Document ID: EPA-HQ-OPP-2009-0032-0010
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2010-05-12T04:00Z

UNITED STATES ENVIRONMENTAL PROTECTION AGENCY

WASHINGTON, D.C.  20460

	OFFICE OF PREVENTION, PESTICIDE

	AND TOXIC SUBSTANCES

  SEQ CHAPTER \h \r 1 MEMORANDUM

Date:  		3/31/2010

SUBJECT:	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.

PC Code:  129098	DP Barcodes:  D360840 and D366506

Decision Nos.:  404254 and 412727	Registration No.: 71512-1

Petitions:  8E7506, 9E7570, and 9F7571	Regulatory Action:  Section 3
Registration

Risk Assessment Type:  NA	Case No.:  NA

TXR No.:  NA	CAS No.:    SEQ CHAPTER \h \r 1 79622-59-6

MRID Nos.: 47631001, 47631002, and

                      47756601 through 47756611	40 CFR:  §180.574

		              									

FROM:  	Douglas Dotson, Ph.D., Chemist

		Risk Assessment Branch II

		Health Effects Division (7509P)	  SEQ CHAPTER \h \r 1 

		

THROUGH:	William Drew, Chemist

		Richard Loranger, Ph.D., Senior Scientist

		Risk Assessment Branch II

		Health Effects Division (7509P)

TO:		Laura Nollen/Barbara Madden, RM Team 5, RIMUERB

		John Bazuin/Tony Kish, PM Team 22, Fungicide Branch

		Registration Division (7505P)		  SEQ CHAPTER \h \r 1   SEQ CHAPTER \h
\r 1 

		

  SEQ CHAPTER \h \r 1 This document was originally prepared under
contract by Dynamac Corporation (1901 Research Boulevard, Suite 220;
Rockville, MD 20850 and 1910 Sedwick Rd., Building 100, Suite B; Durham,
NC 27713).  The document has been reviewed by the Health Effects
Division (HED) and revised to reflect current Office of Pesticide
Programs (OPP) policies.

Executive Summary

Fluazinam
(3-chloro-N-[3-chloro-2,6-dinitro-4-(trifluoromethyl)phenyl]-5-(trifluor
omethyl)-2-pyridinamine) is a member of the phenyl-pyridinamine class of
compounds.  Fluazinam is a non-systemic, preventive, contact fungicide
with a multi-site mode of action.  It disrupts the production of energy
at several metabolic sites within the fungal cell.  Fluazinam is a
protectant fungicide.  That is, 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.

The Agency has received three separate tolerance petitions to establish
permanent tolerances for fluazinam residues of concern in/on various raw
agricultural and animal commodities.  This document summarizes the
available residue chemistry data in support of these tolerance
petitions.  The end-use product (EP) relevant to all petitions is
OMEGA® 500F (EPA Reg. No. 71512-1), a flowable concentrate (FlC)
formulation containing 40% fluazinam (equivalent to 4.17 lb ai/gal).  

In PP#8E7506, the Interregional Research Project No. 4 (IR-4), on behalf
of the Agricultural Experiment Stations of Arizona, California,
Colorado, Georgia, Idaho, New York, Ohio, Tennessee, and Texas has
requested registration of fluazinam for use on lettuce (head and leaf)
and Onion, bulb, subgroup 3-07A.  The proposed use of OMEGA® 500F on
lettuce is for a single broadcast or banded foliar application or soil
drench application at crop thinning at up to 1.0 lb ai/A.  The proposed
use for the bulb onion subgroup is for a maximum of six broadcast foliar
applications during vegetative development at up to 0.52 lb
ai/A/application for a maximum seasonal rate of 3.1 lb ai/A.  The
minimum retreatment interval (RTI) is 7 days for onions, and both uses
only allow for applications using ground equipment.  The minimum
preharvest intervals (PHIs) are 50 days for head lettuce, 30 days for
leaf lettuce, and 7 days for onions.  In conjunction with these uses,
IR-4 has proposed the establishment of permanent tolerances for residues
of fluazinam in/on the following commodities:

Lettuce, head 	0.02 ppm

Lettuce, leaf 	2 ppm

Onion, bulb, subgroup 3-07A 	0.15 ppm

Also included in PP# 8E7506 is a request by IR-4 to revise the
established 7.0-ppm tolerance for fluazinam residues in/on the Bushberry
subgroup (13B) and other bushberry type crops (aronia berry, buffalo
currant, Chilean guava, European barberry, highbush cranberry, edible
honeysuckle, jostaberry, juneberry, lingonberry, native currant, salal,
and sea buckthorn).  These tolerances were originally established under
PP# 6E7137 (Memo, D335640, W. Drew, 8/22/07).  IR-4 is requesting that a
single tolerance be established for the updated Bushberry subgroup

(13-07B) for residues of fluazinam at a level of 4.5 ppm.  This proposed
tolerance would then replace the multiple tolerances on the various
bushberries.

In PP# 9E7570, IR-4, on behalf of the Agricultural Experiment Stations
of California, Georgia, Texas, and Washington, has requested the
registration of fluazinam for use on carrots.  The proposed use includes
a maximum of four directed band applications prior to or during disease
development at up to 0.52 lb ai/A/application, with a 7-day RTI, for a
maximum seasonal rate of 2.1 lb ai/A.  Applications may be made using
ground equipment.  A 7-day PHI is proposed.  Concurrently, IR-4 has
proposed the establishment of the following permanent tolerance for
residues of fluazinam:

Carrot, roots	0.8 ppm

In PP# 9F7571, ISK Biosciences Corporation requested the registration of
fluazinam for use on apples.  The proposed use includes a maximum of ten
broadcast foliar applications prior to or during disease development at
up to 0.45 lb ai/A/application, with a 7-day RTI, for a maximum seasonal
rate of 4.50 lb ai/A.  Applications may be made using ground and aerial
equipment.  A 28-day PHI is proposed.  In conjunction with this use, ISK
Biosciences has proposed the establishment of permanent tolerances for
residues of the fungicide fluazinam and its metabolite AMGT in or on the
following commodities:

Apples	1.7 ppm

Apple, pomace, wet	5.0 ppm

In addition, ISK Biosciences has proposed tolerances for residues of
fluazinam and its metabolites DAPA and AMPA in the following livestock
commodities: 

Cattle, fat	0.03 ppm

Cattle, kidney	0.03 ppm

Cattle, liver	0.03 ppm

Cattle, meat	0.03 ppm

Cattle, meat byproducts	0.03 ppm

Goat, fat	0.03 ppm

Goat, kidney	0.03 ppm

Goat, liver	0.03 ppm

Goat, meat	0.03 ppm

Goat, meat byproducts	0.03 ppm

Horse, fat	0.03 ppm

Horse, kidney	0.03 ppm

Horse, liver	0.03 ppm

Horse, meat	0.03 ppm

Horse, meat byproducts	0.03 ppm

Milk	0.03 ppm

Sheep, fat	0.03 ppm

Sheep, kidney	0.03 ppm

Sheep, liver	0.03 ppm

Sheep, meat	0.03 ppm

Sheep, meat byproducts	0.03 ppm

Permanent tolerances have been established for residues of fluazinam
in/on plant commodities at levels ranging from 0.01 to 7.0 ppm [40 CFR
§180.574(a)(1)], and a tolerance has also been established for residues
of fluazinam and its metabolite AMGT on wine grapes at 3.0 ppm (40 CFR
§180.574(a)(2)).  No tolerances have been established for livestock
commodities.

There are currently no established Codex, Canadian, or Mexican maximum
residue limits (MRLs) for fluazinam on apples, carrots, lettuce, onions,
or berry crops.  Therefore, there are no harmonization issues for the
requested tolerances.

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 fluazinam per se 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 fluazinam residues 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 for fluazinam.  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
regulatory interest in animal commodities were determined by HED to be
parent plus the metabolites AMPA and DAPA, and their sulfamate
conjugates.

An adequate gas chromatography with electron capture detection (GC/ECD)
method (6148-94-0170-MD-001) is available for determination of fluazinam
residues in crop matrices.  The method was deemed to be suitable for
tolerance enforcement after it successfully completed the routes
required for an enforcement method including radiovalidation,
independent laboratory validation (ILV), and tolerance method validation
(TMV).  Samples collected from the magnitude of the residue and storage
stability studies were analyzed for residues of fluazinam using various
GC/ECD methods based on, and revised slightly from, the plant
enforcement method.  The GC/ECD methods are adequate for data collection
based on acceptable method recovery data.  The lowest limit of method
validation (LLMV) was 0.01 ppm in all tested commodities except in
carrots where the LLMV was 0.02 ppm.

An enforcement method for determination of AMGT, a residue of concern in
wine grapes, is available.  The registrant requested that the blueberry
submission serve as the ILV for the method.  The range of fortification
levels and the percent recovery values are acceptable.  As a result, HED
considers the AMGT analytical portion of the blueberry field trial study
to be adequate, and that it can serve as the ILV for the analytical
method for AMGT residues in plant commodities.

PTRL Method 1676W was the data-collection method used for the analysis
of samples collected from the dairy cattle feeding study.  The method is
written for LC/MS/MS analysis of fluazinam, AMPA, and DAPA in bovine
milk and tissues, and for GC/MS analysis of DAPA in milk only.  The
limit of quantitation was 0.01 ppm for each analyte in milk, muscle,
fat, liver and kidney.  Preliminary method validation data showed that
Method 1676W adequately recovered  residues of fluazinam, AMPA, and DAPA
in milk, muscle, and fat.  However, poor and/or marginal recoveries were
obtained for AMPA and DAPA residues in kidney and liver.  Method 1676W
was marginally validated by an independent laboratory during the third
trial and following slight modifications to make the method more rugged.
 For the purpose of developing an animal enforcement method,
radiovalidation data are required to determine whether the method can
detect aged residues.  HED forwarded Method 1676W to ACB/BEAD for a
tolerance method validation (TMV) because the method encountered
difficulties in the ILV (e.g., three attempts for successful validation
and/or marginal recoveries with wide variability and high coefficient of
variation).  The method was forwarded to ACB/BEAD for method validation.
 ACB/BEAD reported that the lab that performed the ILV made major
modifications to the original method.  As a result, HED requested that
the registrant submit a revised method that incorporated the revisions
made by the ILV lab.  ISK Biosciences revised the method and
re-submitted it.  ACB/BEAD performed a cursory review of the revised
method.  The registrant incorporated the modifications made by the ILV
lab.  However, the revision eliminated the cleanup steps that were in
the original method, and the only recovery data for the revised method
were data provided by the ILV lab.  ACB/BEAD was concerned that the
elimination of the cleanup steps could lead to dirty samples which
would, in turn, lead to analytical problems, such as interference. 
ACB/BEAD performed no further validation of the method.  As the revised
method contains major revisions to the original method, and ACB/BEAD
believes it is very likely that the revised method still is not adequate
for sample analysis or tolerance enforcement, HED recommends that the
registrant submit an ILV of the revised method.

Adequate FDA   SEQ CHAPTER \h \r 1 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.  Multiresidue method testing data are required for the
regulated metabolites, AMPA and DAPA, in animals.

Analytical reference standards for fluazinam, AMPA, and DAPA are
currently available in the EPA National Pesticide Standards Repository;
however an analytical standard for the AMGT metabolite is not currently
available.

Adequate storage stability data are available to support the storage
conditions and durations of samples collected from the magnitude of the
residue studies on apples, carrots, lettuce, and onions.  As residues of
fluazinam were found to be stable in tested crop matrices, storage
stability corrections do not need to be applied to the recommended
tolerances for fluazinam residues in/on apples, carrots, lettuce, and
onions.  Storage stability data for the fluazinam metabolite AMGT in
apple commodities were also submitted, and showed that residues in wet
apple pomace were reasonably stable for 12 months, but declined to an
average corrected recovery of 58-63% at the 19-month interval, 61% at
the 32-month interval, and 33% at the 37-month interval.  The submitted
storage stability data for bovine milk, meat, and meat byproducts showed
mixed results.

Based on the poor recoveries of metabolites AMPA and DAPA from liver and
kidney and the mixed storage stability results, the residue data from
the cattle feeding study are not adequate to satisfy data requirements. 
The feeding study data (adjusted for residue decline) along with the
calculated dietary burdens for beef cattle (0.04 ppm) and dairy cattle
(1.27 ppm) indicate that tolerances are needed for the combined residues
of fluazinam and its metabolites DAPA and AMPA in the fat and meat
byproducts of cattle and other ruminants at 0.05 ppm to support the
proposed uses on apples and carrots and the established tolerances on
potatoes and peanuts.  However, no tolerances are required for milk and
meat of ruminants, as the expected combined residues in these matrices
are below the combined method limit of quantitation (LOQ) of 0.03 ppm
for the regulated compounds.  Based on the transfer coefficients for
livestock tissues and the relatively low dietary burden for swine of
0.003 ppm for fluazinam, tolerances for hog commodities are not needed. 
A poultry feeding study is not required at this time because of the low
dietary burden for poultry (0.005 ppm).

With the exception of the use of adjuvants, the submitted field trial
data for fluazinam on head lettuce, leaf lettuce, bulb onions, carrots,
and apples are acceptable and support the proposed uses.  Adequate
numbers of trials were conducted in the appropriate geographic regions
at ~1x the maximum proposed rate using a representative FlC formulation,
and the appropriate samples were collected from each test at the
proposed PHI.  However, no residue data were provided on AMGT residues
in lettuce.  Although AMGT residue data on lettuce are still required
for purposes of dietary risk assessment, the available data would
support tolerances of 0.02 ppm for head lettuce, 2.0 ppm for leaf
lettuce, 0.20 ppm for the bulb onion (subgroup 3-07A), 1.5 ppm for
apple, and 0.7 ppm for carrot.  The tolerance spreadsheet (Guidance for
Setting Pesticide Tolerances Based on Field Trial Data SOP) was used for
determining appropriate tolerance levels.

