### UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON , D

. C

. 20460 OFFICE O F CHEM IC A L SA FET Y AND POLLUTION PREVENTI ON
MEMORANDUM Date: 09/ 1 7 / 20 1 5 Su bj ect: Pronamide (Propyza mid e). Su mm ary of Analytical Ch e mi s try and R es idue Data to Support Re g i s tration Review.

Fro m : PC Code: 1 0170 1 Deci sio n No.: 476677 Petiti on No.: NA Risk Assessment Type: NA TX R No.: NA MRlD No.: See Below DP Barcode: D42708 I and 04 279 14 Registratio n No .: NA Regulato ry Action: Reg istration Re v iew Case No.: 4092 C AS No.: 23950-58-5 40 CF R : 180 .317

### ~0 ldeli z Neg r6 n- En carnac i 6 n , Ph.D. , C he mi s t**~~** Ri s k Assess me nt Bran ch V Health Effects Di v is ion ( HED ; 7509P) Throug h: Danett e Drew, Se nior C h emist

## ~-**~** Risk Assess me nt Bran ch V ( HED ; 7509P ) and Thomas B loe m ,

### m~**~~** Elizabeth H o lm a n , Ph.D. , Chem is t

### ~ft"- 

## »-lScienc e A dvi so ry Co uncil for C h e mi s try (C hemSAC) HED (7509P)

To: Kev in Coste ll o , Ri s k Manager Pro duct Re-reg is tration Di v i sion (7505 P)

Summary of S ubmitted R es idu e C hemistry S tu dies OCS PP 860 MRID Number Tit l e Series Guide lin e 860. 1340 46413406 Recd , R

. (2002) I nd e p end ent L abora tory Val id at ion of Dow Ag r oScic nc es LLC Method GR M 02.2 1

- Det erm inati o n of R esidues of Prop yzamide in Anima l Ti ss ue s by Gas Ch r omat og raph y with Nega ti ve- I on C h em ical Io nization Mass Spec trom e try. 860.1380 4965440 1 I luang. R .: Fleming. C. (2005) Waiver R eques t for a r:rozen S torage Stab ility S tudy on Mc1abolites of Pronamid e.

Page I of20Pronamide Summary of Analytical Chemistry and Residue Data Barcode 0427081



### Tab l e of Co nte nts 1 .0 Exec ut i ve S um1n ary .......................................... .. .. .. .. .. .. .. .. .. ... . . ... ............................ .. ..... .. . .. 3 2.0 Regu latory Recommendations

............................................................................................ 4 2.1 D ata Deficiencie s / Data Needs

....................... .. ...... .. .. .. .. .. ............................. . ................... 4 2

. 2 Tolerance Considerations

................................................................................................. 5 2.2.1 Enforcement Analytical Method

............................................................... .. .............. 5 2

. 2

. 2 Recommended Tolerances

........................................................................................ 5 2.2.4 Internat ional Harmonization

.............. .. .. .. .. .. .. .. .. .. . ... . .. .. .. ............................. .. ........... 7 3.0 In troduct ion .............. .. . .. .. .. .. . .. .. .. ... .......... .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. ...................... .. .. .. . .. . .. .. . ..... 8 3. 1 C hem ical Ide nti ty
.. . .. . ... . ............................... . .... . .. .. .. .. . ... .. . .............................. . .. .. ............ 

## g 3.2 Physica l / Chemical Characteristics

................................................................................... 8 3.3 Pes ti c id e Use Pattern/Directions for Use (860.1200)

....................................................... 9 4.0 Metabolite/ Degradate Residue Profile

................. .. .. .. .. .. ................................................... I 0 5.0 Residue Profil e .................................................................................................................. 11 5. 1 Res idu e A n a l yti cal Methods (860.1340)

.. .. .. .. .. ................. ........................ ...... .. ............. 11 5.1. 1 Mu lt i -Residue Methods (860. 1 360)

........................................................................ 11 5.1.2 T o le ranc e E nforc e ment Methods

............................................................................ 11 5.1.4 Submi tt a l of Ana lytical Reference Standards (860 .1650 )

...................................... 13 5

. 2 Storage Stabi l ity (860.13 80 )

............... .. . ..................................................... ................... 1 3 5.3 Re sid ue D ata
..... ....................................................... .. .. ...... . .. ......................................... 15 5.3 .1 Crop Fi e ld T ri a ls (860.1500)

.................... .................... .......................................... 15 5.3

. 2 Fi e ld Rotat io n al Crops (860.1900)

............................................ .. .. . .. ............... ....... 16 5

. 3

. 3 Proce ssed Food and Feed (860.1520)

............................................... .......... .. ........ .. 1 6 5.3.4 Meat , Mi l k , Poultry and Eggs (860.1480)

.......... .. .................................. . ............... 16 5.3.4.1 D ietary Burden

. ... ............................................................................................ 16 5

. 3

. 5

. Fo od Hand lin g (860 .1 460)

...................................................................................... 17 5.3.6 Water , F is h , and Ir rigated Crops (860 .1 400)

...... ...... . ................ ............................. 17 5

. 4 F ood R es idue Profile

.......... ...... .. .. .. .. ........................................................ .. . .. .. .. . .. .......... 18 Appe ndi x A. Inte rn at ional R es idu e Limit s Tab le ................ .. ...................................................... 19

Pa ge 2 of20Pronamide Summary of Anal ytical Chemistry and Residue Data Barcode 0427081


### 1.0 Exec utiv e S ummary This residue chemistry chapter was prepared to support registration review of pronamide, and reflect c hang es in the residue c he mi stry database since the 2009 scop ing docwn e nt. Pronamid e , also call ed propyzamide , [3 , 5-d ich loro- N-( I , I -dimethyl-2-propynyl)benzamide] is a se lective , systemic , pre-and post-emergence herbicide registered for the control of grasses and broadleaf weeds. It is a soi l active systemic herbicide w ith uptake by s u sceptible weeds occurring through the roots. The qualitative na ture of the residue in primary crops ( i. e. alfalfa and lettuce) as well as rotational crops is understood. The terminal residues of concern in plants and li ves tock commodities, and in drinking water , are pronamide and its metabolites co ntainin g the 3 , 5- dichlorobenzoyl moiety. Tolerances are estab li shed in 40 CF R §180.3 17 (a) , (c) and (d) for the combined res idues of propyzamide and its metabolites containi ng the 3 , 5 -dichl orobenzoy l moiety (calc ulated in parent eq uivalent s)

