                    UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
                               WASHINGTON, D.C.  20460
                                          
                                       
	OFFICE OF CHEMICAL SAFETY
	AND POLLUTION PREVENTION
                                                                               


MEMORANDUM

Date:  		9/7/2012

SUBJECT:	Fluazinam.  Petitions for the Establishment of Tolerances and Registration of New Uses on Soybeans, the Melon Subgroup (9-A), and the Pepper/Eggplant Subgroup (8-10B).  Summary of Analytical Chemistry and Residue Data


PC Code:  129098
DP Barcodes:  D397721, D398888, D399475
Decision Nos.:  459107, 459565, 461245
Registration Nos.:  71512-1
Petitions:  1E7959, 8E7506, 2F7977
Regulatory Action:  Section 3 Registration
Risk Assessment Type:  Single Chemical
                    Aggregate
Case No.:  7013
TXR No.:  NA
CAS No.:  79622-59-6
MRID Nos.:  48704501, 48704502, 
                       48707301, 48712601 
40 CFR:  §180.574


		              									
FROM:  	 Douglas Dotson, Ph.D., Chemist
		 Risk Assessment Branch II
		 Health Effects Division (7509P)	
		
THROUGH:	 Dennis McNeilly, Chemist
		 Risk Assessment Branch II
		 Health Effects Division (7509P)

TO:		 Andrew Ertman/Barbara Madden, RM Team 5, RIMUERB
		 Dominic Schuler/Tony Kish, PM Team 22, Fungicide Branch
		 Registration Division (7505P)		
		


Executive Summary

Fluazinam (3-chloro-N-[3-chloro-2,6-dinitro-4-(trifluoromethyl)phenyl]-5-(trifluoromethyl)-2-pyridinamine) is a member of the phenyl-pyridinamine class of compounds.  It is a non-systemic, preventive, contact fungicide with a multi-site mode of action.  When applied to plants, it remains primarily on the plant surface, it is not taken up to any extent by the plant, and it is not translocated within the plant.  Fluazinam operates by disrupting the production of energy at several metabolic sites within the fungal cell.  

Tolerances have been established in 40 CFR §180.574(a)(1) for residues of fluazinam in/on plant commodities at levels ranging from 0.01 in or on Brassica leafy vegetables (Crop Group 5) to 7.0 ppm in or on the Bushberry subgroup (13-07B).  A tolerance has been established in 40 CFR §180.574(a)(2) for residues of fluazinam and its metabolite AMGT on wine grapes at 3.0 ppm (40 CFR).  Finally, a tolerance of 0.05 ppm has been established for fluazinam and its metabolites AMPA and DAPA and their sulfamate conjugates in or on the fat and meat byproducts of cattle, goats, horses, and sheep.

The registrants, ISK Biosciences and the Interregional Research Project No. 4 (IR-4) submitted two petitions to establish tolerances for fluazinam in/on soybeans and the commodities of the melon and pepper/eggplant subgroups (9-A and 8-10B, respectively).  In addition, the registrant submitted residue data for the metabolite, AMGT (see Table 2 for chemical name and structure), on lettuce in order to fulfill a condition of registration cited in a previous HED risk assessment.  This document summarizes the available residue chemistry data in support of the tolerance petitions.  The end-use product (EP) relevant to all petitions is OMEGA[(R)] 500F (EPA Reg. No. 71512-1), a flowable concentrate (FlC) formulation containing 40% fluazinam (equivalent to 4.17 lb ai/gal).  

In PP# 2F7977, ISK Biosciences requested the registration of fluazinam for use on soybeans.  In conjunction with this petition, the registrant proposed tolerances of 0.01 ppm for soybean seed and 0.02 ppm for soybean hulls.  In PP# 1E7959, IR-4 requested the registration of fluazinam for use on the commodities of the melon and pepper/eggplant subgroups.  In conjunction with this petition, IR-4 proposed tolerances of 0.08 ppm for melons and 1.5 ppm for peppers and eggplant.    

In a previous tolerance petition, PP#8E7506, IR-4 requested registration of fluazinam for use on both head and leaf lettuce.  In conjunction with this use, a tolerance of 0.02 ppm was established for head lettuce and a tolerance of 2.0 ppm was established for leaf lettuce.  HED cited the following data deficiency with respect to the lettuce field trials:  "Samples from the lettuce field trial need to be analyzed for residues of AMGT.  The residue data should also be supported by data depicting the frozen storage stability of AMGT in the lettuce samples.  HED recommends that these data be made a condition of registration" (D360840, D. Dotson, 3/31/2010).  IR-4 submitted the requested data.  The data are adequate.  Residues of AMGT were below the LOQ of 0.01 ppm in all field trial samples.  

Use Directions

The registrant submitted adequate use directions for the application of fluazinam to soybeans as well as the commodities of the melon and pepper/eggplant subgroups.  The use directions are sufficient to allow for evaluation of the submitted residue data relative to the proposed use patterns.  The available field trial data support the proposed use directions for the crops addressed in this summary document.

Nature of the Residue

The nature of fluazinam residues in plants is adequately understood based on metabolism studies on potatoes, peanuts, and grapes.  The Agency previously concluded that the residue of concern in peanuts and root and tuber vegetables is parent fluazinam for purposes of both dietary risk assessment and tolerance enforcement.  For all other crops, the residues of concern include both fluazinam and its metabolite AMGT for purposes of dietary risk assessment.  For setting tolerances, the residues of concern in plants include fluazinam and AMGT for wine grapes, but only fluazinam for all other plant commodities.

The nature of the residue in rotational crops is adequately understood.  Based on the absence of metabolites containing the intact fluazinam nucleus, HED has determined that the residue of concern in rotational crops is the parent compound, and that rotational crop tolerances are not required.  The available confined field rotational crop study supports the 30-day plant-back interval (PBI) that is currently listed on the label for crops without direct uses of fluazinam.

The nature of the residue in livestock is also understood based on acceptable goat and hen metabolism studies.  The fluazinam residues of concern in livestock commodities for tolerance enforcement and risk assessment were determined by HED to be parent plus the metabolites AMPA and DAPA, and their sulfamate conjugates.

Analytical Methods

Enforcement Method

An adequate GC/ECD method (6148-94-0170-MD-001) is available for enforcing fluazinam tolerances on plant commodities.  The method was adequately radiovalidated and underwent a successful ILV trial.  The method was forwarded to BEAD's Analytical Chemistry Branch (ACB) for a TMV trial, and was subsequently determined to be suitable as a tolerance-enforcement method.  The method LOQ is 0.01 ppm for fluazinam in plant commodities.  An adequate enforcement method for determination of AMGT is also available.  The method is an HPLC/UV method entitled "Method Evaluation for the Analysis of AMGT in Grapes."  The registrant submitted an adequate ILV for the method.  The range of fortification levels and the percent recovery values were acceptable.

Data-Collection Methods

Samples collected in the cantaloupe and pepper field trials, as well as in the soybean field trials and storage stability study, were analyzed for residues of fluazinam and AMGT using LC/MS/MS methods that were based on the GC/ECD plant enforcement method.  The methods are adequate for data collection based on acceptable method recovery data included in the reviewed studies.  The fortification levels used in the method validation trials bracketed the expected field residues.  The LOQ was 0.01 ppm in all tested commodities.  The LOD was 0.0045 ppm in cantaloupes and peppers and 0.003 ppm in soybean seed, forage, and hay.  In all trials, the data-collection method was validated prior to and in conjunction with the analysis of the field trial samples.
Multiresidue Methods

Adequate FDA multiresidue method (MRM) testing data are available for fluazinam and its metabolite AMGT.  As fluazinam is partially recovered through Sections 302, 303, and 304 of PAM Volume I, the MRMs can serve as a confirmatory procedure for residues of fluazinam.  The FDA MRM methods are not suitable for determining residues of AMGT.

Storage Stability

The registrant submitted storage stability data for soybeans.  The data are adequate and indicate that residues of fluazinam and AMGT are relatively stable in/on soybean forage, hay, and seed stored frozen for up to 153 days (5.0 months).