For dietary risk assessment, HED will calculate AMGT residues in lettuce
using a ratio of 0.35:1 for AMGT:fluazinam residues (ChemSAC Memo,
meeting of 2/6/08).  The samples from the lettuce field trials still
need to be analyzed for residues of AMGT.  AMGT is not a residue of
concern for risk assessment for carrots or onions; therefore, for these
commodities it does not need to be included in the dietary exposure
assessment.  AMGT residues were measured in apples and the combined
residues of parent and AMGT need to be included in the dietary exposure
assessment.

After reevaluating the available residue data for blueberries, HED
concludes that the residue data for fluazinam from the blueberry field
trials are distributed lognormally.  As a result, the recommended
tolerance for the revised Bushberry subgroup 13-07B is 7.0 ppm, which is
equivalent to the current tolerance for Subgroup 13B.

The apple processing study is not adequate to satisfy data requirements.
 In apples, the metabolite AMGT is a residue of concern for risk
assessment.  In the processing study there were no quantifiable residues
of AMGT in raw apples.  In several of the field trials that were
performed, however, raw apples contained AMGT residues at, or above, the
limit of quantitation.  In a few cases, residues of AMGT were comparable
to those of the parent fluazinam.  Canadian and U.S. residue chemistry
guidelines both stipulate that in the processing study, if residues in
the raw agricultural commodity (RAC) are below the LOQ, but quantifiable
residues occurred in the RAC in field trials performed at the maximum
requested label rate, that the processing study should be conducted at
exaggerated rates in order to ensure that quantifiable residues will be
present in the RAC.  Quantifiable residues are needed in the RAC in
order to determine a reliable concentration factor.  As a result, the
processing study needs to be repeated at a sufficiently high rate that
residues of both parent and AMGT are quantifiable in the RAC.  In the
event that the potential for phytotoxicity exists, the application rate
does not need to be greater than the rate at which phytotoxicity occurs.

  SEQ CHAPTER \h \r 1 Regulatory Recommendations and Residue Chemistry
Deficiencies

No major deficiencies were noted in the current petitions that would
preclude establishing permanent tolerances for fluazinam residues of
concern on head lettuce; leaf lettuce; Onion, bulb, subgroup 3-07A; and
Bushberry, subgroup 13-07B.  In addition, the deficiency listed below
for lettuce field trials needs to be resolved as a condition of
registration.  A revised Section F is needed as detailed below
(860.1550).

Because of major data deficiencies, HED does not recommend in favor of
the establishment of tolerances for apples or carrots.  HED also does
not recommend in favor of tolerances for animal commodities.  The data
deficiencies associated with the apple and carrot petitions are
discussed below.

For PP# 8E7506, pending submission of residue data for AMGT on lettuce
as a condition of registration, the submitted data support the proposed
uses of OMEGA® 500F on lettuce and Onion, bulb, subgroup 3-07A along
with the establishment of tolerances for residues of fluazinam in/on:

Lettuce, head 	0.02 ppm

Lettuce, leaf 	2.0 ppm

Onion, bulb, subgroup 3-07A 	0.20 ppm

The petition also supports revising the existing Bushberry subgroup 13-B
tolerance to a Bushberry subgroup 13-07B tolerance at 7.0 ppm along with
deletion of existing individual tolerances on various berries as
detailed in Table 13.

List of Deficiencies

860.1200 Directions for Use

 	•	The use directions for all the subject crops (lettuce, onions,
bushberries, apples, and carrots) should include a prohibition against
addition of adjuvants to the spray mixture.

The following deficiency applies to both existing and proposed uses of
fluazinam

860.1650 Submittal of Analytical Reference Standards

	•	An analytical reference standard for the AMGT metabolite is not
available.  RD should request that the registrant submit this standard
to the EPA’s National Pesticide Repository as soon as possible.

List of Deficiencies by Petition 

PP# 8E7506 (Lettuce, Onion, Bushberry Subgroup)

860.1500 Crop 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.

860.1550 Proposed Tolerances

	•	The registrant should submit a revised section F in which a
tolerance of 0.2 ppm is proposed for Onion, bulb, subgroup 3-07A, and a
tolerance of 7.0 ppm is proposed for Bushberry, subgroup 13-07B.

PP#s 9F7570 (Carrots) and 9F7571 (Apples)

860.1340 Residue Analytical Methods

•	At the present time, an analytical method is not available for
enforcement of fluazinam tolerances in animal commodities.  The
registrant should submit an ILV of the revised method.  This method
needs to be adequate for analysis of parent fluazinam as well as the
metabolites AMPA and DAPA in animal commodities for which tolerances
will be established.

•	If the registrant resolves the data deficiencies associated with the
apple and carrot petitions and re-submits the tolerance petitions,
multiresidue method testing data will be required for the regulated
metabolites, AMPA and DAPA, in animals.

860.1480 Meat, Milk, Poultry, and Eggs

	•	The residue data from the cattle feeding study are not adequate to
satisfy data requirements.  To support tolerances for apples and
carrots, the registrant needs to submit a new cattle feeding study in
which a method with better recoveries of AMPA and DAPA from liver and
kidney was used.  When this study is repeated, the samples need to be
analyzed as soon as possible after sacrifice in order to minimize
degradation of residues.

860.1520 Processed Food and Feed (Applies Only to Apple Petition,
9F7571)

	•	The apple processing study is not adequate to satisfy data
requirements.  The processing study needs to be repeated at a
sufficiently high rate that residues of both parent and AMGT are
quantifiable in the RAC.  In the event that the potential for
phytotoxicity exists, the application rate does not need to be greater
than the rate at which phytotoxicity occurs.

Note to RD:

HED recommends that 40CFR §180.574(a)(1) be amended by replacing the
tolerance expression with the following:  “Tolerances are established
for residues of fluazinam
(3-chloro-N-[3-chloro-2,6-dinitro-4-(trifluoromethyl)phenyl]-5-(trifluor
omethyl)-2-pyridinamine), 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
fluazinam.”  HED further recommends that 40CFR §180.610(a)(2) be
amended by replacing the tolerance expression with the following: 
“Tolerances are established for residues of fluazinam, 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 fluazinam and its metabolite AMGT
(3-[[4-amino-3-[[3-chloro-5-(trifluoromethyl)-2-pyridinyl]amino]-2-nitro
-6-(trifluoromethyl) phenyl]thio]-2-(beta-D-glucopyranosyloxy) propionic
acid).”

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 1, and the physicochemical
properties of fluazinam are summarized in Table 2.  

o- 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-(tri
fluoromethyl)pyridine

Table 2.   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/cm3

	Water solubility (25ºC)	(pH buffered to 5) 0.131 mg/L

(pH buffered to 7) 0.157 mg/L

(pH buffered to 9) 3.384 mg/L

	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 (Pa)	2.3 x 10-5 at 25ºC

1.3 x 10-4 at 35ºC

6.7 x 10-5 at 45ºC

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

	Octanol/water partition coefficient, Log (KOW)	1.08 x 104 (Log Kow =
4.03)

	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 to ISK Biosciences in the U.S. for use on food/feed crops. 
This EP is Omega® 500F (EPA Reg. No. 71512-1), which is a 4.17 lb/gal
FlC formulation.  It is registered for use on ginseng, peanuts,
potatoes, turnip greens, Crop Group 5 (Brassica leafy vegetables),
Subgroup 6A (except pea), Subgroup 6B (except pea), Subgroup 6C (except
pea), the Bushberry Subgroup (13B), and other bushberries. 

IR-4 is proposing to add uses on carrot, lettuce, and members of the
Bulb onion subgroup, 3-07A, to the label for the 4.17 lb/gal FlC
formulation for control of fungal diseases.  Proposed supplemental
labels were provided for these new uses.  The proposed use directions
are summarized in Table 3.

ISK Biosciences is proposing to add use on apple to the product label
for the 4.17 lb/gal FlC formulation.  The proposed use directions are
summarized in Table 3.

Table 3.   Summary of Directions for Use of Fluazinam.

Applic. Timing; Type; and Equipment	Formulation

[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 Limitations1

Apple

Broadcast foliar applications prior to or during disease development;

Ground and aerial equipment 	4.17 lb/gal FlC

[71512-1]	0.33-0.45	10	4.50	28	Apply in a minimum of 5 gal/A (both
ground and aerial equipment).  The minimum RTI is 7 days.

Bulb Onion Subgroup 3-07A

Broadcast foliar applications prior to or during disease development;

Ground equipment 	4.17 lb/gal FlC

[71512-1]	0.52	6	3.12	7	Apply in a minimum of 5 gal/A.  The minimum RTI
is 7 days.

Carrot

Directed band to the crop prior to or during disease development;

Ground equipment 	4.17 lb/gal FlC

[71512-1]	0.52	4	2.08

(implied)	7	Apply in a minimum of 30 gal/A.  The minimum RTI is 7 days.

Lettuce (Head and Leaf)

Broadcast or banded  foliar application or  soil drench application at
thinning;

Ground equipment	4.17 lb/gal FlC

[71512-1]	0.52-0.99	1	1.0	50 - head lettuce;

30 - leaf lettuce	Apply in a minimum of 50 gal/A.  

1  A 48-hour restricted entry interval is specified.  

The current (and proposed) product label for the 4.17 lb/gal FlC
formulation specifies that applications are allowed through the
following types of irrigations 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, with the
exception of the use of adjuvants.  As the crop field trials did not
include adjuvants, the use directions for all proposed crops should
include a prohibition against the addition of adjuvants to the spray
mixture.

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 regulatory interest in primary crops are listed in Table
4.

Table 4.   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 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);
however, HED has not reviewed it.  In the event that the registrant
resolves the data deficiencies associated with the apple petition and
re-submits it, the apple metabolism study should be reviewed.

860.1300 Nature of the Residue - Livestock

MARC Decision Memo, D272624, W. Cutchin, 4/23/01 

Residue Chemistry Memo D257115, W. Cutchin, 5/21/01

 was only a minor component (≤2.7% TRR) of the [14C]-residues in
poultry tissues and eggs, and was not detected in ruminant tissues or
milk.  The residues of concern in livestock are listed in Table 5.

Table 5.   Fluazinam Residues of Concern in Livestock

Matrix	Residues Included in 

Risk Assessment	Residues Included in

Tolerance Expression

Ruminant	Parent fluazinam, AMPA, and DAPA, and their sulfamate
conjugates	Parent fluazinam, AMPA, and DAPA, and their sulfamate
conjugates

Poultry	Parent fluazinam, AMPA, and DAPA, and their sulfamate conjugates
Parent fluazinam, AMPA, and DAPA, and their sulfamate conjugates

 

860.1340 Residue Analytical Methods

Residue Chemistry Memo D335640, W. Drew, 8/22/07

Residue Chemistry Memo D212612, G. Herndon, 9/5/95

TMV Results Memo, D266802, P. Golden, 6/22/01

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 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 enforcement method for determination of AMGT, a residue of concern in
wine grapes, is available.  HED recommended in a previous memorandum
(Memo, D335640, W. Drew, 8/22/07) that a previously submitted HPLC/UV
method based on Method Evaluation for the Analysis of AMGT in Grapes
(MRID 45593101) undergo an ILV trial, and potentially, a TMV trial by
the ACB.  This method was used to measure AMGT residues in the blueberry
field trials that the registrant performed.  The method procedure and
fortification recoveries were discussed in the DER for MRID 46986701.

  

Concurrent recoveries were measured in samples fortified with AMGT at
0.020 ppm, 0.100 ppm, 0.200 ppm, and 1.00 ppm (1x, 5x, 10x, and 50x the
LLMV).  Individual recoveries of AMGT (n = 35) ranged from 58 to 125%
(see TABLE 6, below).  At each fortification level, average recoveries
were within the generally recognized acceptable range (70-120%) except
for 2 samples spiked with 0.020 ppm AMGT (125 and 65% recovery) and 3
samples spiked with 0.100 ppm AMGT (58, 68, and 68% recovery).  Overall,
peaks were well-defined and symmetrical in all chromatograms.  Three
untreated control samples showed identifiable peaks at or near the
retention time for AMGT (ranging from 0.0333 to 0.1307 ppm).  Treated
sample chromatograms showed analyte peaks within the area of analytical
interest, and no carryover from controls.  Calibration curves were not
provided for AMGT.  Treated and control blueberry samples were stored
frozen (at -21 ± 7°C) for durations of up to 229 days before AMGT
analysis.  A concurrent freezer storage stability study was conducted
in/on blueberry samples fortified with AMGT at 0.15 ppm.  Residues in
the concurrent freezer storage stability samples remained stable for
intervals of 251 days.  Therefore, there are no concerns with the
stability of residues over time in this study.

TABLE 6.	Summary of Method Validation and Concurrent Recoveries of AMGT
from Blueberries.

Crop [Matrix]	Recovery Type 1	Spike Level

(ppm)	Sample Size (n)	Recoveries

(%)	Mean ± Std. Dev. (%)

AMGT

Blueberry [Berry]	MV	0.020	3	110, 95, 100	102  ± 8

	MV	0.100	3	95, 81, 94	90 ± 8

	MV	1.00	3	110, 110, 110	110 ± 0

	CR	0.020	5	1252, 65, 70, 70, 110	88 ± 28

	CR	0.100	27	93, 76, 81, 82, 71, 58, 84, 82, 78, 79, 74, 72, 1003, 733,
70, 88, 683, 683, 1103, 703, 80, 90, 89, 83, 84, 82, 88	80 ± 11

	CR	1.00	1	77	77

	CR	0.200	2	95, 80	88 ± 11

1. MV = Method Validation; CR = Concurrent Recovery.

2. High recovery might be due to a slight interference in the peak.

3. After interference found in the untreated sample is subtracted.

The registrant requested that the blueberry submission serve as the ILV
for the method.  The range of fortification levels and the percent
recovery values are acceptable.  As a result, HED considers the AMGT
analytical portion of the blueberry field trial study to be adequate,
and that it can serve as the ILV for the analytical method for AMGT
residues in plant commodities.  