. Pronamide tolerances li s ted in section (a) range from 0.02 ppm for certain livestock co mmoditi es to 1 0.0 ppm for non-grass animal feeds group 18 and alfalfa seed. Regional tolerances are l isted in section (c) for pronamide o n field pea seed (0.05 ppm) and rhubarb (0. 1 ppm). To lerances are listed in section (d) for inadvertent residues of pronamide on cerea l grain forage, hay and straw, group 1 6. Adequate e nforc eme nt method s are avail able for the determination of residues in/on plant and liv estock commodi ti es. The H E D scoping document for pronamide registration review includes a summary of the deficiencies of the residue chemistry database ( 0. Drew , 0 363904, 06/17/ 2009). The data deficiencies that were identified included th e following: ( 1) Additional residue data fo r dri ed winter peas , and the vines and ha y of winter peas were required. A magnitude of residue study has s ince been submitted and reviewed (MRID 46376601 ). (2) Removal of a tolerance for leaf lettuce was required. (3) Further opti mi ze/ improve the revised livestock enforcement method (TR 34-91-68) to y ie ld acceptab le recoveries at a fortificat ion l eve l eq ual to es tabli s h ed liv es to ck tolerances, and to submit bridging independent laboratory validation data. A li vestock ana l ytical method study was submitted (MRI D 46413406) and forwarded to EPA ' s Analytica l Chemistry Branch on 6118/ 07 for a Tolerance Method Va l idation (TMV). (4) Additional confirmatory storage stabi li ty data for the regulated pronamide metabolites on a l falfa, app les , grapes, lettuce , and peaches or p l um s were requ ired but have not ye t been s ubmitted. Interregional Research Project No. 4 has submitted crop fie ld trial data for winter peas (I. Negr6n-Encarnaci6n , 0383372 , 09/17/2015). The s ubmitted magn i tu d e of the residue data are inadequate to satisfy data requir e me nt s because of insufficient numb e r of tria ls. Although inadequate, the results of the pea fie ld tria l s i ndicate that the maxim u m residues of pronamide convertible to 3 , 5-dichlo robenzoa te were 0.98 ppm in/on fresh pea vines, 2.40 ppm in/on dry pea hay , and 0.08 ppm in/on dry peas following a single broadcast foliar appl icati o n of pronamide at 1.48 lb ai/A (lx). Because residues are above th e tolerance c urrentl y estab li s hed for field pea seed (0.05 ppm) 

### it is recommended to estab li s h an interim tolerance of 0. 1 0 ppm. Moreover, tolerances are not estab li shed for pea vines and dry pea hay because a feeding/graz i ng restriction is currently included in the labels. This restric ti on is impractical and unenforceable and may be removed from the labels

. Interim tolerances of 1.5 ppm and 3.0 ppm a r e recommended for pea v in es and dry pea hay, respective ly. The tolerance for leaf lettuce was removed from the 40 CFR 180. 317 and the use was removed

P age 3 of20Pronamide Summary of Analytical Chemistry and Residue Data Barcode 0427081

from the labels. However , magnitude of residue studies were submitted by Dow Agrosciences and the IR-4 project to re-establish a tolerance for leaf lettuce (I. Negr6n-Encarnaci6n , D424598 , 01 / 15 / 20 1 5)

. A tolerance of2 ppm was recommended provided that the label was modified to add additional instructions for split applications

. Dow AgroSciences submitted an Independent Laboratory Va lidation (ILV) study in support of the livestock commodity enforcement analytical method (MRID 46413406). This GC/ MS method (Method ORM 02.21) has been adequately vali dated by the petitioner and has undergone successful ILV (W

. Donovan , D3 l 2924, 06 / 18/2007). Storage stabi l ity data were requ ired for the metabolites of pronamide on mi l k, lettuce, apples , p lums , grapes and alfaJfa. A waiver request for the storage stability study was submitted by the registrant. HED recommends waiving the requirement for additional confirmatory storage stabi l ity data for the regulated pronamide metabolites on a lfalfa, apples, grapes, lettuce , and peaches or plums , winter pea and milk. Based on the metabolism studies with lettuce and alfalfa , and with ruminants the main metabolites of pronamide contain the 3,5-dichlorobenzoyl moiety and are expected to be converted to methy l-3,5

- dich lorobenzoate by the current enforcement methods

. If degradates of these metabolites are formed in frozen storage conditions , it is likely that these are converted to the common moiety by the enforcement method. A reassessment of the dietary burden was performed to consider res idues in pea feedstuff commodities, include maximum res idues instead of tolerance level residues, and to use the most current version of Tab l e l (OPPTS Test Guidelines 860.1000) released in June 2008. The reestimated dietary burdens are l ower or similar to those used to establish the tolerances in livestock commodities; therefore, the cur r ent to lerances are adequate to cover potential residues of pronamide in these.



### 2.0 Regu latory Reco mm e nda tion s The chemistry database of pronamide is adequate to support re - registration provided that the data needs (Section 2.1 ), and recommended tolerances (Section 2.2) and label modifications (Section 2

. 2

. 2) are cons idered. Simi lar ly , the new use in leaf lettuce is supported provided that to lerance recommendations and label amendments are followed

. The standards of pronamide and methyl 3,5-dichlorobenzoate will expire in September 2015. In addition, the standard of 3,5-dichlorobenzoic acid is needed to support the new enforcement method for livestock commodities

. The registrant should submit these standards to the EPA National Pesticide Standards Repository as detailed in secti on 5.1.4

. 

