### UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON, D.C

. 20460

MEMORANDUM

DATE: Novem ber 21, 2014 OFFICE OF CHEMICAL SAFETY AND POLLUTION PREVENTION



### SUBJECT: Cyazofamid. Human Health Assessment Scoping Document in Support of Registra t ion Review.

PC Code: 085651 Decision No. : 495518 Petition No.: NA Risk Assessment Type: NA
TXRNo .: NA MRIDNo.:NA DP Barcode: D422931 Registration No .: NA Regulatory Action: Registra t ion Review Case No.: NA CAS No.: 120116

- 88-3 40 CFR: § 180

. 60 I

FROM: Cass i Walls, Senior

## Biologist**~** ;2..

## l...J:°'~ Nancy Dodd, Chemist

## ~**~** Whang Phang

, Senior

## To**xicologist /¥~**  Kris t in Rury, Biolo gis t

## V-~ Risk Assessment Branch III

### (RA**~** He a l th Effec ts Division (7509P) Office of Pesticide Programs

THROUGH: Christine Olinger, Branch

# Chie~ 

# & /;

Risk Assessment Branch III (RAB3)

## o~-­ Health Effe cts Division (7509P) Office of Pesticide Programs

TO: Jose Gayoso Chemical Review Manager Risk Management and Implementation Branch II (RMIB2) Pest ic ide Re-Evaluation Division (PRD) (7508P) Office of Pesticide Programs

E xecutive Summary

Attached is the Health Effects Division's (HED's) human health ri s k assessment scoping document for the fungicide cyazofarnid to support Registration Review. To evaluate th e scope of work necessary to support Registration Review

, HED has considered recent risk assessments for cyazofamid, updates to i ts toxicity, exposure and usage databases, and the latest Agency science policy and risk assessment methodologies. The most recent risk assessment for cyazofamid was conducted in 2012 for numerous new uses (D397213, N. Tsaur, 06/28/2012).

Page 1 of 25

### Cyazofamid is currently registered for use on numerous agricu l tural crops; golf course turf, sod farms, seed farms, college and professional sports fields; residential and commercial lawns; and ornamentals in greenhouses, fields, and resident i al areas.

The toxico l ogy database for cyazofamid is complete and adequate for risk assessment. On January 8, 2015, HED's Hazard and Science Policy Council (HASPOC), based on a weight of evidence approach considering all the available hazard and exposure information granted a waiver for the subchronic neurotoxicity and the immunotoxicity studies that are required under the 2007 40 CFR t oxicology data requirements (TXR No. 0056341 ).

Based on the toxicology database, cyazofamid has no consistent target organ of toxicity

. In repeated dose studies, the kidney was the target organ of cyazofamid toxicity in rats. In dogs, there were no major toxicity findings. Based on the available data

, there are no concerns for neurotoxicity, immunotoxicity, or mutagenicity. Cyazofamid is classified as "not likely to be carcinogenic to humans" based on the l ack of evidence of carcinogenicity in both the rat and the

mouse studies.

Based on the avai l able data

, toxicity endpoints and points of departure (PODs) have been selected for acute dietary (for females 13-49 years of age), chronic dietary, short

- and intermediate-term incidenta l oral, short

- and intermediate-term dermal (for adults), and short- and intermediate-term inhalation exposure scenarios. Since no observed effects could be attributed
to a single dose, an acute endpoint for acute dietary exposure was not estab li shed for the general population. Dermal absorption is 37%. T he 1 

### OX Food Quality Protection Act (FQPA) Safety Factor was removed (i.e, IX) because there are no residual uncertainties for pre- and post

- natal toxicity and there are no residual uncertainties in the exposure database.

The residue chemistry database is sufficient to support the current registrations of cyazofamid. The nature of the residue in plants and livestock has been adequately defined. Adequate field trial/processing data, rotational crop data, enforcement methods are available. The storage stab ili ty data are under review.

New dietary exposure and aggregate risk assessments are not anticipated during Registration Review, unless there are significant increases to the drinking water exposure estimates that need
to be incorporated

.

New residential hand l er and post-application exposure and risk assessments are not anticipated to be needed during Registration Review

. However, occupational handler and post-application
exposure and risk assessments are needed during Registration Review to account for recent changes in exposure assessment assumptions, including body weight

, amount treated, and transfer coefficients.

The Agency will examine the need for spray drift and volatilization assessments during Registration Review for cyazofamid based on the potential for exposure to bystanders from agricultura l applications.

Page 2 of25

### Page 3 of 25

I ntroduction

Cyazofamid is a fungicide which belongs to a no vel chemical class based on the cyanoimidazole moiety. Cyazofamid is a broad spectrum fungicide active against oomycete fungi (such as Phytophthora, Plasmopara, Pseudoperonospora, and Pythium ) and Plasmodiophoromycetes ( Plasmodiospora brassicae ). It spec ifically interferes with the cytochrome bc 1 

### complex (ubiquinol cytochrome c oxidoreductase) in the mitochondrial respiratory chain of oomycetes fungi.

The most recent human health risk assessment for cyazofamid was finalized in 20 12 for

numerous new use s ( D 397213

, N

. Tsaur
, 0 6/28/12 ). Cyazofamid is currently registered on a variety of food crops including, potatoes, basil, peppers, cucumber, squash, eggplant, tomatoes, broccoli, cabbage, and ornamental plants and turf. Homeowners may apply cyazofamid to residential turf and ornamental plants

. Cyazofamid end use products are formulated as soluble
concentrate s

. Applications can be made with ground, aerial, chemigation, and hand he ld spray equipment

. Refer to Attachment 1 (Table 1) for the chemical identi ty of cyazofamid

. Hazard Identification/Toxicology

The toxicology database for cyazofamid is adequate for risk assessment, e xcept for the immunotoxicity and subchronic neurotoxicity studies. All other toxicity data requirements for conventional pesticide registration under the current 40 CFR §158.500 have been satisfied. All studies needed to assess susceptibility concerns, such as dev elopmental and reproduction studies,

were submitted. On January 8, 2015, t he HASPOC granted waiver for the subchronic neurotoxicity and the immunotoxicity studies ( TXR#0056341). The acute neurotoxicity study tested up to the limit dose (2000 mg/kg) did n ot yield any neurotoxic effect, and the existing data did not suggest cyazofamid to affect the parameters generally related to immunotoxicity.

Cyazofamid belongs to a chemical class based on the cyanoimidazole moiety. It specifically interferes with the cytochrome bc 1 

### complex (ubiquinol cytochrome c oxidoreductase) in the mitochondrial respiratory chain of oomycetes fungi. The mechanism of toxicity in mammal s is not clear. However, the toxicology database ind icates cyazofamid has relatively low toxicity via all routes of administration (see Attachment 5 for the complete toxicity profile tables for cyazofamid ). In some studies the adverse effects were seen at relatively high dose levels, some of which were eith er approaching the limit dose or at the limit dose. Cyazofamid has no specific target organ of toxicity. For example in rats, cyazofamid was found to produce kidney effects characterized by an increase in the incidence of basophilic kidney tubules in the 2 8

- day and 90

- day oral toxicity studies. However, similar kidney finding was not observed following life time exposure at doses approaching the limit dose (1000 mg/kg/day). I n the mouse carcino genicity study, skin lesions were found following repeated life time oral dosing at the limit dose (1000 mg/kg/day). In the dog 1

- year study , the overall toxicity produced by cyazofamid seemed to be limited to an increase in the incidence of parathyroid cysts in the h igh dose males (1000 mg/kg/day).