Field Trials

The submitted field trial data for fluazinam on soybeans, cantaloupes, and peppers are acceptable and support the proposed uses.  Adequate numbers of trials were conducted in the appropriate geographic regions at ~1x the maximum proposed rate, and the appropriate samples were collected from each test at the proposed PHI.  The data support the establishment of tolerances of 0.01 ppm on soybean seed, 0.07 ppm for melons, and 0.09 ppm for peppers and eggplant.  The OECD MRL calculation procedures were used for determining appropriate tolerance levels.  The conditional AMGT residue data for lettuce and the supporting storage stability data are acceptable.  

Processing Studies

The submitted soybean processing study is adequate.  Following application of fluazinam at a 1x application rate, residues of fluazinam and AMGT were each below the LOQ (<0.01 ppm) in/on all samples of soybean seed (RAC) and AGF; therefore, processing factors were not calculated for AGF.  Following application of fluazinam at a 10x application rate, fluazinam residues concentrated in hulls (5.3x) and in refined oil (1.1x), but did not concentrate in meal (processing factor of <0.22x).  Residues of AMGT were below the LOQ in/on seed, hulls, meal, and refined oil; therefore, processing factors were not calculated.  The observed concentration factors were less than the theoretical concentration factors.

Analytical Reference Standards

Analytical reference standards for fluazinam, AMPA, DAPA, and AMGT are currently available in the EPA National Pesticide Standards Repository.  The analytical standards for AMPA and DAPA are due to expire in June and July of 2017, respectively.  The standard for AMGT is due to expire in December 2012.  By that date, the registrant will need to submit a new standard or an updated certificate of analysis for the current AMGT standard.




MRL Harmonization

There are currently no Codex or Mexican maximum residue limits (MRLs) for fluazinam on melons, bell peppers, non-bell peppers, or soybeans.

There are no Canadian MRLs for melons or peppers.  The petition for melons and peppers is being evaluated jointly by the U.S. and Canada's PMRA.  Both countries are using the OECD MRL calculation procedures to determine the recommended tolerances.  For this reason, the MRLs for melons and peppers will be harmonized between the two countries.  There is a Canadian MRL of 0.10 ppm for edible-podded soybean.  There is no need to harmonize this MRL with HED's recommended tolerance of 0.01 ppm for soybean seed because edible-podded soybean and soybean seed are different commodities.  Edible-podded soybean is an immature seed that includes the pod.  It is likely to have a higher residue level than soybean seed, which is the dried seed alone.


Regulatory Recommendations and Residue Chemistry Deficiencies

HED has examined the residue chemistry database for fluazinam.  Pending submission of a revised Section F, there are no residue chemistry issues that would preclude granting full registration for the requested use of fluazinam on soybeans, the melon subgroup, and the pepper/eggplant subgroup.  The recommended tolerances are listed in Table 1.

Table 1.  Tolerance Summary for Fluazinam
Commodity as Proposed by Registrant
                           Proposed Tolerance (ppm)
                          Recommended Tolerance (ppm)
Comments; Correct Commodity Definition
Melon subgroup 9A
                                     0.08
                                     0.07
                                       
Pepper/eggplant subgroup 8-10B
                                     0.10
                                     0.09
                                       
Soybean, Seed
                                     0.01
                                     0.01
Soybean, seed
Soybean, Hulls
                                     0.02
                                     0.05
Soybean, hulls

860.1550 Proposed Tolerances
      
   * The registrant needs to submit a revised Section F in which the tolerances and correct commodity definitions recommended in Table 1 are proposed.


Background

Fluazinam is a contact, multi-site fungicide with activity against a number of plant pathogenic fungi.  It disrupts the production of energy at several metabolic sites within the fungal cell.  Fluazinam is a protectant fungicide that remains primarily on the plant surface.  It is not taken up to any extent by the plant, and is not translocated within the plant.  The nomenclature for fluazinam and its metabolites AMGT, AMPA, and DAPA is presented in Table 2, and the physicochemical properties of fluazinam are summarized in Table 3.  
Table 2.   Fluazinam Nomenclature
Compound
                                       
Common name
Fluazinam
Company experimental name
IKF-1216
IUPAC name
3-chloro-N-(3-chloro-5-trifluoromethyl-2-pyridyl)-α,α,α-trifluoro-2,6-dinitro-p-toluidine
CAS name
3-chloro-N-[3-chloro-2,6-dinitro-4-(trifluoromethyl)phenyl]-5-(trifluoromethyl)-2-pyridinamine
CAS registry number
79622-59-6
End-use product (EP)
Omega[(R)] 500F Fungicide (EPA Reg. No. 71512-1)
Compound
                                       
Common name
AMGT
Chemical name
3-[[4-amino-3-[[3-chloro-5-(trifloromethyl)-2-pyridinyl]amino]-2-nitro-6-(trifluoromethyl) phenyl] thio]-2-(beta-D-glucopyranosyloxy) propionic acid)
Compound
                                       
Common name
AMPA
Chemical name
2-(6-amino-3-chloro- α,α,α.-trifluoro-p-toluidino)-3-chloro-5-(trifluoromethyl) pyridine
Compound
                                       
Common name
DAPA
Chemical name
3-chloro-2-(2,6-diamino-3-chloro-α,α,α.-trifluoro-p-toluidino)-5-(trifluoromethyl)pyridine


Table 3.   Physicochemical Properties of the Technical Grade Fluazinam 
Parameter
Value
References 
Melting Point/Range
                                  115-117ºC
DP# 335640, 8/22/07, W. Drew
pH
                                     5.85

Density (25ºC)
                                 1.02 g/cm[3]

Water Solubility (25ºC)
                         0.131 mg/L (pH buffered to 5)
                         0.157 mg/L (pH buffered to 7)
                         3.384 mg/L (pH buffered to 9)

Solvent Solubility (g/L at 20ºC)
acetone: 853
dichloromethane: 675
ethyl acetate: 722
ethyl ether: 231
hexane: 8
methanol: 192
octanol: 41
toluene: 451

Vapor Pressure at 25ºC
                             1.73 x 10[-][7] mm Hg
                                2.3 x 10[-5] Pa

Dissociation Constant, pKa
                   7.22 in 50% ethanol/water (v/v, average)

Octanol/Water Partition Coefficient
                         Log Kow = 4.03 (Kow = 10,800)

UV/visible absorption spectrum (λmax, nm)
                                  238 at pH 5
                               239, 342 at pH 7
                           260, 343, 482 at pH>10



860.1200 Directions for Use

There is currently a single end-use product (EP) for fluazinam registered in the U.S. for use on several food and feed crops.  This product is Omega[(R)] 500F (EPA Reg. No. 71512-1), which is a 4.17 lb/gal FlC formulation.  IR-4 is proposing to add uses on soybeans, the melon subgroup (9A) and the pepper/eggplant subgroup (8-10B).  Proposed supplemental labels were provided for these new uses.  The proposed use directions are summarized in Table 4.




Table 4.  Summary of Directions for Use of Fluazinam
                       Applic. Timing, Type, and Equip.
                                Formulation[1] 
                                 Applic. Rate 
                                 (lb ai/A)†
                          Max. No. Applic. per Season
                     Max. Seasonal Applic. Rate (lb ai/A)
                                  PHI (days)
                        Use Directions and Limitations
                            Melons (Crop Group 9A)
Broadcast; Ground, Chemigation (Sprinkler System).
OMEGA(R) 500F 
40% ai, Soluble Concentrate 
(4.17 lb ai/gal)
                           0.78 lb ai/A (24 fl oz/A)
                                       6
                                 4.68 lb ai/A 
                                 (144 fl oz/A)
                                      30
Re-treatment Interval = 7-10 days. 
Apply when disease symptoms first appear.
                          Peppers (Crop Group 8-10B)
Broadcast; Ground, Chemigation (Sprinkler System), Soil Drench (at transplanting)..
OMEGA(R) 500F 
40% ai, Soluble Concentrate 
(4.17 lb ai/gal)
                           0.78 lb ai/A (24 fl oz/A)
                                       6
                                 4.68 lb ai/A 
                                 (144 fl oz/A)
                                      30
Re-treatment Interval = 7-14 days.
Apply as a soil drench at transplanting. Foliar applications should begin 7 days after transplanting.
                                    Soybean
Apply at early bloom to full bloom stage of development.
Broadcast; Aerial, Ground.
OMEGA(R) 500F 
40% ai, Soluble Concentrate 
(4.17 lb ai/gal)
                           0.52 lb ai/A (16 fl oz/A)
                                       2
                                 1.04 lb ai/A 
                                 (32 fl oz/A)
                              Growth Stage R3[3]
Re-treatment Interval = 10-14 days (at early pod formation).
For preventive treatment or low disease pressure use low rate.
Do not feed hay from treated field to livestock or allow livestock to graze on treated areas.
Do not apply after early pod formation.
[1]  EPA Reg. No. 71512-1
[2] Maximum Single Application Rate (lb ai/A) = (1 pint/A)(16 fl oz/1 pint) (4.17 lb ai/gal)(1 gal/128 fl oz) = 0.52 lb ai/A.
[3]  Do not apply after growth stage R3, early pod formation.


