A cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 1 of 17 **UNITED STA TES ENVIRONMENTA**  **L PROTECTION A GENCY** 

**WA SHINGTON, D.C. 20460**  **Office of Chemical Safety and Pollution Prevention MEMORANDUM**  **Date :**  February 23 , 2012 **SUBJECT:** Acibenzolar - S - Meth yl . Petition for the Establishment of Permanent Tolerances on Lo w Growing Berries (Subgroup 1 3 - 07 G ). Summary of Anal yt ical Chemistry

and Residue Data. **PC Code:** 0 61402 **DP Barcode s**  **:** 386364 **Decision No s**  **.:** 443561 **Registration No. :**  100 - 922 **Petition No s**  **.:** 0E7818 **Regulatory Action:** Amended Section 3 **Risk Assessment Type :**  NA **Case No.:** NA **T XR No.:**  NA **CAS No.:** 135158 - 54 - 2 **MRID Nos.:** 4 83328 01 **40 CFR:** §180.561 **FROM:** Thurston G. Morton, Chemist

Risk Assessment Branch 4 Health Effects Division (7509P) **THROUGH:** Susan V. Hummel, Chemist/Senior Scientist

Elissa Reaves PhD, Branch Chief Risk Assessment Branch 4 Health Effects Division (7509P) **TO:** Becky Daiss, Risk Assessor

Risk Assessment Branch 4 Health Effects Division (7509P)

And Sidney Jackson (RM 05 ) Risk Integration, Minor Use & Emergency Response Branch Registration Division (7505P) **MRID Summary Table MRID No.**  **Study Type Comments**  48332801 860.1500 Straw berry New DER; 48332801.der.docA cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 2 of 17 **Executive S ummary**  Acibenzolar - S - methyl, [benzo(1,2,3)thiadiazole - 7 - carbothioic acid - S - methyl ester], is a sy stemic chemical and is in a class of chemicals known as the benzothiadiazoles. It is a “plant activator” that acts to protect the plant against invading pathogens b y stimulating the plant’s natural defense mechanisms. It is currently registered for use on bulb vegetables, leafy vegetables, Brassica leafy vegetables, fruiting vegetables, cucurbits, and tobacco for the control or suppressi on of downy mildew, black rot, white rust, bacterial leaf spot and speck, and blue mold. This document summarizes the available residue chemistry data submitted by the petitioner in support of proposed new use on food crops. Under PP# 0 E 7818 , the Interregional Research Project No. 4 (IR - 4), on behalf of the Agricultural Experiment Stations of CO, NM, UT, ID, and WA, is proposing to amend the product label for Actigard® (EPA Reg. No. 100 - 922), a water - dispersible granule (WG) formulation containing 50% acibenzolar - S - methyl , to add a new use on the low growing berry crop subgroup 1 3 - 07 G . Actigard® is proposed for multiple foliar applications on the low growing berry subgroup using ground or aerial equipment at a maximum of 0.0 23 lb ai /A per application, a maximum seasonal rate of 0. 188 lb ai/A, and a 0 - day pre - harvest interval (PHI). Concurrentl y, IR - 4 requests the establishment of a permanent tolerance for “the combined residues of the fungicide acibenzolarS - methyl, benzo(1,2,3)thia diazole - 7 - carbothioic acid - S - methyl ester”, in/on:

Low growing Berry

Crop Subgroup 13 - 07G .............. 0.15 ppm Tolerances have been established under 40 CFR §180.561 for residues of acibenzolar- S - methyl. Permanent tolerances have been established under §180.561(a) for spinach (1.0 ppm), tomato paste (3.0 ppm), Brassica leafy vegetables, group 5 (1.0 ppm), fruiting vegetables (1.0 ppm), onion, bulb, subgroup 3 - 07A (0.1 ppm), vegetable, cucurbit, group 9 (2.0 ppm) and leafy vegetables (0.25 ppm), and for bananas (0.1 ppm) as an import tolerance. There are no established Codex, Mexican, or Canadian maximum residue limits (MRL s) for acibenzolar - S - methyl in/on any commodity . The nature of the residues in plants has been adequately delineated based on metabolism studie s with lettuce, tomato, rice, wheat, and tobacco. The HED MARC has determined that the residues of concern in plants for tolerance expression are residues convertible to benzo(1,2,3)thiadiazole - 7 - carbox yl ic acid (CGA - 210007), expressed as acibenzolar - S - me thyl. For purposes of risk assessment, residues of the 4 - and 5 - hydrox y metabolites (CGA - 324041 and CGA - 323060) of CGA - 210007 should also be considered. For this purpose, the toxicity of CGA - 324041 and CGA - 323060 should be considered to be equivalent to that of the parent compound acibenzolar - S - methyl. These residues occur in significant quantities in lettuce and tomato. At this time, acibenzolar - S - methyl does not have common metabolites with other agrochemicals. The residues of concern for drinking water are the same as those listed in the tolerance expression; that is, residues convertible to CGA - 210007. There are no livestock feedstuffs associated with the proposed use on the low growing berry subgroup . Therefore, no livestock metabolism data, enfo rcement methods, storage stability data, or feeding studies are required to support th is petition .A cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 3 of 17 HPL C/UV Method AG - 6 17 A is available for tolerance enforcement. This method has undergone a successful tolerance method validation by the Anal yt ical Chemistry Branch of BEAD. The method was forwarded to FDA for inclusion in PAM Volume II. Briefl y, the method consists of an initial hydrol ys is with NaOH to convert acibenzolar - S - methyl to CGA 210007 followed b y methanol extraction. Residues are then diluted w ith HCl and purified b y a series of solid - phase extraction steps. Prior to HPLC/UV anal ys is, residues are partitioned into ethyl acetate, dried down, and re - dissolved in phosphoric acid. This method has a limit of quantitation (L OQ) of 0.02 ppm. The method includes optional detection via HPL C/MS, giving a means of residue confirmation. Strawberry samples from the submitted field trial studies were anal yz ed for residues of acibenzolar - S - methyl using Method AG - 6 71 A, an L C/MS method adapted with modificati on from the enforcement method. The methods are adequate for data collection based on acceptable method verification and/or concurrent method recoveries. With respect to FDA’s multiresidue methods, acibenzolar - S - methyl yielded adequate response to GL C an al ys is and was completely recovered through the Protocol D and E multiresidue methods. Partial recovery

of CGA - 210007 was achieved via Protocol B after derivatization. The number and locations of field trials are in accordance with OPPTS Guideline 860.15 00 for strawberry as the representative crop of the low growing berry subgroup 1 3 - 07 G . The trials conducted reflect the proposed maximum seasonal rate and pre - harvest interval ( PHI ) , and there are adequate storage stability data to validate sample storage conditions and duration. The majority of the tank mixes contained an adjuvant. The results for strawberry indicate that following application of the test formulation according to the proposed use rate and PHI ( 0 - day) , the maximum residues of acibenzol ar - S - methyl (i.e., residues convertible to CGA - 210007) were 0.0 87 ppm in/on strawberry . The residue data was entered into the OECD MRLCalculator . T he available strawberry field trial data are deemed adequate to support a tolerance of 0.1 5 ppm for residu es of acibenzolar - S - methyl in/on the low growing berry subgroup 1 3 - 07 G . Adequate confined rotational crop data have been submitted previously . Based on the available field rotational crop data for wheat, turnip, and lettuce, it has been determined that tolerances are not needed for rotational crops provided a 30 - day plant - back interval is established for all crops not listed as target crops. The existing rotational crop restrictions on the Actigard® label are adequate. **Regulatory Recommendations and Res idue Chemistry Deficiencies**  HED has examined the residue chemistry database for acibenzolar - S - methyl . Pending submission of a revised Section F (see requirements under Proposed Tolerances), there are no residue chemistry

issues that would preclude granti ng a Section 3 registration for the requested use of acibenzolar - S - methyl on crop subgroup 1 3 - 07 G , or the establishment of a tolerance for residues of acibenzolar - S - methyl, as follow s :

Berry , low growing, subgroup 13 - 07G ............0.15 ppmA cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 4 of 17 **Note to Regi stration Division**  : Tolerances have been established under 40 CFR §180.561 for residues of acibenzolar - S - methyl. HED continues to recommend that the appropriate tolerance expression for acibenzolar - S - methyl read as follows. Tolerances are established f or residues of acibenzolar - S - methyl, benzo(1,2,3)thiadiazole - 7 - carbothioic acid - S - methyl ester, including its metabolites and degradates, in or on the commodities in the table below. Compliance with the tolerance levels specified below is to be determined by

measuring only

those acibenzolar - S - methyl residues convertible to benzo(1,2,3)thiadiazole - 7 - carbox yl ic acid (CGA - 210007), expressed as the stoichiometric equivalent of acibenzolar - S - methyl, in or on the commodity . A human health risk assessment is forthcoming. 860.1550 Proposed Tolerances For the low growing berry subgroup petition, a revised Section F is required to remove reference to “combined” residues in the tolerance expression . 860.1 6 50 Submittal of Analy tical Reference Standards Analy tical standards for acibenzolar - S - methyl (CGA 245704) and its metabolite CGA 210007 are currentl y available in the EPA National Pesticide Standards Repository (personal communication with Theresa Cole, ACB, 6/21/11). acibenzolar metabolite (CGA210007), expir ation date 1/31/2012 acibenzolar S - methyl (CGA245704), expiration date 3/1/2013 If new standards are being submitted, they should be sent to the Anal yt ical Chemistry L ab, which is located at Fort Meade, to the attention of either Theresa Cole or Thuy Ngu yen at the following address: USEPA National Pesticide Standards Repository /Anal yt ical Chemistry