Data-collection Methods

Fluazinam

Samples collected from the magnitude of the residue and storage
stability studies were analyzed for residues of fluazinam using GC/ECD
methods revised from the plant enforcement method.  The GC/ECD 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
LLMV was 0.01 ppm in all tested commodities, except in carrots, where
the LLMV was 0.02 ppm.

AMGT

Samples of apples and apple processed commodities were also analyzed for
residues of AMGT using an HPLC/UV method similar to that reviewed in
Memo, D335640, W. Drew, 8/22/2007.  The HPLC/UV method is adequate for
data collection based on acceptable method verification and concurrent
method recoveries.  The validated method LOQ was 0.01 ppm in all apple
matrices.

Animal commodities

DER Reference:	47756602.der.doc (Includes method validation data from
MRID 47756605)

ISK Biosciences submitted PTRL Method 1676W (MRID 47756602) entitled
“Fluazinam, AMPA and DAPA Method in Milk and Tissues as Described in
‘Magnitude of Fluazinam Residues in Bovine Tissues and Milk from a
28-Day Feeding Study.’”  Method 1676W was the data-collection method
used for the analysis of samples collected from a companion cattle
feeding study (MRID 47756605).  As a result of a June 30, 2007 meeting
between HED and ISK representatives to discuss concerns about the
analytical approach for the feeding study, the method has incorporated a
mild acid hydrolysis step to convert the sulfamate conjugates to free
AMPA and DAPA.  The method is written for LC/MS/MS analysis of
fluazinam, AMPA, and DAPA in bovine milk and tissues, and for GC/MS
analysis of DAPA in milk only.  The reported method LOQ was 0.01 ppm for
each analyte in milk, muscle, fat, liver, and kidney.  The method is
described briefly below.

For milk, a sample was mixed with HCl, and residues were extracted with
methanol.  The extract was concentrated, and residues were mixed with
water and sodium chloride for partitioning with dichloromethane (DCM) or
hexane for LC/MS/MS analysis for fluazinam, AMPA, and DAPA residues, or
for GC/MS analysis of DAPA residues.  For muscle, residues were
extracted with acetonitrile (ACN)/water, with acetic acid for fluazinam
and without acetic acid for AMPA and DAPA.  After concentration and/or
dilution, the extracts were cleaned up through a solid phase extraction
(SPE) column prior to LC/MS/MS analysis.  For fat, residues were
extracted with ACN/water with acetic acid, then again with ACN.  The
combined extract was diluted for partitioning with ACN-saturated
cyclohexane.  The ACN phase was evaporated and reconstituted for
LC/MS/MS analysis.  For analysis of liver and kidney using the
non-hydrolysis method, residues were extracted with ACN/water with
acetic acid, then again with ACN/water.  The combined extract was
diluted, concentrated, and mixed in water with sodium chloride for
partitioning with DCM prior to LC/MS/MS analysis.  For analysis of liver
and kidney using the hydrolysis method, residues were extracted with
ACN/water.  The extract was diluted and concentrated prior to the
addition of HCl (held for 1 hour at 37°C).  Water and sodium chloride
were added for partitioning with hexane (liver) or ethyl acetate
(kidney).  The hexane phase was concentrated and reconstituted for
LC/MS/MS analysis.  The LC/MS/MS method cites two transition ions for
each analyte in order to provide confirmation.

Method validation data, generated by PTRL West, Inc. (Hercules, CA),
were included in the cattle feeding study.  Milk was fortified with
fluazinam, AMPA, and DAPA at 0.010 and 0.10 ppm each.  Muscle, liver,
and kidney were fortified with the same analytes at levels of 0.01 and
0.05 ppm each.  Fat was fortified with fluazinam, AMPA, and DAPA at
0.01, 0.02, 0.1, and 0.5 ppm each.  Overall, the results showed that
Method 1676W adequately recovered residues of fluazinam, AMPA, and DAPA
in milk, muscle, and fat.  Method recoveries in these matrices were
generally within the acceptable recovery range of 70-120%.  However,
recoveries in liver and kidney were low and declined with an increase in
fortification level.  For hydrolyzed liver, overall mean recoveries of
fluazinam, AMPA, and DAPA were 87%, 68%, and 38%, respectively, at a
fortification level of 0.01 ppm.  Overall mean recoveries of fluazinam,
AMPA, and DAPA were 79%, 65%, and 34%, respectively, at a fortification
level of 0.05 ppm.  For hydrolyzed kidney, overall mean recoveries of
fluazinam, AMPA, and DAPA were 96%, 75%, and 31%, respectively, at a
fortification level of 0.01 ppm.  Overall mean recoveries of fluazinam,
AMPA, and DAPA were 63%, 39%, and 9%, respectively, at a fortification
level of 0.05 ppm.  The petitioner reported that extensive method
development work was performed on liver in an effort to obtain
acceptable recoveries for DAPA residues.

An ILV of Method 1676W was performed by ADPEN Laboratories, Inc.
(Jacksonville, FL) using two fortification levels:  at the LOQ (0.01
ppm) and 10x the LOQ.  Although minor modifications of the method for
some matrices were made to make it more rugged, the ILV was completed
within three trials.  The performing laboratory concluded that, based on
acceptable recoveries of fluazinam, AMPA, and DAPA in milk, beef fat,
and beef liver down to a level of 0.01 ppm for each compound, Method
1676W, with minor modifications, is a suitable method for use on bovine
tissues and milk.  In the first trial, the method was successfully
validated for fluazinam and AMPA in milk, for AMPA in beef fat, and for
AMPA in the liver non-hydrolysis procedure.  In the second trial,
analysis of fluazinam and DAPA in beef fat was successfully completed
after communication with the sponsor and the method developers.  In the
third trial, fluazinam, AMPA, and DAPA in milk and non-hydrolyzed liver
were successfully completed after minor method modification.

The method was forwarded to ACB/BEAD for method validation.  ACB/BEAD
reported that the lab that performed the ILV made major modifications to
the original method.  As a result, HED requested that the registrant
submit a revised method that incorporated the revisions made by the ILV
lab.  ISK Biosciences revised the method and re-submitted it.  ACB/BEAD
performed a cursory review of the revised method.  The registrant
incorporated the modifications made by the ILV lab.  However, the
revision eliminated the cleanup steps that were in the original method,
and the only recovery data for the revised method were data provided by
the ILV lab.  ACB/BEAD was concerned that the elimination of the cleanup
steps could lead to dirty samples which would, in turn, lead to
analytical problems, such as interference.  ACB/BEAD performed no
further validation of the method.  As the revised method contains major
revisions to the original method, and ACB/BEAD believes it is very
likely that the revised method still is not adequate for sample analysis
or tolerance enforcement, HED recommends that the registrant submit an
ILV for the revised method.

Conclusions.  An adequate GC/ECD method (6148-91-0170-MD-001) is
available for determination of fluazinam residues in crop matrices.  The
method has been deemed suitable for tolerance enforcement because it has
successfully completed the routes required for an enforcement method,
including radiovalidation, ILV trial, and TMV trial.

An enforcement method for determination of AMGT in plants is available. 
AMGT is a residue of concern for tolerance expression in wine grapes. 
The registrant requested that the blueberry submission serve as the ILV
for the method.  The range of fortification levels and the percent
recovery values are acceptable.  As a result, HED considers the AMGT
analytical portion of the blueberry field trial study to be adequate,
and that it can serve as the ILV for the analytical method for AMGT
residues in plant commodities.  

At the present time, an analytical method is not available for
enforcement of fluazinam tolerances in animal commodities.  The
registrant should submit an ILV of the revised method.  This method
needs to be adequate for analysis of parent fluazinam as well as the
metabolites AMPA and DAPA in animal commodities for which tolerances
will be established.

860.1360 Multiresidue Methods

Parent Fluazinam

Reference:  Residue Chemistry Memo, D257115, W. Cutchin, 5/21/01

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.  Data depicting
the recovery of AMGT through FDA MRMs were submitted in support of the
apple petition.

Metabolite AMGT

DER Reference:	47756603.der.doc

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 study was
conducted by Colorado Analytical R&D Corporation (Colorado Springs, CO).

 

AMGT does not have an N-methylcarbamate structure and was not found to
be naturally fluorescent in testing under Protocol A.  In Protocol B
testing, the methyl ester of AMGT was not recovered from Florisil;
therefore, no additional work was performed under Protocol B.

Protocol C testing of the compound indicated that further testing using
Protocols D, E, and F was required for all test substances.  The
Protocol C results indicated that AMGT was detected using GC with
electron capture detection (ECD) and nitrogen phosphorous detection
(NPD).  

Metabolite AMGT was not recovered from grapes (non-fatty matrix)
fortified at 0.05 and 0.25 ppm using Protocol D (DB-1 column with NPD)
because of matrix interference.  AMGT was not recovered from any of the
Florisil screens in Protocol E, and Protocol F testing was not
performed.  

These data indicate that FDA multiresidue methods are not suitable for
determining residues of the fluazinam metabolite AMGT.  The submitted
data will be forwarded to U.S. FDA for further evaluation.  

Conclusions:  Adequate FDA   SEQ CHAPTER \h \r 1 MRM testing data are
available for fluazinam and 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. 
Multiresidue method testing data are required for the regulated
metabolites, AMPA and DAPA, in animals.

860.1380 Storage Stability

Residue Chemistry Memo:  D257115, W. Cutchin, 5/21/01

Residue Chemistry Memo:  D335640, W. Drew, 8/22/07

Plant commodities  

The storage durations and conditions of crop samples from the submitted
magnitude of the residue and processing studies are presented in Table
7.

Table 7.   Summary of Storage Conditions and Durations of Samples from
Crop Field Trials.  

Matrix 	Storage Temperature  (°C)	Actual Storage Duration (days)
Interval of Demonstrated 

Storage Stability (days) for Fluazinam

Apple, RAC 	-24.4 to -6.0	40-291	1,097

Apple, processed commodities	-20 to -18	758-914	1,094-1,096

Carrot	-22 to -4	450	470 1

Lettuce, head	<0	64-229	419 1

Lettuce, leaf	<0	42-214	419 1

Onion	<-6	70-418	429 1

1  Storage stability determined from concurrent field trial samples.

s for up to 18 months, but declined by 35% in whole potatoes and ≥70%
in granules within 18 months.  Residues declined by 40% in wet peel
within 5 months.  Residue declines during storage have also been
observed for the following crops:  70% decline in cabbage by 18.3
months, 55% decline in mustard greens by 19 months, 30% decline in
ginseng by 11.4 months, 45% decline in dried beans by 10 months, and a
40% decline in lima beans by 15 months.  To account for dissipation of
residues during frozen storage, HED incorporated correction factors into
the recommended tolerances for fluazinam residues for certain crop
commodities.

To support the current lettuce, onion, and carrot field trial data,
storage stability studies were conducted in conjunction with the field
trials.  For each crop, control samples of the respective matrices were
fortified with fluazinam at 1.0 ppm and placed in storage under the same
frozen conditions as the treated samples.  No zero-day analyses were
conducted on the fortified samples placed into storage.  Rather,
triplicate stored samples were analyzed along with a control sample and
freshly fortified samples at an interval reflecting the maximum storage
duration from the field trials.  Average corrected recoveries of
fluazinam following frozen storage were 73% from lettuce after 13.8
months, 86% from onions after 14.1 months, and 73% from carrots after
15.5 months.  Although no 0-day analysis was conducted, the data
indicate that fluazinam is stable under frozen storage conditions for up
to 14 months in lettuce and onion and 16 months in carrot.

Storage Stability Data for Apple Fruit, Juice, and Wet Pomace

Storage stability data for fluazinam and its metabolite AMGT in apple
fruit, juice, and wet pomace were submitted in support of the apple
petition.

DER Reference:	47756604.der.doc

ISK Biosciences submitted storage stability data for fluazinam and its
metabolite AMGT in apple fruit, juice, and wet pomace.  Homogenized
untreated samples of fruit, juice, and wet pomace were spiked with
fluazinam or AMGT at 0.50 ppm, and stored frozen (-20 to -18ºC) for up
to 37 months.  Samples were analyzed initially (after 0 or 1 day) and
following storage for 1, 2, 3, 6, 12, 18, 30, and 36 months.

Samples of apple fruit, juice, and wet pomace were analyzed for residues
of fluazinam and AMGT.  The GC/ECD method used for fluazinam analysis is
similar to the current enforcement method for fluazinam in crops.  In
addition, the HPLC/UV method used for AMGT is similar to the method
described in “Method Evaluation for the Analysis of AMGT in Grapes”
(Memo, D335640, W. Drew, 8/22/2007).  The methods were adequate for data
collection based on acceptable concurrent method recoveries.  The
reported “limit of detection” (LOD) was 0.025 ppm for each analyte
in apple fruit and wet pomace, and 0.05 ppm for each analyte in apple
juice.

  SEQ CHAPTER \h \r 1 Residues of fluazinam appear to be relatively
stable in/on apple fruit, juice, and wet pomace stored frozen for up to
36 months.  Residues of AMGT were relatively stable in/on apple fruit
and juice stored frozen for up to 36-37 months.  Residues of AMGT
declined slightly in apple fruit to an average corrected recovery of 69%
at the 36-month interval.  However, AMGT residues in/on apple wet pomace
were relatively stable for only 12 months of frozen storage.  Residues
of AMGT declined in wet pomace to an average corrected recovery of
58-63% at the 19-month interval, 61% at the 32-month interval, and 33%
at the 37-month interval.

