UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
                               WASHINGTON, D.C.  20460

	OFFICE OF CHEMICAL SAFETY
	AND POLLUTION PREVENTION

MEMORANDUM

Date:  		September 23, 2011

SUBJECT:	Fluopyram.  Application for Section 3 Registration for Use on Apple, Banana (Import only), Dried Beans, Cherry, Grape (Wine Production only), Peanut, Pistachio, Potato, Sugar Beet, Strawberry, Tree Nuts Crop Group 14, Watermelon, and Rotational Crops Alfalfa, Canola, Cotton, Cereal Grains Crop (Except Rice) Group 15, and Forage, Fodder, and Straw of Cereal Grains Crop (Except Rice) Group 16.  Summary of Analytical Chemistry and Residue Data.

PC Code:  080302
DP Barcodes:  D387587, 389081
Decision No.:  402628
Registration No.:  264-RNTI
Petition No.:  8F7463
Regulatory Action:  Section 3 Registration
Risk Assessment Type:  NA
Case No.:  NA
TXR No.:  NA
CAS No.:  658066-35-4
MRID No.:  See MRID Summary Table
40 CFR:  To be determined
		              									Ver.Apr.08

FROM:	Leung Cheng, Chemist
		Risk Assessment Branch III
		Health Effects Division (7509P)

THROUGH:	Paula Deschamp, Chief
		Risk Assessment Branch III
		Health Effects Division (7509P)

TO:		Cynthia Giles-Parker, Chief
		Fungicide Branch
		Registration Division (7505P)

MRID Summary Table
MRID No.
Study Type
Monograph Section; Comments

1[st] Entry Monograph
2[nd] Entry Monograph
47372524
860.1300 Grape
B.7.1.1

47372525
860.1300 Grape
B.7.1.1

47372526
860.1300 Bean
B.7.1.3

47372527
860.1300 Bean
B.7.1.3

47372528
860.1300 Potato
B.7.1.2

47372529
860.1300 Potato
B.7.1.2

47372530
860.1300 Red pepper
B.7.1.4

47372531
860.1300 Red pepper
B.7.1.4

47372532
860.1300 Laying hen
B.7.2.1

47372533
860.1300 Laying hen
B.7.2.1

47372534
860.1300 Lactating goat
B.7.2.2

47372535
860.1300 Lactating goat
B.7.2.2

47372536
860.1300 Cell cultures
B.7.1.5

47372537
860.1340 Crop commodities
B.5.2.1

47372538
860.1340 Crop commodities
B.5.2.1

47372539
860.1340 Extraction efficiency
B.5.2.1

47372540
860.1340 Livestock commodities
B.5.2.2

47372541
860.1340 Livestock commodities
B.5.2.2

47372542
860.1340 Livestock commodities
B.5.2.2

47372543
860.1340 Crop commodities
B.5.2.1; reviewed by Germany

47372544
860.1340 Crop commodities
B.5.2.1; reviewed by Germany

47372545
860.1340 Crop commodities
B.5.2.1; reviewed by Germany

47372546
860.1340 Crop commodities
B.5.2.1

47372547
860.1340 Waiver request (radiovalidation)
B.5.2.1

47372548
860.1360 Multiresidue Methods
B.5.2

47372549
860.1380 Crop commodities
B.7.6.2

47372550
860.1380 Orange
B.7.6.2

47372601
860.1480 Cattle
B.7.8.1

47372602
860.1480 Hen
B.7.8.2

47372603
860.1500 Fruiting vegetable
B.5.2.1, B.7.6.1
B.7.6.1
47372604
860.1500 Grape
B.7.6.1

47372605
860.1500 Strawberry
B.7.6.1, B.7.7.2

47372606
860.1500 Grasses

B.7.6.1
47372608
860.1500 Tomato (Europe)
B.7.6.1

47372610
860.1500 Tomato (Europe)

47372615
860.1500 Strawberry (Europe)
B.7.6.1

47372622
860.1520 Grape
B.7.7.2

47372623
860.1520 Tomato
B.7.7.2

47372624
860.1520 Strawberry (Europe)
B.7.7.2

47372625
860.1520 Strawberry (Europe)

47372626
860.1520 Raisin (Europe)
B.7.7.2

47372627
860.1520 Wine (Europe)
B.7.7.2

47372628
860.1520 Wine (Europe)

47372629
860.1520 Tomato (Europe)
B.7.7.2

47372630
860.1520 Tomato (Europe)

47372631
860.1850 Confined rotational crops
B.7.9.1

47372632
860.1850 Confined rotational crops
B.7.9.1

47372633
860.1900 Field rotational crops (Europe)
B.7.9.2

None assigned; Document No. M-296625-01
860.1900 Field rotational crops (Europe)

None assigned; Document No. M-296652-01
860.1900 Field rotational crops (Europe)

None assigned; Document No. M-296671-01
860.1900 Field rotational crops (Europe)

47567010
860.1500 Sugar beet and turnip

B.7.6.1
47567011
860.1500 Root vegetables except sugar beet

B.7.6.1
47567012
860.1500 Potato

B.7.6.1
47567013
860.1500 Canola

B.7.6.1
47567014
860.1500 Field and sweet corn

B.7.6.1
47567015
860.1500 Wheat and sorghum

B.7.6.1
47567016
860.1500 Soybean

B.7.6.1
47567017
860.1500 Hops

B.7.6.1
47567018
860.1500 Sunflower

B.7.6.1
47567019
860.1500 Peanut

B.7.6.1
47567020
860.1500 Tree nuts

B.7.6.1
47567021
860.1500 Dry bulb onions

B.7.6.1
47567022
860.1500 Green onions

B.7.6.1
47567023
860.1500 Leafy vegetables

B.7.6.1, B.7.7.2
47567024
860.1500 Artichoke (globe)

B.7.6.1
47567025
860.1500 Head and stem Brassica

B.7.6.1, B.7.7.2
47567026
860.1500 Leafy Brassica greens

B.7.6.1, B.7.7.2
47567027
860.1500 Edible-podded legume vegetables

B.7.6.1
47567028
860.1500 Succulent shelled pea and bean

B.7.6.1
47567029
860.1500 Dried shelled pea and bean

B.7.6.1
47567030
860.1500 Cucurbit vegetables

B.7.6.1
47567031
860.1500 Citrus fruit

B.7.6.1
47567032
860.1500 Pome fruit

B.7.6.1
47567033
860.1500 Stone fruit

B.7.6.1
47567034
860.1500 Caneberry

B.7.6.1
47567035
860.1500 Bushberry

B.7.6.1
47567036
860.1500 Banana

B.7.6.1
47567037
860.1500 Herbs

B.7.6.1
47567101
860.1500 Dill seed

B.7.6.1
47567111
860.1520 Apple

B.7.7.2
47567112
860.1520 Orange

B.7.7.2
47567113
860.1520 Plum

B.7.7.2
47567114
860.1520 Peanut

B.7.7.2
47567115
860.1500/1520 Wheat

B.7.7.2
47567116
860.1520 Canola

B.7.7.2
47567117
860.1500/1520 Field corn

B.7.7.2
47567118
860.1520 Potato

B.7.7.2
47567119
860.1500/1520 Soybean

B.7.7.2
47567120
860.1520 Sugar beet

B.7.7.2
47567121
860.1520 Sunflower

B.7.7.2
47567122
860.1520 Cotton

B.7.7.2
47567123
860.1900 Limited field rotational crop study

B.7.9.2
47567124
860.1900 Rotated alfalfa

B.7.9.2
47567125
860.1900 Rotated cotton

B.7.9.2

This document was originally prepared under contract by Dynamac Corporation (1901 Research Boulevard, Suite 220; Rockville, MD 20850).

The document has been reviewed by the Health Effects Division (HED) and revised to reflect current Office of Pesticide Programs (OPP) policies.

Executive Summary

Fluopyram (AE C656948) is a new fungicide with a broad spectrum of activity against numerous fungal diseases, including Ascomycetes, leaf spots, molds, scab, Alternaria, Septoria, Monlinia, Botrytis, Sclerotinia, and powdery mildews, in a wide range of crops and plants.  Fluopyram represents a new class of chemistry (pyridylethylamide), and its biochemical mode of action has been shown to rely on the inhibition of the enzyme succinate dehydrogenase (complex II) within the fungal mitochondrial respiratory chain, thus blocking electron transport.  Fluopyram is a Group 7 fungicide.

In 2008, Bayer CropScience submitted a petition proposing the establishment of tolerances for residues of fluopyram (N-[2-[3-chloro-5-(trifluoromethyl)-2-pyridinyl]ethyl]-2-(trifluoromethyl)benzamide) in/on the following crop commodities:
      Alfalfa, forage 	0.25 ppm
      Alfalfa, hay 	0.80 ppm
      Almond, hulls 	8.0 ppm
      Apple, wet pomace 	2.5 ppm
      Artichoke 	2.0 ppm
      Banana 	1.0 ppm
      Beet, sugar, roots 	0.10 ppm
      Berry, lowgrowing, subgroup 13-07G 	2.0 ppm
      Brassica, head and stem, subgroup 5A 	3.0 ppm
      Brassica, leafy greens, subgroup 5B 	35 ppm
      Bushberries, subgroup 13-07B 	10 ppm
      Caneberries, subgroup 13-07A 	5.0 ppm
      Citrus, oil 	10 ppm
      Corn, sweet, kernel plus cob with husk removed 	0.10 ppm
      Cotton, gin byproducts 	0.05 ppm
      Cotton, undelinted seed 	0.10 ppm
      Fruit, citrus, group 10 	1.0 ppm
      Fruit, pome, group 11 	1.0 ppm
      Fruit, small, vine, climbing, except fuzzy kiwifruit, subgroup 13-07F 	2.0 ppm
      Fruit, stone, group 12 	2.0 ppm
      Grain, cereal, forage, fodder and straw, group 16, except rice; forage 	8.0 ppm
      Grain, cereal, forage, fodder and straw, group 16, except rice; hay, straw and stover 	14 ppm
      Grain, cereal, forage, fodder and straw, group 16, except rice; aspirated fractions		50 ppm
      Grain, cereal, group 15, except rice and sweet corn 	3.0 ppm
      Grape, raisin 	3.5 ppm
      Grass, forage, fodder and hay, group 17; forage 	80 ppm
      Grass, forage, fodder and hay, group 17; hay 	30 ppm
      Herbs, subgroup 19A, fresh 	50 ppm
      Herbs, subgroup 19A, dried 	260 ppm
      Hop, dried cones 	100 ppm
      Nut, tree, group 14 (including pistachio) 	0.05 ppm
      Okra 	8.0 ppm
      Oilseed, group 20, except cotton 	5.0 ppm
      Onion, bulb, subgroup 3-07A 	0.30 ppm
      Onion, green, subgroup 3-07B 	20 ppm
      Peanut 	0.05 ppm
      Peanut, hay 	50 ppm
      Pepper, non-bell 	8.0 ppm
      Potato, processed potato waste 	0.15 ppm
      Soybean, aspirated fractions 	70 ppm
      Soybean, forage 	8.0 ppm
      Soybean, hay 	30 ppm
      Soybean, hulls 	0.40 ppm
      Soybean, seed 	0.30 ppm
      Spices, except black pepper, subgroup 19B 	100 ppm
      Vegetable, cucurbit, group 9 	1.0 ppm
      Vegetable, foliage of legume, except soybean, subgroup 7A; forage 	30 ppm
      Vegetable, foliage of legume, except soybean, subgroup 7A; hay 	75 ppm
      Vegetable, foliage of legume, except soybean, subgroup 7A; vines 	16 ppm
      Vegetable, fruiting, except non-bell pepper, group 8 	1.0 ppm
      Vegetable, leafy, except Brassica, group 4 	35 ppm
      Vegetable, leaves of root and tuber, group 2 	30 ppm
      Vegetable, legume, edible podded, subgroup 6A 	2.0 ppm
      Vegetable, legume, succulent shelled, subgroup 6B 	0.20 ppm
      Vegetable, pea and bean, dried shelled (except soybean), subgroup 6C 	0.50 ppm
      Vegetable, root and tuber, except sugarbeet, subgroup 1B 	0.50 ppm
      Vegetable, tuberous and corm, subgroup 1C 	0.05 ppm

In addition, Bayer has proposed tolerances for residues of fluopyram and its metabolite 2-(trifluoromethyl)benzamide, expressed in parent equivalents, in the following livestock commodities:
      Cattle, fat 	0.10 ppm
      Cattle, meat 	0.10 ppm
      Cattle, meat byproducts, except liver 	0.10 ppm
      Cattle, liver 	1.2 ppm
      Eggs 	0.10 ppm
      Goat, fat 	0.10 ppm
      Goat, meat 	0.10 ppm
      Goat, meat byproducts, except liver 	0.10 ppm
      Goat, liver 	1.2 ppm
      Hog, fat 	0.01 ppm
      Hog, meat 	0.01 ppm
      Hog, meat byproducts, except liver 	0.01 ppm
      Hog, liver 	0.15 ppm
      Horse, fat 	0.10 ppm
      Horse, meat 	0.10 ppm
      Horse, meat byproducts, except liver 	0.10 ppm
      Horse, liver 	1.2 ppm
      Milk 	1.2 ppm
      Poultry, fat 	0.05 ppm
      Poultry, meat 	0.03 ppm
      Poultry, meat byproducts 	0.20 ppm
      Sheep, fat 	0.10 ppm
      Sheep, meat 	0.10 ppm
      Sheep, meat byproducts, except liver 	0.10 ppm
      Sheep, liver 	1.2 ppm

However, due to a subsequent cancer classification of fluopyram by HED CARC, the registrant submitted a letter (Subject: Withdrawal of certain pending new crops for the active ingredient Fluopyram, 8-March-2011) withdrawing tolerance proposals and registration requests for the following crops:

"CG 1B Root veg and 1C Tuberous and corm veg (except potatoes and sugarbeet)
CG2 Leaves of root and tuberous veg
CG3-07A&B Bulb veg
CG4 Leafy veg
CG5 Brassica
CG6A Edible legumes
CG6B Succulent beans and peas
CG6C (part) Dried peas (and some dried beans)
CG7 Foliage of legume veg
CG8 Fruiting veg
CG10 Citrus
CG11 Pome fruit (except apple)
CG13-07A&B Caneberries and Bushberries
CG13-07F Vine fruit (except wine grapes)
CG13-07G Low growing berries (except strawberry)
CG15 Cereal Grains (except for rotational purposes)
CG16 Forage Cereals (except for rotational purposes)
CG17 Grasses grown for forage or seed
CG18 Non grass animal feeds
CG19 Herbs and Spices
CG20 Oilseeds (except canola)
Hops, Globe artichoke, Christmas Trees, Turf and Ornamentals"

In addition, the registant submitted a revised label specifying uses only on the following crops: apple, cucurbit vegetables, dried beans (selected), peanut, potato, stone fruit, sugar beet, strawberry, tree nuts and wine grapes.  The revised label reduces the initially proposed seasonal maximum use rates on dried beans, potato, stone fruit, and sugar beet, restricts the use on grapes to wine grapes only, and increases the pre-harvest interval (PHI) for apples to 7-day from the initially proposed PHI of 0 day.  The revised label continues to allow crop rotation to alfalfa and cotton, but restricts crop rotation to canola, cereal grains (except rice), and soybean only after application of fluopyram to dried beans and potato.  A revised Section F has not been submitted.