Cyazofamid has a low order of acute toxicity via the oral, dermal, and inhalation routes of
exposure. It produces minimal but reversible eye irritation, is a slight dermal irritant, a nd is a

### Page 4 of 25

weak dermal sensitizer. Cyazofamid is classified as “not likely to be carcinog enic to humans” based on the lack of evidence of carcinogenicity in both the rat and the mouse studies.

The database for FQPA assessment is complete. The pre

- and post

- natal toxicology database for cyazofamid includes rat and rabbit developmental toxicity studies and a two

- generation reproduction toxicity study in rats. There was no evidence of increased susceptibility following
pre

- natal exposure to r abbits at the Limi Dose (1000 mg/kg/day) or following pre and post natal exposure to rats at the Limit Dose in a two generation reproduction study. In the pre

- natal developmental toxicity study in rats, evidence for increased susceptibility manifested as i ncreased incidences of fetuses with bent ribs was at the Limit Dose in the absence of any maternal toxicity. The concern for this effect, however, is low since the effects were seen only at the Limit Dose (i.e, no dose response) and the observed effects ar e reversible anomaly rather than frank malformation. Thus, there are no residual uncertainty for pre

- natal toxicity. Consequently the 10X FQPA Safety Factor is removed (i.e, 1X).

Refer to Attachment 4 for a summary of the toxicity endpoi nts and PODs for c yazofamid and Attachment 5 for the complete toxicity profile tables for cyazofamid.

Conclusions: The toxicology database of cyazofamid is adequate for risk assessment. Additional toxicity data are not required during Registration Review.

Dietary Expos ure
The nature of the residue in plants and livestock has been adequately defined

. Adequate metabolism studies on grape, potato, tomato, goat and hen are available. The residues of concern for the dietary risk assessment and the tolerance expression are parent and CCIM in plants, and parent only in rotational crops. The residues of concern in water are parent, CCIM, CCIM

- AM, and CTCA. No livestock tolerances have been established.

A dequate enforcement methods are available. Cyazofamid and the met abolite CCIM are completely recovered (>80% recovery) in non

- fatty commodities using FDA’s Multi r esidue Protocol D (without cleanup). The multiresidue method can be used as an enforcement method. In addition, a high performance liquid chromatography/ultra violet ( HPLC/UV ) detection method (MRID 45409020; revised in MRID 46550401) is acceptable for use as a single analyte enforcement method provided a confirmatory method such as liquid chromatography with tandem mass spectrometry ( LC/MS/MS ) is used.

Adequate field trial and processing data are available to support the established tolerances. The storage stability data are currently under review.

Adequate rotat ional crop data are available. A confined accumulation in rotational crops study indicated that c yazofamid and structurally related metabolites are not expected to be present at a level greater than 0.01 ppm in/on three representative rotational crop commodities at a plantback interval of 30 days. The study supports the 30

- day plantback interva l currently on the labels. Based on the confined rotational crop stud y, a field accumulation in rotational crops study is not required and rotationa l crop tolerances are not needed.

### Page 5 of 25

The registered uses fall under 40 CFR §180.6(a)(3) for livestock commodi ties, i.e., although it is not possible to establish with certainty whether finite residues will be incurred, there is no reasonable expectation of finite residues in livestock commodities. For ruminants, this conclusion is based on a goat metabolism stud y and a low calculated dietary burden. For poultry and swine, there are no livestock feed items associated with the registered uses ( Table 1 Feedstuffs , June 2008).

Acute and chronic dietary (food and drinking water) exposure assessments were conducted. The assessments were conservative, i.e., based on the assumption that residues in all crops were at the tolerance levels and that 100% of the crops were treated. The acute assessment was conducted for females 13

- 49 years old only, since an acute endpoint was not identified for the general US population. Both the acute estimated drinking water concentration (EDWC) of 136.2 ppb and the chronic EDWC of 133.5 ppb were mo deled using Tier 2 PRZM/EXAMS ( Pesticide Root Zone Model/Exposure Analysis Modeling System) for residues in surface water from use on turf and ornamentals, which yielded the highest acute and chronic EDWCs considering all registered/proposed uses. The acu te and chronic dietary assessments show that the acute and chronic dietary risk estimates are below HED’s level of concern (i.e., <100% of the acute or chronic population adjusted doses (aPAD or cPAD). The acute dietary risk estimate for females 13

- 49 yea rs old at the 95 th 

### percentile of exposure was 2.5% of the aPAD. For the general US population, the exposure for food and drinking water utilized <1% of the cPAD. The chronic dietary risk estimate for the highest reported exposed population subgroup, chil dren (1

- 2 years old), utilized 1.5% of the cPAD.

A cancer dietary risk assessment was not conducted since cyazofamid is classified as “not likely to be carcinogenic to humans.”

Conclusions: A new dietary exposure assessment is not anticipated to be needed during Registration Review unless there are significant increases to the drinking water exposure estimates

. Residential Exposure

Cyazofamid is currently registered for use in residential settings on turf and ornamental pla nts. There is potential for short

- term residential exposure to cyazofamid during handling (mixing, loading, and applying) and post

- application activities.

Residential Handlers The Cyazofamid 400SC label (EPA Reg. No. 71512

- 13) indicates that it is al lowed for use on turf areas associated with golf courses, sod farms, seed farms, college and professional sports fields, residential and commercial lawns, and on ornamental plants in landscapes and those grown in commercial greenhouses and nurseries. Whil e the label s ays “application by homeowners to residential turf is prohibited,” the label does not identify this product as a restricted use; therefore, a residential handler exposure assessment was previously performed to be protective of potential homeow ner exposure. Previous residential exposure and risk assessments for cyazofamid were conducted at the maximum registered application rate and

### Page 6 of 25

incorporated the revised 20 12 Residential SOPs (K. Rury, D3 97215, 06/18/2012). Applications can be made using ha ndheld spray equipment to turf and ornamentals. Residential handler total (dermal + inhalation) margins of exposure (MOEs) ranged from 2,400 to 59,000.

Residential Post

- Application Short

- term dermal and incidental oral exposure and risk estimates wer e reassessed using the 2012 Residential SOPs in the last cyazofamid ORE assessment (K. Rury, D3 97215, 06/18/2012). A dermal risk estimate for children was not conducted because there is no dermal toxicity up to the limit dose of 1,000 mg/kg/day in a 28

- da y dermal toxicity study; therefore, in the absence of hazard identified for this population, a risk assessment is not necessary. However, a dermal risk assessment for adults is required to protect for effects on the developing fetus because developmental toxicity (increased incidence of bent ribs) was seen at 1,000 mg/kg/day. Chemical

- specific DFR studies on tomatoes, squash, and grapes were submitted for a previous action on cyazofamid. These studies were reviewed and found to be acceptable for risk

as sessment. Furthermore, a turf transferrable residue (TTR) study using the modified California roller is available so the residue values have been incorporated into the most

- recent post

application exposure assessment to treated turf in place of default as sumptions and inputs for the initial fraction of residue available for transfer.