### 2 . 1 D ata D efi ci encies/D at a N eeds The submitted field tria l data for field pea commodities are inadequate (I. Negr6n

- Encarnaci6n, D383372 , 09 / 17/ 2015). Field trial data from two additional field trials for pea vines and hay and four additional field trials for pea seed are required. Storage stability data will be required if samples to be co ll ected from the requested field pea fie l d tria ls are stored frozen for a period

Page 4 of20Pronamide Summary of Analytical Chemistry and Residue Data Barcode 04 270 81 long e r than three years. 

### 2.2 Tolerance Co n s iderations 2.2.1 Enforc eme nt A nalytical Method Residue Che mi stry C hapt er for TRED ( J. Morale s, 28-FE B-200 2; 0 275536) Plant : An adequate residue ana l ytical method is available for tolerance enforcement of residues in/on plant commodities. The PAM II li sts a GLC/E CD m etho d , designated as Method I , which conve11s residue s of pronamide and its metabolites to methyl 3 , 5-dich lorobenzoate (MDCB). Method I was successfully radiovalidated using aged samples from the alfalfa and lettuce metabolism studies. 

### Livestock: The 4/16/ 93 Residue Chapter reported that a revised version of the existi ng livestock enforcement method (Method I in PAM Volume 

### 11 ), was submitted and resulted in increased method efficiency. Several modifications and an ILV were required and submitted by the registrant. Current l y, a n ILY of a gas chromatography method with negat ive-ion c hemical ioni zati o n mas s spec trom e try d e t ection (GC / MS) , Method GRM 02.21 , for the determination of residues of pronamide and metabolites co nt ain ing th e 3,5-dic hlorob enzoy l moiety as 3,5- dichJorobenzoic acid (3,5 -DCBA ) in milk , bovine liver , kidney , mu scle, and fat, and eggs and poultry muscle and fat is available. In general, reco veries we re within the acceptable range of 70- 120%, and fortification lev e ls ( 0.01 , 0.05 , 0.25, and 0.50 ppm) bracketed the established tolerances for li vestock commodities. The limit of quantitation (LOQ) was 0.0 I ppm. The reported limit of detection was 0.003 ppm. This method is cons id ered adequate to enforce tolerances for re s idues of pronamide in/on livestock commodities. 

### 2.2.2 Recommended Tolerances The current tolerance expression fo r pronamide (40C FR 

### § 180.317 ) is adequate for plant commodities, and includes both coverage and compliance stateme nt s for enforcement purposes. Recently , a tolerance of2 ppm ha s been recommended in s upport of a peti tion for the use of pronamide on leaf lettuce (I. Negron

- E ncarna c io n , 0424598 , 09/ 17/2 015). Moreover, based on limited field tria l data to support the use of pronam id e on field pea , HED recommends increasing the to lerance for fi e ld pea seed from 0.05 ppm to 0.10 ppm , and establishing tolerances of 1.5 ppm and 3.0 ppm for field pea vines and dr y pea hay , respectively ( I. Negr on-Encarnacion, 0 3833 72, 09117 12015). The current tolerance expression is not adequate to enforce tolerance s in livestock commod ities as the tolerance e n fo rceme nt m ethod converts residues of pronamide to 3 , 5-dichlorobenzo ic acid instead of methyl-3 ,5 -dichlorobenzoate. Ba sed on this , HE D recommends to modify the 40 CfR 

### § 180.317 (a) General in two sec tions, ( 1) and (2). The first section may includ e the current tolerance expression for plant s fo ll owed by a table with the tolerances for plant commodities while the second sec tion may co ntain the tolerance express ion below followe d w ith a table with the tole ra nce s for livestock commodities. The reco mmend ed tolerance expression for sect ion (2) is: " Toleranc es are establis hed for residues of the herbicide prop yzamide , includin g its metabolites and degradates , in or on t he
Page 5 of20Pronamide S umm ary of An a lyt i ca l C hem i s t

ry and Residue Data Ba r code 042708 1


### commoditi es in th e fo ll ow in g tab le . Com pli ance w ith t he to lerance leve ls sp ecifie d in th e tabl e be low is to be dete rmine d by m eas urin g o nl y tho se p ro pyz amid e r esi du es convertibl e to 3,5- di c hl orobenzoic ac id, ex presse d as the sto ic hiom et ri c eq ui vale nt o f pro pyzamid e, 3,5- dichl oro N-(1 , 1- d im eth yl

-2

- pro p yn y l) be nzam id e, in or o n the co mm o d ity."

Tabl e 2.2.2 . To lerance Re a ssessm e nt Summary for Prona mid e Commod ity Establ ished I P ro p osed* Recommended Comments; Correct Commodity To lerance (ppm) Tolerance ( ppm) Definition To lera nce to be L isted Under 40 CF R §I 80.3 I 7(a)( l ) A l fa l fa, see d 10.0 10.0 A n im al feed, nongrass , 1 0.0 10.0 group 18 Apple 0. 1 0 .1 Art ic hoke , g lobe 0

. 01 0

. 0 1 B lackbe rry 0.05 0.05 B lueberry 0.05 0.05 Boysenberry 0

. 05 0.05 En di ve 1. 0 1. 0 Fru it, sto ne , group 12 0 .1 0.1 G rape 0. 1 0. 1 Lettuce, h ead 1. 0 1.0 Lettuce, leaf 1. 0* 2

. 0 Pear 0.1 0 .1 Rad icchio 2.0 2.0 Raspberry 0.05 0.05 To l erance to be Listed Unde r 40 CFR § 180.3 1 7(a)(2) Cattle, fat 0.2 0.2 Cattle, k id ney 0.4 0.4 Cattle , li ve r 0