In conjunction with this petition, Bayer is requesting registration of Fluopyram 500 SC Fungicide, a 500 g/L (4.16 lb/gal) suspension concentrate (SC; equivalent to a flowable concentrate) formulation [EPA File Symbol No. 264-RNTI].

The registrant withdrew additional pending uses in its letter of 8 August 2011 which states that

"The crops being withdrawn from these actions are as follows;
CG9 Cucurbits (except watermelon)
CG12 Stone fruit (except cherry)

The following crops remain pending for registration
Potatoes
Sugarbeet
CG6C - Dried beans (select dried beans  -  see list below*)
Watermelon
Apple
Cherry
Grapes (grown for wine production only)
Strawberry
CG14 Tree nuts (including Pistachio)
Peanuts
CG15 Cereal Grains - for rotational purposes only
CG16 Forage of cereal grains - for rotational purposes only
Canola - for rotational purposes only
Soybean (&forage) - for rotational purposes only
Alfalfa - for rotational purposes only
Cotton - for rotational purposes only
Banana - for import tolerance only

*Dried beans include Dried Shelled Bean, Bean (Lupinus spp., includes grain lupin,
sweet lupin, white lupin, and white sweet lupin), Bean (Phaseolus spp., includes field
bean, kidney bean, lima bean (dry), navy bean, pinto bean, tepary bean), Bean
(Vigna spp., includes adzuki bean, blackeyed pea, catjang, Crowder pea, moth
bean, mung bean, rice bean, Southern pea, Urd bean,) Other Beans [Broad bean
(dry), chickpea, Guar, Lablab bean, Lentil.]"

The nature of the residue in plants is adequately understood based on metabolism studies with grape, potato, bean, and red pepper.  The nature of the residue in rotational crops is adequately understood based on confined rotational crop studies with rotated Swiss chard, turnip, and wheat.  The metabolic pathway of fluopyram is similar in primary and rotational crops, and mainly involves hydroxylation of parent compound leading to AE C656948-7-hydroxy and AE C656948-8-hydroxy, conjugation of the hydroxylated parent compound, mainly with sugars, and cleavage of the molecule leading to AE C656948-benzamide, AE C656948-pyridyl-acetic acid (AE C656948-PAA), and AE C656948-pyridyl-carboxylic acid (AE C656948-PCA).  HED has determined that the residue of concern in plant commodities (primary and rotational crop) for tolerance enforcement is fluopyram, and for risk assessment is fluopyram in all crops except in legumes and oilseed crops where AE C656948-benzamide is also included (finite levels of the benzamide were only found in beans, peas, rapeseed and rapeseed forage in the EU field trials).

The nature of the residue in livestock is adequately understood based on metabolism studies with goats and hens.  Metabolism of fluopyram in livestock mainly involves hydroxylation of the parent compound to AE C656948-7-hydroxy and AE C656948-8-hydroxy, elimination of water from compounds hydroxylated in the ethylene bridge leading to AE C656948-Z/E-olefines, cleavage of the molecule leading to AE C656948-benzamide and AE C656948-PAA, and conjugation of the hydroxylated parent compound, mainly with glucuronic acid.  For tolerance enforcement, HED has determined that the residues of concern are fluopyram and AE C656948-benzamide.  For risk assessment, the residues of concern in ruminants are fluopyram and its metabolites AE C656948-benzamide, AE C656948-E-olefine, AE C656948-Z-olefine and AE C656948-7-hydroxy, and in hens are fluopyram and its metabolites AE C656948-benzamide, AE C656948-E-olefine, and AE C656948-Z-olefine.

The German multiresidue method DFG Method S 19, a gas chromatography with mass selective detection (GC/MSD) method, has been proposed for the enforcement of tolerances for fluopyram residues in/on crop commodities, and a high performance liquid chromatography (HPLC) method with tandem mass spectrometry detection (MS/MS), Method 01079, has been proposed for the enforcement of tolerances for residues of fluopyram and AE C656948-benzamide in livestock commodities.  The validated limit of quantitation (LOQ) is 0.01 ppm for each analyte in each matrix.  The proposed enforcement method for plant commodities (DFG Method S19) and livestock commodities (Method 01079) are deemed adequate as enforcement methods.  Adequate HPLC/MS/MS methods were used for data collection for crop and livestock commodities.  The FDA multiresidue methods of PAM Vol. I are suitable for the determination of fluopyram in non-fatty matrices (using Section 302), but are not suitable for detection of AE C656948-benzamide residues.

Storage stability data have been submitted for residues of fluopyram and metabolites AE C656948-benzamide, AE C656948-PAA, AE C656948-PCA, AE C656948-7-hydroxy, and AE C656948-methyl-sulfoxide (AE 1344122) in/on lettuce, dry pea seed, orange, rape seed, and wheat grain, respectively representing water-, protein-, acid-, oil-, and starch-containing commodities.  Under frozen conditions, residues of fluopyram and its metabolite AE C656948-benzamide are stable for at least 36 months in water-, starch-, protein-, oil-, and acid-containing materials, and residues of AE C656948-PCA and AE C656948-methyl-sulfoxide are stable for at least 36 and 24 months, respectively, in protein-, oil-, and acid-containing materials.  Residues of AE C656948-PAA are stable for at least 36 months in water-, starch-, protein-, and oil-containing materials, and for 24 months in acid-containing materials.  Furthermore, residues of AE C656948-7-hydroxy are stable for at least 36 months in water- and starch-containing materials and for at least 24 months in protein-, oil-, and acid-containing materials.

Based on the livestock metabolism studies which did not show a change in the metabolic profile after 10 months of frozen storage for eggs, muscle and fat of hens and 6 months of frozen storage for milk, fat, kidney and liver of goats, and since the egg, milk, and tissue samples from the livestock feeding studies were stored frozen and analyzed within 30 days of collection, HED concludes that storage stability data for the livestock matrices are unnecessary.

Adequate field trial data are available to support the proposed uses on apple, cucurbit vegetables, dried beans, peanut, potato, stone fruit, strawberry, sugar beet, tree nuts, and wine grapes.  Application rates used in the residue field trials conducted on dry beans, cherry, and sugar beet were more than 25% higher than the corresponding proposed maximum seasonal use rates, therefore, the proportionality principle had been applied when estimating the tolerances for these crops using the NAFTA calculator.

The petitioner proposed a tolerance for "nut, tree, group 14 (including pistachio)" at 0.05 ppm.  Separate tolerances must be proposed for the tree nut crop group and pistachio.  The available data indicate that 0.05 ppm is an appropriate level for these tolerances.

The petitioner has proposed a tolerance and submitted supporting field trial data for banana.  The petitioner is not proposing use of fluopyram on banana in the U.S.  The petitioner must submit information pertaining to the proposed use of fluopyram on bananas to be imported to the U.S.

Processing studies have been submitted for many crops including apple, canola, field corn, cotton, peanut, plum, potato, soybean, sugar beet, and wheat.  The available processing data indicate that tolerances for fluopyram residues are needed for wet apple pomace and processed potato waste.  No tolerances are needed for the remaining processed commodities.

Adequate cattle and poultry feeding studies have been submitted.  The data indicate that tolerances for livestock commodities (egg, milk, and the fat, meat, and meat byproducts of cattle, goat, hog, horse, poultry, and sheep) are needed to support the proposed fluopyram uses.

The confined rotational crop studies are adequate.  An adequate limited field rotational crop study has been submitted.  The study indicates the potential for quantifiable fluopyram residues in/on crops rotated 8 months following fluopyram application.  Adequate extensive field rotational crop studies with alfalfa and cotton have been submitted to support the proposed 14-day plantback interval (PBI) for these crops, pending one repeat field trial with rotated cotton.  The tolerances for alfalfa and cotton commodities must be revised to be specified in terms of indirect or inadvertent residues of fluopyram.

The revised label states "... following potato or dried beans these crops may be planted immediately: canola, cereal grains (except rice), corn and soybean.  Do not plant all other crops for one year following the last application."  However, extensive field rotational crop data for the specified rotated crops are not available.  In the absence of sufficient rotational crop data, the registrant proposed to use the highly conservative target crop residue data for setting rotational crops tolerances.  The Chemistry Science Advisory Council (ChemSAC) understands that the selection of an intermediate level between the confined accumulation/ limited field rotational crop data and primary crop data for the target rotated crops would discourage potential misuse (i.e., direct foliar application) and provide adequate maximum residue levels for legal uses according to label instructions (2/9/2011 meeting).  Pending extensive field rotational crop data, HED recommends rotational (inadvertent residue) crop tolerances be set at half of the calculated primary crop tolerances with a PBI of 30 days: canola seed at 1.8 ppm, soybean seed at 0.10 ppm, soybean forage at 4.0 ppm, soybean hay at 15 ppm, cereal grains except rice group 15 at 1.5 ppm, and forage at 4.0 ppm, hay at 6.5 ppm, straw at 7.0 ppm, stover at 6.0 ppm of cereal grains except rice group 16.

Since the confined accumulation data reflect a maximum PBI of 280 days when TRRs (total radioactive residues) in some crop matrices still exceed 0.01 ppm, rotation to crops other than those discussed above is not supported.  The proposed label should state that crop rotation is prohibited except for the following crops: apple, cherry, dried beans, peanut, potato, sugar beet, strawberry, tree nuts, watermelon, alfalfa, cotton, canola, cereal grains except rice, and soybean.

For harmonization with Codex, the wine grape tolerance should be raised from 1.4 ppm to 2.0 ppm, hog meat byproducts from 0.45 ppm to 0.70 ppm, and milk tolerance from 0.06 ppm to 0.07 ppm.

Regulatory Recommendations and Residue Chemistry Deficiencies

HED has examined the residue chemistry database for fluopyram.  Pending submission of a revised Section B (see requirements under Directions for Use), a revised Section F (see requirements under Proposed Tolerances), and analytical reference standards (see requirements under Submittal of Analytical Reference Standards), there are no residue chemistry issues that would preclude granting conditional Section 3 registration for the requested uses of fluopyram, or establishment of tolerances for residues of fluopyram.  It is recommended that tolerances be established for residues of the fungicide fluopyram (N-[2-[3-chloro-5-(trifluoromethyl)-2-pyridinyl]ethyl]-2-(trifluoromethyl)benzamide), including its metabolites and degradates, in or on the commodities below.  Compliance with the tolerance levels specified below is to be determined by measuring only fluopyram in or on the commodity.

      Almond, hulls	8.0 ppm
      Apple	0.30 ppm
      Apple, wet pomace	0.60 ppm
      Banana	1.0 ppm
      Bean, dry	0.09 ppm
      Beet, sugar, root	0.04 ppm
      Cherry	0.60 ppm
      Grape, wine	2.0 ppm
      Nut, tree, group 14	0.05 ppm
      Peanut	0.02 ppm
      Pistachio	0.05 ppm
      Potato	0.02 ppm
      Potato, processed potato waste	0.08 ppm
      Strawberry	1.5 ppm
      Watermelon	1.0 ppm

It is recommended that tolerances be established for residues of the fungicide fluopyram (N-[2-[3-chloro-5-(trifluoromethyl)-2-pyridinyl]ethyl]-2-(trifluoromethyl)benzamide), including its metabolites and degradates, in or on the commodities below.  Compliance with the tolerance levels specified below is to be determined by measuring only the sum of fluopyram and its metabolite 2-(trifluoromethyl)benzamide, calculated as the stoichiometric equivalent of fluopyram, in or on the commodity.

      Cattle, fat	0.11 ppm
      Cattle, meat	0.15 ppm
      Cattle, meat byproducts	1.1 ppm
      Egg	0.25 ppm
      Goat, fat	0.11 ppm
      Goat, meat	0.15 ppm
      Goat, meat byproducts	1.1 ppm
      Hog, fat	0.05 ppm
      Hog, meat	0.05 ppm
      Hog, meat byproducts	0.70 ppm
      Horse, fat	0.11 ppm
      Horse, meat	0.15 ppm
      Horse, meat byproducts	1.1 ppm
      Milk	0.07 ppm
      Poultry, fat	0.20 ppm
      Poultry, meat	0.15 ppm
      Poultry, meat byproducts	0.60 ppm
      Sheep, fat	0.11 ppm
      Sheep, meat	0.15 ppm
      Sheep, meat byproducts	1.1 ppm

It is recommended that tolerances be established for residues of fungicide fluopyram (N-[2-[3-chloro-5-(trifluoromethyl)-2-pyridinyl]ethyl]-2-(trifluoromethyl)benzamide), including its metabolites and degradates, in or on the commodities below.  Compliance with the tolerance levels specified below is to be determined by measuring only fluopyram in or on the commodity.

      Alfalfa, forage	0.45 ppm
      Alfalfa, hay	1.1 ppm
      Canola, seed 	1.8 ppm
      Cotton, gin byproducts	0.05 ppm
      Cotton, undelinted seed	0.01 ppm
      Grain, cereal, group 15, except rice	1.5 ppm
      Grain, cereal, forage, fodder and straw, group 16, except rice; forage	4.0 ppm
      Grain, cereal, forage, fodder and straw, group 16, except rice; hay, straw and stover	7.0 ppm
      Soybean, forage	4.0 ppm
      Soybean, hay	15 ppm
      Soybean, seed	0.10 ppm

The tolerance for banana should include a footnote stating "No U.S. registrations as of [date of FR notice]."

A human health risk assessment is forthcoming.

860.1200 Directions for Use

   *	The petitioner must submit proposed labels (with English translations if needed) for fluopyram uses on bananas in any countries from which the treated bananas may be imported into the U.S.  The available data would support an application pattern of six applications of an SC formulation at 0.100 kg ai/ha/application (0.089 lb ai/A/application) with a 7-day RTI and a 0-day PHI.

   *	The proposed label must be amended to specify spray application volumes for wine grapes of 50 GPA for ground equipment.

   *	The proposed labels should be amended to clarify that the unit for application rates is fluid ounces per acre.

   *	For wine grapes, the proposed label should be amended to include a statement stating that "Do not treat grapes such as Thompson Seedless and Concord which may be used for purposes other than for wine."