Post

- application MOEs were not of concern for any of the registered residential uses of cyazofamid. The short

- term adult dermal post

- application MOEs range from 660 to 52, 000. The short

- term inci dental oral MOEs for children 1 to < 2 years old range from 3,400 to 570,000.

Spray Drift or Volatilization Residential bystander exposures resulting from off

- site transport (e.g., spray drift or volatilization) may occur as a result of applications of cyazofamid . The potential for spray drift will be evaluated for each pesticide during the Registration Review process which ensures that all uses for that pesticide will be considered concurrently. The approach is outlined in the revised (2012) Standard Operating Procedures for Residential Risk Assessment (SOPs)

– Residential Exposure Assessment Standard Operatin g Procedures Addenda 1: Consideration of Spray Drift. This document outlines the quantification of indirect non

- occupational exposure to drift. In terms of volatilization, the agency has developed a Volatilization Screening Tool and a subsequent Volatil ization Screening Analysis ( http://www.regulations.gov/#!docketDetail;D=EPA

- HQ

- OPP

- 2014

- 0219 ). During Registration Review, the agency will utilize this analysis to deter mine if data (i.e., flux studies, route

- specific inhalation toxicological studies) or further analysis is required for cyazofamid

.

Conclusions: There are registered residential uses of cyazofamid that have been assessed using the maximum application rate s and revised 2012 Residential SOPs. Therefore, a residential handler and post

- application assessment is not required during Registration Review. However, the Agency will examine the need for a volatilization assessment, as well as evaluate the potential cyazofamid exposure from spray drift during Registration Review.

### Page 7 of 25

Aggregate Risk Assessment

Aggregate assessments consider exposures from food, drinking water, and residential uses. Since there are residential uses of cyazofamid , aggregate risk consi der s residential handler and post

- application exposures, food

, and drinking water

.

In the most recent human health risk assessment ( D 397213

, N. Tsaur , 0 6 / 28 /20 12 ), an acute aggregate risk assessment , which is equivalent to the acute dietary risk assessment, was not of concern to HED. Only females 13

- 49 years old were assessed

, since an acute endpoint of concern was not identified for the general population.

A s hort

- term aggregate risk assessment was conducted for adults and children. For adults , the pathways leading to aggregate exposure include oral dietary (background), dermal

, and inhalation (primary) routes. F or children, the pathways leading to aggregate exposure include oral dietary (background), incidental oral

, and dermal (primary) rout es. The adult and children short

- term aggregate dietary and residential MOEs are 390 and 2,20 0, respectively (LOC = 100)

. Chronic exposure from the residential pathway is not anticipated based on the current cyazofamid use pattern. Therefore, the chronic aggregate risk (food and drinking water) is equivalent to the chronic dietary (food and drinking water) exposure estimate , which was not of concern to HED.

Conclusions : Aggregate exposure to cyazofamid is currently not of concern t o HED. N ew aggregate assessments are not anticipated to be needed during Registration Review , unless changes in uses, procedures, or EDWCs occur

. Occupational Exposure

Cyazofamid is registered for use on numerous agricultural crops; on golf course turf , sod farms, seed farms, college and professional sports fields, and residential and commercial lawns; and on ornamentals in greenhouses and fields . There is potential for short

- and intermediate

- term occupational exposure to cyazofamid during handling (mixing, loading, and applying) and post

application activities.

Occupational Handlers The registered cyazofamid labels require occupational handlers to wear long sleeved shirts, long pants, shoes, socks, and chemical resistant gloves. A pplications can be made using ground, aerial, chemigation, and handheld spray equipment. Although some of the updated occupational exposure assessment policies and body weight assumptions were incorporated into previous cyazofamid exposure assessments, up dated occupational handler and post

- application assessments will be needed for occupational handlers of cyazofamid for the following use sites: carrots, cucurbits, grapes, hops, ornamentals, and turf. However, an updated occupational handler exposure asse ssment on sod would be protective of the assessment for cyazofamid use on carrots, cucurbits, and ornamentals, but an additional exposure assessment on grapes and hops would be needed to account for exposure from airblast application methods. All current occupational handler MOEs are not of concern (MOE > 100).

### Page 8 of 25

Occupational Post

- Application Occupational post

- application short

- and intermediate

- term dermal and inhalation exposures are possible for workers contacting treated crops. Dislodgeable foliar residue (DFR) studies are available for cyazofamid, and were incorporated into the most recent exposure assessment (K. Rury , D297215, 06/18/2012 ). That assessment incorporated changes in transfer coefficients and body weight assumptions; however, previous post

- application assessments for carrots, cucurbits, grapes, hops, ornamentals, and turf did not incorporate updated exposure assessment policies. Therefore, an updated occupational post

- application exposure assessment will be required during Registratio n Review for cyazofamid use on carrots, cucurbits, grapes, hops, ornamentals, and turf. All current occupational post

- application MOEs are not of concern (MOE > 100).

Conclusions: There is sufficient information available to assess occupational handler and post

application exposures. There are also acceptable DFR and TTR studies available to evaluate the post

- application exposure to cyazofamid. Updated occupational handler and post

- appl ication exposure assessments will be conducted during Registration Review for cyazofamid use on carrots, cucurbits, grapes, hops, ornamentals, and turf

. Public Health and Pesticide Epidemiology Data

Based on the low frequency and severity of incident cases reported for cyazofamid in both the OPP Incident Data System (IDS) and the Centers for Disease Control and Prevention/National Institute for Occupational Safety and Health (CDC/NIOSH) Sentinel Event Notification System for Occupational Risk

- Pesticides (SENSOR), there does not appear to be a concern at this time that would warrant further investigation

. For the Main IDS, from January 1, 2009 to August 12, 2014, there were 0 incident s reported for the single chemical only in the database. In Aggregate IDS, fr om January 1, 2009 to August 12, 2014, there were 2 inciden ts reported involving cyazofamid . These incidents were classified as minor severity. Overall, there are few incidents involving cyazofamid reported to IDS.

A query of the Sentinel Event Notification System for Occupational Risk (SENSOR)

- Pesticides database for 1998

- 2010 identified 0 cases involving cyazofamid

.

The Agricultural Health Study (AHS) is a high quality

, prospective epidemiology study evaluating the link between pesticide use and various health outcomes including cancer. The AHS includ es private and commercial pesticide applicators and their spouses

. If there are AHS findings relevant to a particular pesticide going through registration review, the Agency will ensure they are considered in the problem formulation/scoping phase of the p rocess and, if appropriate, fully reviewed in the risk assessment phase of the process

. The AHS includes information on use of 50 different pesticide active ingredients commonly used in agriculture. Cyazofamid is not included in the AHS, and therefore, this study does not provide information for this report.

The Agency will continue to monitor the incident information and if a concern is triggered, additional analysis will be included in the risk assessment.