. 4 0.4 Cattle, meat 0

. 02 0

. 02 Cattle, mea t byp ro d ucts, 0

. 02 0.02 except kid ney and live r Egg 0.02 0.02 Goat, fat 0

. 2 0

. 2 Goat, kidney 0.4 0.4 Goat, li ver 0.4 0.4 Goat, meat 0

. 02 0

. 02 Goat, meat bypro d ucts, 0.02 0.02 except kidney and live r Hog, fat 0.2 0.2 Hog , kidney 0.4 0.4 Hog , li ve r 0.4 0.4 H og, meat 0.02 0

. 02 Hog , meat bypro d ucts, 0.02 0.02 excep t ki dn ey and l iver



### P age 6 of20Pronamide S umm ary of Analytica l C hem is try and Resid ue Data Barcode 04 2708 1

Horse , fat 0.2 0.2 H orse, kidney 0.4 0.4 Hors e , liver 0.4 0.4 Horse, meat 0.02 0.02 Horse , meat byproducts, 0.02 0.02 excep t kidney and liv er Milk 0

. 02 0.02 Poultry , fat 0.02 0.02 Poultry , li ve r 0.2 0.2 Poultry, meat 0.02 0

. 02 Poultry , meat b y product s, 0.02 0.02 e xcept liver Sheep , fat 0.2 0.2 Sheep

. kidne y 0.4 0.4 S heep , liver 0.4 0.4 Sheep , meat 0.02 0.02 S heep , meat byproduct s, 0.02 0.02 ex cept kidne y and liver To lerance Lis ted Und er 40 CF R §I 80.3 I7(c) Pea , field , seed 0.05 0.10 1 Tolerances To B e Li sted Under 40 CF R § I 80.3 I 7(c) Pe a , field , hay 

### -- 3.0 1 Pea , field , vines


### -- 1. 5 ' 1 To be re-evaluated up on s ubmission of adequa te magnitude of residue s tudies.

2.2.4 Internati ona l Harmonization Where possible , E P A enco ura ges the harmonization of the US To leran ces and Maximum Residue Lim i ts ( MRL s) in key ex port marke ts. A tabl e with th e US tol e rances , and the Canadian and Codex MRLs for pronamid e is pro v id e d in A pp e ndi x A. C urrentl y , Mexican o r Co de x MRLs for pronamide have not bee n estab li s he d. T he US and Canadian tolerance ex pres s i o n s are harmonized fo r m eas urement of pronamid e , includin g metabolite s containing the 3 , 5- dichlorobenzoate moi e ty. The Canadian MRLs for apple, blueberry and p ear are harmonized wi th the US to le ranc es. T he Canadian MRL for l e ttu ce i s 1 ppm w hile th e recomm e nded US tolerance is 2 ppm. T he propose d u se pattern do not s upport harmonized to lerance leve ls. An a lte rnativ e u se pattern that supports harm on ization was r ecomme nded , refer to Sec tion 6.0 of 

### 0424598. 2.3 Labe l Recommendations T he c urre nt labels fo r pronamide includ e plant back inte rval s for other crops than those recommended in the 2002 TRED. Rotation to other c rops than leafy vege table s crop group 

### 4 , root and tuber vegetables crop gro up 

### 1 , and ce real g rain s is allowed by following a 365-day 

### PBI. Feeding/grazing restr ictio ns on the labe ls can be removed as potential r es idues of pronamide

Page 7 o f 20Pronamide S ummary of Analytical Chemistry and Residue Data Barcode 0427081


### in/on field pea feedstuff and livestock commodities have been evaluated, and these restrictions are ge nerally considered impractical and/or unenforceable. For the new use on leaf lettuce , it i s recommended that detailed instruction s for split applications are i nc luded in the s uppl eme ntal labe l , i .e. part of the maximum allowab le application rate can be initiall y applied and the balance of the maximum allowab le application rate can be made up to 10 days later , and th e maximum all owab le total rates and corresponding PHi s fo r sp li t applications shou ld follow the recommendations for regular applications. HED recommended a tolerance of2 ppm to support the proposed use pattern on leaf lettuce ; however, an alternative use pattern that supports the tolerance proposed by the registrant 

### (1 ppm) was recommended as well. If this a lternative us e pattern i s desired , the label needs to be modified accordingly. 3 . 0 Introduction Pronamide (also refe rred to as propyzamide) is a selective, systemic, pre-and post-emergence herbicide registered for the contro l of grasses and broad leaf weeds in several food and feed crops as well as woody ornamentals , Christmas tree s, nursery stocks , lawns , turf , and fall ow land. Pronamide is also known by the trade name Kerb®

. 3 . 1 C hemical Identi ty

Tabl e 3. 1 Pronamide Nomenclature. Compound 0

# N~ Cl**~** 

# I H CH 3**~CH ~** 

Cl Common name Pronamide or Pronamide Company experimental name Not applicable I UPAC name 3 , 5-dichloro-N-(1 , 1-dimethylpropynyl)benzamide CAS name 3 , 5-dichloro-N-( I , l- dimethyl-2-propynyl)benzamide CAS registry number 23950-58 -5 End-use product (EP) 50% WP formu lation (Ke rb ® 50-W Herbicide; EPA Reg. No. 62719-397)



### 3.2 Ph ys ical/Ch e mical C h a racte ri s tic s Propyzamide is read il y so lu b le in water and i s not expected to vo l ati li ze significantly due to the compound ' s relatively low vapor pressure of 8.5 x 10- 5 

### torr. The physicochemical properties of pronam ide are in Tab le 3

. 2

.