   *	Also, with the exception of the banana field trials and one grape field trial, no spray adjuvants were used in any of the crop field trials.  However, HED has recently reviewed bridging field trials conducted on strawberry, grape, cucumber, bulb onion, potato, lettuce, tomato, peach, apple and almond with and without addition of 0.125% v/v of a non-ionic surfactant (Induce(R)) and concluded that the addition of the NIS had no significant effect on the residue level concentration on these crops.  Therefore, the product label must be amended to specify that non-ionic surfactants at 0.125% v/v may be added to spray mixtures.

   *	The label, under the rotational crop restrictions, must be amended to specify that canola, cereal grains except rice, and soybean may be replanted 30 days following fluopyram application on dry beans and potato only, and that other crops not specified on the label may not be replanted following fluopyram-treated crops.

860.1550 Proposed Tolerances

   *	The proposed tolerances for the following commodities are too high: reduced tolerances must be proposed: apple wet pomace (0.60 ppm), peanut (0.02 ppm), and processed potato waste (0.08 ppm), sugar beet root (0.04 ppm), and cotton undelinted seed (0.01 ppm).

   *	Separate tolerances must be proposed for the following commodities: apple (0.30 ppm),  cherry (0.60 ppm), dried beans (0.09 ppm), pistachio (0.05 ppm), strawberry (1.5 ppm), and watermelon (1.0 ppm).

   *	The proposed tolerances for alfalfa and cotton commodities must be revised to be specified in terms of indirect or inadvertent residues of fluopyram.  In addition, the proposed tolerances for alfalfa forage and hay must be revised to 0.45 ppm and 1.1 ppm, respectively.

   *	Separate indirect or inadvertent residues of fluopyram must be proposed for the following commodities: canola, seed (1.8 ppm); grain, cereal, group 15, except rice (1.5 ppm); grain, cereal, forage, fodder and straw, group 16, except rice, forage (4.0 ppm); grain, cereal, forage, fodder and straw, group 16, except rice, hay, straw and stover (7.0 ppm); soybean, seed (0.10 ppm); soybean, forage (4.0 ppm); and soybean, hay (15 ppm).

   *	Most of the proposed tolerances for livestock commodities are too low.  The petitioner must propose increased tolerances as specified in Table 30.

   *	The proposed tolerances should be revised to reflect the levels and correct commodity definitions as specified in Table 30.

860.1650 Submittal of Analytical Reference Standards

   *	Analytical reference standard of AE C656948-benzamide must be supplied.

HED recommends that conversion of conditional registration to unconditional registration for the requested uses may be considered upon submission of the following outstanding residue chemistry data.

860.1900 Field Accumulation in Rotational Crops

   *	Submission of the repeat cotton trial due to crop failure, and extensive field rotational crop field trials conducted on canola, cereal grains (except rice), and soybean.

Background

Fluopyram is a new fungicide with a broad spectrum of activity against numerous fungal diseases in a wide range of crops and plants.  Fluopyram represents a new class of chemistry (pyridylethylamide), and its biochemical mode of action has been shown to rely on the inhibition of the enzyme succinate dehydrogenase (complex II) within the fungal mitochondrial respiratory chain, thus blocking electron transport.  Fluopyram is a Group 7 fungicide that Bayer initially has proposed for use on the following crops:  artichoke (globe); Brassica leafy vegetables; bulb vegetables; canola; cereal grains (except rice); citrus; cucurbit vegetables; fruiting vegetables; ginseng; grapes and small vine fruits (except fuzzy kiwifruit); grasses for forage/feed (including grasses grown for seed); herbs and spices (except black pepper); hops; leafy vegetables; legume vegetables; okra; peanut; pome fruit; potato and other root, tuberous, and corm vegetables; small berries (caneberries and bushberries); stone fruit; strawberry and other low growing berries; sunflower; and tree nuts, including pistachio.  Subsequently, the registrant revised the list of crops (as indicated on page 7).  The chemical structure and nomenclature of fluopyram are presented in Table 1, and the physicochemical properties of the technical grade of fluopyram are presented in Table 2.  This petition represents the first food/feed uses of fluopyram proposed in the U.S. and Canada.

The chemical names and structures of fluopyram metabolites are presented in Appendix I.

Table 1.	Fluopyram Nomenclature.
Chemical structure

Common name
Fluopyram
Company experimental name
AE C656948
IUPAC name
N-[2-[3-Chloro-5-(trifluoromethyl)-2-pyridyl]ethyl]-α,α,α-trifluoro-o-toluamide
CAS name
N-[2-[3-Chloro-5-(trifluoromethyl)-2-pyridinyl]ethyl]-2-(trifluoromethyl)benzamide
CAS registry number
658066-35-4
End-use product (EP)
Fluopyram 500 SC Fungicide (4.16 lb/gal SC formulation; EPA File Symbol No. 264-RNTI)
Chemical structure of AE C656948-benzamide metabolite (included in the tolerance expression for livestock commodities)

                         2-(Trifluoromethyl)-benzamide

Table 2.	Physicochemical Properties of Fluopyram.
Parameter
Value
Reference
Melting point
118 ºC
MRID 47372240
pH
6.6 (1% aqueous solution; pure substance)
MRID 47372236
Density
1.53
MRID 47372242
Water solubility (20 °C)
Distilled water (pH 6.7) = 16 mg/L
pH 4 = 15 mg/L
pH 7 = 16 mg/L
pH 9 = 15 mg/L
MRID 47372247
Solvent solubility (g/L at 20 °C)
heptane	0.66
toluene	62.2
dichloromethane	>250
methanol	>250
acetone	>250
ethyl acetate	>250
dimethyl sulfoxide	>250
MRID 47372248
Vapor pressure
1.2 x 10-6 Pa at 20 ºC
3.1 x 10-6 Pa at 25 ºC
2.9 x 10-4 Pa at 50 ºC
MRID 47372249
Dissociation constant (pKa)
0.5 (23 ºC)
MRID 47372243
Octanol/water partition coefficient Log(KOW)
3.3
MRID 47372245
UV/visible absorption spectrum
Peak max at 270 nm
MRID 47372237

860.1200 Directions for Use

The petitioner has submitted a draft specimen label for the end-use product containing fluopyram listed in Table 3.  The product is an SC formulation.  A summary of the proposed use patterns on food and feed crops is detailed in Table 4.

Table 3.	Summary of Proposed End-Use Products.
Trade Name
                                File Symbol No.
                             ai (% of formulation)
                               Formulation Type
Target Crops
Label Date
Fluopyram 500 SC Fungicide/ LUNA(R)  PRIVILEGE
                                   264-RNTI
                              41.5 [4.16 lb/gal]
                                      SC
Apple, cherry, dried beans, wine grape, peanut, potato, strawberry, sugar beet, tree nuts, and watermelon.
08/15/11

Table 4.	Summary of Directions for Use of Fluopyram.
Applic. Timing; Type; and Equipment
                                  Formulation
                             [EPA File Symbol No.]
                                 Applic. Rate
                                   (lb ai/A)
                          Max. No. Applic. per Season
                          Max. Seasonal Applic. Rate
                                   (lb ai/A)
                                      PHI
                                    (days)
                       Use Directions and Limitations[1]
Apple
Postemergence;
Foliar spray;
Ground, or chemigation
                                4.16 lb/gal SC
                                  [264-RNTI]
                                  0.078-0.222
                                      NS
                                     0.445
                                       7
The proposed RTI is 7-14 days.
Cherry
Postemergence;
Foliar spray;
Ground, aerial, or chemigation
                                4.16 lb/gal SC
                                  [264-RNTI]
                                     0.092
                                      NS
                                     0.183
                                       0
The proposed RTI is 5-7 days.
Dried shelled beans:  bean (Lupinus spp., includes grain lupin, sweet lupin, white lupin, and white sweet lupin), bean (Phaseolus spp., includes field bean, kidney bean, lima bean (dry), navy bean, pinto bean, tepary bean), bean (Vigna spp., includes adzuki bean, blackeyed pea, catjang, cowpea, Crowder pea, moth bean, mung bean, rice bean, southern pea, urd bean), other beans (broad bean (dry), chickpea, guar, lablab bean, lentil)
Postemergence;
Foliar spray;
Ground, aerial, or chemigation
                                4.16 lb/gal SC
                                  [264-RNTI]
                                     0.134
                                      NS
                                     0.268
                                      14
The proposed RTI is 7-14 days.  Do not feed hay or threshings or allow livestock to graze in treated areas.
Peanut
Postemergence;
Foliar spray;
Ground, aerial, or chemigation
                                4.16 lb/gal SC
                                  [264-RNTI]
                                  0.182-0.222
                                      NS
                                     0.445
                                       7
The proposed RTI is 14 days.  Do not feed hay or threshings or allow livestock to graze in treated areas.
Potato
Postemergence;
Foliar spray;
Ground, aerial, or chemigation
                                4.16 lb/gal SC
                                  [264-RNTI]
                      0.130-0.178 (ground, chemiga-tion)
                                0.092 (aerial)
                                      NS
                         0.356 (ground, chemiga-tion)
                                0.275 (aerial)
                                       7
The proposed RTI is 5-7 days.  The grazing of livestock in treated areas within 7 days of application is prohibited.
Strawberry
Postemergence;
Drip/ chemigation, greenhouse spray
                                4.16 lb/gal SC
                                  [264-RNTI]
                                     0.222
                                      NS
                                     0.445
                             0 (drip appli-cation)

                             1 (green-house uses)
The proposed RTI is 5-7 days.  Applications may be made to strawberries grown in a greenhouse of 10 acres or larger.
Sugar beet
Postemergence;
Foliar spray;
Ground, or chemigation
                                4.16 lb/gal SC
                                  [264-RNTI]
                                     0.111
                                      NS
                                     0.222
                                       7
The proposed RTI is 5-7 days.
Tree nuts:  almond, beech nut, Brazil nut, butternut, cashew, chestnut, chinquapin, filbert (hazelnut), hickory nut, macadamia nut (bush nut), pecan, pistachio, walnut [including black and English (Persian) walnuts]
Postemergence;
Foliar spray;
Ground, aerial, or chemigation
                                4.16 lb/gal SC
                                  [264-RNTI]
                                  0.104-0.222
                                      NS
                                     0.445
                                      14
The proposed RTI is 7-14 days.
Watermelon: watermelon (includes hybrids and/or varieties of Citrullus lanatus)
Postemergence;
Foliar spray;
Ground, or drip/chemigation
                                4.16 lb/gal SC
                                  [264-RNTI]
                                  0.078-0.222
                                      NS
                                     0.445
                                0 (spray uses)

                             3 (green-house uses)

                            7 (drip appli-cations)
The proposed RTI is 5-14 days.  Applications may be made to cucurbits grown in a greenhouse of 10 acres or larger.
Wine Grapes:  varieties only such as but not limited to these varieties: Chardonnay, Cabernet sauvignon, Syrah, Merlot, Pinot Noir, and Zinfandel
Postemergence;
Foliar spray;
Ground, or chemigation
                                4.16 lb/gal SC
                                  [264-RNTI]
                                  0.078-0.222
                                      NS
                                     0.445
                                       7
The proposed RTI is 12-21 days.
[1]  RTI = Retreatment interval.  All applications are to begin preventatively and continue as needed (when multiple applications are permitted).

The label specifies that for field and vegetable crops, applications are to be made in >=10 GPA for ground equipment and >=5 GPA for aerial equipment; for tree crops, applications are to be made in >=50 GPA for ground equipment and >=15 GPA for aerial equipment.  Application volumes for grapes are not specified.

The label specifies that applications may be made as foliar, soil, or soil-less mix applications to cucurbit vegetables and strawberry grown in greenhouse of 10 acres or larger.

The label indicates that the product is compatible with most commonly used fungicides, herbicides, insecticides, and foliar nutrient products.  No tank mix partners are specified, but the label recommends that before applying any tank mixture, safety to the crop should be confirmed on a small portion of the crop to be treated.

The label specifies that to limit the potential for development of disease resistance, no more than two sequential applications of fluopyram or any Group 7 fungicide may be made before rotating with a fungicide from a different group.

The following PBIs are specified on the proposed label for EPA File Symbol No. 264-RNTI:  14 days for alfalfa and cotton; and 0 days for apple, cherry, cucurbit vegetables, dried beans, peanut, potato, stone fruit, sugar beet, strawberry, tree nuts, and wine grapes.  Furthermore, following potato or dried bean these crops may be planted immediately: canola, cereal grains (except rice), corn and soybean.  Do not plant all other crops for one year following the last application.

A 12-hour restricted entry interval (REI) is specified for EPA File Symbol No. 264-RNTI.

Conclusions.  The use directions are adequate to allow evaluation of the residue data relative to the proposed use in the U.S.  The proposed maximum seasonal rate and minimum RTIs are supported by the submitted field trial data.  The proposed minimum PHIs are supported by the submitted field trial data.

No North American greenhouse data were submitted.  Use of fluopyram on cucurbit vegetables and strawberry grown in a greenhouse is supported by greenhouse data collected in Europe.

No proposed use information was provided for bananas.  The petitioner must submit proposed labels (with English translations if needed) for fluopyram uses on bananas in any countries from which the treated bananas may be imported into the U.S.  The available data would support an application pattern of six applications of an SC formulation at 0.100 kg ai/ha/application (0.089 lb ai/A/application) with a 7-day RTI and a 0-day PHI.

The proposed label must be amended to specify spray application volumes for wine grapes of 50 GPA for ground equipment.  In addition, the proposed labels should be amended to clarify that the unit for application rates is fluid ounces per acre.

For wine grapes, the proposed label should be amended to include a statement stating that "Do not treat grapes such as Thompson Seedless and Concord which may be used for purposes other than for wine.

      Also, with the exception of the banana field trials and one grape field trial, no spray adjuvants were used in any of the crop field trials.  However, HED has recently reviewed bridging field trials conducted on strawberry, grape, cucumber, bulb onion, potato, lettuce, tomato, peach, apple and almond with and without addition of 0.125% v/v of a non-ionic surfactant (Induce(R)) and concluded that the addition of the NIS had no significant effect on the residue level concentration on these crops.  Therefore, the product label must be amended to specify that non-ionic surfactants at 0.125% v/v may be added to spray mixtures.

Adequate data have been submitted to support the proposed PBIs of 14 days for alfalfa and cotton.  No extensive field rotational crop data were submitted to support the proposed PBIs of 0 day for canola, cereal grains, and soybean.  In the absence of sufficient rotational crop data and based on the foliar crop data, the label must be amended to specify that canola, cereal grains, and soybean may be replanted 30 days following fluopyram application.  Based on the confined accumulation and limited field rotational crop data, there is a potential for quantifiable residues of fluopyram in/on rotational crops at PBIs of beyond 12 months.  Therefore, all other crops not on the label may not be replanted following fluopyram-treated crops.