Storage Stability Data for Cattle Milk, Meat, and Meat Byproducts

Samples collected from the dairy cow feeding study (MRID 47756605) were
held under frozen storage conditions prior to residue analysis.  The
maximum storage durations of samples, from collection to analysis, were
146 days for milk, 185 days for cream, 179 days for skim milk, 157 days
for muscle, 203 days for fat, and 255 days for liver and kidney.  A
storage stability study was conducted to validate sample storage
conditions and durations.  The results showed mixed results.  In milk,
cream, and skim milk, residues of fluazinam, AMPA, and DAPA were found
to be stable for 183 days.  In muscle:  (i) fluazinam was stable for 1
day, but declined after 164 days to an average corrected recovery of
55%; (ii) AMPA was stable for 2 days, but declined after 161 days to an
average corrected recovery of 41%; and (iii) DAPA was stable for 2 days,
but declined after 161 days to an average corrected recovery of 10%.  In
fat:  (i) fluazinam and AMPA were stable for 205 days; and (ii) DAPA
showed relative instability with average corrected recoveries of 64%
after 205 days.  In liver and kidney:  (i) fluazinam was unstable,
average recoveries in liver were 26-34% after 210 days, and 41-44% in
kidney after 218 days; (ii) AMPA was stable in liver for 80/89 days, but
not in kidney where average corrected recoveries were 53-58% after 218
days; and (iii) DAPA is not stable with average corrected recoveries of
43-52% in liver after 209 days and 17-25% in kidney after 218 days.

Conclusions.  Adequate storage stability data are available to support
the storage conditions and durations of samples collected from the
magnitude of the residue studies on apples, carrots, lettuce, and
onions.  As residues of fluazinam were found to be stable in tested crop
matrices, storage stability corrections do not need to be applied to the
recommended tolerances for fluazinam residues in/on apples, carrots,
lettuce, and onions.  Storage stability data for the fluazinam
metabolite AMGT in apple commodities were also submitted and showed that
residues in apple wet pomace were reasonably stable for 12 months in
apple wet pomace, but declined to an average corrected recovery of 33%
at the 37-month interval.  The submitted storage data for bovine milk,
meat, and meat byproducts showed mixed results.  Because of major data
deficiencies, HED is not recommending in favor of tolerances on apple
and carrot commodities.  It is very possible that the registrant will
resolve the deficiencies and re-submit the tolerance petitions.  As a
result, HED has determined the maximum reasonable dietary burdens for
livestock based on the assumption that tolerances will be established
for apples and carrots.

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 wet apple pomace and carrot culls.  The
livestock feedstuffs associated with registered crops include peanut
meal and processed potato waste.  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 considered for dietary burden calculations.  The
maximum reasonable dietary burdens of fluazinam to livestock are
presented in Table 8.  The calculated dietary burdens of fluazinam are
0.04 ppm for beef cattle, 1.27 ppm for dairy cattle, 0.005 ppm for
poultry, and 0.003 ppm for swine.

Table 8.   Calculation of Dietary Burdens of Fluazinam Residues to
Livestock.

Feedstuff1	Type1	% Dry Matter2	% Diet2	Tolerance (ppm)	Dietary
Contribution (ppm)3

Beef Cattle

R not registered	R	--	15	--	--

Potato, processed waste	CC	15	30	0.02	0.04

CC not registered	CC	--	50	--	--

PC not registered	PC	--	5	--	--

TOTAL BURDEN	--	--	100	--	0.04

Dairy Cattle

R not registered	R	--	45	--	--

Apple, wet pomace	CC	40	10	5.0	1.25

Potato, processed waste	CC	15	10	0.02	0.013

CC not registered	CC	--	25	--	--

Peanut, meal	PC	85	10	0.02	0.002

TOTAL BURDEN	--	--	100	--	1.27

Poultry

CC not registered	CC	--	75	--	--

Peanut meal	PC	85	25	0.02	0.005

TOTAL BURDEN	--	--	100	--	0.005

Swine

CC not registered	CC	--	85	--	--

Peanut meal	PC	85	15	0.02	0.003

TOTAL BURDEN	--	--	100	--	0.003

1  R:  Roughage; CC:  Carbohydrate concentrate; PC:  Protein
concentrate.

2  OPPTS 860.1000 Table 1 Feedstuffs (June 2008).  

3  Contribution = ((tolerance) ( (% DM)) x (% of diet) for beef and
dairy cattle

    Contribution = ((tolerance) x (% of diet)) for poultry and swine. 

Dairy Cattle Feeding Study

DER Reference:	47756605.der.doc

Fourteen lactating Holstein cows were divided into four treatment
groups.  Three groups were dosed orally with gelatin capsules containing
fluazinam at target levels of 2.5 ppm (three cows), 7.5 ppm (three
cows), and 25 ppm (six cows) in the feed (dry weight basis) for 28
consecutive days.  The fourth group, consisting of two cows, served as a
control.  Based on feed consumption data, the actual dosing levels were
2.91, 8.72, and 28.84 ppm.  Fluazinam was weighed directly into gelatin
capsules, and the capsules were administered using a balling gun, once
per day after the morning feeding.  One control cow and three cows dosed
at 25 ppm were reserved to investigate the depuration of residues in
cow’s milk and tissues.  

Milk was collected twice daily   SEQ CHAPTER \h \r 1 on Days 0, 1, 3, 4,
5, 6, 7, 10, 13, 16, 19, 22, 25, and 28.  In addition, extra milk
samples from one control animal and three cows from the 2.91- and
28.84-ppm dose groups were collected on Study Days 13 and 28 to produce
data on skim milk and cream.  Cows from the three treatment groups were
sacrificed ≤ 24 hours after the final dose administration.  One cow
from the depuration group was sacrificed on Study Days 30, 32, and 36. 
At sacrifice, samples of liver, kidney, fat (subcutaneous, abdominal,
and perirenal), and muscle (leg and loin) were collected from each
animal.

Samples of milk plus cream, skim milk, muscle, liver, kidney, and fat
were analyzed using PTRL Method 1676W entitled “Fluazinam, AMPA and
DAPA Method in Milk and Tissues as Described in Magnitude of Fluazinam
Residues in Bovine Tissues and Milk from a 28-Day Feeding Study,”
which was included in Appendix B of the feeding study.  This method has
not been reviewed by the PMRA or the USEPA.  The method is written for
LC/MS/MS analysis of fluazinam, AMPA, and DAPA in bovine milk and
tissues, and for GC/MS analysis of DAPA in milk only.  The method has a
mild acid hydrolysis step for analysis of some matrices (milk, liver,
and kidney) to convert the sulfamate conjugates to free AMPA and DAPA. 
The method limit of quantitation (LOQ) was 0.01 ppm for each analyte in
milk and tissues.  The initial validation of Method 1676W showed that it
adequately recovered residues of fluazinam, AMPA, and DAPA in milk and
fat.  In muscle, recoveries in most were acceptable.  Mean recoveries of
fluazinam, AMPA, and DAPA at the 0.01 ppm spiking level were 82%, 96%
and 90%, respectively.  Mean recoveries of fluazinam, AMPA, and DAPA at
the 0.05 ppm spiking level were 66%, 95%, and 95%, respectively. 
Recoveries of AMPA and DAPA in liver and kidney were low.  Recoveries of
fluazinam from non-hydrolyzed kidney samples were also low.  The results
of concurrent method validation also showed similar low recoveries of
the metabolites in liver and kidney.  Mean recoveries of AMPA ranged
from 48 to 93% in hydrolyzed liver and from 36 to 75% in hydrolyzed
kidney.  Mean recoveries of DAPA ranged from 35 to 51% in hydrolyzed
liver and from 9 to 33% in hydrolyzed kidney.  Because of low
recoveries, the petitioner corrected residues in cow liver and kidney
for concurrent method recoveries <70%.   According to the petitioner,
correspondence with ISK’s Central Institute in Japan indicates that,
based on their experience with a variety of matrices, DAPA is easily
oxidized and easily absorbed to protein and organic matter.  This
information is consistent with the observed instability and the
difficulty experienced with recovering DAPA from some matrices.  

Samples were held under frozen storage conditions prior to residue
analysis.  Refer to the storage stability section of this memorandum for
information pertaining to sample storage duration as well as supporting
storage stability data.  As Method 1676W gave very poor recovery for
AMPA and DAPA in liver and kidney, the method needs to be revised and
the feeding study needs to be repeated.  When the feeding study is
repeated, the samples need to be analyzed as soon as possible after
sacrifice in order to minimize degradation of residues.  In muscle,
fluazinam declined by 45%, AMPA by 59%, and DAPA by 90% after 161-164
days.  In fat, fluazinam and AMPA were stable while DAPA declined by 36%
after 205 days.  In liver, fluazinam declined by as much as 74% after
210 days, AMPA was stable, and DAPA declined by 57% after 209 days.  In
kidney, fluazinam declined by 59% after 218 days, AMPA declined by 47%,
and DAPA declined by 83% after 218 days.

The maximum residues of concern in cow milk and tissues at the dose
level closest to the maximum reasonably balanced diet (MRBD),
uncorrected and corrected for storage stability recoveries, are
summarized in Table 9.  Residues in milk increased until about Day 5 and
were essentially stable beyond that time.

Table 9.   Results of a Dairy Cattle Feeding Study with Fluazinam.

Matrix	Feeding Level 1	Maximum Uncorrected Residues for Storage Decline
Maximum Corrected Residues for Storage Decline 2

Parent	AMPA	DAPA	Total	Parent	AMPA	DAPA	Total

Milk (Days 0-28)	8.72	<0.01	<0.01	<0.01	<0.03	--	--	--	--

Cream	28.84	<0.01	0.0582	0.1200	<0.19	--	--	--	--

Skim milk	28.84	<0.01	<0.01	<0.01	<0.03	--	--	--	--

Muscle (loin)	28.84	<0.01	0.0101	0.0179	<0.04	<0.018	<0.025	<0.179
<0.222

Muscle (round)	28.84	<0.01	<0.01	<0.01	<0.03	<0.018	<0.024	<0.100	<0.142

Liver (hydrolysis method)	2.91	<0.01	<0.01	<0.01	<0.03	<0.038	<0.01
<0.023	<0.071

Liver (non-hydrolysis method)	2.91	<0.01	<0.01	<0.01	<0.03	<0.038	<0.01
<0.023	<0.071

Kidney (hydrolysis method)	2.91	<0.01	<0.01	<0.01	<0.03	<0.024	<0.019
<0.059	<0.102

Kidney (non-hydrolysis method)	2.91	<0.01	<0.01	<0.01	<0.03	<0.024
<0.019	<0.059	<0.102

Fat, perirenal	2.91	<0.01	<0.01	<0.01	<0.04	<0.01	<0.01	<0.016	<0.036

Fat, abdominal	2.91	<0.01	0.0169	0.0219	<0.05	<0.01	0.0169	<0.034	<0.061

Fat, subcutaneous	2.91	<0.01	<0.01	0.0107	<0.03	<0.01	<0.01	0.0167
<0.037

1  Dose level closest to the MRBD or only dose level represented.

2   Individual residues were corrected for decline while in storage. 
Correcting residues for decline was made by dividing uncorrected
residues by (100% - % decline).  For milk, cream, and skim milk, no
correction of individual residues was made.  For muscle:  fluazinam
declined by 45%, AMPA by 59%, and DAPA by 90% after 161 days.  For fat: 
fluazinam and AMPA were stable while DAPA declined by 36% after 205
days.  For liver:  fluazinam declined by as much as 74% after 210 days,
AMPA was stable, and DAPA declined by 57% after 209 days.  For kidney: 
fluazinam declined by 59% after 210 days, AMPA declined by 47%, and DAPA
declined by 83% after 218 days.

For the depuration study, total residues were not observed at levels
higher than the combined LOQ (0.03 ppm) by Day 29, one day after dosing
was discontinued.  Total residues in tissues, except fat, of the animals
sacrificed on Days 29, 31, and 35 after the last dose administration
were all below the combined LOQ (0.03 ppm).  In fat of animals
sacrificed on Day 35, total residues were <0.031 ppm in perirenal fat,
<0.053 ppm in abdominal fat, and <0.051 ppm in subcutaneous fat.

Expected Secondary Residues in Milk, Meat, and Meat Byproducts

To determine the need for tolerances for the combined residues of
fluazinam, AMPA, DAPA, and their sulfate conjugates in milk and tissues,
the anticipated secondary residues in cattle matrices were estimated
using transfer coefficient factors calculated from the maximum combined
residues of fluazinam observed at the dose level closest to the
reasonably balanced dietary burden in the dairy cattle feeding study. 
The transfer coefficients (calculated as residue level to feed level
ratios) are presented in Table 10.  The transfer coefficient for each
matrix was then used to calculate the expected secondary residues by
multiplying the transfer coefficient by the calculated dietary burden. 
The expected combined residues and the recommended tolerances based on
expected residues are presented in Table 11.

Table 10.    Residue-Level-to-Feed Ratios (Transfer Coefficients) in
Dairy Cattle Milk and Tissues.

Matrix	Total Residues (Fluazinam + AMPA +  DAPA, and Their Sulfamate
Conjugates) in ppm	

Feeding Level

(ppm)	

Transfer

Coefficient 2

	Uncorrected	Corrected for Storage Decline 1

Milk (Day 0 to Day 28)	<0.03	<0.03	8.72	<0.0034

Cream	<0.19	<0.19	28.84	<0.0066

Skim milk	<0.03	<0.03	28.84	<0.0010

Muscle (loin)	<0.04	<0.22	28.84	<0.0077

Muscle (round)	<0.03	<0.14	28.84	<0.0049

Liver (hydrolysis method)	<0.03	<0.07	2.91	<0.0244

Liver (non-hydrolysis method)	<0.03	<0.07	2.91	<0.0244

Kidney (hydrolysis method)	<0.03	<0.10	2.91	<0.0351

Kidney (non-hydrolysis method)	<0.03	<0.10	2.91	<0.0351

Fat, perirenal	<0.04	<0.04	2.91	<0.0124

Fat, abdominal	<0.05	<0.06	2.91	<0.0210

Fat, subcutaneous	<0.03	<0.04	2.91	<0.0126

1   Individual residues were corrected for decline while in storage. 
For milk, cream, and skim milk, no correction was necessary.

2  Calculated from the maximum corrected residues at the dose level
closest to the MRBD divided by the dose level.

Table 11.    Expected Total Fluazinam Residues of Concern in Meat and
Milk.