Page 8of20Pronamide Summary of Analytical Chemistry and Re sidue Data Barcode D42708 I

Table 3.2. Ph ys i cochc mic a l Properties of Pron a mid e. Parameter Value Ref erence Melting range 155-156°C Pronamid e TRED; J

. Moral es. 28-FE B- 2002 ; DP# : 275536 Water s olubility 15 mg/ L at 25 ° C Pronamide TRED; J. Solven t so lubili ty Dimet hyl s ulfoxide and dimethy l Morales , 28-FE B- 2002 ; formarnide at 33 mg/L ; DP # : 275536 Mesityl oxide , isophorone , methyl ethyl ketone , and cyclohexanone at 20 m g/L ; Methanol , isopropanol , and chlorobenzene at I2- I 5 mg/L; Butyl cellosolve , xylene, acetonitrile, and kerosene at I0 m g/ L; Nitrobenzene and ethylene dichloride at 5 mg/L Vapor pressure 8.50 x I0 · 5 torr at 25 ° C Octanol/ water partition coeffic ient , 3.05-3.27 Pronarnid e TRED; J

. Lo g ( P o\\') Morales. 28-FEB-2002; DP # : 275536 

### 3.3 Pesticide Use Pattern/ Di recti ons fo r Use (860. 1 200) Th ere are two end- u se product labe ls regi stered with pronamide as the active in gredient. The se include liquid, and wettable powder formulations. Pronamide, also known by the trade nam e Kerb ®, may be applied using grou nd spray eq uipment , by soil incorporation , or by aircraft. The single application rates fo r ag ri cu ltur a l u ses range from 0.5 to 4 lb 

### ai l A, and the maximum application rate per year range from 1 to 6 lbs ai/A. The 02 1 28 1 2002 Residue Chemistry Chapter recommended plant back intervals of 30-day for leafy vegetab les (except Brassica vegetable s) ( Crop Group 4); 90-day for root and tuber vegetab les (Crop Group 1 ) ; 360 -day for cereal grains (Crop Group 1 5) and the forage , fodder, and straw of ce real grains (Crop Group 16). Currently, extens iv e rotational crop restrictions, depend e nt on app li cation rate s , are currently estab li shed o n the product labels. The restrictions for root and tuber vegetab le s , leafy vegetab les cro p group 4 , and cerea l grains are considered adequate. However , restrictions for othe r crops such as legume vege tab les and cotton (90 to 1 50 -day PBl) , and brassi ca leafy vegetab les, cucurbit vegetables , fruiting vegetab les , and bulb vege table s (90 to 2 10-day PB ls) are included in the labels. As indicated in the 2002 TRED , these rotational crop re str ict ions are inappropriate based on the results of the reviewed confined and limited rotational field studies. 

### Conclus ions: Appropriate plant back interval s are established for leafy vegetables c rop group 4, root and tuber vegetab les crop group 1 , and cereal grains in the lab e ls of Kerb ® (EPA Reg. Nos. 62719-578 , and 627 19-397 ) and those from Willowood (EPA Reg. Nos. 87290-22, 87290-9 and 87290-3). However, rotation to other crops th an those was not recommended i n the 2002 TRED

. Rotation to other cro ps is allowed by following a 365

- day PBI. The s pecial local ne e d label (E P A R eg . No. 627 1 9-578) specifies si n g le and m aximum app li cation rates of 4 lb s 

### ai lA and 6 lbs 

### ai lA/year, r espect i ve ly, for head l ettuce. This is not s upported by the limited rotational crop study which was conducted at a maximum application rate of 2 lb s ai l A. Ho wever , the lab el
Page 9 of20Pronamide Summary of Analytica l Che mi stry and Residue Data Barcode 0427081 

### specifies that crops other than lettuce should not be planted back for a period of 12 months follow in g app lication. The use on leaf lettuce h as been removed from the l abel accordi n g to previous recomm e ndation s; however , a new u se on leaf lettuce h as been proposed

. Other label modifications have been recommended to support petitions for use of pronamide on leaf lettuce and field pea; refer to D 424598 and 0383372 , re spe ctiv e ly , for detai led information

. These are: (1) For the new u se on leaf lettuce , it i s recommended t h at detail ed in structio n s for split app lications are in cluded in the supplemental labe l , e.g. for split applications part of th e maximum a ll owab le app li cation rate can be initially app li ed and the balance of the maximum allowable application rate can be made up to 10 days later. The maximum allo wab le total rates and correspond in g PHi s for split applications for leaflettuce should be 0.5 , 0 .7 5, 1. 5, and 2.0 lbs ai / A with correspond ing PH i s of 25, 35, 45, and 55 days , respective ly. (2) For t he estab li s hed use on field p ea, the registrant may de lete the ex isting feeding/ grazing restrictions on the l abel since they are impractical a nd/or unenforceable.

4 . 0 Metabolite/Degradate Residue Profile Residue C he mi stry Chapter for TRED (J

. Morales, 02 /28/ 2002 ; 0275536) 

### Primmy Crops: The qualitative nature of the residue in plants is adequately und er s tood on alfalfa , l ettuce and s im ilar crops. The 4/ 1611993 Residue Chapter reported that stud ies with a lfalfa and l ettuce indicate that pronamide is read il y absorbed by p l ants through the root system, translocated upward , and distri buted into the ent ire plant. The degree of trans l ocati on from leaf absorpti on is not appreciable. The te rminal r es idues in a lfa l fa and lettuce consist mainly of the parent compound pronamide (N-(1, l-dimethylpropynyl)-3 ,5- dichlorobenzamide) , glucose conjugates, and g lu cose-mal onyl conj ugates of pronamide, a ll containing the 3,5

- dichlorobenzen e moiety. Metabolism primarily occ u rs via co nju gation to (malonyl) glucose. No evi dence of fragmentat i on or lo ss of the chloro substituent of the aromatic ring was observed

. The termi nal residues of concern are pronamide and its metabolites containing the 3,5- dichlorobenzoy l moiety. 

### Rotational Crops: The qua litative nature of the residue in rotational crops is adequately understood based on an acceptab l e confined rotational crop study (MRID 41462001 and 41913506)

. In this study, uniform l y ring l abeled [ 1 4 

### C]pronamide was app li ed at a rate of4.0 lb ai / A to bare ground plots, and c rop s we re p l anted in the treated areas after 30 d ays, 6 months , and 1 year . The subplots representing eac h p l antback interval (PB I) were allowed to remain fallow until suc h time that lettuce (a leafy vegetab le) , carrots (a root crop) , and wheat (a sma ll grain) were p lanted. The terminal residues of concern in rotated crops following treatment of primary crops w ith pronamid e consist of the parent pronamide and its metabolites bearin g th e 3 , 5

- dichlorobenzoy l moi ety
. 