860.1300 Nature of the Residue - Plants

First Entry Monograph for Fluopyram, Sections B.7.1.1 through B.7.1.5 (MRIDs 47372524-47372531, and 47372536)

The petitioner submitted studies pertaining to the metabolism of fluopyram in grape, bean, potato, and red pepper.  Two different radiolabels were used in the plant studies; the label positions are presented below:

                         [phenyl-UL-[14]C]AE C656948

                        [pyridyl-2,6-[14]C]AE C656948

Numerous metabolites were identified in these studies.  The chemical structures and report names for these metabolites are provided in Appendix I.

Grape

MRID 47372525:  The metabolism was investigated in grape vines following three spray applications with [phenyl-UL-[14]C]fluopyram formulated as a suspension concentrate (SC 500) targeting a maximum annual application rate of 500 g ai/ha. The targeted single application rates amounted to 100 g ai/ha, 200 g ai/ha, and 200 g ai/ha for the first, second, and third application, respectively.  The actual total application rate was 504 g ai/ha (0.45 lb ai/A), 1x the proposed maximum seasonal rate to grapes.  Applications were performed at growth stages BBCH 17-19, 71, and 81. Grapes and leaves were harvested at maturity with a PHI of 18 days for grapes and 19 days for leaves. After the second application, shoots and leaves ("summer cut") were taken for method development.

The total radioactive residues (TRR) amounted to 28.55 ppm in the summer cut, 1.86 ppm in grapes, and 48.06 ppm in leaves. A total of 98.6% and 93.9% TRR in grapes and leaves, respectively, were extracted conventionally using acetonitrile (ACN)/water.

Parent compound and metabolites in the extracts were quantified by radio-HPLC. A total of 98.6% TRR was identified in grapes and 93.8% TRR in leaves.  The TRR consisted nearly quantitatively of unchanged parent compound which represented 1.82 ppm (97.6% TRR) in grapes and 44.11 ppm (91.8% TRR) in leaves.  Apart from parent, the residues consisted of AE C656948-benzamide and AE C656948-7-hydroxy in grapes.  In the leaves at harvest, AE C656948-7- and 8-hydroxy and the glucoside conjugate of the AE C656948-7-hydroxy were identified.

MRID 47372524:  The metabolism was investigated in grape vines following three spray applications with [pyridyl-2,6-[14]C]fluopyram formulated as a suspension concentrate (SC 500) targeting a maximum annual application rate of 500 g ai/ha.  The targeted single application rates amounted to 100 g ai/ha, 200 g ai/ha, and 200 g ai/ha for the first, second, and third application, respectively.  The actual total application rate was 498 g ai/ha (0.444 lb ai/A), 1x the proposed maximum seasonal rate to grapes.  Applications were performed at growth stages BBCH 17-19, 71, and 81.  Grapes and leaves were harvested at maturity with a PHI of 18 days for grapes and 19 days for leaves.  After the second application, shoots and leaves ("summer cut") were taken for method development.

The TRR amounted to 1.7 ppm for grapes and 42.66 ppm in leaves.  A total of 97.1% and 94.7% TRR in grapes and leaves, respectively, were extracted conventionally using ACN/water.

Parent compound and metabolites in the extracts were quantified by radio-HPLC.  A total of 97.0% TRR was identified in grapes and 94.7% in leaves.  The TRR consisted nearly quantitatively of unchanged parent compound which represented 1.63 ppm (95.8% TRR) in grapes and 39.0 ppm (91.3% TRR) in leaves.  Apart from parent, the identified residues consisted of AE C656948-PCA and AE C656948-7-hydroxy in grapes.  In the leaves at harvest, the identified residues were AE C656948-7- and 8-hydroxy, the glucoside conjugate of the AE C656948-7-hydroxy and AE C656948-PCA.

Potato

MRID 47372528:  The metabolism was investigated in potatoes following three spray applications with [phenyl-UL-[14]C]fluopyram formulated as a suspension concentrate (SC 500).  The target application rate for a single application amounted to 167 g ai/ha/application; the actual rates for each of the three applications were 167.3 g ai/ha, 175.6 g ai/ha and 176.0 g ai/ha, resulting in a total actual application rate of 518.8 g ai/ha (0.463 lb ai/A), 1x the proposed maximum seasonal rate to potato.  Applications were performed at growth stages BBCH 16, 55, and 71.  Potato leaves and tubers were harvested at maturity with a PHI of 51 days.

The TRR for tubers (0.008 ppm) were very low while the TRR in leaves were 47.64 ppm.  A total of 96.7% and 99.4% TRR in tubers and leaves, respectively, were extracted conventionally using ACN/water.

Parent compound and metabolites in the extracts were quantified by HPLC.  A total of 77.1% TRR was identified in tubers and 99.2% TRR in leaves.  Parent compound amounted to 0.006 ppm (68.8% TRR) in tubers and 46.69 ppm (98.0% TRR) in leaves.  AE C656948-benzamide (0.001 ppm, 7.1 % TRR) was identified in the extract of tubers by HPLC co-chromatography and also assigned in the extract of leaves (0.23 ppm, 0.5 % TRR).  AE C656948-7-hydroxy was detected in low amounts (<=1.2% TRR) in the extracts of tubers and leaves.

MRID 47372529:  The metabolism was investigated in potatoes following three spray applications with [pyridyl-2,6-[14]C]fluopyram formulated as a suspension concentrate (SC 500).  The target application rate for a single application amounted to 167 g ai/ha/application; the actual rates for each of the three applications were 170.1 g ai/ha, 170.5 g ai/ha and 165.1 g ai/ha, resulting in a total actual application rate of 505.7 g ai/ha (0.451 lb ai/A), 1x the proposed maximum seasonal rate to potato.  Applications were performed at growth stages BBCH 16, 55, and 71.  Potato leaves and tubers were harvested at maturity with a PHI of 51 days.

The TRR in the tubers (0.012 ppm) were very low while in the leaves TRR were 21.67 ppm. Significant portions of the TRR, 95.3% and 99.6% TRR in tubers and leaves, respectively, were extracted conventionally using ACN/water.

Parent compound and metabolites in the extracts were quantified by HPLC. A total of 74.1% TRR was identified in tubers and 99.2% TRR in leaves.  Parent compound amounted to 0.003 ppm (23.2% TRR) in tubers and 21.26 ppm (98.1% TRR) in leaves.  AE C656948-PCA was identified in the extract of tubers and leaves at 49.8% TRR (0.006 ppm) and 0.5% TRR (0.11 ppm), respectively.  AE C656948-7-hydroxy was detected in low amounts (<=1.1% TRR) in the extracts of tubers and leaves.

Bean

MRID 47372527:  The metabolism was investigated in beans following two spray applications with [phenyl-UL-[14]C]fluopyram formulated as a suspension concentrate (SC 500) targeting a maximum annual application rate of 500 g ai/ha (0.446 lb ai/A). The actual rates for each of the two applications were 269 and 259 g ai/ha (0.240 and 0.231 lb ai/A), respectively, resulting in a total actual application rate of about 528 g ai/ha (0.471 lb ai/A), 1x the proposed maximum seasonal rate to legume vegetables.  Applications were performed at growth stages BBCH 51 and 75. The immature raw agricultural commodities (RACs) investigated, green beans and foliage, were collected four days after the second application. The mature RACs, beans (succulent shelled bean) and straw, were harvested 29 days after the last application.  A portion of the mature beans was dried for 11 days and analyzed as edible RAC dry bean seeds.

The TRR in the edible RACs succulent and dry beans were very low and amounted to 0.07 ppm and 0.12 ppm, respectively.  In green beans, foliage, and straw, TRR were 1.40 ppm, 36.66 ppm and 16.55 ppm, respectively.  The major amount of radioactivity (93.9-98.1% TRR) was effectively extracted with ACN/water from all RACs.

Parent compound and metabolites in the extracts were quantified by radio-HPLC.  The identification rate was high in all RACs and ranged from 84% to 98% TRR.

In the edible RACs succulent and dry beans, the active substance was cleaved to a greater extent compared to the other RACs, green beans, foliage, and straw.  In succulent and dry beans, unchanged parent compound represented 11-13% TRR.  In these RACs the main metabolite was AE C656948-benzamide which amounted to 0.04 ppm (51.6% TRR) in succulent beans and 0.08 ppm (64.0% TRR) in dry beans.

In green beans, foliage, and straw, the TRR consisted nearly quantitatively of unchanged parent compound which represented >90% TRR.  AE C656948-benzamide was either not detected or represented <1% TRR in these RACs.

Apart from the active substance and AE C656948-benzamide, AE C656948-7-hydroxy, AE C656948-8-hydroxy, and several conjugates of the hydroxylated active substance were identified.

MRID 47372526:  The metabolism was investigated in beans following two spray applications with [pyridyl-2,6-[14]C]fluopyram formulated as a suspension concentrate (SC 500) targeting a maximum annual application rate of 500 g ai/ha (0.446 lb ai/A). The actual rates for each of the two applications were 265 and 254 g ai/ha (0.236 and 0.227 lb ai/A), respectively, resulting in a total actual application rate of about 519 g ai/ha (0.463 lb ai/A), 1x the proposed maximum seasonal rate to legume vegetables.  Applications were performed at growth stages BBCH 51 and 75. The immature RACs, green beans and foliage, were collected four days after the second application. The mature RACs, beans (succulent shelled bean) and straw, were harvested 29 days after the last application. A portion of the mature beans was dried for 11 days and analyzed as edible RAC dry bean seeds.

The TRR in the edible RACs succulent and dry beans were very low and amounted to 0.17 ppm and 0.31 ppm, respectively.  In green beans, foliage and straw, TRR amounted to 3.88 ppm, 38.53 ppm, and 19.02 ppm, respectively.  The major amount of radioactivity (95.7-99.3% TRR) was effectively extracted with ACN/water from all RACs.

Parent compound and metabolites in the extracts were quantified by radio-HPLC.  A total of 76% to 99% TRR was identified in the RACs.

In the edible RACs succulent and dry beans, the active substance was cleaved to a greater extent compared to the other RACs, green beans, foliage, and straw.  In succulent and dry beans, unchanged parent compound represented 4.8-5.7% TRR.  In these RACs, AE C656948-PAA and AE C656948-PCA were the main metabolites representing 23-33% TRR. A third and minor label-specific metabolite, AE C656948-hydroxyethyl-glycoside, was detected and represented <=3.1% TRR.

In green beans, the TRR consisted quantitatively of unchanged parent compound which represented 99.3% TRR.  In foliage, parent compound amounted to 92.3% TRR and 6 minor metabolites were identified.  In straw, parent compound amounted to 87.1% TRR and 7 minor metabolites were identified.

Apart from the active substance and the three label-specific metabolites, AE C656948-7-hydroxy, AE C656948-8-hydroxy, and several conjugates of the hydroxylated active substance were identified.

Red pepper

MRID 47372531:  The metabolism of the fungicide fluopyram was investigated in red bell pepper following drip irrigation with [phenyl-UL-[14]C]fluopyram formulated as a suspension concentrate (SC 500).  An aqueous dilution of the formulation was applied to the plants growing on stone wool substrate according to practice in professional horticulture. The targeted single application rate amounted to 5 mg ai/plant.  Additionally, an overdose experiment (4x) was conducted at 20 mg ai/plant for method development and to facilitate identification of metabolites. The applications were performed when the fifth to seventh leaf of the main shoot was unfolded (BBCH 15-17).

A plant at an intermediate growth stage was harvested from the 4x experiment 33 days after application.  Fruits were harvested at maturity from plants of the 1x experiment at three time points (55-96 days after application).  The remaining plants were sampled one day after the third harvest of fruits (97 days after application).  The RACs from the overdose experiment were used to facilitate isolation and identification of metabolites.

The TRR in the mature fruits were very low and amounted to 0.038 ppm.  In the rest of the plant, TRR amounted to 3.54 ppm.  The major amount of radioactivity (>96% TRR) was effectively extracted with ACN/water from all RACs.

Parent compound was the major part of the residue in the fruits and the rest of the plant (48.9% and 64.0% TRR, respectively).  The main metabolite was the cleavage product AE C656948-benzamide (16.1% TRR in fruits and 10.0% TRR in remaining plants).  Other metabolites detected (<=9% TRR each) were AE C656948-7-hydroxy and its glucoside conjugate in both fruits and the remaining plants.  In the rest of the plant, AE C656948-8-hydroxy and a malonic acid conjugate of the AE C656948-7-hydroxy-glc were additionally observed at minor amounts (<1% TRR each).

MRID 47372530:  The metabolism of the fungicide fluopyram was investigated in red bell pepper following drip irrigation with [pyridyl-2,6-[14]C]fluopyram formulated as a suspension concentrate (SC 500).  An aqueous dilution of the formulation was applied to the plants growing on stone wool substrate according to practice in professional horticulture.  The targeted single application rate amounted to 5 mg ai/plant.  Additionally, an overdose experiment (4x) was conducted at 20 mg ai/plant for method development and to facilitate identification of metabolites.  The applications were performed when the fifth to seventh leaf of the main shoot was unfolded (BBCH 15-17).

A plant at an intermediate growth stage was harvested from the 4x experiment 33 days after application at BBCH 61.  Fruits were harvested at maturity from plants of the 1x and the 4x experiment at three time points (55-96 days after application).  The remaining plants were sampled one day after the third harvest of fruits (97 days after application).

The TRR in the mature fruits (1x) were very low and amounted to 0.06 ppm.  In the rest of the plant, TRR amounted to 2.34 ppm.  The major amount of radioactivity (>95% TRR) was effectively extracted with ACN/water from all RACs.

In the fruits parent compound amounted to 0.01 ppm (16.2% TRR).  Identified metabolites were the cleavage product AE C656948-PCA and two glycoside isomers of the AE C656948-PAA amounting to 0.026 ppm (43.5% TRR), 0.014 ppm (23.8% TRR) and 0.009 ppm (14.2% TRR), respectively.  Non-conjugated AE C656948-PAA and AE C656948-7-hydroxy were additionally found in the fruits of the 4x overdose experiments.

In the rest of the plant (1x), parent compound accounted for the major part of the residues (1.64 ppm, 70.1% TRR).  Only minor metabolites were detectable (each <=9.2% TRR) and consisted of AE C656948-7-hydroxy and its glucose conjugate, AE C656948-N-oxide, and a higher glucose conjugate of the cleavage product AE C656948-hydroxyethyl.  In the intermediate plant (4x), AE C656948-8-hydroxy was additionally identified.

Additional studies

The petitioner submitted a study of the degradation of fluopyram in plant suspension cell cultures.  The main purpose of this study was to facilitate metabolite identification and to produce radiolabeled reference compounds for the identification of metabolites in the plant and livestock metabolism studies.  Radiolabeled reference standards of the following compounds were generated in the cell culture study:  AE C656948-deschloro-3-OH-glc; AE C656948-7-hydroxy; AE C656948-7-hydroxy-glc; AE C656948-8-hydroxy-glc; AE C656948-hydroxymethyl-benzamide; AE C656948-benzamide; AE C656948-pyridyl-hydroxyethyl; AE C656948-pyridyl-hydroxymethyl; and AE C656948-PCA.