Branch/OPP 701 Mapes Road Fo rt George G. Meade, MD 20755 - 5350A cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 5 of 17 **Background** Acibenzolar - S - methyl has been classified as a reduced - risk compound wi th a mode of action that mimics the natural sy stemic activated resistance response found in most plant species. The chemical structures and nomenclature of acibenzolar - S - methyl and its metabolites CGA - 210007 are presented in Table 1. The phy sicochemical properties of technical grade acibenzolar - S methyl are listed in Table 2. **Table 1. Acibenzolar**  **- S**  **- Methyl**  **Nom enclature**  . Chemical structure

N N S O S C H 3

mo l wt 210 Common name acibenzolar - S - methyl Com pany experimental name CGA - 245704 IUPAC name benzo[1,2,3]thiadiaz ole - 7 - carbothioic acid - S - me thyl ester CAS name S - methyl, 1,2,3 - bensothiadiazole - 7 - carbothioate CAS registry number 135158 - 54 - 2 End - use product (EP) Actigard® 50 WG (EPA Reg. No. 100 - 922) Structure of metabolite CGA - 210007 N N S C O O H

1,2,3 - benzothiadiazole - 7 - carboxylic acid

mo l wt 180A cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 6 of 17 **Table 2. Physicochemical Properties of**  **Acibenzolar -**  **S -**  **Methyl .**  Parameter Value Reference Melting point/range 133 oC DP# 250963, 2/10/99, H. Podall pH 7 . 9 (1% solution in water) at 25 o C Density 1.54 g/cc at 22 o C (bulk density) Water solubility 0.0077 g/L Solvent solubility Methanol 4.2 g/L Acetone 28 g/L Toluene 36 g/L n - Octanol 5.4 g/L n - Hexane 1.3 g/L Ethyl acetate 25 g/L Dichlorome thane 160 g/L European Commission, 6506/VI/99 - final, 5/21/02

Vapor pressure 3.6 x 10 - 6 Torr at 25 oC DP# 250963, 2/10/99, H. Podall Dissociation constant, pKa No dissociation constant in an accessible pH range (1.0 - 9.0) European Commission, 6506/VI/ 99 - final, 5/21/02 Octanol/water partition coefficient, Log(KOW ) Log ( P OW ) 3.1 at 25 oC DP# 250963, 2/10/99, H. Podall UV/visible absorption spectrum 253 nm ε : 14.7 10 3 l mol - 1 cm - 1 288 nm ε : 5.4 10 3 324 nm ε : 7.2 10 3 European Commission, 6506/VI/99 - fi nal, 5/21/02 **860.1200 Directions for Use** The proposed end - use product relevant to this registration action is Actigard® (EPA Reg. No. 100 - 922), a 50% WG formulation. The proposed new uses are summarized in Table 3. **Table 3.**  **Summary of Directions for Use of**  **Acibenzolar -**  **S -**  **me thyl**  **.** Applic. Timing, Ty pe, and Equip. Form

ulation [EPA Reg. No.] Applic.

Rate (lb ai/A) Max. No.

Applic. per Season Max. Seasonal

Applic. Rate (lb ai/A) PHI

(days) Use Directions and

Limitations Lo w growing berries Postemergence Foliar or banded Ground /Aerial 50% WG

[100 - 922] 0.0 16 -

0.0 23 8 (Implied) 0. 188 0 Scout fields regularly for angular leaf spot after plant establishment. Begin applications when disease appears, and continue on a 7 - to 14 - day interval. The following additional use directions were specified for low growing berries : “Apply in sufficient water to ensure adequate coverage. Banded applications can be made early when plants are small (7” proportional band) as long as coverage is adequate. Actigard® should b e applied to health y, actively growing plants. Do not apply prior to thinning or within 5 days of transplanting. Do not apply Actigard® to plants that are stressed due to drought, excessive moisture, cold weather, or herbicide injury , etc. Avoid spray o verlap as injury