Matrix	Dietary Burden

(ppm)	Total Residues 1

(ppm)	Tolerance

(ppm)

Dairy Cattle

Milk (Day 0 to Day 28)	1.27	<0.0043	Not required

Cream	1.27	<0.0084	Not required

Skim milk	1.27	<0.0013	Not required

Muscle (loin)	1.27	<0.0098	Not required

Muscle (round)	1.27	<0.0062	Not required

Liver (hydrolysis method)	1.27	<0.0310	A tolerance of 0.05 ppm is
considered to be a reasonable upper-bound estimate for meat byproducts
based on expected residues in kidney 2.

This tolerance level is also considered to be a reasonable upper-bound
estimate for fat.

Liver (non-hydrolysis method)	1.27	<0.0310

	Kidney (hydrolysis method)	1.27	<0.0446

	Kidney (non-hydrolysis method)	1.27	<0.0446

	Fat, perirenal	1.27	<0.0157

	Fat, abdominal	1.27	<0.0267

	Fat, subcutaneous	1.27	<0.0160

	Swine

Liver	0.003	0.00007	Not required

Kidney	0.003	0.00011	Not required

Muscle	0.003	0.00002	Not required

Fat	0.003	0.00006	Not required

1   Calculated from dietary burden x transfer coefficient from Table 9.

2    According to the July 18, 2007 Minutes of the Chemistry Science
Advisory Council Meeting, the ChemSAC recommended that the guidance
document be revised to include language detailing the use of the highest
residue data for any tissue (liver, kidney, fat, skin, or muscle) to
determine the tolerance for meat byproducts.  A single tolerance on
“meat byproducts” will be recommended based on that highest residue,
and individual tolerances will no longer be set on liver, kidney, or
meat byproducts (except liver and kidney).

Conclusions.  Based on the poor recoveries of metabolites AMPA and DAPA
from liver and kidney and the mixed storage stability results, the
residue data from the cattle feeding study are not adequate to satisfy
data requirements.  The feeding study data (adjusted for residue
decline) along with the calculated dietary burdens for beef cattle (0.04
ppm) and dairy cattle (1.27 ppm) indicate that tolerances might be
needed for the combined residues of fluazinam and its metabolites DAPA
and AMPA in the fat and meat byproducts of cattle and other ruminants at
0.05 ppm to support the proposed use on apples.  The feeding study also
indicates that tolerances probably will not be needed for milk and meat
of ruminants, as the expected combined residues in these matrices are
below the combined method LOQ of 0.03 ppm for the regulated compounds. 
Based on the transfer coefficients for livestock tissues and the
relatively low dietary burden for swine of 0.003 ppm for fluazinam,
tolerances for hogs are not needed.  A poultry feeding study is not
required at this time because of low dietary burden for poultry (0.005
ppm).  To support tolerances for apples and carrots, the registrant
needs to submit a new cattle feeding study in which a method with better
recoveries of AMPA and DAPA from liver and kidney was used.  When this
study is repeated, the samples need to be analyzed as soon as possible
after sacrifice in order to minimize degradation of residues.

860.1500 Crop Field Trials

DER References:	47631001.der.doc (Bulb Onion)

		47631002.der.doc (Leaf and Head Lettuce)

		47755601.der.doc (Carrot)

		47756609.der.doc (Apples:  2006 Trials)

47756606.de1.doc (Apples:  1992-1994 Trials, Includes MRIDs 47756607,
47756608, 47756610, and 47756611)

IR-4 and ISK Biociences Corporation have submitted magnitude of the
residue studies on apples, carrots, lettuce, and onions.    SEQ CHAPTER
\h \r 1 The results from these field trials are discussed below, and the
residue data are summarized in Table 12.  

Table 11.   Summary of Residue Data from Crop Field Trials with
Fluazinam.

Crop Matrix	Total Applic. Rate

(lb ai/A)

[kg ai/ha]	PHI (days)	Residues of Fluazinam (ppm)1

	n	Min.	Max.	HAFT2	Median	Mean	Std. Dev.

Head Lettuce (proposed use = 1.0 lb ai/A total application rate, 50-day
PHI)

Lettuce, head	0.952-1.02	46-52	14	<0.01	0.021	0.016	0.010	0.011	0.010

Leaf Lettuce (proposed use = 1.0 lb ai/A total application rate, 30-day
PHI)

Lettuce, leaf	0.974-1.05	20-32	14	<0.01	1.690	1.570	0.021	0.260	0.010

Onions (proposed use = 3.1 lb ai/A total application rate, 7-day PHI)

Onion, dry bulb	3.11-3.17	6-8	18	<0.01	0.101	0.098	0.014	0.027	0.028

Carrots (proposed use = 2.1 lb ai/A total application rate, 7-day PHI)

Carrot root	2.04-3.03

[2.29-3.40]	6-8	52	<0.02	0.56	0.51	0.10	0.14	0.13

Apples (proposed use = 4.5 lb ai/A total application rate, 28-day PHI)

Apples 3	4.44-4.77

[4.98-5.34]	28-29	24	0.02	1.75	1.75	0.15	0.38	0.58

Apples 4	3.97-5.40

[4.45-6.05]	28-32	16	<0.01	0.06	0.06	0.03	0.03	0.02

1  AMGT was measured in apples and the maximum residue value for this
metabolite was 0.03 ppm.

2  HAFT = highest average field trial result.

3  Results of trials conducted during the 2006 growing season (MRID
47756609).

4  Results of trials conducted during the 1992-1994 growing seasons
(MRIDs 47756606,  47756607, 47756608, 47756610, and 47756611).

Head and Leaf Lettuce.  In fourteen field trials conducted in the United
States in EPA Growing Zones 2, 3, and 10 during 2005-2006, 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 (DAT) for
head lettuce and 20-32 DAT for leaf lettuce.  

Head and leaf lettuce samples were stored at ≤0°C for up to 7.6
months prior to extraction for analysis.  The storage conditions and
duration are supported by the available storage stability data. 
Residues of fluazinam in/on lettuce were determined using an adequate
GC/ECD method (6148-91-0170-MD-001, modified), which has a calculated
LOQ of 0.007 ppm, and a LOD of 0.002 ppm.  The LLMV in/on lettuce was
0.01 ppm.

Following a single foliar application of fluazinam (FlC) to lettuce at a
rate of 0.952-1.05 lb ai/A (1x), residues of fluazinam were <0.01-0.021
ppm in/on 14 head lettuce samples harvested at 46-52 DAT, with only one
sample (0.021 ppm) having residues above the LLMV.  For leaf lettuce,
residues were <0.01-1.69 ppm in/on 14 samples harvested at 20-32 DAT,
with only 2 samples having residues <LLMV.  For head lettuce, average
residues were 0.011 ppm and the highest average field trial (HAFT)
residues were 0.016 ppm.  For leaf lettuce, average residues were 0.26
ppm and the HAFT residue was 1.57 ppm.  No residue decline study was
conducted.  In addition, samples were not analyzed for residues of AMGT,
as required for dietary risk assessment.

Bulb Onion Subgroup (3-07A).  In nine field trials conducted in the
United States in EPA Growing Zones 1, 5, 6, 8, 10, 11, and 12 during
2005-2006, fluazinam (4.17 lb/gal FlC) was applied to bulb onions as six
broadcast foliar applications during vegetative development and bulb
formation at rates of 0.495-0.563 lb ai/A/application and RTIs of 5-8
days, for a total of 3.11-3.17 lb ai/A (1x the proposed maximum seasonal
rate).  Applications were made using ground equipment in volumes of
19-60 gal/A, and no adjuvants were used.  Single control and duplicate
treated samples of onion bulbs were harvested at commercial maturity,
6-8 DAT.  

Onion bulb samples were stored at ≤-6°C for up to 13.9 months prior
to extraction for analysis.  The storage conditions and duration are
supported by the available storage stability data.  Residues of
fluazinam in/on bulb onions were determined using an adequate GC/ECD
method (6148-94-0170-MD-001, modified), which has a calculated LOQ of
0.011 ppm, and a LOD of 0.004 ppm.  The LLMV in/on onion was 0.01 ppm.

Following six broadcast foliar applications of fluazinam (FlC) to onions
at rates totaling 3.11-3.17 lb ai/A (1x), residues of fluazinam were
<0.01-0.101 ppm in/on 18 onion bulb samples harvested at 6-8 DAT, with 7
samples having residues <0.01 ppm.  Average residues were 0.027 ppm and
the HAFT residues were 0.098 ppm.  In the residue decline trial, average
fluazinam residues declined steadily from 0.085 ppm at 0 DAT to <0.01
ppm by 14 DAT.

Carrots:  Thirteen carrot field trials were conducted in the United
States and Canada in NAFTA Growing Zones 1A, 3, 5, 5B, 6, 10, and 11
during the 2005 and 2006 growing seasons.  At each test location, Omega
500F (U.S. trials) or Allegro 500F (Canadian trials), a 4.17 lb/gal
flowable suspension concentrate (FlC) formulation of fluazinam was
applied to carrot plants as four broadcast or directed foliar
applications (five applications were made in one trial) during
vegetative development at rates of 0.492-0.786 lb ai/A/application
(0.551-0.881 kg ai/ha/application) and RTIs of 6 to 9 days, for a total
rate of 2.04-3.03 lb ai/A (2.30-3.40 kg ai/ha; 1.0-1.4x the proposed
maximum seasonal rate).  Applications were made using ground equipment
in volumes of 24-60 gal/A (220-560 L/ha), and no adjuvants were used. 
One untreated plot and two treated plots were used at each site.  Each
treated plot received treatments in similar rates and volumes; however,
one treated plot was irrigated within two hours of each application, and
the other treated plot received no irrigation for at least 24 hours
after each application.  Duplicate treated and untreated samples of
carrot root were harvested at commercial maturity at a 6- to 8-day PHI. 
Additional samples were harvested at 1, 13, and 20-day PHIs at the
Reverside, CA trial site to assess residue decline behavior.

Samples of carrot root were stored frozen for up to 450 days (14.8
months) between harvest and analysis.  Concurrent storage stability data
were provided demonstrating the relative stability of fluazinam in/on
carrot root for up to 470 days (15.5 months) of frozen storage. 
However, zero-day data were not provided.  Storage stability studies
should always 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)).

Residues of fluazinam in carrot root samples were determined using the
gas chromatography/ electron capture detection (GC/ECD) Method
YARL-0605A (Method No. 94-017 in PMRA #1407042).  The modified method
was adequate for data collection based on acceptable method validation
and concurrent recoveries.  The validated limit of quantitation (LOQ; or
the LLMV) was 0.02 ppm.  The calculated LOQ was 0.024 ppm.  The
calculated limit of detection (LOD) was 0.0081 ppm.

Following four foliar applications of the 4.17 lb/gal FlC formulation of
fluazinam to carrots at total rates of 2.04-2.94 lb ai/A (2.29-3.29 kg
ai/ha; 1.0-1.4x application rates) with irrigation within 2 hours of the
last application, residues of fluazinam ranged <0.02-0.56 ppm in/on 26
carrot root samples harvested at a 6- to 8-day PHI.  Residues of
fluazinam ranged from <0.02-0.45 ppm following applications at 2.04-3.03
lb ai/A (2.29-3.40 kg ai/ha; 1.0-1.4x) with irrigation withheld for at
least 24 hours following the last application.  Fluazinam residues from
each treatment pattern were comparable:  the average residues for carrot
samples receiving irrigation within 2 hours or after 24 hours of the
final application were 0.15 and 0.12 ppm, respectively, and the HAFT
residues were 0.51 and 0.43 ppm, respectively.  In the residue decline
trial, average residues of fluazinam in/on carrot roots declined from
the 1-to the 8-day PHI (from 0.67 to 0.15 ppm), increased at the 13-day
PHI (0.36 ppm) and declined slightly at the 20-day PHI (0.34 ppm) for
the early-irrigation treatment pattern.  For the late-irrigation
treatment pattern, average residues declined from the 1- to the 8-day
PHI (from 0.53 to 0.21 ppm), increased at the 13-day PHI (0.45 ppm),
then declined again at the 20-day PHI (0.33 ppm).

Apples (MRID 47756609):  Twelve apple field trials were conducted in the
United States and Canada in NAFTA Growing Zones 1, 5A, 5B, 9, 10, and 11
during the 2006 growing season.  At each test location, consisting of
one untreated and one treated plot, a 4.17 lb/gal FlC formulation of
fluazinam was applied to apple trees (ages 5-38 years) as ten airblast
foliar applications during fruit development at rates of 0.435-0.713 lb
ai/A/application (0.488-0.799 kg ai/ha/application) and RTIs of 6 to 8
days, for a total rate of 4.44-4.77 lb ai/A (4.98-5.34 kg ai/ha).  The
identical product was used in both the U.S. and Canadian trials.  In the
U.S., the name of this product is Omega 500F, and in Canada, the name of
this product is Allegro 500 F.  Applications were made using ground
equipment (tractor or backpack sprayer) in volumes of 51-152 gal/A
(480-1420 L/ha), and no adjuvants were used.  Duplicate treated and
untreated samples of apple fruit were harvested at commercial maturity
at a 28- or 29-day PHI.  At two trials, additional samples were
harvested at 0-, 7-, 14-, and 21-day PHIs to evaluate residue decline.

Samples of apple fruit were stored frozen for up to 144 days (4.7
months) between harvest and analysis.  Adequate storage stability data
are available (refer to the DER for MRID 47756604) for fluazinam and
AMGT to support the storage conditions and durations of apple samples
discussed in this summary document.  These data indicate that residues
of fluazinam and AMGT appear to be relatively stable under frozen
storage in apple fruit for up to 36 months.  

The GC/ECD method used for fluazinam analysis is similar to the current
enforcement method for fluazinam in crops.  In addition, the HPLC/UV
method used for AMGT is similar to the method described in “Method
Evaluation for the Analysis of AMGT in Grapes” (Memo, D335640, W.
Drew, 8/22/2007).  The methods used for fluazinam and AMGT were adequate
for data collection based on acceptable method validation and concurrent
recovery data.  The validated LOQ was 0.01 ppm for each analyte in apple
fruit; the calculated LOQs were 0.011 and 0.017 ppm, respectively, for
fluazinam and AMGT.  The calculated LODs were 0.0031 and 0.0055 ppm for
fluazinam and AMGT, respectively, in apple fruit.

