0056480) and concluded that a repeated exposure inhalation toxicity study is not required for spirotetramat at this time.

### Spirotetramat Human Health Risk Assessment D 413575

Page 10 of 14 

## A.2 Toxic ity Profiles

Table A.2.1. Acute Toxicity Profile

– Spirotetramat Technical.

Guideline No. Study Type MRID(s) Results Toxicity Category

870.1100 Acute oral (rat) 46904527 LD 50 >2000 mg/kg (F) III

870.1200 Acute dermal (rat) 46904529 LD 50 > 2000 mg/kg (M&F) III

870.1300 Acute inhalation (rat) 46904530 LC 50 >4.183 mg/L (M&F) IV

870.2400 Primary eye irritation (rabbit) 46904531 Corneal opacity and iritis (grade 1); cleared by day
8 II

870.2500 Primary dermal irritation (rabbit) 46904532 Negative IV

870.2600 Dermal sensitization (mouse) 46904565 Positive (LLNA) N/A

Table A.2.2. Subchronic and Chronic Toxicity and Genotoxicity Profile

– Spirotetramat Technical.

Guideline
No. Study Type MRID No. (year)/
Classification/Doses Results

870.3100 28

- Day oral toxicity

(mouse) 46904536 (2001) Acceptable/non

guideline 0, 500, or 5000 ppm (equivalent to 0, 136.5, or 1415 mg/kg bw/day [M]) NOAEL = 5000 ppm (1415 mg/kg/day [M]).
LOAEL not observed.

870.3100 90

- Day oral toxicity

(mouse) 46904539 (2005) Acceptable/guideline 0, 70, 350, 1700, or 7000 ppm (equivalent to 0/0, 12.8/16, 59.6/72.4, 300/389, or 1305/1515 mg/kg bw/day [M/F]) NOAEL = 7000 ppm (1305/1515 mg/kg/day [M/F]).
LOAEL not observed.

870.3100 28

- Day oral toxicity

(rat) 46904537 (1998) Acceptable/non

guideline 0, 500 or 5000 ppm (equivalent to 0, 47.3, or 501.8 mg/kg bw/day [F]) NOAEL = 5000 ppm (501.8 mg/kg/day [F]).
LOAEL not observed.

870.3100 90

- Day oral toxicity

(rat) 46904538 (2005) Acceptable/guideline 0, 150, 600, 2500, or 10000 ppm (equivalent to 0/0, 9/11, 26/46, 148/188, or 616/752 mg/kg bw/day [M/F]) NOAEL = 2500 ppm (148/188 mg/kg/day [M/F]). LOAEL = 10000 ppm (616/752 mg/kg/day [M/F]), based on decreased body weight, abnormal spermatozoa and hypospermia i n the epididymis, decreased testicular weight, and testicular degeneration and vacuolation in males, and alveolar macrophages in both sexes.

870.3150 28

- Day oral toxicity

(dog) 46904572 (2004) Acceptable/non

- guideline 0, 100, 400, 1600, or 6400 ppm (equi valent to 0/0, 3/3, 13/12, 42/70, or 104/127 mg/kg/day [M/F]) NOAEL = 1600 ppm (42/70 mg/kg/day [M/F]). LOAEL = 6400 ppm (104/127 mg/kg/day [M/F]) based on decreased thymus size and weight as well as decreased body weight and food consumption, which
resulted in emaciation.

870.3150 90

- Day oral toxicity

(dog) 46904541 (2005) Acceptable/guideline 0, 150, 300, 1200, or 4000/2500

ppm (equivalent to 0/0, 5/6, 9/10, 33/32, or 81/72 mg/kg bw/day) NOAEL = 1200 (32 mg/kg/day) [F] & 2500 ppm (81 mg/kg/day) [M ].
LOAEL = 2500 ppm (72 mg/kg/day) [F] based on decreased food consumption, depressed RBC parameters (red blood cell count, hemoglobin level

. and hematocrit), and thymus atrophy.

870.3200 28

- Day dermal toxicity

(rat) 46904542 (2006) Acceptable/guideline 0, 100, 300, or 1000 mg/kg bw/day (limit dose) NOAEL = 1000 mg/kg/day.
LOAEL not established.

870.3700a Prenatal developmental

(rat) 46904543 (2004) Acceptable/guideline 0, 20, 140, or 1000 mg/kg bw/day Maternal NOAEL = 140 mg/kg/day.
LOAEL = 1000 mg/kg/day based on impaired food consumpti on, transient body weight loss , and reduced final body weight. Developmental NOAEL = 140 mg/kg/day. LOAEL = 1000 mg/kg/day based on reduced fetal weight and increased incidences of malformations and
skeletal variat ions.