may occur. Conclusions . The submitted label is adequate to allow evaluation of the residue data relative to the proposed use rate and PHI on low growing berries .A cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 7 of 17 **860.1300 Nature of the Residue -**  **Plants** DP# 246673, 3/15/00, M. Doherty (PP#8F4974) DP# 258680 , 5 /2 /00, M. Doherty (PP# 9 F 6004 ) No new plant metabolism studies were submitted. Adequate metabolism studies with [phenyl - U 14 C]acibenzolar - S - methyl on lettuce, tomato, rice, wheat, and tobacco have previously been submitted and reviewed. The metabolic pathway for acibenzolar - S - methyl was similar in all tested crops and involves the hydroly sis of the S - methyl ester to form the carboxylic acid metabolite (CGA - 210007). Further hydrox yl ation of CGA - 210007 at the C - 4 and C - 5 positio ns results in the formation of the minor metabolites CGA - 324041 (all crops tested) and CGA 323060 (lettuce and tomato), which may also form acyl sugar conjugates. The HED MARC (11/9/99) determined that the residue s of concern for tolerance expression are residues convertible to benzo(1,2,3)thiadiazole - 7 - carbox yl ic acid (CGA - 210007), expressed as acibenzolar - S - methyl. For purposes of risk assessment, residues of the 3 - and 5 - hydroxy (CGA 324041 and CGA - 323060) metabolites of CGA - 210007 should also be inclu ded and considered as toxic as parent acibenzolar - S - methyl. The metabolism studies on dissimilar crops are adequate to support the requested use s on low growing berries . For the purposes of these petitions, the residue of concern for tolerance setting and risk assessment purposes is considered to be residues of the parent acibenzolar - S - methyl and CGA - 210007, expressed as acibenzolar - S - methyl . **860.1300 Nature of the Residue -**  **Livestock** There are no livestock feedstuffs associated with the proposed use o n the low growing berry subgroup 1 3 - 07 G . Therefore, data requirements for livestock metabolism are not relevant to these tolerance petitions. **860.1340 Residue Analytical Methods** DP# 246673, 3/15/00, M. Doherty (PP#8F4974) DP# 258680 , 5 /2 /00, M. Doherty (PP# 9 F 6004 ) Enforcement method: An adequate method, HPLC/UV Method AG - 6 17 A, is available for tolerance enforcement. This method has undergone a successful tolerance method validation by the Anal yt ical Chemistry Branch of BEAD. The method was forwarded to FDA for inclusion in PAM Volume II. Briefl y, the method consists of an initial hydroly sis with NaOH to convert acibenzolar - S - methyl to CGA - 210007 followed by methanol extraction. Residues are then diluted with HCl and purified by a series of solid - phase extraction steps. Prior to HPL C/UV anal ys is, residues are partitioned into ethyl acetate, dried down, and re - dissolved in phosphoric acid. This method has an L OQ of 0.02 ppm. The method includes optional detection via HPL C/MS, g iving a means of residue confirmation. Data collection methods: Crop samples from the submitted strawberry field trial studies were anal yz ed for residues of acibenzolar - S - methyl using a LC/MS method adapted with modification from the enforcement method AG - 617A. Modifications from Method AG - 671A included using MS detection (confirmatory method) and, therefore, the alkaline par t itioning, C18 SPE and acidic partitioning cleanup steps were not needed. Briefl y, samples of homogenized strawberryA cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 8 of 17 were extrac ted with water:1 N NaOH (9:1, v:v) at 6 5 - 70 oC for 30 minutes. After cooling, methanol was added with shaking for 30 minutes. Following vacuum filtering, the solvent was evaporated to a known volume of 8 - 10 mL , and an equal amount of 1 M HCl was added fo r cleanup through a Chem Elute column; residues were eluted with dichloromethane :ethyl acetate (9:1, v:v). The sample was then evaporated to dryness at 35 C, taken up in 80:20 0.5% formic acid:acetonitrile, sonicated, and filtered prior to analy sis of CGA - 210007 by L C/MS. The column used was Agilent, Zorbax SB - C18. The mobile phase was 0.5% formic acid (mobile phase A) and acetonitrile (mobile phase B). The method was validated at 0.02 ppm. The L C/MS method used to determine residues of acibenzolar - S - m ethyl in/on strawberry was adequatel y validated in conjunction with field sample anal ys es. Concurrent method recoveries from strawberry

fortified with acibenzolar - S - methyl at 0.02 - 2.0 ppm were generally within the acceptable range of 70 - 120% (see Table C. 1); one slightly high recovery

(130%) was obtained at the 0.02 ppm fortification level. Adequate sample calculations and chromatograms were provided. The fortification levels used in the concurrent method recoveries were adequate to bracket expected resi due levels in/on strawberry . Conclusions . The L C/MS methods are adequate for collecting data on residues of acibenzolar - S methyl in/on strawberry . The existing enforcement method (HPL C/UV Method AG - 617A) will be adequate for tolerance enforcement purpos es for low growing berry subgroup . **860.1360 Multiresidue Methods** DP# 246673, 3/15/00, M. Doherty (PP#8F4974) Data depicting the recovery of acibenzolar - S - methyl and its metabolite, CGA - 210007, using FDA Multiresidue Methods, were previously submitted. Acibenzolar - S - methyl yi elded adequate response to GLC analy sis and was completel y recovered through Protocols D and E. Partial recovery of CGA - 210007 was achieved via Protocol B after derivatization . **860.1380 Storage Stability** DP# 246673, 3/15/00, M. Do herty (PP#8F4974) 47375301.der.doc Previously