### Page 9 of 25

C onclusion: Based on the low frequency and l ow severity of incident cases reported for cyazofamid in both IDS and SENSOR

- Pesticides databases , there does not appear to be a concern at this time that would warrant further investigation. The Agency will continue to monitor the incident information an d if a concern is triggered, additional analysis will be included in the risk assessment.

Tolerance Assessment and International Harmonization

Tolerances are established for plant commodities under 40 CFR §180.601 for residues of the fungicide cyazofami d, including its metabolites and degradates. Compliance with the specified tolerance levels is to be determined by measuring only the sum of 4

- chloro

- 2

- cyano

N
, N

- dimethyl

- 5

- (4

- methylphenyl)

- 1 H

- imidazole

- 1

- sulfonamide and its metabolite, 4

- chloro

- 5

- (4

- meth ylphenyl)

- 1 H

- imidazole

- 2

- carbonitrile, calculated as the stoichiometric equivalent of cyazofamid. The tolerances on plants include permanent Section 3 tolerances, a Section 18 emergency exemption tolerance, and a tolerance with regional registration. The plant tolerance levels range from 0.02 ppm to 144 ppm. No livestock tolerances are established.

As shown in Attachment 2, there are no Codex maximum residue limits (MRLs) established for cyazofamid; therefore, there is no harmonization issue with Codex at this time.

The US and Canadian tolerance expressions include the same analytes. For commodities with both US and Canadian tolerances, the Canadian tolerance levels are generally the same as the US tolerance levels, with some exceptions.

Environmental Justice

Potential areas of environmental justice concerns, to the extent possible , w ill be considered in th e registration review , in accordance with US Executive Order 12898 , "Federal Actions to Address Environmental Justice in Minority Populations and Low

- Income Populations," http://www.eh.doe.gov/oepa/guidance/justice/eo12898.pdf ). As a part of every p esticide risk assessment, OPP considers a large variety of consumer subgroups according to well

- established procedures. In line with OPP policy, HED estimates risks to population subgroups from pesticide exposures that are based on patterns of that subgro up’s food and water consumption, and activities in and around the home that involve pesticide use in a residential setting. Extensive data on food consumption patterns are compiled under the NHANES/WWEIA and are used in pesticide risk assessments for all registered food uses of a pesticide. These data are analyzed and categorized by subgroups based on age and ethnic group. Additionally, OPP is able to assess dietary exposure to smaller, specialized subgroups and exposure assessments are performed when co nditions or circumstances warrant. Whenever appropriate, non

- dietary exposures based on home use of pesticide products and associated risks for adult applicators and for toddlers, youths, and adults entering or playing on treated areas post

- application ar e evaluated. Further considerations are currently in development as OPP has committed resources and expertise to the development of specialized software and models that consider exposure to bystanders and farm workers as well as lifestyle and traditional dietary patterns among specific subgroups

.

### Page 10 of 25

Cumulative

Unlike other pesticides for which EPA has followed a cumulative risk approach based on a common mechanism of toxicity, EPA has not made a common mechanism of toxicity finding as
to cyazofamid and any other substances and cyazofamid does not appear to produce a toxic metabolite produced by other substances. For information regarding EPA’s efforts to determine which chemicals have a common mechanism of toxicity and to evaluate the cumulative effects of such chemicals, see the policy statements released by EPA’s Office of Pesticide Programs concerning common mechanism determinations and procedures for cumulating effects from substances found to have a common mechanism on EPA’s website at http://www.epa.gov/pesticides/cumulative/.

Human Studies

The risk assessment s which have been previously conducted for cyazofamid rel ied in part on data from studies in which adult human subjects were intentionally exposed to a pesticide to determine their dermal and inhalation exposure. Many such studies, involving exposure to many different pesticides, comprise generic pesticide exposur e databases such as the Pesticide Ha ndlers Exposure Database (PHED), the Outdoor Residential Exposure Task Force (ORETF), and the Agricultural Reentry Task Force (ARTF) Database

. EPA has reviewed all the studies supporting these multi

- pesticide generic ex posure databases, and has found no clear and convincing evidence that the conduct of any of them was either fundamentally unethical or significantly deficient relative to the ethical standards prevailing at the time the research was conducted. A ll applicable requirements of EPA’s Rule for the Protection of Human Subjects of Research (40 CFR Part 26) have been satisfied , and t here is no regulatory barrier to continued reliance on these studies

.

Data Deficiencies

No toxicological data, residue chemistry, o r occupational and residential exposure data are needed to support the registered uses of cyazofamid at this time.

### Page 11 of 25

References

Recent Memoranda Relevant to Registration Review of Cyazofamid

Author DP/TXR # Date Title

Risk Assessment W. Wassell D292390 4/27/04 Human Health Risk Assessment to Support the Registration of Cyazofamid for use on Potatoes, Tomatoes, Cucurbits, and Imported Wine. First Food Use for Cyazofamid. K. O’Rourke D332426 10/12/06 Human Health Risk Assessment to Support the Registration of Cyazofamid for use on Non

- Residential Turf and Ornamentals. K. O’Rourke D342612 3/19/08 No. 7E7244, DP # Human Health Risk Assessment to Support the Registration of Cyazofamid for Use on Carrot (PC Code: 085651, Petition 342612). N. Tsaur D356837 5/20/09 Cyazofamid

. Human Health Risk Assessm ent for Proposed Uses on Fruiting Vegetables and Okra, Grapes East of the Rocky Mountains, Vegetable Greenhouse Transplants, and Commercial Application on Residential Turf and Residential Ornamentals. N. Tsaur D366114 6/3/09 Cyazofamid. Amendment to D356837, D356878 Human Health Risk Assessment for Proposed Uses on Fruiting Vegetables and Okra, Grapes East of the Rocky Mountains, Vegetable Greenhouse Transplants, and Commercial Application on Resident ial Turf and Residential Ornamentals. N. Tsaur D370671 4/7/10 Cyazofamid. Human Health Risk Assessment for Proposed Uses

on Brassica (Cole) Leafy Vegetables Crop Group 5, Turnip Greens, Spinach, and Hops. B. Hanson D376500 8/3/10 Cyazofamid. Human Health Risk Assessment for Proposed Section 18 Use on Basil. N. Tsaur D397213 6/28/12 Cyazofamid

. Human Health Risk Assessment for Proposed New Uses on Leafy Greens (Crop Subgroup 4A), Succulent

- Podded and Succulent

- Shelled Beans, Basil, Tuberous and Cor m Vegetables (Subgroup 1C), and Fruiting Vegetables (Crop Group 8

- 10) with Updated Residential Risk Estimates of All Existing Residential Uses. Residue Chemistry A. Acierto D323339 2/8/07 PP# 1F06305. Cyazofamid. Data Submitted to Satisfy Conditional Registration Requirements for Use on Cucurbits, Potatoes, Tomatoes, and Imported Wine Grapes. Summary of Analytical Chemistry and Residue Data. A. Acierto D285680 10/1/04 PP# 1F06305. Cyazofamid. Petition for the Establishment of Permanent Tolerances for use on Cucurbits, Potatoes, Tomatoes, and Imported Wine Grapes. Summary of Analytical Chemistry and Residue Data. PC Code: 085601

. DP Barcode: 285680

.