Following ten foliar airblast applications of the 4.17 lb/gal FlC
formulation of fluazinam to apples at total rates of 4.44-4.77 lb ai/A
(4.98-5.34 kg ai/ha; 1.0-1.1x), maximum residues of fluazinam were 1.75
ppm and maximum residues of AMGT were 0.03 ppm.  In the residue decline
trials, residues of fluazinam in/on apple fruit declined from the 0- and
7-day PHIs to the 28- or 29-day PHI.  In one trial, the 0-day samples
were not analyzed.

Apples (MRIDs 47756606, 47756607, 47756608, 47756610, and 47756611): 
Eight apple field trials were conducted in the United States and Canada
in NAFTA growing Zones 1, 1A, 2, 5, and 5A during the 1992, 1993, and
1994 growing seasons.  Three additional trials were conducted in Zones
10 (CA; 1 trial) and 11 (WA; 2 trials); however, these trials were
conducted at lower total application rates (2.17-3.15 lb ai/A) and a
longer PHI (90 days).  At each test location, consisting of one
untreated plot and one treated plot, a 4.17 lb/gal FlC formulation of
fluazinam was applied to apple trees (ages 6-30 years) as 9 to 12 (5 to
7 for the WA and CA trials) airblast foliar applications during fruit
development at rates of 0.43-0.48 lb ai/A/application (0.48-0.54 kg
ai/ha/application) and RTIs of 7 to 23 days.  The total application
rates ranged from 3.97-5.40 lb ai/A (4.45-6.05 kg ai/ha; 0.9-1.2x the
proposed maximum seasonal rate).  At the other three trials, 5 to 7
airblast foliar applications were made at rates of 0.43-0.45 lb
ai/A/application (0.48-0.50 kg ai/ha/application) and RTIs of 4 to 15
days, for a total rate of 2.17-3.15 lb ai/A (2.43-3.53 kg ai/ha). 
Applications were made using ground equipment (tractor-mounted sprayer)
in volumes of 72-184 gal/A (673-1721 L/ha), and no adjuvants were used. 
Two additional plots were established at the PA site, one treated at a
low spray volume (~20-21 gal/A (191-197 L/ha)) and the other at twice
the nominal rate (0.9 lb ai/A (1.0 kg ai/ha)) and a dilute spray volume
(195-198 gal/A (1824-1852 L/ha)).  Duplicate treated and untreated
samples of apple fruit were harvested at commercial maturity at a 28- to
32-day PHI.

Samples were stored frozen for up to 291 days (9.6 months) prior to
determination of residues of fluazinam.  For AMGT analysis, four samples
from the 1992 trials were stored frozen for up to 1189 days (39.1
months) and twenty-two samples from the 1993-1994 trials were stored
frozen up to 797 days (26.2 months).  The submitted storage stability
data for fluazinam and AMGT are adequate to support the storage
durations and conditions of samples from the apple field trials (refer
to the DER for MRID 47756604).  Although apple samples from the 1992
trials were stored longer than 36 months prior to AMGT analysis, the
submitted storage stability data are adequate to support the storage
durations and conditions of samples from the apple field trials as the
majority of samples were stored less than 36 months, and residues in
these samples are comparable.

The GC/ECD method used for fluazinam analysis is similar to the current
enforcement method for fluazinam in crops.  In addition, the HPLC/UV
method used for AMGT is similar to the method described in “Method
Evaluation for the Analysis of AMGT in Grapes” (Memo, D335640, W.
Drew, 8/22/2007).  The methods used for fluazinam and AMGT were adequate
for data collection based on acceptable method validation and concurrent
recovery data.  The validated LOQ was 0.01 ppm for each analyte in apple
fruit.

Following 9 to 12 foliar airblast applications of the 4.17 lb/gal FlC
formulation of fluazinam to apples at total rates of 3.97-5.40 lb ai/A
(4.45-6.05 kg ai/ha), maximum residues of fluazinam were 0.08 ppm and
maximum residues of AMGT were 0.02 ppm.  Residue decline data were not
generated for these trials, but are available in the 2006 apple field
trial data (refer to the DER for MRID 47756609).

In the PA trial performed at a total application rate of 9.0 lb ai/A
(10.1 kg ai/ha), the fluazinam residue levels were 0.08 and 0.10 ppm and
the AMGT residue levels were 0.01 and <0.01 ppm.

Bushberry subgroup 13-07B.  IR-4 submitted a request to revise the
established 7.0-ppm tolerances for fluazinam residues in/on the
Bushberry subgroup 13B and other bushberry type crops (aronia berry,
buffalo currant, Chilean guava, European barberry, highbush cranberry,
edible honeysuckle, jostaberry, Juneberry, lingonberry, native currant,
salal, sea buckthorn).  These tolerances were originally established
under PP# 6E7137 (Memo, D335640, W. Drew, 8/22/07).  IR-4 is requesting
that a single tolerance be established for the Bushberry subgroup 13-07B
for residues of fluazinam at a level of 4.5 ppm.  

No new residue data were submitted in support of the tolerance amendment
request.  The original blueberry residue data for fluazinam per se were
entered into the tolerance spreadsheet.  HED concludes that the residue
data for fluazinam from the blueberry field trials are distributed
lognormally.  Therefore, the recommended tolerance for the bushberry
subgroup is 7.0 ppm, which is equivalent to the current tolerance for
blueberries.  Therefore, the recommended tolerance for the revised
Bushberry subgroup 13-07B is also 7.0 ppm.

Conclusions.  With the exception of the use of adjuvants (see 860.1200),
the submitted field trial data for fluazinam on head lettuce, leaf
lettuce, bulb onions, carrots, and apples are acceptable and support the
proposed uses.  Adequate numbers of tests were conducted in the
appropriate geographic regions at ~1x the maximum proposed rate, and the
appropriate samples were collected from each test at, or close to, the
proposed PHI.  Samples from all the field trials were analyzed for
fluazinam residues using an adequate GC/ECD method, and the storage
stability data support the sample storage durations for these crops. 
However, no residue data were provided on AMGT residues in lettuce. 
Although, AMGT residue data on lettuce are still required for purposes
of dietary risk assessment, the available data support tolerances of
0.02 ppm for head lettuce, 2.0 ppm for leaf lettuce, 0.20 ppm for the
bulb onion (Subgroup 3-07A), 1.7 ppm for apple, and 0.7 ppm for carrot. 
For dietary risk assessment, HED will calculate AMGT residues in lettuce
using a ratio of 0.35:1 for AMGT:fluazinam residues (ChemSAC Memo,
meeting of 2/6/08).  However, the samples from the lettuce field trials
still need to be analyzed for residues of AMGT.

After reevaluating the available residue data for blueberries, HED
concludes that the residue data for fluazinam from the blueberry field
trials are distributed lognormally.  Therefore, the recommended
tolerance for the Bushberry subgroup 13-07B is 7.0 ppm.

860.1520 Processed Food and Feed

DER Reference:	47756606.de2.doc (Includes MRIDs 47756610 and 47756611)

Apples:  In a single crop field trial conducted in NY during the 1993
growing season, a 4.17 lb/gal FlC formulation of fluazinam was applied
to mature apple trees as ten foliar airblast applications at a rate of
0.45 lb ai/A/application (0.50 kg ai/ha/application), with 7- to 14-day
RTIs, for a total rate of 4.50 lb ai/A (5.04 kg ai/ha; 1x the proposed
maximum seasonal rate).  The first application was made when fruit
development was at the green tip growth stage, with subsequent
applications being made up to 6.3-6.6 cm diameter fruit, and all
applications were made using ground equipment in 99-104 gal/A (926-973
L/ha) volumes without an adjuvant.  Mature apples were collected at a
29-day PHI and processed into wet and dry apple pomace, raw apple juice,
and apple cider.

Samples of whole apples and processed commodities were analyzed for
residues of fluazinam and AMGT.   The GC/ECD method used for fluazinam
analysis is similar to the current enforcement method for fluazinam in
crops.  In addition, the HPLC/UV method used for AMGT is similar to the
method described in “Method Evaluation for the Analysis of AMGT in
Grapes.”  These methods were reviewed under DP# 335640, W. Drew,
8/22/07.  These methods are adequate for data collection based on
acceptable method verification and concurrent method recoveries.  The
fortification levels used in method verification and concurrent method
validation were adequate to bracket expected residue levels in/on apple
fruits, the raw agricultural commodity (RAC), wet and dry pomace, juice,
and cider.  The validated method LOQ was 0.01 ppm for each analyte in
all matrices.

Samples of apple fruits were stored frozen for up to 191 days (6.3
months) prior to determination of residues of fluazinam and 805 days
(26.4 months) prior to determination of residues of AMGT.  Samples of
processed commodities were stored frozen for 194-236 days (6.4-7.8
months) prior to fluazinam analysis and for 758-914 days (24.9-30.0
months) prior to AMGT analysis.  The submitted storage stability data
(refer to the DER for MRID 47756604) for fluazinam and AMGT are adequate
to support the storage durations and conditions of samples from the
apple processing study.

The results of the apple processing study are summarized in Table 13. 
Average residues of fluazinam and AMGT were 0.025 ppm and below the LOQ
(<0.01 ppm), respectively, in/on apple fruit (RAC) harvested 29 days
following foliar airblast treatments with the 4.17 lb/gal FlC
formulation at a 1x application rate.  Respective average residues of
fluazinam and AMGT were 0.07 and <0.01 ppm in wet pomace; 0.09 and 0.01
ppm in dry pomace; and <0.01 ppm each in both raw juice and cider.  In
the processing study, residues of fluazinam concentrated in wet pomace
(2.8x) and dry pomace (3.6x), but did not concentrate in juice or cider
(<0.4x in each matrix).  Residues of AMGT did not concentrate in dry
pomace (1x).  Processing factors were not calculated for AMGT in wet
pomace, juice, or cider as the observed residues were <LOQ in both the
RAC and the processed commodity.  The observed processing factors are
less than the theoretical concentration factor of 14x for wet apple
pomace (OPPTS 860.1520, Table 4).

Table 13.   Residue Data from Apple Processing Study with Fluazinam.

Location

(City, State; Year)	Total Rate

(lb ai/A) 

[kg ai/ha]	PHI 

(days)	Commodity	Fluazinam residues

(ppm)

[Avg.]	Fluazinam Processing Factor	AMGT residues

(ppm)

[Avg.]	AMGT Processing Factor1

Williamson, NY; 1993	4.50

[5.04]	29	Fruit (RAC)	0.03, 0.02

[0.025]	--	<0.01, <0.01

[<0.01]	--

	Wet pomace	0.07, 0.07

[0.07]	2.8x	<0.01, <0.01

[<0.01]	NC

	Dry pomace	0.10, 0.08

[0.09]	3.6x	0.012, 0.012

[0.01]	1x

	Raw juice	<0.01, <0.01

[<0.01]	<0.4x	<0.01, <0.01

[<0.01]	NC

	Cider	<0.01, <0.01, <0.01

[<0.01]	<0.4x	<0.01, <0.01, <0.01

[<0.01]	NC

1  NC = Not calculated; residues were below the LOQ (<0.01 ppm) in both
the RAC and the processed fraction.

Conclusions:  The apple processing study is not adequate to satisfy data
requirements.  In apples, the metabolite AMGT is a residue of concern
for risk assessment.  In the processing study there were no quantifiable
residues of AMGT in raw apples.  In several of the field trials that
were performed, however, raw apples contained AMGT residues at, or
above, the limit of quantitation.  In a few cases, residues of AMGT were
comparable to those of the parent fluazinam.  Canadian and U.S. residue
chemistry guidelines both stipulate that in the processing study, if
residues in the raw agricultural commodity (RAC) are below the LOQ, but
quantifiable residues occurred in the RAC in field trials performed at
the maximum requested label rate, that the processing study should be
conducted at exaggerated rates in order to ensure that quantifiable
residues will be present in the RAC.  Quantifiable residues are needed
in the RAC in order to determine a reliable concentration factor.  As a
result, the processing study needs to be repeated at a sufficiently high
rate that residues of both parent and AMGT are quantifiable in the RAC. 
In the event that the potential for phytotoxicity exists, the
application rate does not need to be greater than the rate at which
phytotoxicity occurs.

  SEQ CHAPTER \h \r 1 860.1650 Submittal of Analytical Reference
Standards

Analytical reference standards for fluazinam (expiration date 4/12/2012)
AMPA (expiration date 5/31/2011), and DAPA (expiration date 4/2012) are
currently available in the EPA National Pesticide Standards Repository
(electronic communication D. Wright, ACB, to D. Dotson, HED,
11/18/2009).  An analytical standard for the AMGT metabolite is not
currently available in the EPA National Pesticide Standards Repository. 
The standard 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

Residue Chemistry Memo DP#s 212612, 216941, and 217467; 9/5/95; G.
Herndon

Residue Chemistry Memo DP# 342005, 2/1/08, W. Drew

An adequate confined rotational crop study is available in which barley,
carrots, and lettuce were grown in soil treated with
[phenyl-U-14C]fluazinam or [pyridyl-2,6-14C]fluazinam at 2.0 lb ai/A. 
All three crops were planted at PBIs of 30, 120, and 365 days, and no
crops were grown in the soil during the aging period.  Because of crop
failure, the 30-day PBI barley in the plot treated with
[phenyl-U-14C]fluazinam was replanted at a 68-day PBI.  