The proposed (and current) product label for the 4.17 lb/gal FlC formulation specifies that applications are allowed through the following types of irrigation systems:  center pivot, motorized lateral move, traveling gun, solid set or portable (wheel move, side roll, end tow, or hand move).  The label contains the following rotational crop restrictions:  all crops on the label may be replanted immediately after treatment.  All other crops can be replanted 30 days after the last application.

Conclusions.  The submitted use directions are sufficient to allow for evaluation of the submitted residue data relative to the proposed use patterns.  The available field trial data support the proposed use directions for the crops addressed in this summary document.


860.1300 Nature of the Residue  -  Plants

MARC Decision Memo, DP# 272624, 4/23/01, W. Cutchin
Residue Chemistry Memo DP# 257115, 5/21/01, W. Cutchin
Residue Chemistry Memo DP# 335640, 8/22/07, W. Drew

The nature of the residue in plants is adequately understood, based on acceptable potato, peanut, and grape metabolism studies.  The metabolism of fluazinam appears to be similar in tested crops.  Fluazinam undergoes reduction of one of the nitro groups to an amine, forming AMPA.  AMPA can then be conjugated with glutathione, with subsequent degradation of the glutathione moiety to cysteine.  The AMPA-cysteine conjugate then undergoes transamination, reduction, and conjugation with glucose to form AMGT.  In addition, both rings of fluazinam appear to be labile to ring cleavage, and they subsequently degrade into small fragments that can then be incorporated into a variety of natural plant components.  The residues of concern in primary crops for tolerance enforcement and risk assessment are listed in Table 5.

Table 5.   Fluazinam Residues of Concern in Plants
                                    Matrix
                             Residues Included in 
                                Risk Assessment
                             Residues Included in
                             Tolerance Expression
Primary crop:  grapes
                           Parent fluazinam and AMGT
                           Parent fluazinam and AMGT
Primary crop:  peanuts, bulb vegetables,  and root and tuber vegetables
                               Parent fluazinam
                               Parent fluazinam
Primary crops:  all others
                           Parent fluazinam and AMGT
                               Parent fluazinam
 
The registrant submitted an apple metabolism study (MRID 46991301).  HED has not performed a formal review of the study.  The results of the study indicate that parent fluazinam would be the residue of concern in apples for both risk assessment and tolerance enforcement.  As a result, all residues of concern in apples are included in the current determinations.  


860.1300 Nature of the Residue - Livestock

The nature of the residue in livestock is adequately understood, based on adequate goat and hen metabolism studies.  The metabolism of fluazinam in ruminants and poultry has been discussed previously (D272624, W. Cutchin, 4/23/01, D360840, D. Dotson, 3/31/2010).


860.1340 Residue Analytical Methods

Plant commodities

Enforcement Methods

Fluazinam

An adequate GC/ECD method (6148-94-0170-MD-001) entitled "Fluazinam:  Method for the Analysis in Peanut Nutmeat" (MRID 43521016) is available for enforcing fluazinam tolerances on plant commodities.  The method was adequately radiovalidated and underwent a successful independent laboratory validation (ILV) trial.  The method was forwarded to BEAD's Analytical Chemistry Branch (ACB) for a TMV trial, and was subsequently determined to be suitable as a tolerance-enforcement method (Memo, D266802, P. Golden, 6/22/2001).  For this method, residues of fluazinam are extracted with methanol:acetic acid (100:2, v/v), filtered, diluted with 0.2 N HCl, and partitioned into hexane.  Residues are then further partitioned into 0.5 N NaOH, the aqueous phase is acidified, and residues are partitioned back into hexane.  The resulting hexane fraction is concentrated, and residues are purified using a Florisil column eluted with hexane:ethyl ether (80:20, v/v).  Residues are concentrated to dryness, redissolved in hexane, and analyzed by GC/ECD using external standards for quantitation.  The method LOQ is 0.01 ppm for fluazinam in plant commodities.
AMGT

 An adequate enforcement method for determination of AMGT, a residue of concern in wine grapes, is available.  The method is an HPLC/UV method entitled "Method Evaluation for the Analysis of AMGT in Grapes."  The registrant submitted an adequate ILV for the method.  The range of fortification levels and the percent recovery values were acceptable.
 
Data-collection Methods

Samples collected in the cantaloupe and pepper field trials, as well as in the soybean field trials and storage stability study, were analyzed for residues of fluazinam and AMGT using LC/MS/MS methods that were based on the GC/ECD plant enforcement method.  The methods are adequate for data collection based on acceptable method recovery data included in the reviewed studies.  The fortification levels used in the method validation trials bracketed the expected field residues.  The LOQ was 0.01 ppm in all tested commodities.  The LOD was 0.0045 ppm in cantaloupes and peppers and 0.003 in soybean seed, forage, and hay.  In all trials, the data-collection method was validated prior to and in conjunction with the analysis of the field trial samples.  
 
Soybeans

Homogenized soybean samples were blended with methanol:acetic acid (98:2, v:v) and shaken.  The mixture was filtered, the filter cake was rinsed twice with methanol:acetic acid, and the resulting filtrates were combined and diluted to volume with methanol.  Seed extracts were diluted with acetonitrile (ACN):water (80:20, v:v) for analysis by LC/MS/MS.  Forage and hay extracts were cleaned up using a Waters OASIS HLB extraction cartridge.  Residues were eluted with ACN:water (80:20, v:v), and the eluate was analyzed by LC/MS/MS.

Cantaloupes and Peppers

Residues of fluazinam and AMGT were extracted from cantaloupes using 25 mL of methanol/acetic acid (98/2, v/v).  Samples were shaken at room temperature for 30 minutes, filtered, and then cleaned up using a conditioned Evolute SPE cartridge.  Residues were eluted from the column with 6 mL of acetonitrile/water (80/20, v/v) and analyzed by LC/MS/MS (negative ionization mode for fluazinam; positive ionization mode for AMGT).

Conclusions.  Adequate methods for fluazinam and AMGT are available for enforcement of tolerances in crop matrices.  Adequate methods were used for collection of data in the soybean, cantaloupe, and pepper field trials.  In all trials, the data-collection method was validated prior to and in conjunction with the analysis of the field trial samples.  



860.1360 Multiresidue Methods

Parent Fluazinam

Data depicting the recovery of fluazinam through FDA multiresidue methods (MRM) were submitted and forwarded to FDA for review.  The MRM testing data indicate that fluazinam is partially recovered through Sections 302, 303, and 304 of PAM Volume I, with its recovery being dependent on which Florisil elution system is used.  The MRMs can serve as a confirmatory procedure for residues of fluazinam.
  
Metabolite AMGT

Data depicting the recovery of AMGT through FDA MRMs were submitted in support of the apple petition.  ISK Biosciences submitted multiresidue method testing data for the fluazinam metabolite AMGT following testing and analysis according to FDA Multiresidue Method Test Guidelines in PAM Vol. I (1/94).  The Decision Tree for MRM Testing found in the Multiresidue Protocols was followed in determining the response patterns.  Metabolite AMGT was tested through Protocols A, B, C, D, E, and F.  Testing under Protocol G is not required because AMGT is not a substituted urea.  The multiresidue methods were not found to be suitable for determining residues of AMGT because of poor recovery.


860.1380 Storage Stability

Soybeans:  MRID 48712601

ISK Biosciences submitted the results of a storage stability study with fluazinam and AMGT in soybean matrices.  Untreated samples of soybean forage, hay, and seed were fortified with each analyte separately at a fortification level of 0.10 ppm.  Samples were placed in frozen storage at <-10ºC and analyzed at storage intervals of 0, 37, 62, 125, and 153 days. 