The petitioner also noted that the metabolism of fluopyram in cereal grains after seed treatment was not investigated in a separate metabolism study; the petitioner stated that seed treatment use of fluopyram on cereal grains will be proposed in Europe.  The petitioner concluded that data on the metabolism of fluopyram in cereals following uptake from the soil, from the first rotation of the confined rotational crop studies (see 860.1850), would be equivalent to seed treatment.  The petitioner stated that the highest seed treatment rate in cereals will be 1 g ai/dt [1 g ai/100 kg], equivalent to a worst case application rate of 2.4 g ai/ha (0.0021 lb ai/A).  The application rate used in the confined rotational crop studies was 500 g ai/ha (~200x the seed treatment equivalent rate).  In contrast to the situation of seed treatment, the test substance in the rotational crop studies was evenly applied to the bare ground, and wheat seeds were planted 30 days following application.  The petitioner stated that the results from the laboratory aerobic soil degradation studies with radiolabeled fluopyram show that after 30 days, 81-92% of the applied active substance is still available in soil (i.e., at least 405 g ai/ha from application at 500 g ai/ha).

Conclusions.  The submitted plant metabolism studies are adequate.  The results of the submitted studies with grape, potato, bean, and red pepper indicate that the metabolism of fluopyram is similar in these crops.  The metabolism of fluopyram in plants appears to proceed via hydroxylation of parent compound leading to AE C656948-7-hydroxy and AE C656948-8-hydroxy, conjugation of the hydroxylated parent compound mainly with sugars, and cleavage of the molecule leading to AE C656948-benzamide, AE C656948-PAA, and AE C656948-PCA.  Further conjugation of AE C656948-7-hydroxy-glucose with malonic acid was observed in bean and red pepper; in addition, cleavage of the molecule yielded glycoside conjugates and AE C656948-hydroxyethyl conjugates in bean and red pepper.

The submitted studies are adequate to satisfy plant metabolism data requirements.  The submitted studies represent three dissimilar crop types (fruit/fruiting vegetable, root crop, and a pulse/oil seed), and the parent was found to be the major residue in all four studies.  The residue of concern in plant commodities for tolerance enforcement is fluopyram; the residue of concern in plant commodities except in legumes and oilseed crops for risk assessment is fluopyram, and in legumes and oilseed crops is fluopyram plus C656948-benzamide (ROCKS decision memo, 7/16/2009).

860.1300 Nature of the Residue - Livestock

First Entry Monograph for Fluopyram, Sections B.7.2.1 and B.7.2.2 (MRIDs 47372532-47372535)

There are several livestock feedstuffs associated with the proposed uses of fluopyram.  The petitioner provided livestock metabolism studies with lactating goats and laying hens, which are summarized below.

Hen

MRID 47372532:  In a metabolism study with [phenyl-UL-[14]C]AE C656948, six hens were orally dosed for 14 consecutive days in 24 h intervals with 2.03 mg per kg body weight per day (corresponding to 26.42 mg ai/kg feed) and sacrificed about 24 hours after the last dose.  The dosing level corresponds to ~21x the estimated dietary burden of fluopyram to poultry (see Table 8).  The dose was tolerated without any observable toxicological sign.  Total radioactivity was measured in the excreta and eggs, which were collected daily, as well as in the dissected tissues muscle, fat, liver, and skin collected at sacrifice.  The eggs, muscle, fat, liver, and excreta were analyzed for parent compound and metabolites.

The overall recovery (sum of radioactivity in the excreta, eggs as well as organs and tissues) was 94.8% of the total administered dose.  The majority of the radioactivity (82.7% of the total dose) was detected in the excreta collected before sacrifice.  The excretion was high and started immediately after the first administration; the time course of the excretion was characterized by a relatively constant rate starting at Day 2 until test end.  Radioactivity in eggs accounted for 4.3% of the total dose.  At sacrifice, the compound-related residues in the edible organs and tissues dissected from the bodies amounted to 7.8% of the total dose.  From this, more than half (4.9%) was detected in the muscle.

The TRR values in eggs ranged from 0.462 (Day 1) to 3.901 ppm (Day 14).  An approximately linear increase was observed until Day 7 (3.243 ppm).  After that the residues increased slightly to the test end; however, residues had not plateaued by the end of the dosing period.  The TRR in the eggs collected from the ovary and oviduct (5.771 ppm) were 1.5x the TRR in laid eggs at the test end (3.901 ppm).  TRR were highest in liver (9.536 ppm) and kidney (5.759 ppm), and lower in muscle (3.290 ppm), skin (2.533 ppm), and subcutaneous fat (1.696 ppm).

For analysis of parent compound and metabolites, Day 1-6 and Day 7-14 eggs, muscle, fat, liver, and Day 14 excreta samples from all animals were respectively pooled, and the pooled samples were extracted with ACN/water.  After solid-phase extraction (SPE) purification, the resulting extracts of eggs, muscle, fat, liver, and excreta represented between ca. 92% and 99% TRR in the sample.

The major component in all the edible matrices was the metabolite AE C656948-benzamide, at 68.6% to 98.6% TRR.  Other metabolites identified were AE C656948-Z-olefine (25.9% TRR in fat and <=1.2% TRR in egg, muscle, and liver), AE C656948-E-olefine (<=2.3% TRR in fat and liver), and AE C656948-benzoic acid (0.3% TRR, in liver only).  Parent compound was detected as a minor component only in eggs and fat at <=2.5% TRR.  Identification rates in the organs and tissues ranged from 93% to 99% TRR.

MRID 47372533:  In a metabolism study with [pyridyl-2,6-[14]C]AE C656948, six hens were orally dosed for 14 consecutive days in 24 h intervals with 2.02 mg per kg body weight per day (corresponding to 25.96 mg ai/kg feed) and sacrificed about 24 hours after the last dose.  The dosing level corresponds to ~21x the estimated dietary burden of fluopyram to poultry (see Table 8).  The dose was tolerated without any observable toxicological sign.  Total radioactivity was measured in the excreta and eggs daily, as well as in the dissected tissues muscle, fat, liver, and skin at sacrifice.  The eggs, muscle, fat, liver, and excreta were analyzed for parent compound and metabolites.

The overall recovery (sum of radioactivity in the excreta, eggs as well as organs and tissues) was 95.6% of the total administered dose.  The majority of the radioactivity (94.7% of the total dose) was detected in the excreta collected before sacrifice.  The excretion rate was high and started immediately after the first administration; the time course of the excretion was characterized by a relatively constant rate starting at Day 2 until test end.  Radioactivity in eggs accounted for 0.4% of the total dose.  At sacrifice, the compound-related residues in the edible organs and tissues collected from the hens amounted to 0.5% of the total dose.

The TRR in eggs ranged from 0.047 (Day 1) to 0.321 ppm (Day 8).  A linear increase was observed until Day 8; after that the residues decreased slightly to 0.262 ppm (Days 13 and 14), and the residue plateau was reached. The TRR in the eggs collected from the ovary and oviduct at sacrifice (0.831 ppm) were about 3x the TRR in laid eggs at the test end (0.262 ppm).  The TRR were 0.538 ppm in liver, 0.242 ppm in kidney, 0.498 ppm in subcutaneous fat, 0.152 ppm in skin, and 0.048 ppm in muscle.

For analysis of parent compound and metabolites, Day 1-6 and Day 7-14 eggs, muscle, fat, liver, and Day 14 excreta samples from all animals were respectively pooled, and the pooled samples were extracted with ACN and/or ACN/water (eggs, muscle, liver, and excreta) or with ACN/water and heptane (fat).  After SPE purification, the resulting extracts of eggs, muscle, fat, liver, and excreta represented 43-54%, 58%, 99%, 32% and 92%, respectively, of the TRR. Alternatively, egg pools and liver were extracted after enzymatic digestion to try to solubilize more of the radioactive residues; 64-66% in eggs and 82% in liver was recovered after SPE cleanup of the extract.  Residual solids in egg pool Day 7-14 following conventional extraction and in liver following enzymatic extraction were exhaustively extracted using microwave extraction releasing an additional 51.2% and 7.4% TRR, respectively.  In these samples, remaining nonextractable residues were 1% and 9.7% TRR.  Nonextractable residues in muscle and fat were <0.05 ppm.

The major component identified was the metabolite AE C656948-Z-olefine which accounted for 15.4-19.3% TRR in eggs Day 7-14, 33.0% TRR in muscle, and 70.5% TRR in the fat, but only 4.1% TRR in eggs Day 1-6, and 1.9-3.1% TRR in liver.  The other isomer AE C656948-E-olefine was found at 11.8-13.9% TRR in fat and liver, and 1.0-3.9% TRR in eggs and muscle.  Parent compound was a major residue identified in eggs Day 1-6 at 14.7-17.9% TRR and 12.2% TRR in fat; the parent was identified as a minor residue (<=9.5% TRR) in eggs Day 7-14 and muscle, and was not detected in liver.  AE C656948-PAA and AE C656948-7-hydroxy were only minor metabolites, each present at <=6.4% TRR in eggs and liver.  The absolute levels of the individual metabolites common for both labels (phenyl and pyridyl) were very similar in this study to the levels measured after dosing of [phenyl-UL-[14]C]AE C656948.  Identification rates in the eggs, organs, and tissues ranged from 20% to 95% TRR.

Due to a high amount of matrix components and the low concentration of radioactivity, no HPLC profile of the microwave extracts, which represented 51.2% TRR (0.146 ppm) in eggs and 7.4% TRR (0.039 ppm) in liver, could be recorded.  However, the profile of the egg extract after enzymatic digestion showed neither significant amounts of further metabolites nor different amounts of metabolites compared to the profile of the conventional extract. Based on this result, the petitioner proposed that the radioactivity released by microwave extraction from the solids did not consist of any further unknown metabolites.

Conclusions - Hen.  The submitted phenyl-label hen metabolism study is adequate.  When considered on its own, the pyridyl-label hen metabolism study is not adequate to satisfy data requirements.  Fat was the only matrix for which the majority of the radioactivity was identified (95% TRR); less than 40% TRR was identified in eggs, muscle, and liver.  TRR in the pooled eggs and tissues were at most a quarter of the activity levels found in the phenyl-label study.

However, in consideration of the pyridyl-label study and the phenyl-label study together, HED concludes that adequate hen metabolism data have been submitted.  Based on TRR levels, radioactivity resulting from the pyridyl portion of fluopyram accounted for much less in hen matrices than radioactivity resulting from the phenyl portion, and >=93% TRR was identified in hen matrices in the phenyl-label study.  A larger amount of the dosed radioactivity was excreted in the pyridyl-label study (94.7%) than in the phenyl-label study (82.7%).  In addition, the absolute ppm levels of metabolites that were detected in both studies (parent and the olefines) were very similar.

The most important metabolic reaction in the laying hen was the cleavage of the aliphatic chain, yielding the major metabolite AE C656948-benzamide.  A second major metabolic reaction involved the hydroxylation of the aliphatic chain followed by elimination, yielding the olefines. Hydrolysis of the amide to the parent carboxylic acid, AE C656948-benzoic acid, and oxidative cleavage of the aliphatic chain, yielding AE C656948-PAA, were observed as minor reactions.

Goat

MRID 47372535:  A goat was orally dosed for five consecutive days, at 24 h intervals, with 1.91 mg [phenyl-UL-[14]C]fluopyram per kg body weight per day (corresponding to 46.26 mg ai/kg feed) and sacrificed at about 24 hours after the last dose.  The dosing level corresponds to ~24x the estimated dietary burden of fluopyram to beef cattle and ~14x the dietary burden to dairy cattle (see Table 8).  Total radioactivity was measured in milk (collected each morning prior to dosing and 8 hours later in the evening), plasma, and excreta at timed sampling intervals, and in the edible organs and tissues liver, kidney, muscle, and fat collected at sacrifice. The milk (pooled am and pm samples), edible organs and tissues, and excreta (urine and feces) were analyzed for parent compound and metabolites.

The overall recovery (sum of radioactivity in the excreta, milk as well as organs and tissues) was 93.5% of the total administered dose.  Up to the time of sacrifice, the excretion accounted for about 88.3% of the total dose, with 52.6% in the urine and 35.7% in the feces.  The cumulative urinary and fecal excretion rate was characterized by a linear increase during the whole testing period.

The TRR in plasma, determined over the whole testing period in timed sampling intervals, increased continuously until the test end.  During the 8-hour period after each dosing, a very significant increase was observed.  In the following 16-hour time range until delivery of the next dose, the TRR values increased or decreased only slightly.  This indicated an ongoing absorption of the test item, a rapid distribution of the systemically bioavailable compound-related radioactivity within the body, and a delayed excretion.  A plateau level of plasma concentration was not reached during the observation period.

The time course of TRR in milk samples collected 8 hours (evening milk) and 24 hours (morning milk) after each dosing was similar to that of plasma.  The radioactive residues increased continuously until the test end by a factor of 10 in total; residues did not reach a plateau in milk during the study period.  The TRR in milk samples ranged from 0.045 to 0.454 ppm, and the highest value was detected just before sacrifice.  Pooled samples of morning milk (TRR: 0.276 ppm) and evening milk (TRR: 0.228 ppm) were extracted for metabolism investigations.

In tissues, TRR were highest in liver, at 8.379 ppm.  The TRR for the other organs and tissues, in decreasing order, were 2.295 ppm in kidney, 0.737 ppm in muscle, and 0.399 ppm in fat.  In total, the compound-related residues in all samples were calculated to be about 4.6% of the total dose.

Radioactivity was extracted efficiently (97% to 99% TRR) from milk and fat using conventional solvent extraction.  For muscle, liver, and kidney, 68-77% TRR was extracted using conventional solvent extraction; an additional exhaustive extraction step with microwave assistance at increased temperature was used to solubilize an additional 17-32% TRR from these tissues.  Nonextractable residues were <=6% TRR in all goat matrices.

The major component in all the edible matrices was the metabolite AE C656948-benzamide (49.1% to 97.6% TRR).  Other major compounds identified were AE C656948-Z-olefine and the parent compound, which were found at 13.1% and 18.2% TRR, respectively, in fat.  AE C656948-Z-olefine was identified as a minor residue (<1% TRR) in milk and liver and was not found in muscle and kidney.  The parent compound was identified as a minor component in milk, liver, and kidney, at <=1.7% TRR, and was not detected in muscle. The following additional metabolites were identified as minor residues (<9% TRR each) in goat matrices:  AE C656948-benzamide-SA, AE C656948-7-OH-GA (isomers 1 and 2), AE C656948-di-OH-GA (kidney only), AE C656948-phenol-GA (liver and kidney only), AE C656948-8-OH-GA (isomer 2; liver and kidney only), AE C656948-7-hydroxy, and AE C656948-E-olefine (fat and liver only). Identification rates in milk and tissues ranged 93.5-98.9% TRR.