A. Acierto D343419 3/6/08 Cyazofamid . PP# 7E7244: Petition for the Establishment of a Tolerance for the Combined Residues of Cyazofamid and its Metabolite CCIM, in/on Carrot, Roots. Summary of Analytical Chemistry and Residue Data. A. Acierto D357042 3/18/09 Cyazofamid . Amend Section 3 Registration to Add Uses on Fruiting Vegetables, Okra, Grape (East of the Rocky Mountains), and Greenhouse Vegetable Transplants. Summary of Analytical Chemistry and Residue Data. N. Dodd D396051 4/25/12 Cyazofamid. Petition for the Establishment of Permanent Tolerances and Registration for Use on Basil, Succulent Be ans, the Fruiting Vegetable Group 8

- 10, the Leafy Greens Subgroup 4A,

### Page 12 of 25

Recent Memoranda Relevant to Registration Review of Cyazofamid

Author DP/TXR # Date Title and the Tuberous and Corm Vegetable Subgroup 1C. Summary of Analytical Chemistry and Residue Data. Dietary Exposure Assessment N. Dodd D397214 4/30/12 Cyazofamid Acute and Chronic Aggregate Dietary (Food and Drinking Water) Exposure and Risk Assessments for the Section 3 Registration Action on Basil, Succulent Beans, the Fruiting Vegetable Group 8

- 10, the Leafy Greens Subgroup 4A, and the Tuberous and Corm Vegetabl e Subgroup 1C. Occupational and Residential Exposure Assessment K. Rury D397215 06/18/201 2 Cyazofamid. Occupational Exposure Assessment for Proposed Uses on Leafy Greens, Subgroup 4A; Succulent

- Podded and Succulent

- Shelled Beans; Basil; Vegetable, Tuberous and Corm, Subgroup 1C; and Vegetables, Fruiting, Crop Group 8

- 10 with An Updated Residential Exposure Assessment of All Existing Residential Uses. Incident Report S. Recore D423509 10 / 28 /2014 Cyazofamid : Tier I Review of Human Incidents Toxicology G. Dannan TXR# 005428 04/15/2004 Report of the Hazard Identification Assessment Review Committee K. Rury TXR#0056341 Cyazofamid: Summary of Hazard and Science Policy Council (HASPOC) Recommendations on the requirement of the 28 day inhalation study

.

### Page 13 of 25

Attachment s 1. Chemical Identity Table 2. Physicochemical Properties of Technical Grade Cyazofamid Table 3. Tolerance /MRL Summary Table 4 . Endpoint Selection Table s 5 . Toxicity Profile Tables

### Page 14 of 25 Attachment 1 Chemical Identity Table

Cyazofamid Nomenclature

. Chemical structure

Common name Cyazofamid Company experimental name IKF

- 916
IUPAC name 4

- chloro

- 2

- cyano

- N,N

- dimethyl

- 5

-

p

- tolylimidazole

- 1

- sulfonamide
CAS name 4

- chloro

- 2

- cyano

- N,N

- dimethyl

- 5

- (4

- methylphenyl)

- 1 H

- imidazole

- 1

- sulfonamide CAS registry number 120116

- 88

- 3

Identification of Compounds from the Cyazofamid Metabolism Studies

Common Name/ Code/Matrix Chemical Name Chemical Structure

Cyazofamid/ IKF

- 916 4

- chloro

- 2

- cyano

- N,N

- dimethyl

- 5

- (4

- methylphenyl)

- 1 H

- imidazole

- 1

- sulfonamide

CCIM 4

- chloro

- 5

- p

- tolylimidazole

- 2

- carbonitrile

CCBA 4

- (4

- chloro

- 2

- cyanoimidazol

- 5

- yl)benzoic acid

### Page 15 of 25

Identification of Compounds from the Cyazofamid Metabolism Studies

Common Name/ Code/Matrix Chemical Name Chemical Structure

CCIM

- AM 4

- chloro

- 5

- p

- tolylimidazole

- 2

- carboxamide

CTCA 4

- chloro

- 5

- p

- tolylimidazole

- 2

- carboxylic acid

### Page 16 of 25 Attachment 2 Physicochemical Properties of Technical Grade Cyazofamid Table

Physico c hemical Properties of the Technical Grade Cyazofamid
Parameter Value Reference Melting point (°C) 152.7 PMRA Regulatory Note REG2006

- 05 on Cyazofamid, 12/6/06 pH 4.9 e

- Pesticide Manual, 13 th Edition Density at 20 °C (g/mL) 1.446 Water solubility (at 20 °C) pH Solubility (mg/L) REG2006

- 05
5 7 9 0.121

0.107 0.109 Solvent solubility (at 21.2°C) Solvent Solubility (g/L)
Acetone Ethyl acetate Methanol Dichloromethane Toluene Hexane

n

- Octanol Acetonitrile 2

- Propanol 43.1

16.19 1.74 94.82 6.0 0.03 0.04 29.446 0.43 Vapor pressure at 25°C <1.33 x 10

- 5 Pa Dissociation constant (pK a ) Does not dissociate in water. No pK a in the pH range of 2

- 12. Octanol/water partition coefficient Log K OW = 3.2 K OW = 1.585 x 10 3 UV/visible absorption spectrum pH Λ max (nm)
2 7 10 285

279 287

### Page 17 of 25 Attachment 3 Tolerance/MRL Tabl es
Cyazofamid (085651; Date of Request 11/4/14) Summary of US and International Tolerances and Maximum Residue Limits Residue Definition : US Canada Mexico 1 

### Codex 40 CFR §180.601 (a) General. sum of 4

- chloro

- 2

- cyano

- N

, N

- dimethyl

- 5

- (4

- methylphenyl)

- 1 H

- imidazole

- 1

- sulfonamide and its metabolite, 4

- chloro

- 5

- (4

- methylphenyl)

- 1 H

imidazole

- 2

- carbonitrile 4

- chloro

- 2

- cyano

- N
, N

- dimethyl

- 5

- (4

- methylphenyl)

- 1 H

- imidazole

- 1

- sulfonamide, including t he metabolite 4

- chloro

- 5

- (4

- methylphenyl)

- 1 H

- imidazole

- 2

- carbonitrile None

Commodity Tolerance (ppm) /Maximum Residue Limit (mg/kg) US Canada Mexico 1 

### Codex Basil, dried leaves 90 90 Basil, fresh leaves 30 30
Bean, succulent 0.5 0.5 edible

- podded (jackbeans, moth beans, runner beans, snap beans, sword beans, wax beans, yardlong beans) Bean, succulent shelled 0.08 0.08 succulent shelled ( blackeyed
pea s , broad beans, cowpea s , lima beans, southern pea s) Brassica, head and stem, subgroup 5A 1.2 0.02 b roccoli

, brussels sprouts

, cabbage s, cauliflower s, Chinese broccoli, Chinese mustard cabbages, kohlrabi es, napa Chinese cabbages Brassica, leafy greens, subgroup 5B 12.0