In the study, residues of fluazinam and AMGT were stable in/on soybean forage, hay, and seed stored frozen for up to 153 days (5.0 months).  Lower recoveries of fluazinam and AMGT were obtained in stored samples at the 37- and 62-day intervals than at the 125- and 153-day intervals.  The petitioner assumed that this was an analytical variation because it occurred in all three matrices.  The results of the storage stability studies are presented in Table 6.


TABLE 6.   Stability of  Residues of Fluazinam and AGMT in Soybean Following Storage at <-10C
Commodity
                                    Actual 
                                 Spike Level 
                                     (ppm)
                               Storage Interval
                                     (days)
                              Recovered Residues
                                     (ppm)
                           Mean Recovered Residues 
                                     (ppm)
                                 Mean Recovery
                                      (%)
                              Corrected Recovery 
                                     (%)[1]
                                   Fluazinam
Soybean Forage
                                     0.10
                                       0
                            0.0965, 0.1025, 0.0960
                                    0.0983
                                     98.3
                                     96.4

                                       
                                      37
                            0.0785, 0.0860, 0.0715
                                    0.0787
                                     78.7
                                     83.0

                                       
                                      62
                            0.0755, 0.0760, 0.0670
                                    0.0728
                                     72.8
                                     79.4

                                       
                                      125
                            0.0835, 0.0860, 0.1040
                                    0.0912
                                     91.2
                                     84.4

                                       
                                      153
                            0.0820, 0.0870, 0.1020
                                    0.0903
                                     90.3
                                     86.9
Soybean Hay
                                     0.10
                                       0
                            0.1200, 0.1185, 0.1070
                                    0.1152
                                     115.2
                                     98.0

                                       
                                      37
                            0.0720, 0.0855, 0.0700
                                    0.0758
                                     75.8
                                     85.9

                                       
                                      62
                             0.0730, 0.0680, 0.077
                                    0.0727
                                     72.7
                                     65.8

                                       
                                      125
                            0.0690, 0.0790, 0.0820
                                    0.0767
                                     76.7
                                     76.1

                                       
                                      153
                            0.0775, 0.0805, 0.0800
                                    0.0793
                                     79.3
                                     77.8
Soybean Seed
                                     0.10
                                       0
                            0.0845, 0.0865, 0.0860
                                    0.0857
                                     85.7
                                     102.9

                                       
                                      37
                            0.0780, 0.0730, 0.0865
                                    0.0792
                                     79.2
                                     70.8

                                       
                                      62
                            0.0760, 0.0650, 0.0650
                                    0.0687
                                     68.7
                                     70.1

                                       
                                      125
                            0.0820, 0.0810, 0.0800
                                    0.0810
                                     81.0
                                     81.2

                                       
                                      153
                            0.0740, 0.0730, 0.0725
                                    0.0732
                                     73.2
                                     75.6
                                     AGMT
Soybean Forage
                                     0.10
                                       0
                            0.1075, 0.1195, 0.1180
                                    0.1150
                                     115.0
                                     114.1

                                       
                                      37
                            0.0865, 0.0870, 0.0790
                                    0.0842
                                     84.2
                                     74.3

                                       
                                      62
                            0.0930, 0.0860, 0.0820
                                    0.0870
                                     87.0
                                     80.9

                                       
                                      125
                            0.0835, 0.0805, 0.0920
                                    0.0853
                                     85.3
                                     78.5

                                       
                                      153
                            0.0855, 0.0875, 0.0945
                                    0.0892
                                     89.2
                                     80.0
Soybean Hay
                                     0.10
                                       0
                             0.1175, 0.1160, 0.118
                                    0.1172
                                     117.2
                                     126.7

                                       
                                      37
                            0.0775, 0.0770, 0.0740
                                    0.0762
                                     76.2
                                     69.2

                                       
                                      62
                            0.0770, 0.0705, 0.0760
                                    0.0745
                                     74.5
                                     71.3

                                       
                                      125
                            0.0855, 0.0855, 0.0850
                                    0.0853
                                     85.3
                                     85.8

                                       
                                      153
                            0.0795, 0.0910, 0.0820
                                    0.0842
                                     84.2
                                     79.8
Soybean Seed
                                     0.10
                                       0
                            0.1185, 0.1175, 0.1185
                                    0.1182
                                     118.2
                                     124.1

                                       
                                      37
                            0.0705, 0.0755, 0.0710
                                    0.0723
                                     72.3
                                     66.4

                                       
                                      62
                            0.0535, 0.0565, 0.0585
                                    0.0562
                                     56.2
                                     53.9

                                       
                                      125
                            0.0725, 0.0750, 0.0720
                                    0.0732
                                     73.2
                                     69.4

                                       
                                      153
                            0.0750, 0.0700, 0.0720
                                    0.0723
                                     72.3
                                     77.4
1  Corrected for mean concurrent recovery

Conclusion:  The storage stability data are adequate and indicate that residues of fluazinam and AMGT are relatively stable in/on soybean forage, hay, and seed stored frozen for up to 153 days (5.0 months).


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

Livestock Dietary Burdens

The livestock feedstuffs associated with the proposed uses discussed in this summary document are soybean seed and hulls.  The livestock feedstuffs associated with registered crops include peanut meal, processed potato waste, wet apple pomace, and carrot culls.  Fluazinam is registered for use on Subgroup 6A, the Edible-podded legume vegetables subgroup.  However, the approved label excludes the use on peas; therefore, feedstuffs derived from peas were not included in dietary burden calculations.  The proposed soybean label includes restrictions that prohibit the feeding of soybean forage and hay to livestock.

The maximum reasonable dietary burdens of fluazinam to livestock were calculated for the previous risk assessment.  A tolerance of 0.05 ppm was established for the fat and meat byproducts of cattle, goat, horse, and sheep.  As a result of the establishment of tolerances for soybean seed and hulls, the dietary burden for beef cattle increases from 0.04 to 0.049 ppm, and the dietary burden for dairy cattle increases from 1.27 to 1.28 ppm.  These small increases do not result in a need to increase the tolerances that are established for ruminant commodities.  The dietary burdens for poultry and swine are the same as those calculated for the previous risk assessment.  As a result, no tolerances are needed for poultry and swine commodities.

Conclusions.  The current tolerances for livestock commodities are adequate.


860.1500 Crop Field Trials

DER References:	48712601.DER.doc (Soybeans)
		48704501.DER.doc (Cantaloupe)
		48704502.DER.doc (Peppers)
		

IR-4 and ISK Biociences Corporation submitted magnitude of the residue studies in which fluazinam and AMGT were measured in soybeans, cantaloupes, and both bell and non-bell peppers.  In addition, ISK Biociences submitted magnitude of the residue studies in which AMGT was measured in both head and leaf lettuce.  The results from these field trials are discussed below, and the residue data are summarized in Table 7.  

Soybeans (MRID 48712601)

ISK Biosciences Corporation submitted field trial data for fluazinam on soybeans.  Sixteen field trials were conducted in the United States and Canada during the 2010 growing season in the North American Free Trade Agreement (NAFTA) Growing Zones 2 (NC and VA; 2 trials), 4 (LA and AR; 2 trials), and 5 (IA, IL, IN, MN, MO, ND, OH, SD, WI, and ON; 12 trials).  
Each trial consisted of one untreated plot and one treated plot.  At each trial location, the treated plot received two foliar broadcast applications of a 4.17 lb ai/gal suspension concentrate (SC) formulation of fluazinam (Omega[(R)] 500F for the U.S. trials and Allegro(TM) 500F for the Canadian trial) at 0.49-0.64 lb ai/A/application, for a total seasonal rate of 0.99-1.17 lb ai/A.  Applications were made at 9- to 11-day retreatment intervals, except for the SD trial, where the retreatment interval was 16 days.  Applications were made using ground equipment in spray volumes of 19.2-30.6 gal/A.  No adjuvants were added to the spray mixtures.  Samples were harvested from all trials at 29- to 35-day preharvest intervals (PHIs) for forage and hay and 56- to 94-day PHIs for seed.  Hay samples were allowed to dry in the field for 1-15 days prior to collection, except at one Minnesota trial (06) where samples were allowed to dry in the field for 42 days prior to collection (no explanation provided).  Additional forage and hay samples were harvested at PHIs of 10, 20, and 40 days (NC and OH trials), and additional seed samples were harvested at PHIs of 66, 76, and 95 days (NC; target PHI 87 days) and 36, 46, and 66 days (OH; target PHI 56 days) to assess residue decline.