MRID 47372534:  A goat was orally dosed for five consecutive days, in 24 h intervals, with 2.0 mg [pyridyl-2,6-[14]C]fluopyram per kg body weight per day (corresponding to 44.62 mg ai/kg feed) and sacrificed at about 24 hours after the last dose.  The dosing level corresponds to ~23x the estimated dietary burden of fluopyram to beef cattle and ~13x the dietary burden to dairy cattle (see Table 8).  Total radioactivity was measured in milk (collected each morning prior to dosing and 8 hours later in the evening), plasma, and excreta at timed sampling intervals, and in the edible organs and tissues liver, kidney, muscle, and fat collected at sacrifice. The milk (pooled evening samples), edible organs and tissues, and excreta (urine and feces) were analyzed for parent compound and metabolites.

The overall recovery (sum of radioactivity in the excreta, milk as well as organs and tissues) was 81.9% of the total administered dose.  Up to the time of sacrifice, the excretion accounted for 81.0% of the total dose, with 52.3% found in the urine and 28.6% in the feces.  The cumulative urinary and fecal excretion rate was characterized by a linear increase during the whole testing period.

The TRR in plasma, determined over the whole testing period in timed sampling intervals followed a diurnal course.  During the 8-hour period after each dosing, a significant increase was obtained followed by a decrease measured prior to the delivery of the next dose.  This indicated rapid absorption of the test item, rapid internal distribution of the systemically bioavailable compound-related radioactivity, and rapid elimination from the body.  The radioactive residues in plasma reached a plateau level at about 50-60 hours after the first dosing.

The time course of TRR in the evening and morning milk samples after each dosing showed a similar diurnal pattern as the plasma. The radioactive residues increased significantly during the 8-hour period after each dosing followed by a decrease measured prior to the delivery of the next dose. The TRR in milk reached a plateau at about 32 hours after the first dosing.
The TRR in milk ranged from 0.017 to 0.063 ppm; the highest value was detected at 32 h after the first dose followed by a decrease to 0.026 ppm at sacrifice.  Pooled samples of evening milk (TRR:  0.053 ppm) were extracted for metabolism investigations.

In tissues, TRR were highest in liver, at 1.427 ppm.  The TRR for the other organs and tissues, in decreasing order, were 0.403 ppm in kidney, 0.372 ppm in fat, and 0.042 ppm in muscle.

Radioactivity was extracted efficiently (89% to 97% TRR) from milk (pooled evening samples), muscle, fat and kidneys by conventional solvent extraction and more than 85% TRR from each of these matrices was analyzed for metabolic profiling.  For liver, only 55% TRR was released with conventional solvent extraction; an additional 6.5% TRR was released with further solvent extraction with ACN/aqueous NH3.  An additional exhaustive extraction step with microwave assistance at increased temperature was needed for solubilization of residual radioactivity in liver which remained in the solids after conventional solvent extraction; microwave extraction released 15% TRR.  Even after this extraction step, only 76% TRR in the liver was extracted; approximately 65% TRR was analyzed by chromatography.  Nonextractable residues were <=0.035 ppm (2.9-10.8% TRR) in all goat matrices except liver.  In liver, nonextractable residues were 24.2% TRR (0.346 ppm) following microwave extraction.  Although additional microwave extraction of liver solids with ACN/aqueous NH3 solubilized all of the residues, the extracts could not be chromatographically analyzed due to low radioactivity in the fractions or because of matrix interference.

The parent compound was identified as a major component in milk, muscle, and fat (27.3-46.4% TRR) but was only found as a minor residue in liver (7.7% TRR) and was not found in kidney.  The metabolite AE C656948-Z-olefine was identified as a major metabolite in milk, muscle, and fat, at 12.9-33.7% TRR; it was found at 5.7% TRR in liver and was not found in kidney.  The other isomer AE C656948-E-olefine was found at <5% TRR in milk, muscle, fat, and liver.  AE C656948-7-hydroxy was also found to be a major component of milk, muscle, and fat, at 12.8-21.6% TRR, but a minor residue (<=6% TRR) in liver and kidney.  Other major identified metabolites included AE C656948-7-OH-GA (isomer 1), at 24.2-35.1% TRR in liver and kidney; AE C656948-7-OH-GA (isomer 2), at 16.3% TRR in kidney; and AE C656948-8-OH-GA (isomer 2) at 17.7% TRR in kidney.  These metabolites were also found at <10% TRR in milk, muscle, and liver.  AE C656948-PAA, AE C656948-hydroxyethyl-GA, AE C656948-di-OH-GA, and AE C656948-phenol-GA were only minor metabolites, each present at <=8.6% TRR in liver and kidney.

Conclusions - Goat.  The submitted goat metabolism studies are adequate to satisfy data requirements.  Based on the results of these studies, the petitioner concluded that the metabolism of fluopyram in goats proceeds via the following reactions:
   *	hydroxylation of the ethylene bridge of the molecule resulting in AE C656948-7-hydroxy, AE C656948-8-hydroxy, and a dihydroxylated compound;
   *	hydroxylation of the phenyl ring leading to AE C656948-phenol;
   *	conjugation of the hydroxylated metabolites with glucuronic acid;
   *	elimination of water from compounds hydroxylated in the ethylene bridge leading to AE C656948-Z-olefine and AE C656948-E-olefine (E- and Z-olefine can isomerize into each other);
   *	cleavage of the aliphatic chain to form AE C656948-benzamide and AE C656948-pyridyl-hydroxyethyl;
   *	hydroxylation of AE C656948-benzamide followed by conjugation with sulfate;
   *	conjugation of AE C656948-pyridyl-hydroxyethyl with glucuronic acid;
   *	and oxidation of AE C656948-pyridyl-hydroxyethyl to AE C656948-PAA.

Overall Conclusions - Livestock Metabolism.  The nature of the residue in livestock is adequately understood.  The metabolism of fluopyram in poultry and ruminants was found to be similar.  The residues of concern for tolerance enforcement are fluopyram and AE C656948-benzamide, and the residues of concern for risk assessment are fluopyram and its metabolites AE C656948-benzamide, AE C656948-E-olefine, and AE C656948-Z-olefine (ROCKS decision memo, 7/16/2009).

860.1340 Residue Analytical Methods

A summary of the analytical methods associated with this petition is presented in Table 5.

Table 5.	Summary of Residue Analytical Methods.
Matrix
                                    Analyte
                                  Method No.
                               Method principle
                                      LOQ
                            (fluopyram equivalents)
Crop commodity data generation methods Europe
Crop commodities from storage stability study; bell pepper, tomato, cucumber, melon, and strawberry samples from greenhouse studies
AE C656948
- benzamide
- PCA
- PAA[2]
- 7-hydroxy
- methyl-sulfoxide
                                     00984
                                 HPLC/MS/MS[1]
0.01 ppm for all substances,
excepted for cereal straw (0.05 ppm)
Processed commodities of apple, grape, strawberry, rape seed and tomato
AE C656948
- benzamide
- PCA
- PAA[2]
                                  00984/M001
                                 HPLC/MS/MS[1]
0.01 ppm for all substances
Crop commodity data generation method North America
All crop commodities from U.S./Canada crop field trial, processing, and field rotational crop studies
AE C656948
                                 GM-001-P07-01
                         Modification of  method 00984

                                 HPLC/MS/MS[1]
0.01 ppm
Crop commodity enforcement method + ILV
Validated using lettuce, head; oilseed rape, seeds; wheat, grain; orange, fruit; and peas, seed
AE C656948
                                     S 19
                                  DFG method
                                   GC/MSD[3]
0.01 ppm
Validated using oilseed rape, seeds; wheat, grain; orange, fruit; and cabbage, head
AE C656948
                                      ILV
                                  DFG method
                                     S 19
                                   GC/MSD[3]
0.01 ppm
Livestock commodity data generation method
Cattle and hen feeding studies
AE C656948
- benzamide
- olefine (E-isomer)[4]
- olefine (Z-isomer)[4]
                                     01061
                                 HPLC/MS/MS[1]
- 0.01 ppm for fluopyram and benzamide

- 0.02 ppm for the two olefines[4]
Livestock commodity enforcement method + ILV
Validated using cattle milk, liver, fat, muscle, and kidney;  and hen whole egg
AE C656948
- benzamide
                                     01079
                                  HPLC/MS/MS
0.01 ppm for all substances
Validated using cattle milk, liver, and muscle; and hen whole egg
AE C656948
- benzamide
                              ILV of method 01079
                                  HPLC/MS/MS
0.01 ppm for all substances
Multi-residue method
PAM Multiresidue Analysis
AE C656948
- benzamide
        PAM:  E1 extraction and DG17 detection without Florisil cleanup
                                      GC

[1]  Two different MRM transitions were measured each time.
[2]  AE C656948-PAA sodium salt injected.
[3]  Three ions were monitored in parallel (m/z 223, 173, 396).
[4]  There was inter-conversion between the E and Z isomers of AE C656948-olefine during analysis.  Following fortification with one isomer, both isomers could be separated and determined.  The amount of inter-conversion is dependent on various factors (such as matrix and concentration); therefore, a total recovery (summing both olefins) was calculated and reported.

Crop commodities

First Entry Monograph for Fluopyram, Section B.5.2.1 (MRIDs 47372603, 47372537-47372539, 47372543-47372547)

Bayer has proposed the German multiresidue method DFG Method S 19, for the enforcement of tolerances for fluopyram residues in/on crop commodities.  The method was validated using samples of orange (fruit), oil seed rape (seeds), wheat (grain), lettuce (head) and dry peas (seeds).  The method was not used for analysis in any of the storage stability, crop field trial, processing, or field rotational crop studies submitted with this petition.

DFG Method S 19:  The extraction of fluopyram from orange (fruit) was performed according to extraction module E 3, extraction module E 2 was used for wheat (grain) and dry peas (seeds), extraction module E 7 was used for oil seed rape (seeds), and extraction module E 1 was used for lettuce (head).  Briefly, samples other than oil seeds are mixed with water and then the mixture is extracted with acetone and ethyl acetate:cyclohexane (1:1, v:v).  Oil seeds are extracted with a mixture of acetone and ACN, and the extract is concentrated and redissolved in ethyl acetate:cyclohexane (1:1, v:v).  The organic phase is then cleaned up using module GPC (gel permeation chromatography); the eluate is concentrated and redissolved in ethyl acetate for analysis.  All specimens are analyzed by GC/MSD.  The ions 223, 173 and 396 (used for quantitation) were used in the validation for the determination of fluopyram using GC/MSD.  Quantification was achieved with external standards.

For fluopyram in orange fruit, rape seeds, wheat grain, lettuce (head), and dry pea seeds, the LOQ was 0.01 ppm, and the calculated limit of detection (LOD) was 0.003 ppm.  Analysis of control specimens of orange fruit, rape seeds, wheat grain, lettuce, and dry peas seeds by GC/MSD yielded no residues of fluopyram above the LOD (i.e. 0.003 ppm), indicating that no background level of fluopyram was present in the test system of the study.

The method was adequately validated in orange fruit, rape seed, wheat grain, lettuce, and dry pea seeds at fortification levels of 0.01 and 0.10 ppm.  The average recoveries were within acceptable limits (70-120%), and relative standard deviations (RSDs) were <20% for each fortification level, matrix, and analyte.  Recoveries of fluopyram ranged 68-111% using the quantitation ion (m/z 396).

The method was subjected to independent laboratory validation (ILV) using samples of cabbage (head), wheat (grain), orange (fruit) and oilseed rape (seed) fortified with fluopyram at 0.01 ppm and 0.10 ppm.  The average recoveries were within acceptable limits (70-120%), and RSDs were <20%.

Data collection methods:  Samples of crop commodities from the crop field trial, processing, and field rotational crop studies submitted to support this petition were analyzed using three HPLC/MS/MS methods.  Samples of crop commodities from the storage stability studies and the European greenhouse studies were analyzed using HPLC/MS/MS methods 00984 or 00984-M001.  Samples of crop commodities from the North American crop field trial, processing, and field rotational crop studies were analyzed using HPLC/MS/MS method GM-001-P07-01.  The main difference between the methods is the number of analytes that are determined.  HPLC/MS/MS method GM-001-P07-01 determines residues of fluopyram per se.  Method 00984-M001 determines residues of fluopyram and its metabolites AE C656948-benzamide, AE C656948-PCA, and AE C656948-PAA.  Method 00984 determines residues of fluopyram, metabolites AE C656948-benzamide, AE C656948-PCA, AE C656948-PAA, as well as rotational crop metabolites AE C656948-7-hydroxy and AE C656948-methyl sulfoxide.  The methods are summarized below.

Method 00984:  Briefly, residues of fluopyram and metabolites are extracted from crop samples by two successive extractions using a high speed blender with a mixture of ACN:water (80:20, v:v). After a centrifugation step, the extract volume is adjusted.  The extracts are then diluted ten times by adding the internal standard of each respective substance:  one dilution is performed under acidic conditions, which allows the determination of AE C656948-PCA and AE C656948-methyl-sulfoxide; and in parallel, another dilution is performed under basic conditions, which allows the determination of AE C656948, AE C656948-benzamide, AE C656948-7-hydroxy and AE C656948-PAA sodium (due to the chemical instability of AE C656948-PAA under acidic conditions).  The sample extracts are then separated by HPLC and detected by MS/MS, using one injection in positive electrospray ionization for the determination of AE C656948, AE C656948-benzamide, AE C656948-PAA sodium, and AE C656948-7-hydroxy, and another injection in negative electrospray ionization for the determination of AE C656948-PCA and AE C656948-methyl-sulfoxide under different conditions.  Quantitation is carried out by internal standardization using the following internal standards:  [phenyl-[13]C6]AE C656948; [phenyl-[13]C6]AE C656948-benzamide; [2,6-[13]C,[15]N; carboxylic acid-[13]C]AE C656948-PCA; [acetic acid-1,2-[13]C2/2-d2]AE C656948-PAA sodium; [phenyl-[13]C6]AE C656948-7-hydroxy; and [methyl-[13]CD3]AE C653948-methyl-sulfoxide.  Two mass transitions are monitored for each analyte.  Residues are expressed in parent equivalents.

The LOQ, expressed as parent fluopyram equivalents, is 0.05 ppm for each analyte in wheat straw and 0.01 ppm for each analyte in all other tested sample materials (head lettuce, rape seed, wheat grain, and orange fruit).  The calculated LOD ranged from 0.001 to 0.006 ppm (0.002 to 0.011 ppm for wheat straw).  Apparent residues in the control samples tested were below 0.3xLOQ (at the MRM transition used for quantitation).