Although total radioactive residues (TRR) were greater than 0.01 ppm
in/on all rotational crop RACs (except 365-day PBI pyridine-labeled
carrot roots), no fluazinam or any other metabolite resembling the
intact fluazinam molecule was detected in any rotational crop RAC, from
even the shortest rotation interval (30-68 days).  The majority of the
radioactivity was found in the aqueous soluble fractions of plant
extracts, although some of the barley samples exhibited a high
percentage of bound radioactivity.  The organic fraction of each crop
matrix contained <10% or <0.01 ppm of the TRR.  Radioactivity in aqueous
fractions accounted for up to 75-95% of the TRR (0.06-0.27 ppm) in
phenyl-labeled crops, and up to 27-60% (0.02-0.046 ppm) in
pyridyl-labeled crops.  Analysis of the phenyl-labeled aqueous fractions
indicated that the majority of the radioactivity eluted early (4-5
minutes) in a single band which was identified as TFA.  The
pyridine-labeled aqueous fractions exhibited late-eluting radioactivity
(15-20 minutes), indicating different metabolism pathways between the
two labels.  No metabolites from the pyridine-labeled aqueous fractions
were identified.

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.  HED originally recommended that labels specify
rotational crop restrictions of 70 days for cereal grain and 30 days for
all other crops.  However, in a recent reexamination of the data (Memo,
D342005, W. Drew, 2/01/08), HED concluded that a 30-day PBI is
sufficient for all rotated crops not on the label.  No additional
rotational crop data are required for the proposed uses on carrots,
lettuce, and onions.

860.1550 Proposed Tolerances

For purposes of tolerance expression, HED previously determined that the
residue of concern is fluazinam per se for all plant commodities, except
for wine grapes where the parent compound and the metabolite AMGT are
the residues of concern.  For poultry and ruminants, the fluazinam
residues of regulatory interest include the parent plus the metabolites
AMPA and DAPA, and their sulfamate conjugates.

Tolerances for residues of fluazinam are listed under 40 CFR §180.574. 
Permanent tolerances have been established for residues of fluazinam
in/on plant commodities at levels ranging from 0.01 ppm on turnip greens
and Brassica leafy vegetables to 7.0 ppm in/on the Bushberry subgroup
and other bushberries (40 CFR §180.574(a)(1)).  A tolerance without a
U.S. registration has also been established for residues of fluazinam
and its metabolite AMGT in/on wine grapes at 3.0 ppm (40 CFR
§180.574(a)(2)).

The tolerance expression proposed by IR-4 in PP#8E7506 and PP#9E7570 for
head lettuce, leaf lettuce, carrots, and the Bulb onion Subgroup
(3-07A), is in terms of fluazinam.  The tolerance expression proposed by
ISK Biosciences in PP#9F7571 for apples and apple wet pomace is in terms
of the combined residues of fluazinam and its metabolite AMGT.  However,
only the parent needs to be regulated for these commodities.  Parent and
AMGT are the residues of concern for risk assessment, but not for
tolerance expression.  As a result, if and when ISK Biosciences
resubmits the tolerance petition for apples, the tolerance expression
for apples and apple wet pomace should be defined as fluazinam only. 
The tolerance expression proposed by ISK Biosciences in PP# 9F7571 for
milk, meat, and meat byproducts is appropriate and adequately reflects
the combined residues of fluazinam and its metabolites DAPA and AMPA.

HED recommends that 40CFR §180.574(a)(1) be amended by replacing the
tolerance expression with the following:  “Tolerances are established
for residues of fluazinam
(3-chloro-N-[3-chloro-2,6-dinitro-4-(trifluoromethyl)phenyl]-5-(trifluor
omethyl)-2-pyridinamine), 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
fluazinam.”  HED further recommends that 40CFR §180.610(a)(2) be
amended by replacing the tolerance expression with the following: 
“Tolerances are established for residues of fluazinam, 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 fluazinam and its metabolite AMGT
(3-[[4-amino-3-[[3-chloro-5-(trifloromethyl)-2-pyridinyl]amino]-2-nitro-
6-(trifluoromethyl) phenyl]thio]-2-(beta-D-glucopyranosyloxy) propionic
acid).”

Adequate numbers of field trials at ~1x the proposed use rate are
available for head lettuce, leaf lettuce, bulb onions, carrots, and
apples to support permanent tolerances on these crops.  With the
exception of head lettuce, the Agency’s SOP “Guidelines for Setting
Pesticide Tolerances Based on Field Trial Data” were utilized for
determining the appropriate tolerance level for each commodity (Appendix
II).  The recommended tolerances for these crop commodities are listed
in Table 14.  For head lettuce, the tolerance spreadsheet could not be
used to calculate a possible tolerance because of the large number of
residue values <LOQ (13 of 14 samples).  Therefore, the maximum residue
value (0.021 ppm) was used to determine the appropriate tolerance for
head lettuce of 0.02 ppm.

IR-4 has requested that the Agency reconsider the established 7-ppm
tolerances for members of the Bushberry subgroup 13B, as the established
tolerances are 3.5 times as high as the maximum residue value.  IR-4 has
also requested deletion of the separate tolerances for bushberry crops
that are now covered by the updated Bushberry subgroup 13-07B.  The
existing blueberry residue data for fluazinam were reevaluated according
to the Agency’s SOP “Guidelines for Setting Pesticide Tolerances
Based on Field Trial Data.”  After reviewing the probability plot for
the fluazinam residue data on blueberries, HED has determined that the
residue data are lognormally distributed, and that the recommended
tolerance for the Bushberry subgroup 13-07B should be 7.0 ppm.  In
addition, the separate tolerances for the various bushberry crops can
now be deleted, as all of the listed bushberries are now covered by the
revised Bushberry subgroup 13-07B.

The apple processing study is not acceptable.  In apples, the metabolite
AMGT is a residue of concern for risk assessment.  In the processing
study there were no quantifiable residues of AMGT in raw apples.  In
several of the field trials that were performed, however, raw apples
contained AMGT residues at, or above, the limit of quantitation.  In a
few cases, residues of AMGT were comparable to those of the parent
fluazinam.  Canadian and U.S. residue chemistry guidelines both
stipulate that in the processing study, if residues in the raw
agricultural commodity (RAC) are below the LOQ, but quantifiable
residues occurred in the RAC in field trials performed at the maximum
requested label rate, that the processing study should be conducted at
exaggerated rates in order to ensure that quantifiable residues will be
present in the RAC.  Quantifiable residues are needed in the RAC in
order to determine a reliable concentration factor.  As a result, the
processing study needs to be repeated at a sufficiently high rate that
residues of both parent and AMGT are quantifiable in the RAC.  In the
event that the potential for phytotoxicity exists, the application rate
does not need to be greater than the rate at which phytotoxicity occurs.

The residue data (adjusted for residue decline) from the cattle feeding
study, along with the calculated dietary burdens for beef and dairy
cattle, indicate that tolerances might be needed for the combined
residues of fluazinam and its metabolites DAPA and AMPA in fat and meat
byproducts of cattle and other ruminants at 0.05 ppm.  The feeding study
also indicates that tolerances probably will not be needed for milk and
meat of cattle and ruminants, as the expected combined residues in these
matrices are below the combined method LOQ of 0.03 ppm for the regulated
compounds.  According to the July 18, 2007 Minutes of the Chemistry
Science Advisory Council Meeting, the ChemSAC recommended that the
guidance document be revised to include language detailing the use of
the highest residue data for any tissue (liver, kidney, fat, skin or
muscle) to determine the tolerance for meat byproducts.  A single
tolerance on “meat byproducts” would be recommended based on the
highest residue value, and individual tolerances will no longer be set
on liver, kidney, or meat byproducts (except liver and kidney).

Based on the transfer coefficients for livestock tissues and the
relatively low dietary burden for swine of 0.003 ppm for fluazinam,
tolerances for hogs are not needed.  Tolerances are also not needed for
poultry eggs and tissues at this time because of the low dietary burden
for poultry. 

Tolerances for rotational crops are not required based on the review of
a confined rotational crop study which showed the absence of metabolites
containing the intact fluazinam nucleus.  The existing 30-day PBI for
all rotated crops not on the label is appropriate, and precludes the
need for additional rotational crop data.

There are currently no established Codex, Canadian, or Mexican maximum
residue limits (MRLs) for fluazinam on the apple, carrot, lettuce,
onions or berry crops.  Therefore, there are no harmonization issues for
the requested tolerances.  An International Residue Limit Status Sheet
is attached in Appendix I.

Table 14.  Tolerance Summary for Fluazinam.

Commodity	Proposed  Tolerance (ppm)	Recommended Tolerance (ppm)
Comments; Correct Commodity Definition

PP#8E7506

Bushberry, subgroup 13-07B	4.5	7.0

	Adequate blueberry residue data are available.  The recommended
tolerance for Subgroup 13-07B is the same as that for Subgroup 13B
(i.e., 7.0 ppm)

The established individual tolerances for aronia berry, buffalo currant,
Chilean guava, European barberry, highbush cranberry, edible
honeysuckle, edible jostaberry, Juneberry, lingonberry, native currant,
salal, and sea buckthorn should be deleted because they are included
under the revised Bushberry subgroup, 13-07B.

Onion, bulb, subgroup 3-07A	0.15	0.20	Adequate bulb onion residue data
are available.

Lettuce, head	0.02	0.02	Adequate head lettuce residue data are
available.

Lettuce, leaf	2.0	2.0	Adequate leaf lettuce residue data are available.

PP#9E7570

Carrot, roots	0.8	None	Adequate carrot residue data are available. 
HED’s tolerance generator directs that the tolerance for this data set
should be 0.70 ppm.  

PP#9F7571

Apples	1.7	None	Adequate apple residue data are available.  HED’s
tolerance generator directs that the tolerance for this data set should
be 1.50 ppm.  The USEPA and Canada’s PMRA agreed to establish the
tolerance at 2.0 ppm at such time as the data deficiencies are resolved.

Apple, pomace, wet	5.0	None	Adequate apple processing data are not
available.

Cattle, fat	0.03	None	An adequate cattle feeding study is not available.
 Based on the results of the inadequate study, the maximum expected
residues in meat byproducts and fat is 0.05 ppm.  This value can be used
as a very conservative residue value for the dietary exposure
assessment.

Based on the results of the inadequate feeding study, tolerances will
probably not be needed for milk and meat of cattle, goat, horse, and
sheep.

Cattle, kidney	0.03	Not required

	Cattle, liver	0.03	Not required

	Cattle, meat	0.03	Not required

	Cattle, meat byproducts	0.03	None

	Goat, fat	0.03	None

	Goat, kidney	0.03	Not required

	Goat, liver	0.03	Not required

	Goat, meat	0.03	Not required

	Goat, meat byproducts	0.03	None

	Horse, fat	0.03	None

	Horse, kidney	0.03	Not required

	Horse, liver	0.03	Not required

	Horse, meat	0.03	Not required

	Horse, meat byproducts	0.03	None

	Milk	0.03	Not required

	Sheep, fat	0.03	None

	Sheep, kidney	0.03	Not required

	Sheep, liver	0.03	Not required

	Sheep, meat	0.03	Not required

	Sheep, meat byproducts	0.03	None

	

References

D212612, 216941, and 217467, PP# 2G04099.  Temporary Tolerance Petition
and Experimental Use Permit for Use of Fluazinam on Peanuts;
050534-EUP-E. Submission Dated 1/23/95 in Response to the Memo of G.J.
Herndon Dated 6/19/92, G. Herndon, 9/5/95

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

D257115, PP# 9F5079.  Request for the Use of Fluazinam on Peanuts,
Potatoes, and Wine Grapes.  Evaluation of Analytical Chemistry and
Residue Data, W. Cutchin, 5/21/01

D335640, Fluazinam.  Tolerance Petitions Requesting the Establishment of
Permanent Tolerances (Associated with Section 3 Registration) for Food
Use of the Fungicide on Edible-Podded Beans (Subgroup 6-A, Except Peas),
Shelled Succulent Beans (Subgroup 6-B, Except Peas), Shelled Dried Beans
(Subgroup 6-C, Except Peas), Brassica (Cole) Vegetables (Group 5),
Bushberries (Subgroup 13-B), and Ginseng.  Summary of Analytical
Chemistry and Residue Data, W. Drew, 8/22/07

D342005, Fluazinam.  Request for Label Amendments Adding Aerial
Application of the Fungicide on Potatoes, and Reducing the Plant-Back
Interval (PBI) for All Non-Label Crops from 70 to 30 Days Following Use
of the Fungicide, W. Drew, 2/1/08

  SEQ CHAPTER \h \r 1 Attachments:  

Appendix I - International Residue Limit Status sheet

Appendix II - Tolerance Assessment Calculations

Appendix I	

INTERNATIONAL RESIDUE LIMIT STATUS

Chemical Name: 
3-chloro-N-[3-chloro-2,6-dinitro-4-(trifluoromethyl)phenyl]-5-(trifluoro
methyl)-2-pyridinamine	Common Name:  Fluazinam	X Proposed tolerances

⁯ Reevaluated tolerance

⁯ Other	Date: 8/31/09

Codex Status (Maximum Residue Limits)	U. S. Tolerances

√ No Codex proposal step 6 or above

⁯No Codex proposal step 6 or above for the crops requested	Petition
Numbers:  PP#s 8E7506, 9E7570, and 9F7571

DP #s: 360840 and 366506

Other Identifier:  

Residue definition (step 8/CXL): N/A	Reviewer/Branch:  C. Swartz, RAB2

	Established residue definition:  parent fluazinam for all plant
commodities, except wine grapes; parent fluazinam and AMGT for grapes

Proposed residue definition in PP#s 8E7506 and 9E7570 for plant
commodities:  fluazinam

Proposed residue definition in PP#9F7571 for apple commodities: 
fluazinam and its metabolite AMGT and for livestock commodities: 
fluazinam and its metabolites DAPA and AMPA

Crop (s)	MRL (mg/kg)	Crop(s) 	Proposed

⁯No Limits

√ No Limits for the crops requested	⁯ No Limits

√  No Limits for the crops requested

Residue definition: 
3-chloro-N-[3-chloro-2,6-dinitro-4-(trifluoromethyl)phenyl]-5-(trifluoro
methyl)-2-pyridinamine	Residue definition:  Fluazinam

Crop(s)	MRL (mg/kg)	Crop(s)	MRL (mg/kg)

Potatoes	0.02	Beans	0.1

Potato	0.05

Notes/Special Instructions: S. Funk, 09/02/2009

Appendix II

Tolerance Assessment Calculations

For each of the crops listed below, the Guidance for Setting Pesticide
Tolerances Based on Field Trial Data (SOP), along with the tolerance
spreadsheet (January 2008 version), was used for calculating recommended
tolerances.  As specified in the SOP, the minimum of the 95% upper
confidence limit (UCL) on the 95th percentile and the point estimate of
the 99th percentile was selected as the tolerance value in cases when
the dataset was large (greater than 15 samples) and reasonably
lognormal.  For datasets that were small (≤15 samples) and reasonably
lognormal, the upper bound estimate of the 95th percentile based on the
median residue value was compared to the minimum of the 95% UCL on the
95th percentile and the point estimate of the 99th percentile, and the
minimum value was selected as the tolerance value.  For datasets that
were not lognormal, the upper bound on the 89th percentile was selected
as the tolerance value (distribution-free method).  The rounding
procedures specified in the SOP were also used.