Samples were analyzed for residues of fluazinam and its metabolite AMGT using a high performance liquid chromatography method with tandem mass spectrometric detection (LC/MS/MS) adapted from Ricerca, Inc. analytical methods.  The limit of quantitation (LOQ; determined as the lowest level of method validation, LLMV) was 0.01 ppm for each analyte.  The method was adequate for data collection based on acceptable method validation and concurrent recovery data.  The fortification levels used in concurrent method recovery were adequate to bracket expected residue levels.  Concurrent recoveries and residues in treated samples were not corrected for apparent residues in controls.

Samples were stored frozen (<-10°C) from collection to analysis for 57-150 days (1.9-4.9 months) for forage and hay, and 38-108 days (1.3-3.6 months) for seed.  Samples were analyzed within 0-32 days of extraction.  Adequate storage stability data are available to support the storage conditions and durations of samples from the submitted field trials.

Following two foliar broadcast applications of the 4.17 lb ai/gal SC formulation of fluazinam at 0.99-1.17 lb ai/A, residues of fluazinam in/on soybean forage, hay, and seed, respectively, harvested at a 29- to 35-day PHI (forage and hay) or a 56- to 94-day PHI (seed) were:  0.021-3.725, <0.017-5.207, and <0.01 ppm.  Residues of AMGT were below the LOQ (<0.01 ppm) in/on all samples of soybean forage, hay, and seed.

In the residue decline trials, average residues of fluazinam generally declined with increasing PHI in forage and hay.  Residues of fluazinam in/on seed and residues of AMGT in/on forage, hay, and seed were below the LOQ (<0.01 ppm) in/on all samples; therefore, residue decline could not be assessed.

Melons (MRID 48704501)

IR-4 submitted field trial data for fluazinam on cantaloupes.  A total of 11 cantaloupe field trials were conducted in the United States and Canada during the 2007 growing season encompassing Zones 2 (1 trial each in GA and MD), 5 (2 trials in ON), 5B (1 trial in QC), 6 (2 trials in TX), 9 (1 trial in NM), and 10 (2 trials in AZ, 1 trial in CA).  At each test location, fluazinam (formulated as Omega 500F or Allegro 500F, a suspension), was applied to cantaloupes as 6 foliar broadcast or directed applications at rates of 832  -  958 g a.i./ha/application (0.769  -  0.855 lbs a.i./ha/application), with retreatment intervals of 6  -  9 days, for a total application rate of 5.18  -  5.35 kg a.i./ha/season (4.62  -  4.77 lb a.i./ha/season). Commercially mature cantaloupes were harvested 28-32 days after the last applications.  At Trial NM06 (Las Cruces, NM), cantaloupes were harvested at additional preharvest intervals (PHIs) of 6, 12, 19, and 34 days to assess residue decline behavior.  No adjuvants were used in any of the spray mixtures. 

Cantaloupe samples were analyzed for residues of fluazinam and AMGT using a working method based on analytical method ISK/FLU/08002 (PMRA# 2140647) with a few modifications.  Method ISK/FLU/08002 has been reviewed and deemed adequate for data gathering purposes in potatoes and grapes (PMRA# 2179742).  The validated LOQ for each analyte is 0.01 ppm.  The LOD was reported to be 0.0045 ppm.  The working method was validated prior to sample analysis. Method validation recoveries from cantaloupe samples spiked with 0.01  -  1.0 ppm of fluazinam or AMGT ranged from 77  -  92% and 83  -  97%, respectively.  Method suitability was also verified concurrently during sample analysis.  Concurrent recoveries from pepper samples spiked with 0.01  -  1.0 ppm of fluazinam or AMGT ranged from 84  -  101% and 79  -  102%, respectively.  All recoveries for each analyte fell within the acceptable range of 70  -  120%, indicating that the working method is adequate for the determination of residues of fluazinam and AMGT in/on cantaloupe samples from this study.

Treated cantaloupe samples were stored frozen (-20[o]C) for up to 1180 days prior to extraction. All samples were analyzed within 6 days of extraction.  The freezer storage stability of fluazinam and AMGT residues in/on cantaloupes was assessed concurrently in this study, with control cantaloupe samples spiked with 1.03 ppm of fluazinam or 0.1 ppm of AMGT.  The mean corrected residue recovery (corrected for concurrent recoveries from freshly spiked samples) +- SD of fluazinam in samples stored frozen for 1184 days was 113% +- 3% (n=3).  The mean corrected residue recoveries (corrected for concurrent recoveries from freshly spiked samples) +- SD of AMGT in samples stored frozen for 1035 days and 1055 days were 99% +- 3% (n=3) and 97% +- 6%, respectively.  Although cantaloupe samples were stored frozen for 125 days longer than the demonstrated interval of storage stability for residues of AMGT, given that very good recoveries were observed after a storage period of 1105 days it is not likely that significant residue losses would be observed over the additional storage period.  Furthermore, residues of AMGT were all below the LOQ of 0.01 ppm in all samples whose storage durations ranged from 1111 to 1180 days.  As such, there are no concerns with the stability of residues of fluazinam and AMGT during the frozen storage time period for the cantaloupe samples in this study. 

Total residues of fluazinam and AMGT in/on cantaloupes treated with a suspension formulation, as 6 foliar broadcast or directed applications at total rates of 4.62  -  4.77 lb a.i./ha/season and harvested at PHIs of 27-32 days, ranged from <0.020 - <0.087 ppm.  The residue decline data indicate that total residues of fluazinam and AMGT in/on cantaloupes decline with increasing PHIs.

Peppers (MRID 48704502)

ISK Biosciences submitted field trial data for fluazinam on field peppers (bell peppers and non-bell peppers).  Thirteen field trials were conducted in the United States and Canada during the 2007 growing season. Nine of the trials were conducted using bell peppers and encompassed Zones 2 (1 trial each in NJ and NC), 3 (1 trial in FL), 5 (2 trials in ON), 5B (1 trial in QC), 6 (1 trial in TX), and 10 (2 trials in CA).  The remaining 4 trials were conducted using non-bell peppers and encompassed Zones 3 (1 trial in FL), 5A (1 trial in WI), 6 (1 trial in TX), and 9 (1 trial in NM).

At each test location, fluazinam (formulated as Omega 500F or Allegro 500F, a suspension), was applied to field peppers as 6 applications (2 soil directed applications around the base of the plant + 4 foliar applications) at rates of 0.412  -  0.859 lbs a.i./ha/application, with retreatment intervals (RTIs) of 6  -  53 days, for a total application rate of 3.99  -  4.86 lb a.i./ha/season.  Commercially mature peppers were harvested 28  -  31 days after the last applications.  At Trial 07-NJ16 (Bridgeton, NJ), bell peppers were harvested at additional preharvest intervals (PHIs) of 7, 13, 20, and 35 days to assess residue decline behavior.  No adjuvants were used in any of the spray mixtures.
 
Pepper samples were analyzed for residues of fluazinam and AMGT using a working method based on analytical method ISK/FLU/08002 (PMRA# 2140647) with a few modifications.  Method ISK/FLU/08002 has been reviewed and deemed adequate for data gathering purposes in potatoes and grapes (PMRA# 2179742).  The validated LOQ for each analyte is 0.01 ppm.  The LOD was reported to be 0.0045 ppm.  The working method was validated prior to sample analysis by spiking control samples with 0.01  -  1.0 ppm of fluazinam or AMGT.  Method suitability was also verified concurrently during sample analysis at the same spiking levels for each analyte.  All recoveries for each analyte fell within the acceptable range of 70  -  120%, indicating that the working method is adequate for the determination of residues of fluazinam and AMGT in/on pepper samples from this study.