submitted storage stability data indicate that residues of acibenzolar - S - methyl and CGA - 210007 are relatively stable in a variety of crops. Acibenzolar - S - methyl and CGA - 210007 were stable in/on fortified samp les of cabbage, green and cured tobacco, leaf lettuce, squash, tomato, turnip roots, wheat grain, and wheat straw stored frozen for up to ~21 - 23 months. The storage durations and conditions of samples from the crop field trials submitted to support the subject petitions are presented in Table 4. IR - 4 provided supporting storage stability data in conjunction with the strawberry field trials. Samples of untreated strawberry were fortified with acibenzolar - S - methyl at 0.20 ppm and stored frozen with the fie ld trial samples. No zero - day data were provided. T he concurrent storage stability data indicate that fortified residues of acibenzolar - S - methyl are stable in/on strawberries stored frozen for up to 317 day s.A cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 9 of 17 **Table 4. Summary of Storage Conditions and**  **Durations of Samples from** **Crop Field Trial Studies.**  Matrix Storage Temperature

(°C) Actual

Maximum Storage Duration Interval of Demonstrated Storage Stability Strawberry ≤ - 21 314 days ( 10.3 months) Concurrent storage stability data indicate that resid ues of acibenzolar - S - methyl are stable for up to 317 days in/on frozen strawberrie s. Conclusions . The concurrent storage stability data are adequate t o support the storage durations and conditions of samples o f strawberry from the submitted crop field trial studies. Z ero - day data were not provided with the concurrent storage stability studies. IR - 4 is reminded that storage stability

studies should alway s include a zero - day sampling interval to establish the residue levels present at the time samples are placed into storage [see OPPTS 860.1380(d)(6)(i)]. No corrections for potential decline during storage are needed for any of the residue studies. **860.1400 Water, Fish, and Irrigated Crops** There are no proposed uses that are relevant to this guideline topic. **860.1460 Food Handling** There are no proposed uses that are relevant to this guideline topic. **860.1480 Meat, Milk, Poultry, and Eggs** There are no livestock feedstuffs associated with the proposed uses on strawberry . Therefore, data requirements pertaining to meat, milk, poultry, and eggs are not relevant to th is tolerance petition . **860.1500 Crop Field Trials** IR - 4 has submitted field trials supporting a new use of acibenzolar - s - methyl on strawberries . The results from these studies are summari zed and discussed below. Strawberry DER Reference: 483328 01.der.doc ( Straw berry) IR - 4 has submitted field trial data for acibenzolar - s - methyl on strawberries. A total of ten field trials were conducted in the United States in EPA Regions 1 (NY, 1 trial) , 2 (NC, 1 trial), 3 (FL, 1 trial), 5 (WIand Ontario, 2 trials), 5B (Quebec, 1 trial), 10 (CA, 3 trials), and 12 (OR, 1 trial) during the 2008 growing season. Geographic representation was adequate. At each test location, the treated plots received eight to nine foliar broadcast applications of a water dispersible granule (WG) formulation of acibenzolar- s - methyl (100 - 922, 50%WG) at 0.023 lb ai/A with a 6 to 9 - day retreatment interval (RTI) and a spray volume that ranged from 27 - 59 gal/A (GPA). Samples o f strawberries were harvested at maturity on the day of the last application (0 - day

preharvest interval (PHI)). One site also collected decline samples at 0, 3, 7, 10, and 14 day s after the last application (DALA). An adjuvant was included in the majority ofA cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 10 of 17 tank mixes. Strawberry samples from the submitted field trial studies were anal yz ed for residues of acibenzolar - S - methyl using Method AG - 6 71 A, an L C/MS method adapted with modification from the enforcement method. The limit of quantitation (L OQ) was 0.02 ppm. IR - 4 provided supporting storage stability data in conjunction with the strawberry field trials. Samples of untreated strawberry were fortified with acibenzolar - S - methyl at 0.20 ppm and stored frozen with the field trial samples. No zero - day data were provided. Samples were stored frozen for up to 314 day s. T he concurrent storage stability data indicate that fortified residues of acibenzolar - S - methyl are stable in/on strawberries stored frozen for up to 317 day s. The results from these trials show that residues ranged from 0.020 - 0.087 ppm after eight to nine applications of approximately 0.023 lb ai/A per application of Actigard 50WG for a total seasonal application rate of 0.183 - 0.214 lb ai/A. Residue decline data show that acibenzolar - S methyl residues decrease in/on strawberries with increasing PHIs. **TABLE 6**  **. Summary of Residue Data from**  **Strawberry Field Trials with Acibenzolar**  **- S**  **- me**  **thyl.** Commodity Total Applic.