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Page 11 of 14 Table A.2.2. Subchronic and Chronic Toxicity and Genotoxicity Profile

– Spirotetramat Technical.

Guideline
No. Study Type MRID No. (year)/
Classification/Doses Results

870.3700b Prenatal developmental

(rabbit) 46904544 (2004) Acceptable/guideline 0, 10, 40, or 160 mg/kg bw/day Maternal NOAEL = 4 0 mg/kg/day.
LOAEL = 16 0 mg/kg/day based on sacrifice of
animals in moribund condition and clinical signs Developmental NOAEL = 160 mg/kg/day.
LOAEL not observed.

870.3800 1

- gen. reproduction and

fertility effects (rat)

– range finding 46904571 (2006) Acceptable/non

- guideline 0, 200, 500, 6000, or 10000 ppm (equivalent to 0/0, 10.5/12.8, 27.8/31.4, 320.1/384.1, or 537.9/645.7 mg/kg bw/day [M/F]) Parental/Systemic NOAEL = 500 ppm (27.8 and 31.4 mg/kg bw/day [M/F]).
LOAEL = 6,000 ppm (320.1 and 384.1 mg/kg bw/day [M/F]) based on decreased terminal body weight (F 1 males). Reproductive NOAEL = 500 ppm (27.8 mg/kg bw/day [M]) and 10,000 ppm (645.7 mg/kg/day) [F].
LOAEL = 6,000 ppm (320.1 mg/kg bw/day [M]) based on decreased sperm motility and progression and increased abnormal sperm cells in the F 1 males. Offspring NOAEL = 500 ppm (27.8 and 31.4 mg/kg bw/day [M/F]).
LOAEL = 6,000 ppm (320.1 and 384.1 mg/kg bw/day [M/F]) based on decreased body weight during
lactation in F 1 pups. 870.3800 2

- gen. reproduction and

fertility effects (rat) 46904546 (2006) Acceptable/guideline 0, 250, 1,000, or 6,000 ppm (equivalent to 0/0, 17.2/20, 70.7/82.5, or 419.3/484.7 mg/kg bw/day [M/F]) Parental/Systemic NOAEL = 1000 ppm (70.7/82.5 mg/kg/day [M/F]). LOAEL = 6000 ppm (419.3/484.7 mg/kg/day [M/F])
based on decreases in body weight (F 1 males and females), and food consumption during lactation (P

and F 1

- generation females), and kidney histopathology and decreased kidney weights (F 1 males and females)

. Reproductive NOAEL = 1000 ppm (70.7 mg/kg/day) [M] & 6000 ppm (484.7 mg/kg/day) [F].

L OAEL = 6000 ppm (419.3 mg/kg/day) [M] based on abnormal sperm cells and decreased reproductive performance in the F 1 males. Offspring NOAEL = 1000 ppm (70.7/82.5 mg/kg/day [M/F]). LOAEL = 6000 ppm (419.3/484.7 mg/kg/day [M/F])
based on decreased body weigh t during lactation in
both F 1 and F 2 generations

. 870.4100 Chronic toxicity (1 year;

dog) 46904548 (2006) Acceptable/guideline 0, 200, 600, or 1800 ppm (equivalent to 0/0, 6/5, 20/19, or 55/48 mg/kg/day [M/F]) NOAEL = 200 ppm (6 mg/kg/day) [M] & 1800 ppm (48 mg/kg/day) [F]. LOAEL = 600 ppm (20 mg/kg/day) [M] based on thymus involution [M] and not observed [F].

870.4100 Chronic toxicity

(1 year; rat) 46904547 (2005) Acceptable/guideline 0, 250, 3500, or 7500/12000 ppm (M/F) (equivalent to 0/0, 13.2/18,

1 89/255, or 414/890 mg/kg bw/day [M/F]) NOAEL = 250 ppm (13.2 mg/kg/day) [M] & 3500 ppm (255 mg/kg/day) [F]. LOAEL = 3500 ppm (189 mg/kg/day)[M] based on
dose

- dependent increase in alveolar macrophages; & 12000 ppm (890 mg/kg/day)[F] based on decreased body weight , alveolar macrophages, discoloration of the lung, and yellow/brown staining of the perigenital
area and tail.

870.4200 Carcinogenicity

(rat) 46904549 (2006) Acceptable/guideline 0, 250, 3500, or 7500/12000 ppm (M/F) (equivalent to 0/0, 12.5/16.8, 169/229, or 373/823 mg/kg bw/day [M/F]) NOAEL = 250 ppm (12.5/16.8 mg/kg/day [M/F]). LOAEL = 3500 ppm (169/229 mg/kg/day [M/F]) based on decreased kidney weight and renal tubular
dilation.