Carrot, roots 0.09 0.09 Hop dried cones 10.0 Leafy greens subgroup 4A 10 10 amaranth, arugula, corn salad, dandelion leaves, dock, edible leaved chrysanthemum, endives, fresh chervil leaves, fresh parsley leaves, garden cress, garden purslane , garland chrysanthemum, head lettuce, leaf lettuce, Malabar spinach, New Zealand spinach, orach radicchio, spinach upland cress, winter purslane Turnip, greens 12.0 Vegetable, cucurbit, group 9 0.10 0.1 balsam apples, balsam pears, cantaloupes, chayote fruit, Chinese cucumbers, Chinese waxgourds, citron melons, cucumbers, edible gourds, muskmelons (other than those listed in this item) pumpkins, summer squash, watermelons, west Indian gherkins, winter squash Vegetable, fruiting, group 8

- 10 0.9 0. 02 bell peppers, non

- bell peppers 0.2 tomatoes

### Page 18 of 25

Summary of US and International Tolerances and Maximum Residue Limits Residue Definition : US Canada Mexico 1 

### Codex Vegetable, tuberous and corm, subgroup 1C 0.02 0.02 potatoes

MRLs with NO US Equivalent Fat of cattle 0.02 Fat of goats 0.02 Fat of horses 0.02 Fat of sheep 0.02 Meat byproducts of cattle 0.02 Meat byproducts of goats 0.02 Meat byproducts of horses 0.02 Meat byproducts of sheep 0.02 Meat of cattle 0.02 Meat of goats 0.02 Meat of horses 0.02 Meat of sheep 0.02 Milk 0.02

Completed: M. Negussie; 11 /05/14

1 Mexico adopts US tolerances and/or Codex MRLs for its export purposes.



### ( b) Section 18 emergency exemptions. Time

- limited tolerances are established for residues of the fungicide cyazofamid, including its metabolites and degradates, in or on the commodities in the following table. Compliance with the tolerance levels specified in the following table is to be det ermined by measuring only the sum of cyazofamid, 4

- chloro

- 2

- cyano

- N,N

- dimethyl

- 5

- (4

- methylphenyl)

- 1H

imidazole

- 1

- sulfonamide and its metabolite CCIM, 4

- chloro

- 5

- (4

- methylphenyl)

- 1H

- imidazole

- 2

- carbonitrile, calculated as the stoichiometric equivalent of cy azofamid, resulting from use of the pesticide under FIFRA section 18 emergency exemptions. The tolerances expire and are revoked on the date specified in the table.

Commodity Parts per million Expiration/revocation date Canada
Basil, dried 144 12/31/14 90



### (c) Tolerances with regional registrations. Tolerances with regional registrations are established for residues of the fungicide cyazofamid, including its metabolites and degradates, in or on the commodities in the following table. Compliance with the tolerance levels specified in the following tab le is to be determined by measuring only the sum of 4

- chloro

- 2

- cyano

- N
, N

- dimethyl

- 5

- (4

- methylphenyl)

- 1 H

imidazole

- 1

- sulfonamide and its metabolite, 4

- chloro

- 5

- (4

- methylphenyl)

- 1 H

- imidazole

- 2

- carbonitrile, calculated as the stoichiometric equivalent of cyaz ofamid, in or on the following commodities:

Commodity Parts per million Canada
Grape 1.5 1.2

### Page 19 of 25 Attachment 4

Toxicological Doses and Endpoints for Cyazofamid for Use in Dietary and Non

- Occupational Human Health Risk Assessments

Exposure/ Scenario Point of

Departure Uncertainty/

FQPA Safety Factors RfD, PAD,

Level of Concern for Risk Assessment Study and Toxicological Effects

Acute Dietary (General Population, including Infants and Children) No adverse effects were observed which could be attributed to a single

- dose exposure for the general

population

Acute Dietary (Females 13

- 49 years of age) NOAEL = 100

mg/kg/day UF A = 10x

UF H = 10x FQPA SF= 1x Acute RfD =

1.0 mg/kg/day Rat Prenatal Developmental Toxicity

(MRID 45408933) LOAEL = 1,000 mg/ kg/day based on developmental toxicity findings of increased incidence of bent ribs. Chronic Dietary (All Populations) NOAEL= 94.8

mg/kg/day UF A = 10x

UF H = 10x FQPA SF= 1x Chronic RfD =

0.948 mg/kg/day Mouse Oral Carcinogenicity (MRID

45408932) LOAEL = 985 mg/kg/day based on increased skin lesions. Incidental Oral Short

- Term (1

- 30 days) and Intermediate

- Term (1

- 6 months) NOAEL= 171

mg/kg/day UF A = 10x

UF H = 10x FQPA SF= 1x Residential

LOC for MOE = 100 90

- Day oral toxicity study in rats. LOAEL=

295 mg/kg based on Increased number of basophilic tubules of the kidneys, increased urinary volume, pH, & protein. This toxicity endpoint is also supported by the results of a 28

- day oral dose

- range

- finding s tudy in rats. In this study, at 370 mg/kg/day or above increased incidence of basophilic tubules in the kidneys was found. Dermal Short

Term (1

- 30 days) and Intermediate

Term (1

- 6 months) For CHILDREN:

No toxicity was found at 1000 mg/kg in a 28

- day d ermal toxicity study, therefore, in the absence of hazard identified for this population, a risk assessment is not necessary. For ADULTS: NOAEL = 100 mg/kg/day (Dermal Absorption Rate = 37%) UF A = 10x

UF H = 10x FQPA SF= 1x Acute RfD =

1.0 mg/kg/day Rat Prenatal Developmental Toxicity (MRID

45408933) LOAEL = 1,000 mg/kg/day based on developmental toxicity findings of increased incidence of bent ribs. Inhalation Short

Term (1

- 30 days) and Intermediate

Term (1

- 6 months) No residential exposure is expected via inhalation route.

Cancer (oral, dermal, inhalation) Classification: “Not likely to be Carcinogenic to Humans” based on the absence of significant tumor

increases in two adequate rodent carcinogenicity studies. Point of Departure (POD) = A d ata point or an estimated point that is derived from observed dose

- response data and used to mark the beginning of extrapolation to determine risk associated with lower environmentally relevant human exposures. NOAEL = no observed adverse effect level. L OAEL = lowest observed adverse effect level. UF = uncertainty factor. UF A = extrapolation from animal to human (intraspecies). UF H = potential variation in sensitivity among members of the human population (interspecies). UF L = use of a LOAEL to extrapolate a NOAEL. UF S = use of a short

- term study for long

- term risk assessment. UF DB = to account for the absence of key date (i.e., lack of a critical study). FQPA SF = FQPA Safety Factor. PAD = population adjusted dose (a = a cute, c = chronic). RfD = reference dose. MOE = margin of exposure. LOC = level of concern. N/A = not applicable.

### Page 20 of 25 Summary of Toxicological Doses and Endpoints for Cyazofamid for Use in Occupational Human Health Risk Assessments Exposure/ Scenario Point of

Departure Uncertainty

Factors Level of Concern

for Risk Assessment Study and Toxicological Effects

Dermal Short

Term (1

- 30 days) and Intermediate

Term (1

- 6 months) Oral Study

NOAEL= 100 mg/kg/day (Dermal Absorption Rate = 37%) UF A =10x

UF H =10x Occupational LOC

for MOE = 100 Rat Prenatal Developmental Toxicity (MRID

45408933) LOAEL = 1,000 mg/kg based on developmental toxicity findings of increased incidence of bent ribs.