Rate (lb ai/A) PHI

(days) Residue Levels (ppm)

n Min. Max. HAFT 1 Me dian (STMdR) Mean

(STMR) Std. Dev. Strawberry ~0.188 0 20 0.020 0. 087 0. 085 0.040 0.046 0.022 1 HAFT = Highest Average Field Trial. Conclusions . The petitioner has provided adequate residue data reflecting the maximum proposed use pattern of acibenzo lar - S - methyl on s trawberries . The residue data is supported by adequate storage stability data and an acceptable analy tical method was used for quantitation. The residues of acibenzolar - S - methyl did not exceed the proposed tolerance level of 0.15 ppm in strawberries. The residues ranged from 0. 020 - 0.087 ppm. **860.1520 Processed Food and Feed** There are no processed food/feed commodities associated with the proposed use s on low growing berries . Therefore, data requirements pertaining to processed food an d feed are not relevant to the s e tolerance petition s . **860.1650 Submittal of Analytical Reference Standards** Analy tical standards for acibenzolar - S - methyl (CGA 245704) and its metabolite CGA 210007 are currentl y available in the EPA National Pesticide Stan dards Repository (personal communication with Theresa Cole , ACB, 6/21 / 11 ). acibenzolar metabolite (CGA210007), expiration date 1/31/2012 acibenzolar S - methyl (CGA245704), expiration date 3/1/2013 If new standards are being submitted, they should be sent to the Anal yt ical Chemistry L ab, which is located at Fort Meade, to the attention of either Theresa Cole or Thuy Ngu ye n at the following address:A cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 11 of 17 USEPA National Pesticide Standards Repository /Anal yt ical Chemistry

Branch/OPP 701 Mapes Road Fort Georg e G. Meade, MD 20755 - 5350 (Note that the mail will be returned if the extended zip code is not used.) **860.1850 Confined Accumulation in Rotational Crops** DP# 246673, 3/15/00, M. Doherty (PP#8F4974) Adequat e confined rotational crop studies with acibenz olar - S - methyl have been submitted previously

and reviewed by HED . Following treatment of soil with [ 14 C]acibenzolar - S - methyl at ~0.375 lb ai/A (~1x the maximum seasonal rate to annual crops), total radioactive residues (TRR) accumulated at >0.01 ppm in/on all crop samples from the 30 - and 60 - day plant - back intervals ( PBIs ) . The relative TRR levels among the various RACs were the same at both PBIs, although TRR values were slightl y higher at the 61 - day PBI for each RAC, except radish root. At both PBI s, l evels of radioactivity were lowest in radish roots (0.013 ppm) and highest in wheat forage (0.107 and 0.157 ppm): TRR in the remaining RACs ranged from 0.021 - 0.083 ppm. From the 210 - day

PBI samples, TRR ranged from 0.004 ppm in radish roots to 0.026 ppm in 25% mature wheat forage. In the 30 - and 61 - day

PBI samples, the principal metabolite identified in rotational crops was free CGA - 210007, accounting for 1.3 - 10.5% of the TRR. Other metabolites that were tentatively

identified included the h ydrox yl ated acid metabolites, CGA - 324041 and CGA 323060, which together accounted for <0.8 - 15.9% of the TRR. Trace levels of parent (0.1 - 0.7% TRR) were also detected along with minor unknown metabolites generally accounting for <9% of the TRR. Cellulase and base h ydrol ys es of extracted residues indicated that metabolite CGA 210007 also occurs as an acyl sugar conjugate in rotational crops. Following hydrol ys is, CGA 210007 accounted for 17.3% TRR and 30.4% TRR in wheat forage from the 30 - and 61 - day PBIs, respectivel y. In the 210 - day PBI samples, residues of acibenzolar - S - methyl (< 0.002 ppm) and CGA - 210007 (  0.001 ppm) were identified in all samples. Residues of CGA - 324041 and CGA - 323060 were not identified in any extracts. The confined rotational crop data indicate that the metabolism of [ 14 C]acibenzolar - S - methyl in rotational crops is similar to the metabolism observed in the primary crops (lettuce, tomato, wheat, and rice). HED has determined that acibenzolar - S - methyl + its metabolites convertible to GCA - 210007 + CGA - 324041 + CGA - 323060 comprise the total toxi c residue for acibenzolar - S methyl in rotational crops. As combined levels of these residues exceeded 0.01 ppm in rotational wheat forage from both the 30 and 61 - day PBIs, a limited field rotational crop study for cereal grain crops was required. **860.190 0 Field Accumulation in Rotational Crops**  DP# 246673, 3/15/00, M. Doherty (PP#8F4974) DP# 258680 , 5 /2 /00, M. Doherty (PP# 9 F 6004 )A cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 12 of 17 An adequate limited rotational crop study with wheat, turnips, and lettuce was previousl y submitted. Th e primary