No evidence of carcinogenicity.

870.4200 Carcinogenicity

(mouse) 46904550 (2006) Acceptable/guideline 0, 70, 1700, or 7000/6000 ppm (M/F) (equivalent to 0/0, 10.9/13.7, 263/331, or 1022/1319 mg/kg/day [M/F]) NOAEL = 7000/6000 ppm (1022/1319 mg/kg/day [M/F]).
LOAEL not observed.

No evidence of carcinogenicity.

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Page 12 of 14 Table A.2.2. Subchronic and Chronic Toxicity and Genotoxicity Profile

– Spirotetramat Technical.

Guideline
No. Study Type MRID No. (year)/
Classification/Doses Results

870.5100 Bacterial Gene

Mutation 46904551 (2006) Acceptable/guideline 0, 16, 50, 158, 500, 1581, or 5000 μg/plate +/

- S9 activation Negative.

870.5100 Bacterial Gene

Mutation 46904552 (2002) Acceptable/guideline 0, 16, 50, 158, 500, 1581, or 5000 μg/plate +/

- S9 activation Negative.

870.5300 Mammalian Gene

Mutation 46904553 (2002) Acceptable/guideline 0, 2.5, 5, 10, 20, 30, 40, 50, 60, 70,

or 80  g/mL (

- S9) 0, 20, 40, 60, 80, 92, 100, 108, 116, 120, 124, 132, or 140  g/mL (+S9) Negative.

870.5375 In vitro mammalian

chromosome aberration 46904554 (2002) Acceptable/guideline 0, 10, 12, 24, 30, 48, or 50 μg/mL (

- S9) 0, 20, 40, or 80 μg/mL (+S9) Weakly clastogenic at cytotoxic concentrations only.

870.5375 In vitro mammalian

chromosome aberration 46904 555 (2003) Unacceptable/guideline 0, 30, 50, 70, 90, or 110 μg/mL (

- S9) 0, 40, 60, 80, 100, or 120 μg/mL (+S9) N/A

870.5395 In vivo erythrocyte

micronucleus assay (mouse) 46904556 (2002) Acceptable/guideline 0, 125, 250, or 500 mg/kg bw Negative.

870.5385 In vivo bone marrow

chromosomal aberration assay (mouse) 46904558 (2003) Acceptable/guideline 0, 125, 250, or 500 mg/kg bw Negative.

870.5550 In vivo / in vitro UDS

assay (rat hepatocytes) 46904557 (2003) Acceptable/guideline 0, 1000, or 2000 mg/kg bw Negative.

870.6200a Acute neurotoxicity

screening battery 46904560 (2005) Acceptable/guideline 0, 50, 100, 200, 500, or 2000 mg/kg
bw NOAEL = 100 mg/kg/day. LOAEL = 200 mg/kg/day based on clinical signs (males and females) and decreased motor activity (males).

870.6200b Subchronic

neurotoxicity screening battery 48958802 (2012) Pending formal review.

870.7485 Metabolism and

pharmacokinetics (rat) 46904504 (2006) Acceptable/guideline 2 or 100 mg/kg bw (single) 2 mg/kg bw (repeat) Absorption: 89

- 98% after 48 hrs (no si gnificant differences among low

- dose, high

- dose, and repeated

dose tests).
Distribution: AUC 0

- ∞ (measure of systemic exposure) slightly higher for males than females; <0.2% of administered dose detected in body 48 hrs after

sacr ifice; highest equivalent concentrations detected in liver and kidney. Metabolism: parent compound undetected in urine and feces of all tests; main metabolic reaction was cleavage of the ester group which resulted in formation of the primary metabolite BY I 08330

- enol (53

- 87% of administered dose); all other identified metabolites could be derived from enol; male rats exhibited much higher rates of demethylation of BYI
08330

- enol to BYI 08330

- desmethyl

- enol (25

- 37%) vs. females rats (5

- 10%).
Excretion: 88

- 95% of administered dose eliminated
via urine and 2

- 11% via feces within 48 hrs.

870.7485 Metabolism and

pharmacokinetics 46904561 (2006) Acceptable/guideline At 1000 mg/kg bw: Absorption and excretion less
than low dos e, with 27% of dose excreted in urine

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Page 13 of 14 Table A.2.2. Subchronic and Chronic Toxicity and Genotoxicity Profile

– Spirotetramat Technical.