The method was validated in lettuce (head), rape seed, wheat grain, wheat straw, and orange fruit by fortifying samples with each analyte at the LOQ and 10xLOQ.  The average recoveries were within acceptable limits (70-120%) and RSDs were <20% for each fortification level, matrix, and analyte using both the quantitation ion and the confirmation ion for all analytes except AE C656948-PCA and AE C656948-methyl-sulfoxide.  For AE C656948-PCA in rape seed and AE C656948-methyl-sulfoxide in all tested crops but orange, an interference prevented quantitation at the LOQ at the confirmatory ion transition in all matrices.  Recoveries from all matrices at the quantitation ion ranged 68-122% for fluopyram (1 recovery <70% and 1 recovery >120%), 83-116% for AE C656948-benzamide, 61-108% for AE C656948-PCA (1 recovery <70%), 70-102% for AE C656948-PAA, 80-117% for AE C656948-7-hydroxy, and 81-118% for total residues of AE C656948-methyl sulfoxide.

Adequate concurrent method recovery data for fluopyram, AE C656948-benzamide, AE C656948-PCA, and AE C656948-PAA in lettuce, dry pea seed, orange, rape seed, and wheat grain, and for AE C656948-7-hydroxy in lettuce, dry pea seed, rape seed, and wheat grain were submitted for HPLC/MS/MS Method 00984 with the storage stability study, reflecting fortification levels of 0.01 and 0.2 ppm for each analyte in each commodity.  In addition, adequate concurrent method recovery data were provided for fluopyram, AE C656948-benzamide, AE C656948-PCA, and AE C656948-PAA in bell pepper, tomato, cucumber, melon, and strawberry with the European greenhouse trials; samples were fortified with each analyte at levels ranging 0.01-2 ppm.

Method 00984-M001:  Briefly, residues of fluopyram and metabolites AE C656948-benzamide, AE C656948-PCA, and AE C656948-PAA are extracted from crop samples other than oil using a high speed blender (one extraction) with a mixture of ACN:water (80:20, v:v); residues are extracted from rapeseed oil using liquid/liquid partition with ACN and n-hexane. The ACN/water or ACN extract is filtered, and the volume is adjusted with water (all commodities except rapeseed oil) or water and ACN (rapeseed oil).  For the determination of AE C656948, AE C656948-benzamide, AE C656948-PAA, the extracts are diluted under basic conditions and the internal standards are added.  For the determination of AE C656948-PCA in/on crop commodities other than rape press cake, an aliquot of the extract is acidified and cleaned up by liquid/liquid partition with methyl-t-butyl-ether (MTBE, dilution 2 times).  An aliquot of the MTBE extract is evaporated to dryness and dissolved in the corresponding internal standard solution.  For the determination of AE C656948-PCA in/on rape (press cake), an aliquot of the extract is cleaned up on a Varian BondElut-ENV column and an aliquot of the eluate is diluted with the corresponding internal standard solution and ACN:water (30:70, v:v).

The sample extracts are then separated by HPLC and detected by MS/MS, using one injection in positive electrospray ionization for the determination of AE C656948, AE C656948-benzamide, and AE C656948-PAA sodium, and another injection in negative electrospray ionization for the determination of AE C656948-PCA under different conditions.  Quantitation is carried out by internal standardization.  Two mass transitions are monitored for each analyte.  Residues are expressed in parent equivalents.

The LOQ, expressed as parent fluopyram equivalents, is 0.01 ppm for each analyte in all tested sample materials.  Apparent residues in control samples using the quantitation MRM transitions were generally below 0.3xLOQ, with the exception of AE C656948-benzamide in grape juice (0.004 ppm) and fluopyram in apple dried pomace (0.003 ppm).  It was therefore recommended to use both MRM transitions for analysis and in case of a problem to use the second MRM transition for the quantitation.

The method was validated in apple (fruit, pomace dried, fruit dried, peel rest and sauce), tomato (fruit, juice, puree, and preserve), round cabbage (head and cooking water), grape (bunches of grape, juice, pomace dried, must, and wine), rape (seed, oil, pomace and press cake) and strawberry (fruit, preserve, and jam) by fortifying samples with each analyte at the LOQ and 10xLOQ.  The mean recoveries, for each fortification level, for each matrix, and for all analytes, were within 70-120% with the following exceptions:  AE C656948-PCA in grape juice:  at both 0.01 ppm and 0.1 ppm, a mean recovery of 65% was obtained, with RSD of 11.6% and 0.0% respectively; and AE C656948-PCA in grape must:  at 0.01 ppm, a mean recovery of 61% was obtained, with a RSD of 10.9%.  Relative standard deviations were below 20% for all analytes and sample materials using the quantitation ion, with the exception of fluopyram in grape pomace dried, at the level 0.01 ppm (RSD = 22.3%).

Recoveries from all matrices ranged from 71-122% for fluopyram (1 recovery >120%), 84-115% for AE C656948-benzamide, 54-107% for AE C656948-PCA (9 recoveries <70%, for grape juice, dried pomace, and must), and 65-118% for AE C656948-PAA (2 recoveries <70%).

Adequate concurrent method recovery data for bell pepper and tomato were submitted for HPLC/MS/MS Method 00984-M001 with the European greenhouse submissions, reflecting fortification levels of 0.01-1.0 ppm for fluopyram, AE C656948-benzamide, AE C656948-PCA, and AE C656948-PAA in each commodity.

Method GM-001-P07-01:  Briefly, residues of fluopyram are extracted from homogenized plant matrices using ACN:water (4:1, v:v) by two successive extractions in a high speed blender for at least one minute each.  The combined filtrate is mixed with an isotopically labeled internal standard ([phenyl-[13]C6]AE C656948) and mixed well.  In an additional modification to the original method, an aliquot of the sample extract is passed through a small C18 SPE cartridge, and the eluate is diluted with 0.1% aqueous acetic acid for HPLC/MS/MS analysis.  Quantitation is carried out by internal standardization; the mass transition monitored is m/z 397.10 to 172.95.  The stated LOQ is 0.01 ppm.

No method validation data were provided for this method.  Adequate concurrent method recovery data were submitted for this method with each of the North American crop field trial and processing studies, from crop samples fortified at the LOQ and at least one additional fortification level.  The fortification levels adequately reflected the observed residues in/on all tested raw agricultural and processed crop commodities.

Radiovalidation data:  Bayer submitted radiovalidation data for Method 00984-M001 using samples of grape from the grape metabolism studies.  More than 97% TRR (total radioactive residues) were extracted from [[14]C]fluopyram-treated grapes using the extraction procedures of Method 00984-M001.  The combined residues of AE C656948, AE C656948-benzamide, and AE C656948-pyridyl carboxylic acid accounted for >93% TRR when grape samples were analyzed using the residue analytical method.

No radiovalidation data were provided for Methods 00984 and GM-001-P07-01.  The petitioner requested a waiver of radiovalidation data requirements for these methods, with the following justification.  The extraction procedure used in the residue analytical methods 00984 and GM-001-P07-01 comprised two sequential extractions with ACN:water (8:2; v:v).  The samples of the metabolism studies were also extracted with ACN:water (8:2; v:v):  for the potato, bean, and red pepper metabolism studies, each crop matrix was extracted three times with ACN:water (8:2; v:v); and the samples from the confined rotational crop studies (Swiss chard, turnip roots, turnip leaves, and wheat forage, hay, straw, and grain) were extracted three to four times with ACN:water (8:2; v:v).

The first two extraction steps with ACN:water (8:2; v:v) released:
   *	>89% TRR in all RACs of the target crop metabolism studies
   *	>91% TRR in turnip leaves, turnip roots, Swiss chard, and wheat forage of the confined rotational crop (CRC) studies
   *	>84% TRR in wheat hay and straw of the CRC studies
   *	>=70% TRR in wheat grain of the CRC studies

For wheat hay, grain, and straw, additional radioactivity was released with additional extractions (using microwave conditions for hay and straw or diastase enzyme for grain).  For hay and straw, the microwave extracts showed the same metabolic profiles as the conventional extracts.  For grain, the released radioactivity was characterized as natural compounds (e.g., starch).

No radiovalidation data were submitted for the proposed enforcement method, DFG S 19.  However, the petitioner submitted bridging data comparing the extraction procedures of the data collection methods with the extraction procedures of the proposed enforcement method.  Samples of tomato from the U.S. field trials were separately extracted using acetone (the extraction solvent of the proposed enforcement method) and ACN:water (4:1, v:v; the extraction solvent of the data collection methods).  The extraction of aged fluopyram residues from tomatoes using acetone yielded 101% (average of four samples) of the residues that were extracted using ACN:water (4:1, v:v;), indicating that blending a crop sample in acetone is acceptable for complete extraction of fluopyram residues.

Conclusions.  The submitted residue analytical method data are adequate to satisfy data requirements for crop commodity methods.  Adequate validation data have been provided for the methods used for data collection and the proposed enforcement method.  Adequate ILV data have been submitted for the proposed enforcement method.  Radiovalidation data are not needed since the solvents used in the enforcement and data gathering methods extract the bulk of the TRR in the plant metabolism studies and aged residues from crop field trials.  Because three ions are monitored by GC/MSD in the proposed enforcement method, confirmatory analysis procedures are not needed.  The DFG Method S 19 will be forwarded to FDA for publication in PAM Vol. II.

Livestock commodities

First Entry Monograph for Fluopyram, Section B.5.2.2 (MRIDs 47372540-47372542)

Bayer has proposed an HPLC/MS/MS method for the enforcement of tolerances for fluopyram residues in livestock commodities.  Method 01079 determines residues of fluopyram and its metabolite AE C656948-benzamide in livestock tissues, milk, and eggs.

Method 01079:  Briefly, residues of fluopyram and AE C656948-benzamide are extracted from milk, fat, and egg using a mixture of ACN:water (4:1, v:v).  For muscle, kidney, and liver, samples are extracted twice with ACN:water (4:1, v:v) using microwave heating (120 ºC for 20 minutes).  After filtration, the extracts are cleaned up on a C18 cartridge.  Internal standards of [phenyl-[13]C6]fluopyram and [phenyl-[13]C6]AE C656948-benzamide are added when quantitation is to be performed against internal standard, and the extract is diluted to volume with ACN:water (4:1, v:v).  Aliquots of the extracts are further diluted with a mixture of methanol:water (1:9, v:v) containing 10 mM ammonium formate and 120 μL/L formic acid.  The sample extracts are then analyzed by HPLC/MS/MS.  The residues are determined by external calibration using internal standards or matrix-matched standards.  Two mass transitions are monitored for each analyte.  Residues are expressed in parent equivalents.

The LOQ is 0.01 ppm and the calculated LOD is 0.003 ppm for each analyte in each matrix.  The petitioner stated that residues in control samples were well below 30% of the respective LOQ level.

The method was adequately validated using cattle milk, fat, muscle, liver, and kidney, and hen whole egg fortified with fluopyram and AE C656948-benzamide, each at 0.01 and 0.10 ppm.  The average recoveries were within acceptable limits (70-120%), and RSDs were <20% for each analyte in each matrix using both internal standards and matrix-matched standards.  Recoveries from all matrices using the quantitation ion (both internal and matrix-matched standards) ranged from 69-105% for fluopyram (1 recovery <70%) and 74-108% for AE C656948-benzamide.

The method was subjected to ILV using samples of milk, eggs, beef muscle, and beef liver fortified with fluopyram and AE C656948-benzamide, each at 0.01 and 0.10 ppm.  The average recoveries were within acceptable limits (70-120%), and RSDs were <20% for each analyte in each matrix using both internal standards and matrix-matched standards.

Data collection method:  Samples of livestock commodities from the cattle and hen feeding studies submitted with this petition were analyzed for residues of fluopyram and its metabolites AE C656948-benzamide, AE C656948-olefine (E-isomer) and AE C656948-olefine (Z-isomer) using HPLC/MS/MS Method 01061.  The method is very similar to Method 01079; the main difference is that Method 01061 includes determination of AE C656948-olefines.

Briefly, residues of fluopyram, AE C656948-benzamide, AE C656948-olefine (E-isomer), and AE C656948-olefine (Z-isomer) are extracted from milk, cream, skim milk, hen muscle, hen liver, cattle and hen fat, and egg using ACN:water (4:1, v:v).  For cattle muscle, cattle kidney, and cattle liver, samples are extracted twice with ACN:water (4:1, v:v) using microwave heating (120 ºC for 20 minutes).  After filtration, the extracts are cleaned up on a Mega Bond Elut-C18 cartridge.  Internal standards are added, and the extract is diluted to volume with ACN:water (4:1, v:v).  Aliquots of the extracts are further diluted with a mixture of methanol:water (1:9, v:v) containing 10 mM ammonium formate and 120 μL/L formic acid.  The sample extracts are then analyzed by HPLC/MS/MS.  The residues are determined by external calibration using internal standards.  Internal standards of [phenyl-[13]C6]fluopyram and [phenyl-[13]C6]AE C656948-benzamide are used; for AE C656948-olefine (E-isomer) and AE C656948-olefine (Z-isomer), the fluopyram internal standard was used for quantitation.  Two mass transitions are monitored for each analyte.  Residues are expressed in parent equivalents.

The validation of the two individual olefine isomers, AE C656948-olefine (E-isomer) and AE C656948-olefine (Z-isomer) was not possible due to not repeatable internal conversion (E <--> Z). Therefore, the calculated residue results are presented as the sum of the two isomers (expressed as parent equivalents).

The LOQ is 0.01 ppm for fluopyram and AE C656948-benzamide in all matrices and 0.02 ppm for the total residues of AE C656948-olefine (E-isomer) and AE C656948-olefine (Z-isomer) in all matrices.  The LOD was estimated to be 0.003 ppm for fluopyram, 0.001 ppm for AE C656948-benzamide, and 0.004 ppm for the total residues of AE C656948-olefine (E-isomer) and AE C656948-olefine (Z-isomer) in all matrices.

The method was validated in cattle milk, milk fat, skim milk, liver, muscle, kidney, and fat, and hen whole egg, egg yolk, egg white, muscle, and liver by fortifying each matrix with each analyte at 0.01 ppm and 0.10 ppm.  The average recoveries were within acceptable limits (70-120%), and RSDs were <20% for each fortification level, analyte, and matrix.  Recoveries from all matrices using the quantitation ion ranged from 85-104% for fluopyram, 86-109% for AE C656948-benzamide, and 68-109% for total residues of AE C656948-olefines (E- and Z-isomers; 1 recovery <70%).