Treated pepper samples were stored frozen (-20[o]C) for up to 1249 days prior to extraction.  All samples were analyzed the same day of extraction.  The freezer storage stability of fluazinam and AMGT residues in/on peppers was assessed concurrently in this study, with bell pepper samples being spiked with 1.03 ppm of fluazinam or 0.1 ppm of AMGT.  The mean corrected residue recovery (corrected for concurrent recoveries from freshly spiked samples) +- SD of fluazinam in samples stored frozen for 1241 days was 93 +- 8% (n=3).  The mean corrected residue recovery (corrected for concurrent recoveries from freshly spiked samples) +- SD of AMGT in samples stored frozen for 1106 days was 80 +- 3% (n=3).  Although pepper samples were stored frozen for 143 days longer than the demonstrated interval of storage stability for residues of AMGT, given that good recoveries were observed after a storage period of 1106 days it is not likely that significant residue losses would be observed over the additional storage period.  Furthermore, residues of AMGT were all below the LOQ of 0.01 ppm in all samples whose storage durations ranged from 1165 to 1249 days.  As such, there are no concerns with the stability of residues of fluazinam and AMGT during the frozen storage time period for the pepper samples in this study. 

Total residues of fluazinam and AMGT in/on bell peppers and non-bell peppers treated with a suspension formulation, as two soil directed applications plus 4 foliar applications at a total rates of 3.99  -  4.86 lb a.i./ha/season and harvested at PHIs of 28  -  31 days, ranged from <0.020 - <0.048 ppm for bell peppers and from <0.020  -  0.064 for non-bell peppers.  The residue decline data indicate that total residues of fluazinam and AMGT decline in/on peppers with increasing PHIs.

Lettuce:  Head and Leaf (MRID 48707301)

ISK Biosciences previously submitted the results of 14 field trials on head and leaf lettuce (D360840, D. Dotson, 3/31/2010).  These samples were analyzed for parent fluazinam, but not for the metabolite AMGT (a residue of concern for risk assessment).  HED made the following statement in its original review of the lettuce field trial data:  "Samples from the lettuce field trial need to be analyzed for residues of AMGT.  The residue data should also be supported by data depicting the frozen storage stability of AMGT in the lettuce samples."  This stipulation was necessary because the field trial samples would have been stored for a significant period of time between sampling and analysis.  IR-4 analyzed the stored samples for AMGT and performed a storage stability study.

Seven of the lettuce trials were performed on head lettuce and 7 were performed on leaf lettuce.  The trials were conducted in the U.S. during the 2005 growing season.  One of the head lettuce trials was conducted in EPA Growing Zone 2 (MD), one was conducted in Zone 3 (FL), and 5 were conducted in Zone 10 (4 in CA and 1 in NM).  One of the head lettuce trials was conducted in EPA Growing Zone 2 (GA), one was conducted in Zone 3 (FL), and 5 were conducted in Zone 10 (4 in CA and 1 in NM).

For these trials, fluazinam (4.17 lb/gal FlC) was applied to head and leaf lettuce as a single broadcast foliar application during vegetative development at a rate of 0.952-1.05 lb ai/A (1x the proposed maximum seasonal rate).  Applications were made using ground equipment in volumes of 32-77 gal/A, and no adjuvants were used.  Single control and duplicate treated samples of lettuce leaves or heads, with wrapper leaves attached, were harvested at commercial maturity, 46-52 days after application for head lettuce and 20-32 days after application for leaf lettuce.  

The lettuce samples were analyzed for AMGT residues using the method "Method Evaluation for the Analysis of AMGT in Grapes," Document Number 6268-94-1231-MD-001.  Minor modifications to the analytical procedure were made to improve method performance.  Homogenized samples were blended with acetonitrile/water (4:1, v/v).  After filtration, the sample together with the filter paper was extracted for a second time by shaking with acetonitrile/water (1:1, v/v).  The combined filtrate was rotary evaporated to remove all acetonitrile.  The final extract was adjusted with acetonitrile/water (35.2:67.5, v/v) for LC/MS/MS analysis.  The method was validated at 3 fortification levels (0.02, 0.1, and 1.0 ppm) prior to sample analysis.  Both fortified and unfortified control samples were analyzed concurrently with each sample set to demonstrate the absence of significant interferences and adequate recoveries.  The lowest level of method validation was 0.02 ppm.

In all 28 samples analyzed for AMGT, the residue level was below the LLMV of 0.02 ppm.  For the storage stability study, the performing laboratory analyzed samples that had been stored for 1088 to 1110 days (3.0 to 3.04 years).  This storage interval was equivalent to 50-63% of the actual sample storage duration (4.8 to 5.9 years).  IR-4 reported that additional data will be collected and analyzed under a separate protocol.  Samples were fortified at 0.1 ppm and stored in the same freezer as the field trial samples.  AMGT storage stability ranged from 71-78% (n=7) for samples analyzed at 1088 to 1110 days.  Concurrent recoveries for spikes analyzed along with the storage stability samples were in the range of 82-87%.  HED considers the submitted storage stability data to be adequate to demonstrate that residues of AMGT are sufficiently stable in lettuce to resolve the data deficiency cited in the lettuce tolerance petition.      

Conclusions.  The submitted field trial data for fluazinam on soybeans, cantaloupes, and peppers are acceptable and support the proposed uses.  Adequate numbers of trials were conducted in the appropriate geographic regions at approximately 1x the maximum proposed rate, and the appropriate samples were collected from each test at, or close to, the proposed PHI.  The conditional AMGT residue data for lettuce and the supporting storage stability data are also acceptable.  

TABLE 7.  Summary of Fluazinam Field Trial Residue Data 
Commodity
                                    Analyte
                                     Total
                                     Rate
                                   (lb ai/A)
                                  PHI (days)
                             Residue Levels (ppm)

                                       
                                       
                                       
                                       n
                                  Sample Min.
                                  Sample Max.
                                    LAFT[1]
                                    HAFT[2]
                                    Median
                                     Mean
                                  Std.  Dev.
                                   Soybeans
Soybean Forage
                                   Fluazinam
                                   0.99-1.17
                                     29-35
                                      16
                                     0.021
                                     3.73
                                     0.034
                                     3.06
                                     0.427
                                     0.63
                                     0.74

                                     AMGT
                                       
                                       
                                       
                                   <0.01
                                   <0.01
                                   <0.01
                                   <0.01
                                     0.01
                                     0.01
                                      NA
Soybean Hay
                                   Fluazinam
                                       
                                     29-35
                                      16
                                   <0.017
                                     5.21
                                     0.028
                                     5.21
                                     0.868
                                     1.41
                                     1.48

                                     AMGT
                                       
                                       
                                       
                                   <0.01
                                   <0.01
                                   <0.01
                                   <0.01
                                     0.01
                                     0.01
                                      NA
Soybean Seed
                                   Fluazinam
                                       
                                     56-94
                                      16
                                   <0.01
                                   <0.01
                                   <0.01
                                   <0.01
                                     0.01
                                     0.01
                                      NA

                                     AMGT
                                       
                                       
                                       
                                   <0.01
                                   <0.01
                                   <0.01
                                   <0.01
                                     0.01
                                     0.01
                                      NA
                                  Cantaloupes
Cantaloupe
                                   Fluazinam
                                   5.18-5.35
                                     27-32
                                      22
                                   <0.01
                                     0.077
                                   <0.01
                                     0.048
                                     0.01
                                     0.017
                                     0.015

                                     AMGT
                                       
                                       
                                       
                                   <0.01
                                   <0.01
                                   <0.01
                                   <0.01
                                     0.01
                                     0.01
                                      NA
                          Peppers (Bell and Non-Bell)
Bell Pepper
                                   Fluazinam
                                   4.47-5.45
                                     28-31
                                      18
                                   <0.01
                                     0.038
                                   <0.01
                                     0.030
                                     0.01
                                     0.014
                                     0.007

                                     AMGT
                                       
                                       
                                       
                                   <0.01
                                   <0.01
                                   <0.01
                                   <0.01
                                     0.01
                                     0.01
                                      NA
Non-Bell Pepper
                                   Fluazinam
                                   5.10-5.29
                                     28-31
                                       8
                                   <0.01
                                     0.053
                                   <0.01
                                     0.045
                                     0.01
                                     0.02
                                     0.016

                                     AMGT
                                       
                                       
                                       