crops (leaf lettuce, pepper, squash, or turnips) received four foliar applications of the 50% DF formulation of acibenzolar - S - methyl, with a 7 - day retreatment interval, at 0.094 lb ai/A per application for a total applied rate of 0.376 lb ai/A (3x the maximum proposed seasonal use rate for bulb onion; ~1x the maximum proposed seasonal use rate for cucurbit vegetables ). Following harvest of the primary crops, rotational crops of winter or spring wheat, leaf lettuce, and turnips were planted

 30, 60, and 180 day s after the final treatment. Residues of acibenzolar - S - methyl and its metabolites containing the CGA - 210007 moiety were nondetectable (<0.02 ppm) in all rotational crop matrices (wheat forage, hay , straw and grain ; turnip tops and roots; and leaf lettuce) from the 30 - day PBI. While tolerances are not needed for rotational crops, a plant - back restriction of not less than 30 days is required based on the results from the confined rotational cro p study . A 30 - day plant - back interval for all crops not listed as target crops on the acibenzolar - S - methyl label was required. The recommended rotational crop restrictions agree with the established rotational crop restrictions for the 50% WG formulation (see 860.1200 Directions for Use). **860.1550 Proposed Tolerances** Tolerances are currentl y listed in 40 CFR §180.561 for residues of acibenzolar - S - methyl, (benzo(1,2,3)thiadiazole - 7 - carbothioic acid - S - methyl ester). The HED MARC has determined that the residues of concern in plants for tolerance expression are residues convertible to benzo(1,2,3)thiadiazole - 7 - carbox yl ic acid (CGA - 210007), expressed as acibenzolar - S - methyl. **Note to Registration Division** : Tolerances have been established under 40 CFR § 180.561 for residues of acibenzolar - S - methyl. HED continues to recommend that the appropriate tolerance expression for acibenzolar - S - methyl read as follows. Tolerances are established for residues of acibenzolar - S - methyl, benzo(1,2,3)thiadiazole - 7 - carb othioic acid - S - methyl ester, including its metabolites and degradates, in or on the commodities in the table below. Compliance with the tolerance levels specified below is to be determined b y measuring only those acibenzolar - S - methyl residues convertible t o benzo(1,2,3)thiadiazole - 7 - carbox yl ic acid (CGA - 210007), expressed as the stoichiometric equivalent of acibenzolar - S - methyl, in or on the commodity . The results for strawberry indicate that following application of the test formulation according to the p roposed use rate and PHI (0 - day) , the maximum residues of acibenzolar - S - methyl (i.e., residues convertible to CGA - 210007) were 0.0 87 ppm in/on strawberry . The residue data was entered into the OECD MRLCalculator . T he available strawberry field trial dat a are deemed adequate to support a tolerance of 0.1 5 ppm for residues of acibenzolar - S - methyl in/on the low growing berry subgroup 1 3 - 07 G . The proposed tolerances are listed in Table 7, along with the tolerance level recommended by HED.A cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 13 of 17 There are no es tablished Codex, Mexican, or Canadian maximum residue limits for acibenzolar S - methyl in/on any

commodity . **Table 7**  **. Tolerance Summary for**  **Acibenzolar -**  **S -**  **me thyl.**  Commodity Established Tolerance (ppm) Proposed

Tolerance (ppm) Recommended

Tolerance (ppm) Comments; Correct

Commodity Definition **Tolerances to be listed**  **under 40 CFR §180. 561**  **(a):** B erry , low

growing subgroup 1 3 - 07 G -- 0. 15 0.1 5 **References** DP#: 250963 Subject: TGAI Product Chemistry Review /Action: 100 EPA Reg./File Symbol No.: 100 - OER Chemical: CGA - 245704 Technical; 1,2,3 - benzothiadiazole - 7 - carbothioic acid - S - me thyl ester Com pany: Novartis Crop Protection Inc. From : H. Podall To: C. Giles - Parker/M. Rodriguez Dated: 02/10/99 MRIDs: 44537001 - 03 DP#: 259930 Subject: Acibenzolar - S - methyl: Briefing Memorandum for the HED Metabolism Assessment Review

Committee. From : M. Doherty To: G. Kramer and HED Metabolism Assessment Review Committee Dated: 11/3/99 MRIDs: None DP#: 246673 Subject: PP8F4974: Request for Use of Acibenzolar - S - me thyl on Leafy Vegetables and Fruiting