Extraction efficiency/Radiovalidation:  The petitioner provided data demonstrating the extraction efficiency of the livestock commodity methods using samples from the hen and goat metabolism studies reflecting dosing with [phenyl-UL-[14]C]fluopyram.  The extraction procedures of the livestock commodity analytical methods are very similar to those used in the metabolism studies.  For matrices other than goat muscle, liver, and kidney, the petitioner provided data showing the %TRR extracted from each matrix in the metabolism studies with a single extraction (the procedures of the analytical method).  For goat muscle, liver, and kidney, where the extraction procedures differed from the ones used in the analytical method, radiovalidation experiments were performed within the metabolism study, using the procedures of the analytical method.  The extraction efficiencies ranged from 84-97% for all matrices.  These data are adequate to satisfy radiovalidation data requirements for the livestock commodity methods.

Conclusions.  The submitted data are adequate to satisfy data requirements.  For the livestock commodity methods, adequate validation data were provided for each analyte.  The enforcement method includes adequate confirmatory procedures.  Adequate data have been provided to satisfy radiovalidation requirements.

860.1360 Multiresidue Methods

First Entry Monograph for Fluopyram, Section B.5.2 (MRID 47372548)

Bayer CropScience submitted data on the testing of fluopyram and its metabolite AE C656948-benzamide using the multiresidue method protocols of FDA PAM Volume I, third edition.  Fluopyram and AE C656948-benzamide were tested according to Protocols A, C, D, and F of the FDA PAM Vol. I testing procedures.

Protocol A of the PAM Vol. I testing procedures is not suitable for the detection of either fluopyram or AE C656948-benzamide because neither compound is an N-methyl carbamate and neither compound is naturally fluorescent.  Because the compounds are not acids or phenols, they were not tested under Protocol B.  Protocol C was tested as per protocol guidelines.  Fluopyram and AE C656948-benzamide (DG7 Level II) were deemed suitable for GC analysis and further testing using Protocols D and F was conducted according to the decision tree.  Protocol G was not tested because the compounds are not substituted ureas.

Using Protocol D, adequate recovery of fluopyram was obtained using E1 extraction with no Florisil cleanup; recoveries were 90-111% from samples of whole apple fortified at 0.10 and 0.80 ppm.  However, fluopyram could not be recovered through the Florisil column cleanup procedures; therefore, Protocols E and F are not appropriate for fluopyram.

AE C656948-benzamide could not be recovered through the Florisil column cleanup procedures of Protocol D or F; therefore, no further testing of this compound was conducted.

The data indicate that the FDA multiresidue methods are suitable for detection and enforcement of fluopyram in non-fatty matrices by Protocol D (Section 302), using E1 extraction and DG17 detection without Florisil cleanup, but are not suitable for detection of AE C656948-benzamide residues.

Conclusions.  The submitted data are adequate to satisfy data requirements.

860.1380 Storage Stability

First Entry Monograph for Fluopyram, Section B.7.6.2 (MRIDs 47372549, 47372550, 48239911, 48239912, and 48239913)

Bayer CropScience submitted three storage stability studies: one study with head lettuce, wheat grain, dry pea seed and rape seed; one study with orange fruit; and one study with dry pea, rape seed and orange fruit.

One study was conducted to investigate the stability of residues of fluopyram and its metabolites AE C656948-benzamide, AE C656948-PCA, AE C656948-PAA, and AE C656948-7-hydroxy under freezer conditions at ~<=-18 °C.  Samples of lettuce (head), wheat grain, dry pea seed, and rape seed were spiked with 0.20 ppm each of fluopyram, AE C656948-benzamide, and AE C656948-PAA (as the sodium salt); samples of dry pea seed and rape seed were spiked with 0.20 ppm of AE C656948-PCA; and samples of lettuce and wheat grain were spiked with 0.20 ppm of AE C656948-7-hydroxy.  The fortified samples were stored in plastic containers at ~<=-18 °C for nominal storage intervals of 0, 3, 6, 13, 18, 24 and 36 months.

After a deep-freezer storage period of 36 months, the average corrected recovery values (corrected for average concurrent recovery) ranged from 106-110% for fluopyram, 93-99% for AE C656948-benzamide, 97-106% for AE C656948-PCA, 79-102% for AE C656948-PAA and 102-107% for AE C656948-7-hydroxy.  These data indicate that fluopyram and its metabolites AE C656948-benzamide and AE C656948-PAA are stable for at least 36 months at ~<=-18 °C in/on lettuce, wheat grain, rape seed, and dry pea seed; that metabolite AE C656948-PCA is stable for at least 36 months at ~<=-18 °C in/on dry pea seed and rape seed; and that metabolite AE C656948-7-hydroxy is stable for at least 36 months at ~<=-18 °C in/on lettuce and wheat grain (see Table 6).

Another study was conducted to investigate the stability of residues of fluopyram and its metabolites AE C656948-benzamide, AE C656948-PCA, and AE C656948-PAA in orange under freezer conditions at ~<=-18 °C.  Samples of orange fruit were spiked with 0.20 ppm of each analyte and stored in plastic containers at ~<=-18 °C for nominal storage intervals of 0, 4, 6, 12, 18, 24 and 36 months.  Mean recovery values after corrected for mean concurrent recoveries were 100% for AE C656948, 103% for AE C656948-benzamide, 102% for AE C656948-PCA, and 60% for AE C656948-PAA.  Other than AE C656948-PAA, recoveries measured for the remaining 3 compounds at the 7 storage intervals in orange were all >90%.  For AE C656948-PAA, recoveries declined gradually from103% at 0 month to 69% at 24 months and 60% at 36 months.

Also submitted were stability data for AE 1344122 (AE C656948-methyl-sulfoxide) and BCS-AA 10065 (AE C656948-7-hydroxy) in dry pea seed, rape seed and orange under deep freeze storage for up to 24 or 25 months.  Samples of dry pea seed, rape seed and orange fruit were spiked with 0.20 ppm of each analyte and stored at ~<=-18 °C for nominal storage intervals of 0, 4, 6, 12, 18, and 24 months.  Mean recovery values after corrected for mean concurrent recoveries ranged from 92-93% for the methylsulfoxide metabolite and from 97-102% for the 7-hydroxy metabolite.  These data indicate that AE 1344122 and BCS-AA 10065 are stable for at least 24 months under frozen conditions in dry pea seed, rape seed and orange fruit.

At each storage interval, residues of fluopyram and its metabolites were determined using high performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) Method 00984.  The mean of the concurrent recoveries were within the acceptable range of 70-110% for all matrices and for all fortification levels.  In the control samples, the residues of all compounds were below the LOQ (<0.01 mg/kg for each analyte expressed as AE C656948 parent).

Table 6.                Storage Stability of Fluopyram and its Metabolites in Months

Head lettuce
- Water
Wheat grain
- Starch
Dry pea
- Protein
Rape
- Oil
Orange
- Acid
Fluopyram
                                      36
                                      36
                                      36
                                      36
                                      36
Benzamide
                                      36
                                      36
                                      36
                                      36
                                      36
PCA

                                      36
                                      36
                                      36
PAA
                                      36
                                      36
                                      36
                                      36
                                      24
7-OH
                                      36
                                      36
                                      24
                                      24
                                      24
Sulfoxide

                                      24
                                      24
                                      24

The storage durations and conditions of samples from the crop field trial, processing and field rotational crop studies submitted to support this petition are presented in Table 7.

Table 7.	Summary of Storage Conditions and Durations of Samples from Crop Field Trial, Processing, and Field Rotational Crop Studies.
Matrix
                          Storage Temperature  (°C)
                        Actual Storage Duration (days)
Primary Crop Raw Agricultural Commodities[1]
Potato, tuber
                                     <0
                                    312-623
Sugar beet, root
                                     <0
                                    338-573
Sugar beet, tops
                                     <0
                                    498-573
Dried bean, seed
                                     <0
                                    331-439
Soybean, seed
                                     <0
                                    249-518
Soybean, forage
                                     <0
                                    490-581
Soybean, hay
                                     <0
                                    498-588
Cucurbit vegetable, group 9
  Cucumber
                                     <0
                                    146-280
  Cucumber - greenhouse (Europe)
                                     <0
                                    176-338
  Muskmelon
                                     <0
                                    160-219
  Muskmelon - greenhouse (Europe)
                                     <0
                                    119-313
  Summer squash
                                     <0
                                    203-282
Apple
                                     <0
                                    64-451
Stone fruit, group 11
  Cherry
                                     <0
                                    528-562
  Peach
                                     <0
                                    179-546
  Plum
                                     <0
                                    290-543
Grape
                                    <-15
                                    49-401
Strawberry
                                     <0
                                    78-231
Strawberry - greenhouse (Europe)
                                    <=-18
                                    217-271
Tree nut, group 14
  Almond
                                     <0
                                    591-655
  Almond, hull
                                     <0
                                    576-640
  Pecan
                                     <0
                                    501-564
Cereal grain, group 15, except rice
  Corn, field, grain
                                     <0
                                    214-390
  Sorghum, grain
                                     <0
                                    258-374
  Wheat, grain
                                     <0
                                    23-411
  Corn, sweet, K+CWHR
                                     <0
                                    373-437
Forage, fodder and straw of cereal grain, group 16, except rice
  Corn, field, stover
                                     <0
                                    248-372
  Corn, forage
                                     <0
                                    238-409
  Sorghum, forage
                                     <0
                                    263-386
  Sorghum, stover
                                     <0
                                    285-391
  Wheat, forage
                                     <0
                                    56-380
  Wheat, hay
                                     <0
                                    62-453
  Wheat, straw
                                     <0
                                    50 -411
Banana
                                     <0
                                    234-343
Canola, seed
                                     <0
                                    151-517
Peanut
                                     <0
                                    367-593
Peanut, hay
                                     <0
                                    538-583
Processing/Residue Reduction Studies
Apple processed commodities
                                    <-15
                                    233-239
Canola processed commodities
                                     <0
                                    18-158
Corn processed commodities
                                     <0
                                     47-66
Cotton processed commodities
                                     <0
                                     8-11
Peanut processed commodities
                                     <0
                                    33-112
Plum processed commodities
                                    <-13
                                    282-285
Potato processed commodities
                                     <0
                                    308-312
Soybean processed commodities
                                    <-4
                                     34-61
Sugar beet processed commodities
                                     <0
                                    98-101
Wheat processed commodities
                                     <0
                                     4-17
Field Rotational Crop Studies
Alfalfa, forage
                                     <0
                                    104-328
Alfalfa, hay
                                     <0
                                    104-325
Cotton, undelinted seed [2]
                                     <0
                                    140-220
Cotton, gin byproducts
                                     <0
                                    179-220
Mustard greens
                                     <0
                                    333-405
Turnip, tops
                                     <0
                                    329-405
Turnip, roots
                                     <0
                                    332-405
Wheat, forage
                                     <0
                                    389-405
Wheat, hay
                                     <0
                                    331-353
Wheat, straw
                                     <0
                                    309-329
Wheat, grain
                                     <0
                                    304-325
[1]  From crop field trial and processing studies.
[2]  From field rotational crop and processing study.

Conclusions.  The submitted data are adequate to satisfy data requirements.

The crop commodities used in the storage stability studies represent commodities containing water (lettuce), starch (wheat grain), protein (dry pea seed), oil (rape seed), and acid (orange).  It is concluded that, upon frozen storage, residues of fluopyram and its metabolite AE C656948-benzamide are stable for at least 36 months in water-, starch-, protein-, oil-, and acid-containing materials, and residues of AE C656948-PCA and AE C656948-methyl-sulfoxide are stable for at least 36 and 24 months, respectively, in protein-, oil-, and acid-containing materials.  It is also concluded that residues of AE C656948-PAA are stable for at least 36 months in water-, starch-, protein-, and oil-containing materials, and for 24 months in acid-containing materials.  Furthermore, residues of AE C656948-7-hydroxy are stable for at least 36 months in water- and starch-containing materials and for at least 24 months in protein-, oil-, and acid-containing materials.

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

First Entry Monograph for Fluopyram, Sections B.7.8.1 and B.7.8.2 (MRIDs 47372601 and 47372602)

There are livestock feedstuffs associated with the 2011 revised proposed uses of fluopyram, and the following ones are considered important:  alfalfa hay, alfalfa meal, almond hulls, canola meal, field/sweet corn forage/silage, sorghum grain, sugar beet molasses, and wheat milled byproducts (email, J. Stokes, 5/16/2011).  In estimating the livestock dietary burdens, residue values in alfalfa hay and meal are based on the extensive field rotational crop data, and almond hulls and sugar beet molasses are based on the foliar (primary) crop data.  Since the proposed label permits limited crop rotation to canola, cereal grains, corn, and soybean with which there are feed items, and since extensive field rotational crop data are not available, the residue values used for canola meal, field/sweet corn forage/silage, sorghum, and wheat milled byproducts are based on half of the primary crop values (see ChemSAC minutes, 6/29/2011 and p. 88): 2.76 ppm for field corn forage/silage (see Appendix III, Figure III-18: maximum residue=5.52 ppm), 1.62 ppm for sorghum grain, 0.30 ppm for wheat milled byproducts, and 0.075 ppm for canola meal (Appendix III, Figure III-2; since this is a blended item, the average value of 0.51 ppm in seed was used to estimate residue level in canola meal, adjusted with a processing factor of 0.3x as indicated in Table 26).  As a result, the estimated dietary burdens of fluopyram to livestock, as presented in Table 8, are conservative.  The estimated dietary burdens are 1.90 ppm for beef cattle, 3.39 ppm for dairy cattle, 1.24 ppm for poultry, and 1.33 ppm for swine.  Bayer submitted feeding studies with cattle and poultry, which are summarized below.

Table 8.	Calculation of Dietary Burdens of Fluopyram Residues to Livestock.
Feedstuff
                                    Type[1]
                                % Dry Matter[2]
                                   % Diet[2]
                             Maximum Residue (ppm)
                         Dietary Contribution (ppm)[3]
Beef Cattle: 15% R, 80% CC, 5% PC
Corn, field, forage/silage
                                       R
                                      40
                                      15
                                     2.76
                                     1.04
Sorghum, grain, grain
                                      CC
                                      86
                                      40
                                     1.62
                                     0.75
Wheat, milled byproducts
                                      CC
                                      88
                                      30
                                     0.30
                                     0.10
Beet, sugar, molasses
                                      CC
                                      75
                                      10
                                     0.026
                                    0.0035
Canola, meal
                                      PC
                                      88
                                       5
                                     0.075
                                    0.0043
TOTAL BURDEN
                                      --
                                      --
                                      100
                                      --
                                     1.90
Dairy Cattle: 45% R, 45% CC, 10% PC
Almond, hulls
                                       R
                                      90
                                       5
                                     6.12
                                     0.34
Corn, sweet, forage/silage
                                       R
                                      48
                                      40
                                     2.76
                                      2.3
Sorghum, grain, grain
                                      CC
                                      86
                                      35
                                     1.62
                                     0.66
Beet, sugar, molasses