                                   <0.01
                                   <0.01
                                   <0.01
                                   <0.01
                                     0.01
                                     0.01
                                      NA
                                    Lettuce
Head Lettuce
                                     AMGT
                                   0.95-1.02
                                     46-52
                                      14
                                   <0.02
                                   <0.02
                                   <0.02
                                   <0.02
                                     0.02
                                     0.02
                                      NA
Leaf Lettuce
                                     AMGT
                                   0.97-1.05
                                     20-32
                                      14
                                   <0.02
                                   <0.02
                                   <0.02
                                   <0.02
                                     0.02
                                     0.02
                                      NA
[1]  LAFT = lowest average field trial
[2]  HAFT = highest average field trial result.
NA = not applicable


860.1520 Processed Food and Feed

Soybeans (MRID 48712601)

ISK Biosciences Corporation submitted a processing study for fluazinam on soybean.  In one crop field trial conducted in Louisiana, soybeans grown in separate plots received two foliar broadcast applications of a 4.17 lb ai/gal suspension concentrate (SC) formulation of fluazinam (Omega(R) 500F).  Applications were made at a rate of 0.52-0.53 lb ai/A/application for a total seasonal rate of 1.05 lb ai/A to produce seed for generation of aspirated grain fractions (AGF), and at an exaggerated rate (10x field rate) of 5.30-5.34 lb ai/A/application for a total seasonal rate of 10.64 lb ai/A to produce seed for generation of processed fractions.  Applications were made at an 11-day retreatment interval (RTI) using ground equipment in spray volumes of 19.2-23.6 gal/A.  Adjuvants were not used at either plot.  Soybean seed was harvested from the plots at a 90-day preharvest interval (PHI) and processed into AGF, hulls, meal, and refined oil using simulated commercial procedures.  Processing was conducted by GLP Technologies (Navasota, TX).  The registrant provided adequate descriptions of the processing procedures, including material balance summaries.

Samples were analyzed for residues of fluazinam and its metabolite AMGT using a high performance liquid chromatography method with tandem mass spectrometric detection (LC/MS/MS) adapted from Ricerca, Inc. analytical methods.  The limit of quantitation (LOQ; determined as the lowest level of method validation, LLMV) was 0.01 ppm for each analyte.  The method was adequate for data collection based on acceptable method validation and concurrent recovery data.  The fortification levels used in method validation and concurrent method recovery were adequate to bracket expected residue levels (within an order of magnitude for hulls).  Concurrent recoveries and residues in treated samples were not corrected for residues in controls.

Samples were stored under ambient conditions (field sites) and frozen (<=12°C at the processing facility and <-10°C at the analytical laboratory) from harvest/processing to analysis.  Processing took place within 71 days of harvest.  Sample analyses were completed within 77-117 days (2.5-3.8 months) for seed (RAC) and 46 days (1.5 months) for the processed commodities.  All samples were analyzed within 1-9 days of extraction.  Adequate storage stability data are available to support the storage conditions and durations of samples from the submitted processing study.

Following application of fluazinam at 1.05 lb ai/A, residues of fluazinam and AMGT were each below the LOQ (<0.01 ppm) in/on all samples of soybean seed (RAC) and AGF; therefore, processing factors were not calculated for AGF.  Following application of fluazinam at 10.64 lb ai/A, average residues of fluazinam were 0.044 ppm in/on seed, 0.231 ppm in hulls, 0.046 ppm in refined oil, and below the LOQ (<0.01 ppm) in meal.  Residues of fluazinam concentrated in hulls (5.3x) and refined oil (1.1x), but did not concentrate in meal (processing factor of <0.22x).  Residues of AMGT were below the LOQ in/on seed, hulls, meal, and refined oil; therefore, processing factors were not calculated.
 
The processing factors calculated in this study were less than the maximum theoretical concentration factors for soybean of 11.3x for hulls, 2.2x for meal, and 12.0x for oil (based on separation into components; OPPTS 860.1520, Table 3).

TABLE 8.  Residue Data from Soybean Processing Study with Fluazinam.
RAC
                              Processed Commodity
                                  Total Rate
                                   (lb ai/A)
                                      PHI
                          Residues (ppm)[1] [Average]
                             Processing Factor[2]

                                       
                                       
                                       
                                   Fluazinam
                                     AMGT
                                   Fluazinam
                                     AMGT
Soybean
                                 Seed (RAC)[3]
                                     1.05
                                      90
                               ND, ND [<0.01]
                               ND, ND [<0.01]
                                      --
                                      --

                                      AGF
                                       
                                       
                         <0.01, <0.01 [<0.01]
                               ND, ND [<0.01]
                                     NC[4]
                                      NC

                                 Seed (RAC)[3]
                                     10.64
                                      90
                             0.043, 0.044 [0.044]
                               ND, ND [<0.01]
                                      --
                                      --
                                       
                                     Hulls
                                       
                                       
                             0.242, 0.220 [0.231]
                               ND, ND [<0.01]
                                     5.3x
                                      NC
                                       
                                     Meal 
                                       
                                       
                         <0.01, <0.01 [<0.01]
                               ND, ND [<0.01]
                                   <0.22x
                                      NC
                                       
                                  Refined oil
                                       
                                       
                             0.047, 0.044 [0.046]
                               ND, ND [<0.01]
                                     1.1x
                                      NC
[1]  The LOQ was 0.01 ppm for all analytes in all matrices.  Per trial average residues were calculated by the study reviewer using the LOQ for all residues reported as <LOQ.
[2]  Processing Factor = [Measured residue for analyte in the processed fraction] / [Measured residue for analyte in the seed RAC]. 
[3]  Values for bulk seed samples shipped to the processor.
[4]  NC = Not calculated; residues were below the LOQ in both the RAC and the processed fraction.

Conclusions:  The submitted soybean processing study is acceptable.  Following application of fluazinam at 1.05 lb ai/A, residues of fluazinam and AMGT were each below the LOQ (<0.01 ppm) in/on all samples of soybean seed (RAC) and AGF; therefore, processing factors were not calculated for AGF.  Following application of fluazinam at 10.64 lb ai/A, average residues of fluazinam were 0.044 ppm in/on seed, 0.231 ppm in hulls, 0.046 ppm in refined oil, and below the LOQ (<0.01 ppm) in meal.  The processing data indicate that residues of fluazinam may concentrate in hulls (5.3x) and may concentrate slightly in refined oil (1.1x), but do not concentrate in meal (processing factor of <0.22x).  The observed concentration factors were less than the theoretical concentration factors.  Residues of AMGT were below the LOQ in/on seed, hulls, meal, and refined oil; therefore, processing factors were not calculated.  

An acceptable method was used for residue quantitation, and the study is supported by adequate storage stability data.

860.1650 Submittal of Analytical Reference Standards

Analytical reference standards for fluazinam, AMPA, DAPA, and AMGT are currently available in the EPA National Pesticide Standards Repository (electronic communication, T. Cole to D. Dotson, 7/18/2012).  The analytical standards for AMPA and DAPA are due to expire in June and July of 2017, respectively.  The standard for AMGT is due to expire in December 2012.  The registrant will need to submit new standards or updated certificates of analysis for the current standards by their respective expiration dates.

The standards should be sent to the Analytical Chemistry Lab, which is located at Fort Meade, to the attention of Theresa Cole at the following address:

	USEPA
	National Pesticide Standards Repository/Analytical Chemistry Branch/OPP
	701 Mapes Road
	Fort George G. Meade, MD  20755-5350

The mail will be returned if the extended zip code is not used.
860.1850/1900 Confined and Field Accumulation in Rotational Crops

HED previously reviewed an adequate confined rotational crop study (D360840, D. Dotson, 3/31/2010).  Based on the absence of metabolites containing the intact fluazinam nucleus, HED concluded that the residue of concern in rotational crops is the parent compound, and that rotational crop tolerances are not required for fluazinam.  The 30-day PBI on the label is sufficient for all rotated crops not on the label.  No additional rotational crop data are required for the proposed uses on soybeans, melons, or peppers.


860.1550 Proposed Tolerances

The residue of concern for tolerance enforcement is parent fluazinam for all plant commodities, except for wine grapes, for which the parent compound and the metabolite AMGT are the residues of concern.  For poultry and ruminants, the residues of concern for tolerance enforcement are the parent compound plus the metabolites AMPA and DAPA, and their sulfamate conjugates.