Vegetables. Evaluation of Analytical Methods and Residue Chemistry Data. From : M. Doherty To: C. Giles - Parker/M. Rodriguez Dated: 3/15/00 MRIDs: 44014255, 44537051 - 63, 44828008, 44828025, 44993301 - 02A cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 14 of 17 DP#: 258680 Subject: PP 9F06004: Request for Use of Acibenzolar - S - methyl on Brassica Leafy Vegetables and

Bananas (imported). Evaluation of Analytical Methods and Residue Chemistry Data. From : M. Doherty To: C. Giles - Parker/M. Rodriguez Dated: 5/2/00 MRIDs: 44828004 - 07, 4482800 9 - 24 DP#: 250386 Subject: PP#s 8F4984, 8F5031, and 0F6141. Tolerance Petitions for the Use of Pymetrozine on Cotton,

Hops, Pecans, Leafy Vegetables (Except Brassica Vegetables), Head and Stem Brassica, Leafy Brassica Greens, Turnip Greens, Cucurbits, an d Fruiting Vegetables. Evaluation of Residue Chemistry and Analytical Methodology. From : D. Dotson To: Daniel Peacock/Deborah McCall Dated: 11/19/01 MRIDs: 44572208, 44647906 - 08, 44753301, 44753302, 44830401, 45091001 - 04, 45105301, 45301301 Attachments: International Residue Limit Status sheet Appendix 1: Tolerance Assessment CalculationsA cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 15 of 17 **A ppendix I**  **. International Residue Limits.** 



### Acibenzolar - s - me thyl (061402; 03/2 9 /2011) Summary of US and Internati onal Tolerances and Maximum Residue Limits Residue Definition : US Canada Mexico 2 Codex 3 40 CFR 180. 561 : Plant s : acibenzolar - S - methyl, benzo(1,2,3)thiadiazole - 7 - carbothioic acid S - methyl ester, including its metabolites and degradates, in or on the commodities in the table below. Compliance with the tolerance levels specified below is to be determined by measuring only those acibenzolar - S - methyl residues convertible to benzo(1,2,3)thiadiazole - 7 - carboxylic acid (CGA - 210007), expressed as the stoichiometric equivalent of acibenzolar - S methyl, Livestock: Not established 1,2,3 -benzothiadiazole - 7 - carbothioic acid S - methyl ester, including the metabolite 1,2,3 - benzothiadiazole - 7 - carboxylic acid None

Tolerance (ppm) /Maximum Residue Limit (mg/kg) US Canad a Mexico 2 Codex Berry , low growing, subgroup 13 - 07G 0.15

Completed: M. Negussie; 03/31/2011 1 Includes only commodities of interest for this action. Tolerance values should be the HED recommendations and not those proposed by the applicant. 2 Mexico adopts US tolerances and/or Codex MRLs for its export purposes. 3 * = absent at the limit of quantitation; Po = postharvest treatment, such as treatment of stored grains. PoP = process ed postharvest treated commodity, such as processing of treated stored wheat. (fat) = to be measured on the fat portion of the sample. MRLs indicated as proposed have not been finalized by the CCPR and the CAC.A cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 16 of 17 **Appendix I**  **I. Tolerance Assessment Calculat ions.**  **Berry, low growing, subgroup 13 -**  **07G** The dataset used to establish a tolerance for acibenzolar - S - methyl on the Berry , low growing, subgroup 13 - 07G consisted of field trial data for strawberry (the representative crop of Berry , low growing, subgroup 13 - 07G ), representing application rates of 0. 188 lb ai/A (8 applications at 0.0 235 lb ai/A/application) with a 0 - day PHI. As specified b y the SOP, the field trial application rates and PHIs are within 25% of the maximum label application rate and minimum label PHI , respectivel y. The residues values used to calculate the tolerance are provided below . Using the OECD MRL Calculator , the recommended tolerance is 0.15 ppm for Berry , low growing, subgroup 13 - 07G . **Residues (mg/kg) n**  0.021 1 0.024 1 0.026 1 0.029 1 0.039 1 0.045 1 0.0625 1 0.063 1 0.065 1 0.085 1A cibenzolar - s - Methyl Summary of Analytical Chemistry and Residue Data DP#: 3 86364

Page 17 of 17 **Acibenzolar s methyl**  **Berry, low growing, subgroup 13 -**  **07G US**  **0 -**  **day Total number of data (n)**  10 **Percentage of censored data** 0% **Number of non -**  **censored data** 10 **Lowest residue** 0.021 **Highest residue** 0.085 **Median residue** 0.042 **Mea n**  0.046 **Standard deviation (SD)** 0.022 **Correction factor for censoring (CF)** 1.000 **Proposed MRL estimate**

**- Highest residue**  0.085

**- Mean + 4 SD**  0.133

**- CF x 3 Mean**  0.138 **Unrounded MRL** 0.138 **Rounded MRL 0.15**