Dermal Long Term (>6 months) NOAEL=

94.8 mg/kg/day (Dermal Absorption Rate = 37%) UF A =10x

UF H =10x Occupational LOC

for MOE = 100 18

- Month Mouse Oral Carcinogenicity (MRID

45408932) LOAEL = 985 mg/kg/day based on increased skin lesions.

Inhalation Short

Term (1

- 30 days) and Intermediate

Term (1

- 6 months) NOAEL=

100 mg/kg/day UF A =10x

UF H =10x Occupational LOC

for MOE = 100 Rat Prenatal Developmental Toxicity (MRID

45408933) LOAEL = 1,000 mg/kg based on developmental toxicity findings of increased incidence of bent ribs.

Inhalation Long

Term (>6 months) NOAEL=

94.8 mg/kg/day UF A =10x

UF H =10x Occupational LOC

for MOE = 100 18

- Month Mouse Oral Carcinogenicity (MRID

45408932) LOAEL = 985 mg/kg/day based on increased skin lesions. Cancer (oral, dermal, inhalation) Classification: “Not likely to be Carcinogenic to Humans” based on th e absence of significant tumor

increases in two adequate rodent carcinogenicity studies.

Point of Departure (POD) = A data point or an estimated point that is derived from observed dose

- response data and used to mark the beginning of extrapolation to determine risk associated with lower environmentally relevant human exposures. NOAEL = no ob served adverse effect level. LOAEL = lowest observed adverse effect level. UF = uncertainty factor. UF A = extrapolation from animal to human (intraspecies). UF H = potential variation in sensitivity among members of the human population (interspecies). UF L = use of a LOAEL to extrapolate a NOAEL. UF S = use of a short

- term study for long

- term risk assessment. UF DB = to account for the absence of key date (i.e., lack of a critical study). MOE = margin of exposure. LOC = level of concern. N/A = not ap plicable.

### Page 21 of 25 Attachment 5

Toxicity Profile for Cyazofamid

Acute Toxicity of Cyazofamid Technical

Guideline No. Study Type MRID No. Results Toxicity Category

870.1100 Acute Oral

– Rat 45408910 LD 50 > 5000 mg/kg [M/F] IV

870.1200 Acute Dermal

– Rat 45408911 LD 50 > 2000 mg/kg [M/F] III

870.1300 Acute Inhalation

– Rat 45408912 LC 50 > 5.5mg/L [M/F] IV

870.2400 Primary Eye Irritation

–

Rabbit 45408913 Grade 2 conjunctival redness and discharge that resolved by 24 hours IV

870.2500 Primary Skin Irritation

– Rabbit 45408914 Very slight to well defined erythema that cleared by 7 days III

870.2600 Dermal Sensitization

(Guinea Pig Maximization test) 45408915 Positive (weak sensitizer)

-

Toxicity Profile of Cyazofamid Technical.

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

870.3100 28

- Day dose

- range finding study in rats 45408927 Acceptable/non

- guideline 0, 50, 500, 5000, & 20,000 ppm Male: 0, 3.8, 38.5, 370, & 1488 mg/kg/day Female: 0, 3.6, 37.1, 389, & 1535 mg/kg/day NOAEL = 38.5 [M] mg/kg/day LOAEL = 370 [M] mg/kg/day based on dose related increase in the incidence of basophilic kidney tubules in males.

870.3100 90

- Day oral toxicity in
rats 45408928 (1999) Acceptable/guideline Males: 0, 10, 50, 500, or 5,000 ppm 0, 0.6, 2.9, 29.5, or 294.5 mg/kg/day Females: 0, 50, 500, 5,000, or 20,000 ppm 0, 3.3, 33.3, 337.6, or 1,359.0 mg/kg/day NOAEL = 29.5 [M] mg/kg/day LOAEL = 295 [M] mg/kg/day based on increased number of “basophilic kidney tubules,” and increased urinary volume, pH, and protein.

870.3150 90

- Day oral toxicity in dogs 45408929 (2001) Acceptable/guideline 0, 40, 200, or 1,000 mg/kg/day NOAEL = 1,000 [M/F] mg/kg/day LOAEL = not observed.

### Page 22 of 25 Toxicity Profile of Cyazofamid Technical.

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

870.3200 28

- Day dermal toxicity in rats 45408930 (1997) Acceptable/guideline 0, 250, 500, or 1,000 mg/kg/day NOAEL = 1,000 [M/F] mg/kg/day LOAEL = not observed.

870.3465 28

- Day inhalation toxicity Waived (HASPOC; TRX# 0056341; 7/11/12)

870.3700a Prenatal developmental in rats 45430404 (1999) Acceptable/guideline 0, 30, 100, or 1,000 mg/kg/day Maternal NOAEL = 1,000 mg/kg/day LOAEL = not observed Developmental NOAEL = 100 mg/kg/day LOAEL = 1,000 mg/kg/day based on increased incidence of bent ribs.

870.3700b Prenatal developmental in rabbits 45408934 (1999) Acceptable/guideline 0, 30, 100, or 1,000 mg/kg/day Maternal NOAEL = 1,000 mg/kg/day LOAEL = not observed Developmental NOAEL = 1,000 mg/kg/day LOAEL = not observed

870.3800 Reproduction and fertility effects in rats 45408935 (1998) Acceptable/guideline 0, 200, 2,000, or 20,000 ppm F 0 M: 11

- 22, 108

- 213, and 1114

- 2185 mg/kg/day F 0 F: 14

- 21, 137

- 219, and 1416

- 2179 mg/kg/day F 1 M: 11

- 27, 107

- 267, and 1117

- 2714 mg/kg/day F 1 F: 14

- 27, 135

- 267, and 1451

- 2678 mg/kg/day Parental/Systemic NOAEL = 1114/ 1416 [M/F] mg/kg/day LOAEL = not observed Reproductive NOAEL = 1114/ 1416 [M/F] mg/kg/day LOAEL = not observed Offspring NOAEL = 1114/ 1416 [M/F] mg/kg/day LOAEL = not observed

870.4100a Chronic toxicity in rats 45408936 (1999) Acceptable/guideline Males: 0, 10, 50, 500, or 5,000 ppm 0, 0.35, 1.7, 17.1, or 171.1 mg/kg/day Females: 0, 50, 500, 5,000, or 20,000 ppm 0, 2.0, 20.2, 207.8, or 856.0 mg/kg/day NOAEL = 171/ 856 [M/F] mg/kg/day LOAEL = not observed.

870.4100b Chronic toxicity in dogs 45408931 (2001) Acceptable/guideline 0, 40, 200, 1,000 mg/kg/day NOAEL = 200 [M/F] mg/kg/day LOAEL = 1,000 [M/F] mg/kg/day based on increased cysts in parathyroids in males.