Guideline
No. Study Type MRID No. (year)/
Classification/Doses Results

(rat) 2 or 1000 mg/kg bw (single) after 24 hours (18% in feces); plasma radioactivity slightly higher than in liver and kidney; these results are consistent with saturation of cellular transport mechanisms. Tissue radioactivity decreased from 1 to
24 hours post dose. Metabolism profile qualitatively
similar to that of the low

- dose group; BYI 08330

- enol was most prominent metabolite; similar to low

- dose group, BYI 08330

- desmethyl

- enol levels greater in urine than in plasma and organs.

870.7600 In vivo dermal

penetration (rat) 46904563 (2006) Acceptable/guideline 100, 15, or 5 μg ai/cm 2 (OD150 formulation) Dermal absorption = 10%.

870.7800 Immunotoxicity

(rat) 48525901 (2011) Acceptable/guideline 500, 2500, or 12,000 ppm (equivalent to 33, 164, or 795 mg/kg/day) The functional immunotoxicity NOAEL is 12,000 ppm (equivalent to 795 mg/kg/day, the highest dose tested); the LOAEL was not established.

T he systemic toxicity LOAEL for s pirotetramat in male rats is 12,000 ppm (equivalent to 795 mg/kg bw/day) based on decreased body weights, lowered food consumption, lower thymus weight, and observations of atrophic thymus. The NOAEL for systemic toxicity is 2500 ppm (equivalent to 164 mg/kg bw/day) Special study Male reproductive

toxicity 46904569 (2005)
Ac ceptable/non

- guideline 1000 mg/kg bw/day (3, 10, 21, or 41 days) Primary testicular effects on or after day 10 were degeneration of round and elongating spermatids (stage 7

- 8 and 9

- 14, respectively), decreased sperm count, and increased numbers of aberrant /abnormal spermatozoa in the epididymis.

Table A.2.3. Toxicity Profile

– Metabolites.

Guideline
No. Study Type MRID No. (year)/
Classification /Doses Results

Exploratory study 10

- day gavage (enol

metabolite) 46904601 (2006) Acceptable/non

- guideline 800 mg/kg bw/day No treatment related effects; only limited parameters measured; h istopathology not evaluated.

Special study 21

- day gavage (enol

metabolite) 47070901 (2006) Acceptable/non

- guideline 800 mg/kg bw/day Clinical signs of toxicity and testicular/spermatotoxicity.

870.5100 Bacterial Gene

Mutation (ketohydroxy metabolite) 46904594 (2005) Acceptable/guideline 0, 16, 50, 158, 500, 1581, or 5000 μg/plate +/

- S9 activation Negative.

870.7485 Metabolism and

Pharmacokinetics (rat) (ketohydroxy metabolite) 46904595 (2006) Acceptable/guideline 2 mg/kg bw (single) Absorption: ≥55% after 48 hrs. Distribution: highest concentrations detected in GI tract, liver, and kidney; <0.2% of administered dose detected in body 48 hrs post dose. Metabolism: parent compound undetected in urine and trace amounts in feces; main metabolic reaction was oxidative demethylation of cyclohexyl

- O

- methyl group to form desmethyl

- ketohydroxy metabolite (15% of administered dose); all other identified metaboli tes could be derived from desmethyl

- ketohydroxy metabolite.
Excretion: 54% of administered dose eliminated via
urine and 44% via feces within 48 hrs.

870.5100 Bacterial Gene

Mutation (desmethyl

ketohydroxy metabolite) 46904597 (2006) Acceptable/guideline 0, 16, 50, 158, 500, 1581, or 5000 μg/plate +/

- S9 activation Negative.

### Spirotetramat Human Health Risk Assessment D 413575

Page 14 of 14 Table A.2.3. Toxicity Profile

– Metabolites.

Guideline
No. Study Type MRID No. (year)/
Classification /Doses Results

870.5100 Bacterial Gene

Mutation (dihydroxy metabolite) 46904599 (2005) Acceptable/guideline 0, 16, 50, 158, 500, 1581, or 5000 μg/plate +/

- S9 activation Negative.

870.7485 Metabolism and

Pharmacokinetics (rat) (enol glucoside metabolite) 46904602 (2006) Acceptable/non

- guideline 0.1 mg/kg bw (single; 1 rat) Absorption: 54% after 48 hrs. Distribution: plasma concentrations peaked 4 hrs post dose; 1% of administered dose det ected in body 48 hrs post dose. Metabolism: parent compound detected in feces (21%); main metabolite was enol (64% of administered dose).
Excretion: 53% of administered dose eliminated via
urine and 44% via feces within 48 hrs.

870.5100 Bacterial Gene

Mutation (monohydroxy metabolite) 46904604 (2005) Acceptable/guideline 0, 16, 50, 158, 500, 1581, or 5000 μg/plate +/

- S9 activation Negative.