### Livestock: The qualitative nature of the residue i n livestock is adeq uate l y understood. T he 4/ 16/ 93 Residue C hapter reported stud ies invo l vin g lactatin g goats and laying hen s indicate that the primary route of eliminati o n is by exc reti o n (urine and feces). M inim a l residu es were distributed to goat and poultry muscle

. The major metabolites i n the eggs , li ver , and fat of poultr y are pronamide an d 3,5 dichlorobenzoic acid. The major metabolites in the mi lk , fat,

P age 10 of20Pronamid e Summary of Analytical Chemistry a nd Re sid ue Data Barcode D42708 I

muscle , and li ve r of goats are pronamide , 3 , 5 -di chlorobenzoic acid, and compounds co ntainin g the 3,5 -di ch lorobe n zoyl m oiety. T h e metabo li c p athway in vo lves m odi fi ca tion of th e a l iph a tic portio n of pronamide. The ter min a l resid ues of concern in li vestoc k co mm o di t i es a re pronarnide and its metabolites containin g the 3 , 5-dichlorobenzoyl moiety. The re sid u es of concern fo r pro n amid e are dep i cted in Tabl e 4.3.

Tab le 4.3. S umm ary o f Pronam id e Metabo lites a nd D eg r a d ates t o be included in the Ri sk As sess men t a nd To le ranc e Ex press io n Matrix Residues in c luded in Ri sk R es idues in c lu ded in Assessment To lerance Exore ss ion Plants P rimary Crop R

otational Crop pronamide and its m e tabo lites pronamide and its metabolites Livestock Ruminant containing th e 3,5- contai nin g th e 3 , 5- Poultry dich lorobenzoy l moi ety dich lorobenzoyl moiety Drinking Water Not Aoolicable 

### Conclus ions

. For purp oses of reg i s trati o n rev iew, no add i ti onal plant m e tabo l is m s tudies are required; however, because the availab le metabolism s tudie s were only co ndu cted on alfalfa and lettuce, HED may reques t additional me tabolism stud ies in the future s hould the petitioner seek for additiona l uses on o th e r crop gro up s .


### 5

. 0 Re s i d u e P rofil e 5 .1 Re s idu e Ana ly tical Method s (860. 1340) 5.1.1 Multi-Residu e Me thod s (860

. 1360)

. Residue Chem istry Chapter for TRED (J. Morale s, 02/ 28/ 2002; D275536 ) The 10 / 99 FD A PESTDATA ( P AM Volume I, Appendix) indicates that pronamide is co mpl e tely recove red ( > 80%) u s in g MRMs P AM Volume 

### I Sect io n s 302 ( Luke method ; Pro toco l D ) but is o nl y partia ll y recove re d using Method 303 (Mi ll s, Onley, and Gaith er method ; P rotocol E). MRM testing data for pronamidc and a me tabo li te containing th e 3,5

- dic hl orobenzoyl moiety are also availab le (MRID 43493203) ; these data have been forwarded t o FDA for review 

### (W

. Donovan, 0340389, 06 / 06/ 2007). 

### 5.1.2 To l era nce E nforcement Method s Re s idue C h emis try Chapter for TRED ( J

. Morale s , 02 /28/ 2002; D2755 36) 

### Plants: An adequate residue analyt ical m ethod i s availab le fo r tolerance enfo rcement. The PAM II li sts a GL C/ EC D method , designated as Method I , which conve rt s res idu es of pro n amid e and its metabo lites to MDCB

. Thi s me th od consist s of hydr o l ys is of t he p are n t compo und and its metabolites b y re flu xing wit h methanol and su lfur ic acid. The h ydro lyzed products are methylated , and the methyl ester reaction product i s steam dist ill e d. Th e es ter is extracted from the disti ll ate w it h petroleum ether followed by an add i tion a l Flo r isil co l umn clean up . R es idu e is determin ed by gas li qu id c hrom a t ograp hy u s in g an e lectron cap ture d e t ec to r. Two co lumn s can be used; one may be used for confirmatory purpose s. Duri ng Age ncy va lidation of this meth od

Page 1 1 of 20Pronamide S umm ary of An a l yt i ca l Che mi s t ry and Residue Data Barcode 0 427081

u s in g le ttuc e as the s ub st rate , reco ve ri es were 80-8 5% at th e 2-p pm fortification l eve l, and 8 2.5

- 92.5% at the 4-ppm le ve l. Th e li s ted detec tion limit is 0.2 ppm for plant co mmoditie s . Metho d I was successfu ll y rad io validated u s in g aged sam pl es from the alfalfa and lettuce me taboli s m studies. 