870.4200 Carcinogenicity rats 45408936 (1999) Acceptable/guideline NOAEL = 171/ 856 [M/F] mg/kg/day LOAEL = not observed. No evidence of carcinogenicity

### Page 23 of 25 Toxicity Profile of Cyazofamid Technical.

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

870.4300 Carcinogenicity mice 45408932 (1999) Acceptable/guideline NOAEL = 94.8 [M] mg/kg/day LOAEL = 985 [M] mg/kg/day based on increased incidence of skin lesions including hair loss, body sores, dermatitis, ulceration, and acanthosis. No evidence of carcinogenicity

Gene Mutation 870.5100 Bacterial reverse mutation assay 45408937 (1998) Acceptable/ guideline (ai) Negative ± S9 up to 5,000 μ g/plate by standard plate and tube preincubation (not cytotoxic but there was precipitation at ≥1,500 μg/plate.

Gene Mutation 870.5300 Mammalian cell culture 45409002 (1998) Acceptable/ guideline (ai) Negative ± S9 up to cytotoxic and precipitating concentration of 100 μ g/mL

Cytogenetics 870.5375 Chromosomal aberrations 45409003 (1996) Acceptable/ guideline (ai) Negative ± S9 for clastogenic/aneugenic activity up to cytotoxic and precipitating 200 μ g/mL

Cytogenetics 870.5395 Micronucleus test on

mouse 45409004 (1996) Acceptable/ guideline (ai) Negative up to the highest dose tested (limit dose) 2,000 mg/kg

Other Effects 870.5500 Bacterial DNA repair test (Rec

- assay) 45409005 (1998) Acceptable/ guideline (ai) Negative ± S9 up to limit of solubility at 8,000 μ g/disc

870.6200a Acute neurotoxicity screening battery 45434601 (2000) Acceptable/guideline 0, 80, 400, or 2000 mg/kg NOAEL = 2000 [M/F] mg/kg/day LOAEL = not observed. The HIARC concluded that the slight increase ( ∼ 45%) in motor activity at day 14 among the mid

- and high

dose males is marginal and should not be considered an adverse finding.

870.6200b Subchronic neurotoxicity screening battery Study not available.

870.6300 Developmental neurotoxicity Study not available.

### Page 24 of 25 Toxicity Profile of Cyazofamid Technical.

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

870.7485 Metabolism and pharmacokinetics in
rats 45409006

- 45409009 and 45434602 (1998

- 1999) Acceptable/guideline Dose: 0.5, 1,000, or 14 day 0.5 mg/kg repeat non

- labeled followed by 0.5 mg/kg [phenyl

- U

14 C]

- or [ 14 C

- Imidazole] IKF

- 916. Biliary excretion study: 0.5 or 1,000 mg/kg [phenyl

- U

14 C]

- or [ 14 C

Imidazole] IKF

- 916. There was rapid absorption (irrespective of dose t cmax = 0.25

- 0.5 hrs) and rapid elimination at the low dose (t 1/2 4.4

- 5.8 hrs) while there was saturated absorption with prolonged elimination (t 1/2 of 7.6

- 11.6 hrs) at the high

dose. The extent of absorption (as per cent of administered dose) was highly dose

- dependent being nearly 75% at the low dose and only about 5% at the high dose. Both the u rine and feces were major routes of excretion at the low dose with most of the urinary radioactivity being a metabolite named CCBA (4

- (4

- chloro

- 2

- cyanoimidazol

- 5

- yl)benzoic acid). The biliary elimination was highly variable at the low dose ( ∼ 12

- 39% of the administered low dose) and negligible (<2%) in the high

- dose groups. Urinary or biliary excretion in the high

- dose groups was low (each ∼ 2%) with most of the radioactivity being CCBA. Irrespective of the dosing regimen, most of the recovered fecal radi oactivity was unchanged parent compound; the major fecal metabolites were CCBA and 4

- chloro

- 5

-

p

- tolylimidazole

- 2

- carbonitrile (CCIM) each of which being less than 5% of the administered dose. Tissue burdens at t 1/2 , t max , and at 168 hours post dose indica ted rapid clearance and low tissue burdens suggesting little or no bioaccumulation or sequestration.

870.7600 Dermal penetration Study not available.

Special study: In Vitro Metabolism of IKF

- 916 and CCIM in Blood and Stomach Contents MRID 45409010 (1999) Acceptable/non

- guideline The in vitro biodegradation of [ 14 C

- Bz]IKF

- 916 or [ 14 C

Bz)CCIM by rat blood or rat stomach contents was examined. The results indicated that, during the 60

- minute incubation period, IKF

- 916 underwent degradation to CCIM ( ∼ 3 0%) in blood but was stable in stomach contents while CCIM was unaffected in either medium. The results also suggest that CCIM could be produced in rats following IKF

- 916 entering the blood compartment but not necessarily in the stomach compartment.

### Page 25 of 25 Toxicity Profile of Cyazofamid Technical.

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

Spec ial study: In Vitro Metabolism of CCIM in S9 Fraction MRID 45409011 (1999) Acceptable/non

- guideline The in vitro metabolism of 14 C

- CCIM was followed up to five minutes in the presence of rat liver S9. The results suggested that CCIM was rapidly and completely metabolized within 5 minutes to at least three metabolites with the major one being CHCN (4

- chloro

5(4

- hydroxymethyl

- phenyl)

- imidazole

- 2

- carbonitrile) which accounted for 77

- 94% of the radioactivity. Lower concentrations were found of CCBA (3

- 1 3%) and one or more unknown metabolites. It was concluded that CCIM produced in the intestines or blood would be rapidly and completely metabolized by the liver.

Special study: In Vitro Study to Identify Metabolites Absorbed Through the Intestinal Mucosa After Incubation with IKF

916 and CCIM MRID 45409012 (1999) Acceptable/non

- guideline Using in vitro isolated rat intestinal segments containing [ 14 C]IKF

- 916 or [ 14 C]CCIM, 2.6% of IKF

- 916 and 10.4% of CCIM was transferred from the lumen (mucosal side) to t he exterior (serosal side) of the gut segment. Of the IKF

- 916 that was absorbed and transferred to the buffer solution, approximately 60% had undergone biotransformation/degradation to CCIM, CCBA and three additional products. Also, CCIM appeared to be d egraded to two metabolites not observed for intestine preparations containing IKF

- 916. Based on the findings of this study, cyazofamid seems to be poorly absorbed from the intestinal lumen although its conversion to CCIM or other metabolites may increase overall transfer across the gut wall.

Special study: Comparative metabolism of [ 14 C]IKF

- 916 or [ 14 C]CCIM in rats MRID 45409013 (1999) Acceptable/non

- guideline Rats were given a single gavage dose of 0.46 mg/kg of [ 14 C

- Bz]IKF

- 916 or 0.33 mg/kg of [ 14 C

- Bz]CCIM and, after 30 minutes, blood, liver, stomach (plus contents), and duodenum (plus contents) were collected and analyzed for radioactivity and metabolites. Following a single gavage dose of CCIM, the metabolites CCBA and CHCN, in addition to uncha nged CCIM, were detected in the plasma and liver. The same metabolites, as well as CCIM, were also detected in plasma and liver following administration of IKF

- 916. Therefore, orally administered IKF

- 916 is converted to CCIM and subsequently to CCBA and C HCN.