Adequate numbers of field trials at ~1x the proposed use rate are available for cantaloupe, bell peppers, non-bell peppers, and soybeans to support the establishment of tolerances on these crops.  Tolerances do not need to be recommended for soybean forage and hay because the label contains a restriction that prohibits the feeding of forage and hay to livestock.
The OECD MRL calculation procedures were used to determine the recommended tolerances, which are listed in Table 9.  OECD MRL calculation procedures were not used for soybean seed because all of the residue values were below the LOQ of 0.01 ppm.  HED is recommending in favor of a tolerance of 0.01 ppm for soybean seed.

There are currently no Codex or Mexican maximum residue limits (MRLs) for fluazinam on melons, bell peppers, non-bell peppers, or soybeans.

There are no Canadian MRLs for melons or peppers.  The petition for melons and peppers is being evaluated jointly by the U.S. and Canada's PMRA.  Both countries are using the OECD MRL calculation procedures to determine the recommended tolerances.  For this reason, the MRLs for melons and peppers will be harmonized between the two countries.  There is a Canadian MRL of 0.10 ppm for edible-podded soybean.  There is no need to harmonize this MRL with HED's recommended tolerance of 0.01 ppm for soybean seed because edible-podded soybean and soybean seed are different commodities.  Edible-podded soybean is an immature seed that includes the pod.  It is likely to have a higher residue level than soybean seed, which is the dried seed alone.

An International Residue Limit Status Sheet can be found in Attachment 1. 


References

D272624, Fluazinam.  Decision by Metabolism Assessment Review Committee (MARC), W. Cutchin, 4/23/01

D360840, Fluazinam.  Petitions for the Establishment of Tolerances and Registration of New Uses on Apples, Carrots, Lettuce, and Bulb Onion Subgroup (3-07A), and a Request for a Reduced Tolerance on Bushberry Subgroup (13-07B).  Summary of Analytical Chemistry and Residue Data, D. Dotson, 3/31/2010

D387187, Fluazinam.  Human Health Risk Assessment for the Section 3 Registration Action on Turfgrass, D. Dotson, et al., 2/7/2012





Attachments:
  
Attachment 1:  International Residue Limit Status sheet
Attachment 2:  Tolerance Assessment Calculations

                  Attachment 1:  International Residue Limits

                         Fluazinam (129098; 7-10-2012)
                                       
Summary of US and International Tolerances and Maximum Residue Limits 
Residue Definition:  
US
Canada
Mexico[2]
Codex[3]
40 CFR 180.574:

Plants:  Fluazinam, 3-chloro-N-[3-chloro-2,6-dinitro-4-(trifluoromethyl)phenyl]-5-(trifluoromethyl)-2-pyridinamine





Commodity[1]
Tolerance (ppm) /Maximum Residue Limit (mg/kg)

                                      US
                                    Canada
                                   Mexico[2]
                                   Codex[3]
Soybean, Seed
                                     0.01
0.10 ppm:  Edible-podded soybean


Soybean, Hulls
                                     0.02



Melon subgroup 9A
                                     0.07



Pepper/Eggplant
Subgroup 8-10B
                                     0.09




                                       



Completed:  M. Negussie;  07/16/2012 
[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 = processed 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.




                                        
               Attachment 2:  Tolerance Assessment Calculations

Soybeans

HED's current policy is to use OECD MRL calculation procedures to determine recommended tolerances.  In the case of soybeans, all of the residue values were below the LOQ of 0.01 ppm.  When all residue values are below the LOQ, HED recommends in favor of establishing the tolerance at the LOQ.  Therefore, HED recommends in favor of a tolerance of 0.01 ppm for soybean seed.

The average processing factor for soybean hulls is 5.25x.  Multiplying this value by the soybean seed LOQ of 0.01 ppm yields a value of 0.0525 ppm.  HED recommends in favor of a tolerance of 0.05 ppm for soybean hulls.
 
Melon Subgroup (9A)

For melons, the OECD MRL Calculator was used for calculating recommended tolerances.  Average cantaloupe field trial values were entered into the calculator.  The dataset used to establish tolerances on the melon subgroup consisted of field trial data representing applications at rates of 4.6-4.8 lb ai/A (six applications at approximately 0.78 lb ai/A/application) with a 27-32 day PHI.  Using the MRL calculator, the recommended tolerance is 0.07 ppm.  

Pepper/Eggplant Subgroup (8-10B)

For peppers, the OECD MRL Calculator was used for calculating recommended tolerances.  Average field trial values were entered into the calculator.  Field trial data for bell and non-bell peppers were entered separately and the higher of the two recommended tolerances (i.e., that for the non-bell peppers) is the one that HED is recommending as the tolerance for the entire subgroup.  The dataset used to establish tolerances on the pepper/eggplant subgroup consisted of field trial data representing applications at rates of 4.6-4.7 lb ai/A (six applications at approximately 0.78 lb ai/A/application) with a 28-31 day PHI.  Using the MRL calculator, the recommended tolerance is 0.09 ppm.  The MRL calculator recommends a tolerance of 0.05 ppm for the bell peppers.  As stated above, the recommended tolerance for the non-bell peppers is being recommended as the tolerance for the entire subgroup because it is the higher of the two values.  




                    Fluazinam:  OECD MRL Calculator Results


OECD MRL Calculator Results for the Melon Subgroup (9A) and the Pepper/Eggplant Subgroup (8-10B)
Compound
 
                                   Fluazinam
                                   Fluazinam
                                   Fluazinam
Crop
 
                                 Bell Peppers
                               Non-Bell Peppers
                                  Cantaloupes
Region / Country
 
                                US & Canada
                                US & Canada
                                US & Canada
GAP
 
                             4.47-5.45 kg a.i./ha
                            (3.99 - 4.86 lb a.i./A)
                              PHI = 28 - 31 days
                            5.10 - 5.29 kg a.i./ha
                            (4.55 - 4.72 lb a.i./A)
                              PHI = 28 - 31 days)
                            5.18 - 5.35 kg a.i./ha
                            (4.62 - 4.77 lb a.i./A)
                              PHI = 27 - 32 days
Total number of data points (n)
 
                                       9
                                       4
                                      11
Percentage of censored data
 
                                      33%
                                      50%
                                      45%
Number of non-censored data
 
                                       6
                                       2
                                       6
Lowest residue
 
                                     0.010
                                     0.010
                                     0.010
Highest residue
 
                                     0.030
                                     0.045
                                     0.048
Median residue
 
                                     0.011
                                     0.013
                                     0.011
Mean
 
                                     0.014
                                     0.020
                                     0.017
Standard deviation (SD)
 
                                     0.007
                                     0.017
                                     0.012
Correction factor for censoring (CF)
 
                                     0.778
                                     0.667
                                     0.697
- Highest residue
 
                                     0.030
                                     0.045
                                     0.048
- Mean + 4 SD
 
                                     0.041
                                     0.087
                                     0.063
- CF x 3 Mean
 
                                     0.034
                                     0.041
                                     0.036
Unrounded MRL
 
                                     0.041
                                     0.087
                                     0.063
                                       
Rounded MRL
 
                                     0.05
                                     0.09
                                     0.07
                                       
 
 
                                       
            High uncertainty of MRL estimate due to small dataset.
                                       
 
 
                               Residues (mg/kg)
                               Residues (mg/kg)
                               Residues (mg/kg)
 
 
                                     0.010
                                       *
                                     0.016
                                       
                                     0.020
                                       
 
 
                                     0.019
                                       
                                     0.010
                                       *
                                     0.010
                                       *
 
 
                                     0.010
                                       *
                                     0.045
                                       
                                     0.011
                                       
 
 
                                     0.010
                                       *
                                     0.010
                                       *
                                     0.010
                                       *
 
 
                                     0.015
                                       
                                       
                                       
                                     0.010
                                       *
 
 
                                     0.015
                                       
                                       
                                       
                                     0.048
                                       
 
 
                                     0.030
                                       
                                       
                                       
                                     0.010
                                       *
 
 
                                     0.011
                                       
                                       
                                       
                                     0.024
                                       
 
 
                                     0.010
                                       
                                       
                                       
                                     0.014
                                       
 
 
                                       
                                       
                                       
                                       
                                     0.021
                                       
 
 
                                       
                                       
                                       
                                       
                                     0.010
                                       *

                                       