### Liv estock: The 4116 / 93 Residue Chapter r e p o rt ed that a revised vers io n of the existing liv es tock e n fo rcemen t method (Me th o d I in PAM Volume II based o n GC / EC) , whi c h includes an alkaline h ydro ly s i s to re lease bound residues , was s ubmitted and res ulted in increased method efficiency. The C hapter req uired an ind e p e nd e nt laboratory validati o n ( l LV) of the rev ise d method prior to met hod val id a tion by the Agency. An ILV of a gas chromatography me thod with negati ve -ion chemical io ni zation ma ss s pectrometr y detection (GC / M S) , M e th o d ORM 0 2.2 1 , for the d e termination of res idu es of pronamid e and me tabolite s containing th e 3,5-dic hl orobenzoyl moi e t y as 3 , 5-d ic hl orobenzoic ac id (3,5-DCBA) in pou ltry fa t a nd bov in e li ver was submitted ( MR JD 46413406). In addition to the ILY report , the s ubmi ss ion in cl ud ed a complete method description of th e ori g inal method , along with s upportin g me th o d validation data for milk , bovine li ve r, kidne y , muscle , and fat , and eggs and poultr y mu scle a nd fat. Fat samples were ex tracte d w i th ethy l acetate and centrifuged. An aliquot of the ex tract was evaporated to dryness , red isso lved in 1 , 4-dioxane , and hydro lyzed b y adding 9 N s ulfuri c ac id and heatin g at 100 ° C. Samp les of milk , eggs, and tissues other than fat were ext racted and hydrolyzed by add in g 4 N sodi um h ydrox ide and heatin g at 80 ° C , followed by seve ra l s teps includin g a second hydrolysis procedu re. Durin g hyd r o lys i s , bound residue s of propyzamide and metabo lites were furt her s ol ubili z ed and hydro l yzed to 3 , 5-dichlorobenzoyl acid. This me th od was forwarded to E PA ' s Analytical C hemi stry Branch on 6/18 / 07 for a T o lerance Method Validati o n (TMY); however , cur rent procedures do not requi re a TMV. T h e met hod was reviewed to determine if is ap p rop ri a t e as an enfo rc e ment method. T he method as written was adeq uatel y valid ated and the sampl e ana lysis (excl udin g overnight incubation , and aut omated GC an a lys i s) was conducted in less th a n 24 hours. In ge n e ral , recoveries we re within th e acceptable rang e of 70-120% , and fortification leve ls (0.0 1 , 0.05 , 0.25 , and 0.50 ppm ) bracke t e d th e es tabli s hed tolerances for liv es to ck co mmoditi es. T he limit of quantitation (LOQ , d e te rmined as the l owes t limit of me thod valid ation; LLMV) wa s 0

. 01 ppm . Th e re porte d limit of d e tec tion was 0.003 ppm. The reg i s trant indi cated that confirmation i s by compar iso n of th e re te ntion time (GC) as well as the p ea k a rea r atio res ultin g from se lected ion monitorin g fo ll ow in g MS ana lys i s in the negative c he mi ca l ioni zation m o d e . Additiona l confirmatory d ata wi ll not b e required as Me thod I in PAM Vo lume II (base d o n GC /EC) may be u sed for co nfirm at io n . No r adiova lid at io n data was pro v ided

. Part of the rad ioac ti v ity in the he n metabolism stud y was liberated using ace ton itril e o r methanol (M RID 40782201). A seco nd study was conducted to further characteri ze/ identify the no n - extractable residue. In this study , base hydrolysis was shown to re lease most of the residue in liver in c reas in g from 48 % using methanol and aceton itril e t o 92% using base h y drol ys i s (MRJD 42614201 ). This stu d y also indicates that the e n fo rcement method by that tim e was m od ifi ed to incorporate a base hydrolysis step , resu ltin g in sign ifi cant increase in recoveries , 40% to 66% , 59% to 74% , and 65 % to 91 % for breast mu scle , thigh muscle , and li ve r , respectively. 

### Conclus ions. E n fo rce m e nt methods are avai lab le to e nforce th e tolerances of pronarnide o n plants and li vestock commodities. Becau se the e nfor ce me nt m e th od for li vestock in volves base

Page 12 of20Pronamide Summary of Analytical Chemistry and Residue D a t a Barcode 042 7081 hydrolysis extraction and the res ults of the secon d metabolism study with hen which showed good recoveries whe n using ba se hydrol ysis the radio va lidation of the enforcement meth od fo r livestock can be waived.



### 5 . 1. 4 S ubmitta l o f A naly ti ca l Refe re n ce S tanda rd s (86 0 . 1650 ) Re fe re nce standards for pro n amide are curre n tl y ava il able in the standard repos itory (perso na l communication w i th Gregory Verd in (05/ 11 /2 015); howeve r, th e exp i ration dates are 09 /22/2 015 for Pronamide and 09/ 30 / 2015 for methyl 3 , 5-d ichlorobenzoate (MDCB). 

### In addit ion, the standard of 3 , 5-dichlorobenzoic acid is n eeded to support the new enforcement method for livestock commod i ties. These standards s hould be sent to the Analytical Chemistry Branch in Ft. Meade who i s th e owne r of this re p os itory. T he ad dress to submit standa rds is: USEPA Nationa l Pesticide Sta ndard s Repository/ Ana lytical Chemistry Bran ch/O P P 70 I Mapes Road Fort George G. Meade , MD 20755-5350 The full 9 digit zip code is mandatory or the mail will be returned to yo u . 5.2 

### S to rag e S ta bility (8 60.1380) Residue Chemistry Chapte r for TRED (J. Morales , 02 /28/ 2002; 0275536) Waiver Request for Storage Stability Study (MRJD 4965440 I) 

### Plants: The TRED of pronamide su mmari zes the re s ult s of a three-year storage stability study (MR ID 4472270 I). The sub m itted data i nd icate that pronami d e 

### p e r se ( th e predominant residue identified in plant metabolism s tudies ), as m eas ured by a s pecific GC method for pronamide , i s reasonably stable under frozen sto rage conditions for up to three years in/on alfalfa, apples , grapes , lettuce, and peaches (p l um surrogate). Additional confirmatory storage stabi li ty data for the r eg ulated pro namide metabo l ites on a lfa lfa, app l es, grapes, lettuce , and peac h es or plum s was required. Limited sto rage stabi l ity data for pronamide is available i n pea commodities. Storage s tability data subm itted in conjunction with pea field tr ia l indicate that residue s of pronamide are stable under frozen sto rage for up to 98 days for pea vines, 95 days for pea ha y , and 84 days for pea seed wh i ch do not covers the storage duration of the crop field trial samples. 