Apple

The dataset used to establish a tolerance for fluazinam on apple
consisted of field trial data representing application rates of
3.97-5.40 lb ai/A (9-12 applications at 0.43-0.71 lb ai/A/application)
with PHIs of 28-32 days.  As specified in the SOP, the trial application
rates and PHIs are within 25% of the maximum label application rate and
minimum label PHI, respectively.  The residue values used to calculate
the tolerance for apple are provided in Table II-1.

All but 4 of the 42 field trial sample results for fluazinam in/on apple
were above the LLMV (LLMV = 0.01 ppm).  The apple dataset was large (42
samples).  The result from the approximate Shapiro-Francia test
statistic (Figure II-2) indicated that the assumption of lognormality
should be rejected.  Visual inspection of the lognormal probability plot
(Figure II-1) provided in the spreadsheet confirmed that the dataset is
not lognormal.  Using the tolerance spreadsheet, the recommended
tolerance is 1.5 ppm for apple.



Table II-1.	Fluazinam Residues in/on Apples Following Nine to Twelve
Foliar Applications of Fluazinam (FlC) at Rates Totaling ~4.5 lb
ai/A/season.

Regulator:	EPA	EPA

Chemical:	Fluazinam	Fluazinam

Crop:	Apple	Apple

PHI:	28-32 Days	28-29 Days

Total App. Rate:	3.97-5.40 lb ai/A	4.44-4.77 lb ai/A

Submitter:	ISK Biosciences Corporation	ISK Biosciences Corporation

MRID Citation:	MRIDs 47756607, 47756608, 47756610, and 47756611	MRID
47756609

	Residues of Fluazinam (ppm) 1

	0.050	0.160

	0.030	0.160

	0.050	0.160

	0.03	0.160

	0.05	0.030

	0.060	0.020

	0.030	0.090

	0.040	0.150

	0.010	0.150

	0.010	0.120

	0.010	0.150

	0.010	0.120

	0.020	1.680

	0.040	1.390

	0.020	0.050

	0.040	0.020

	0.080	0.170

	0.080	0.090

	0.020	1.310

	0.030	1.490

0.150

0.130

0.050

0.300

0.120

0.170

1	Residue values <LLMV (0.01 ppm) are listed in bold.

Figure II-1.  Lognormal probability plot of fluazinam field trial data
for apple.

 

 Figure II-2.  Tolerance spreadsheet summary of fluazinam field trial
data for apple.

Bulb Onions (Subgroup 3-07A)

The dataset used to establish a tolerance for fluazinam on the bulb
onion subgroup consisted of field trial data for bulb onions
representing application rates of 3.11-3.17 lb ai/A (6 applications at
~0.52 lb ai/A/application) with PHIs of 6-8 days.  As specified by the
SOP, the trial application rates and PHIs are within 25% of the maximum
label application rate and minimum label PHI, respectively.  The residue
values used to calculate the tolerance for bulb onion are provided in
Table II-2.

As 7 of the 18 field trial sample results for bulb onion were <LOQ,
maximum likelihood estimation (MLE) procedures were used to impute the
censored values for onions.  The censored dataset for onions, which
consisted of 18 samples, was then entered into the tolerance
spreadsheet.  Visual inspection of the lognormal probability plot
(Figure II-3) and the results from the approximate Shapiro-Francia test
statistic (Figure II-4) indicate that the dataset is reasonably
lognormal.  

The recommended tolerance is 0.20 ppm for bulb onion, which is the
representative crop for the bulb onion subgroup 3-07A.  

Table II-2.	Fluazinam Residues in/on Onions Following Six Foliar
Applications of Fluazinam (FlC) at Rates Totaling ~3.12 lb ai/A/season.

Regulator:	EPA

Chemical:	Fluazinam

Crop:	Bulb Onion (MLE)

PHI:	6-8 days 

App. Rate:	3.11-3.17 lb ai/A

Submitter:	IR-4

MRID Citation:	MRID 47631001

	Residues (ppm)1

	0.002	0.015

	0.003	0.016

	0.004	0.021

	0.005	0.031

	0.006	0.033

	0.007	0.035

	0.009	0.045

	0.013	0.095

	0.013	0.101

1  Residue values <LLMV (0.01 ppm) are listed in bold.

Figure II-3.  Lognormal probability plot of fluazinam field trial data
for bulb onion.

Figure II-4.  Tolerance spreadsheet summary of fluazinam field trial
data for bulb onion.

Bushberry subgroup (13-07B)

Based on previously reviewed blueberry data (PP# 6E7137; D335640, W.
Drew, 8/22/07), a 7.0 ppm tolerance was established on the Bushberry
subgroup 13B and several related bushberry crops.  To assess IR-4’s
request to lower the bushberry tolerance, HED has reexamined the
original blueberry residue data supporting the tolerance (Table II-3). 
Only the fluazinam residue data were included, as fluazinam per se is
the residue of concern for tolerance assessment.  

The data were entered into the tolerance spreadsheet.  The results from
the approximate Shapiro-Francia test statistic (Figure II-6) indicated
that the assumption of lognormality should not be rejected.  Therefore,
the recommended tolerance for the Bushberry subgroup 13-07B is 7.0 ppm.

Table II-3.	Fluazinam Residues in/on Blueberries Following Six Foliar
Applications of Fluazinam (FlC) at Rates Totaling ~3.12 lb ai/A/season.

Regulator:	EPA

Chemical:	Fluazinam

Crop:	Blueberry

PHI:	23-32 Days

App. Rate:	3.8-4.1 lb ai/A

Submitter:	IR-4

MRID Citation:	MRID 46986701

	Residues (ppm)1

	0.064	0.64

	0.070	0.68

	0.074	0.70

	0.12	1.0

	0.13	1.2

	0.16	1.2

	0.17	1.5

	0.42	1.6

	0.45	1.6

	0.49	1.7

	0.50	2.0

	0.55

	

Figure II-5.  Lognormal probability plot of fluazinam field trial data
for blueberry.

Figure II-6.  Tolerance spreadsheet summary of fluazinam field trial
data for blueberry.

 

Carrot

The dataset used to establish a tolerance for fluazinam on carrot
consisted of field trial data representing application rates of
2.04-3.03 lb ai/A (4-5 applications at 0.492-0.786 lb ai/A/application)
with PHIs of 6-8 days.  Each field trial consisted of two treatment
plots (Treatments 2 and 3).  Both plots received treatments in similar
rates and volumes; however, Treatment 2 was irrigated within 2 hours of
each application, and Treatment 3 received irrigation after 24 hours of
each application.  As specified by the SOP, the trial application rates
and PHIs are within 25% of the maximum label application rate and
minimum label PHI, respectively.  The residue values used to calculate
the tolerance for carrot are provided in Table II-4.

All but 4 of the 52 field trial sample results for fluazinam in/on
carrot were above the LLMV (LLMV = 0.02 ppm).  The carrot dataset was
large (52 samples).  Visual inspection of the lognormal probability plot
(Figure II-7) and the results from the approximate Shapiro-Francia test
statistic (Figure II-8) indicated that the carrot dataset was reasonably
lognormal.  Using the tolerance spreadsheet, the recommended tolerance
is 0.7 ppm for carrot.

瑹㶴<

瑹㶴<

瑹㶴<

 

0

I

J

Q

S

T

[

]

_

x

y

~

Ý

Þ

\

]

^

_

y

 

¼

õ

ø

;

>

瑹㶴<ᄀÞ

ß

ó

ô

õ

ø



#

%

9

:

>

->

?

S

T

 

¡

Ã

Ø

Ý

 h

2>

hÞ

hÞ

hÞ

hÞ

hÞ

hÞ

hÞ

hÞ

 h

h±

h†

h

	ü

	ü

	ü

	ü

	ü

hÄ

 h†

h†

 h†

²

²

²

²

²

²

²

h

&

&

&

&

&

&

&

&

&

&

&

  h

h

h

h

h

 h

h

  h

%

  h

hU

hU

$

$

hÇ

hÇ

hÇ

h±

 

`„6ÿgd±

'

@

C

t

µ

Â

Ô

 ha

hU

hU

态ú摧墔¤

yt

yt

yt

yt

yt

yt

yt

hU

hU

hU

yt

yt

yt

yt

yt

yt

yt

h

hU

hÇ

hU

hU

hU

hU

hU

hU

hU

$

$

h

hU

hU

hU

hÇ

hU

hU

hU

gdU

h

h

 h±

h±

h±

h

 hªl

h

H*

b	@

¶

b	@

¶

H*

b	@

¶

b	@

¶

b	@

¶

 h~

h~

h~

 h

 h

h

 h

 h

$

$

$

 h 

kd

 h 

 h 

$

$

 h 

 h¶

㄀Ĥ摧㜓C

 h¶

 h€

 h€

$

 h 

Carrot (Treatment 3; irrigated after 24 hours of each application)

PHI:	6-8 Days	6-8 Days

Total App. Rate:	2.04-2.94 lb ai/A	2.04-3.03 lb ai/A

Submitter:	IR-4	IR-4

MRID Citation:	MRID 47756601	MRID 47756601

	Residues of Fluazinam (ppm) 1

	0.040	0.050

	0.060	0.060

	0.340	0.240

	0.390	0.280

	0.020	0.020

	0.020	0.020

	0.120	0.030

	0.120	0.040

	0.120	0.100

	0.140	0.110

	0.050	0.170

	0.060	0.190

	0.110	0.410

	0.120	0.450

	0.070	0.050

	0.190	0.070

	0.460	0.160

	0.560	0.250

	0.100	0.090

	0.100	0.090

	0.090	0.080

	0.110	0.090

	0.220	0.020

	0.230	0.020

	0.080	0.040

	0.090	0.050

1	Residue values <LLMV (0.02 ppm) are listed in bold.

Figure II-7.  Lognormal probability plot of fluazinam field trial data
for carrot.

Figure II-8.  Tolerance spreadsheet summary of fluazinam field trial
data for carrot.

Head Lettuce

The dataset used to assess a tolerance for fluazinam residues on head
lettuce consisted of field trial data representing application rates of
0.952-1.02 lb ai/A with PHIs of 46-52 days.  As specified by the SOP,
the field trial application rates and PHIs are within 25% of the maximum
label application rate and minimum label PHI, respectively.  Because all
but one of the residue values for head lettuce were <LOQ (<0.01 ppm),
the tolerance spreadsheet could not be used to calculate a possible
tolerance (Table II-5).  Therefore, the maximum residue value (0.021
ppm) was used to determine the tolerance.  

Table II-5.	Fluazinam Residues in/on Head Lettuce Following a Broadcast
Foliar Application of Fluazinam (FlC) at a Rate of ~0.99 lb ai/A/season.

Regulator:	EPA

Chemical:	Fluazinam

Crop:	Head Lettuce

PHI:	46-52 days 

App. Rate:	 0.952-1.02 lb ai/A

Submitter:	IR-4

MRID Citation:	MRID 47631002

	Residues (ppm)1

	0.01	0.01

	0.01	0.01

	0.01	0.01

	0.01	0.01

	0.01	0.01

	0.01	0.01

	0.01	0.021

1  Residue values <LLMV (0.01 ppm) are listed in bold.

Leaf Lettuce

The dataset used to assess a tolerance for fluazinam residues on leaf
lettuce consisted of field trial data representing application rates of
0.974-1.05 lb ai/A with PHIs of 20-32 days.  As specified in the SOP,
the field trial application rates and PHIs are within 25% of the maximum
label application rate and minimum label PHI, respectively.  The residue
values used to calculate the tolerance are provided in Table II-6.  

The leaf lettuce dataset, which is relatively small (14 samples), was
entered into the tolerance spreadsheet.  Field trial sample results were
above the LLMV (0.01 ppm) for 12 of 14 samples.  Visual inspection of
the lognormal probability plot (Figure II-9) and the result from the
approximate Shapiro-Francia test statistic (Figure II-10) indicated that
the assumption of lognormality should be rejected.  

Using the tolerance spreadsheet, the recommended value is 2.0 ppm for
leaf lettuce.  

Table II-6.	Fluazinam Residues in/on Leaf Lettuce Following a Broadcast
Foliar Application of Fluazinam (FlC) at a Rate of ~0.99 lb ai/A/season.

Regulator:	EPA

Chemical:	Fluazinam

Crop:	Leaf Lettuce

PHI:	20-32 days

App. Rate:	0.974-1.05 lb ai/A

Submitter:	IR-4

MRID Citation:	MRID 47631002

	Residues (ppm)1

	0.01	0.021

	0.01	0.022

	0.011	0.038

	0.017	0.150

	0.019	0.159

	0.020	1.45

	0.020	1.69

1  Residue values <LLMV (0.01 ppm) are listed in bold.

Figure II-9.  Lognormal probability plot of fluazinam field trial data
for leaf lettuce.

Figure II-10.  Tolerance spreadsheet summary of fluazinam field trial
data for leaf lettuce.

Fluazinam	Summary of Analytical Chemistry and Residue Data	DP#:  360840

Page   PAGE  1  of   NUMPAGES  54