### Livestock: The 4/ 1 6 / 93 Re si du e Chapter repo rted that pronamide is stab le in poultry eggs and tissues under frozen conditions for up to 34 months. For the purposes of reregi stratio n , the Chapte r concluded that the avai lable stability data for poultry commodities may be tra n slated to satisfy the storage s tabi li ty data requ i rement s for rumina n t tissues. T he Chapter, however, re quir ed addit ional storage stabi l ity data for milk. In the TRED, submitted data indicate that pronamide 

### per se , as measured by a specific GC method for pronamide , is reasonably stable under frozen storage conditions for up to three years in mi lk. Samples of l ivestock commodities that were used for to lerance reassessment were stored frozen for interva l of less t han t hree years. However, analysis of metabo l ites containing the 3,5-d ich lorobenzoy l moiety was not included in the study

. Additional confirmatory storage stability data for the regulated pronamide metabol i tes on milk was required. This data has not ye t been submitted.
Page 13 of20Pronamide Summary of Analytical Chemistry and Residue Data Barcode 0427081



### Waiver request A waiver for the storage stability study for the metabolites of pronamide was requested (MRID 49654401 ). As the reg istrant describes, the existing frozen storage stabil ity data indicates the two key structural components of pronamide, the 3,5 -di chlorobenzoyl moiety and the alkyl side cha in , are stable in the presence of plant and milk matrices under frozen storage conditi on. In the plant and animal metabolism studies , pronamide was observed to be slowly metabolized through a series of simple oxidative reactions that take place on the alky l side c h ain of the molecule. While the alkyl s id e chain is susceptible to metabolism via oxidation and sugar conjugation , metabolites showing degradat ion of the 3,5-dich lorobenzoy l moiety were not observed. Therefore , it is proposed to be structurall y stable and resistant to metabolism in crop and animal tissues , i.e. cellular enzymes in living plant and animal tissues only affect the metabolism and transformation of the alkyl s ide chain , but not the stable 3,5-dich lorobenzoyl structure of pronamide and its metabolites. While data is not available to specifically address the storage stabil ity of each individual pronamide metabolite or the relative stab ili ty of the various forms of the alkyl s id e chain of the metabolites under frozen sto rage cond ition , it is indicated that the 3,5- dichlorobenzoy l moiety is stable. Because the enforcement analytical method measures all the re s idues convertible to the 3 , 5-dichlorobenzoyl moiety it is stated that the metabolites containing such moiety can be accurately determined regardless of the relative stability of the various a lkyl side chains.

Cl 0 H CH, 

# 0 11 I -C-N**-~-C1CH** Cl CH, ProMmlcle 

# + Cl O 

# 0-A-R

Cl m1tabollta <0nt1lnln13,S · dlchloroblnzoyl moiety
Cl 0



# 0-!- R

Cl**~OJCHa** Cl 0 CH 1 0M Cl 0 M ell, 

### V'i. CHa (\._.(.' 

## )ai.**o-~ -~-~** **-COOH** ct CM, RH· Z'6M ,)---' "'w CM, Rff
..25111 Cl CMi RH· l46 SO Cl O H CM, Cl 0 H C. 1 Cl 0 H 

# r

0 11 1 

# 0

. 1, 

# 0 .1.**-C-11-~-x** °'•**-c-· ~-~**  CH,OH 

## -c -•-' -x · COOH Cl CM, R H · 245IO Cl CMo RH · 21702 Ct at, Rlt-2760 Cl 0 H 

# l"'" Cpl 0 M Clfa Cl 

# i M r;MtM 

# O JI I Ill 

# p 11! -C·ll·**'-~** CH,OH**-C-11-~ -CH** **,CH,OH** -**-11-lji-~~COON** CH OH CH, CH, Cl 1 RH · 2652l C R H· 26J17 RH · 27U4 Cl 

### y H CHa Clo**~** Cl 0 " CM,

### -c-~-~-cH,cooH - 

## "°"**/-\ -~-~** **-~-C•H,OH** Cl**~...** RH
-26059 Cl OCBA 

### ).J**~.** RH · 24"& Cl + Sup r conJupte of the mebbolltes Figure 5.2 Structu re of pronamide and its metabol ite s co ntaining t he 3 , 5-dich lorobenzoyl moiety. Sugar conjugation occu r s on the a l ky l side chain

. Excerpt from registrant wa iver request (MRID 49654401)

. 

### Conclusions: HED recommends waiving the requirement for additional confirmatory storage stabi li ty data for the regulated pronamide metabolites on a lfalfa, app les , grapes, lettuce , and peaches or plums, and milk. Based on the metabolism studies with lettuce and alfalfa the main metabolites of pronamide containing the 3 , 5-dichlorobenzoyl moiety are RH25891 (m-g),
Page 14 of20Pronamide Summary of Analytical Chemistry and Residue Data Barcode 0427081


### RH26521 (m-g) , RH26521 (g) and RH24848 (m-g), where m refers to malonyl and g to g lu cose conjugates. The main metabolites of pronam id e on milk are compound I (RH

- 24644), II (RH24580) , V (RH-26377) , DCBA, X (unclear from best copy avai lab le) , and XII (RH-27414). The structures of these metabolites are in Pronamide Response to Registration Standard (J.B

. Stokes, 08/16/1989) and in MRIDs 40494801 (goat), 40494802 (lettuce) , and 40494803 (alfa lfa)

. All these metabolites are expected to be converted to methyl-3 , 5-dich lorobenzoate by the c urrent enforcement methods. Metabolism studies and the enforcement method sample preparation procedures suggest that the 3,5 -di ch l orobenzoyl moiety is quite stable to plant and animal enzymes , and to base and/or acid hydrolysis. 

### If degradates of these metabolites are formed in frozen storage conditions, it is li kely that these are converted to the common moiety by the enforcement method.



### 5.3 Residue Data 5.3.1 Crop Field Trials (860.1500) Field Pea Crop field trial data were required for field pea in the 2009 scoping document. These data were submitted by the IR-4 project. The residue chemistry memo summarizing the data and HED's recommendations is available as D383372 (I. Negron

- Encarnacion , 09117 / 2015). In summary , the submitted magnitude of the residue data are inadequate to satisfy data requirements because of insufficient number of trials. A minimum of three trials is required for fie ld pea forage and ha y with Austrian Winter pea as the preferred cultivar; and field pea seeds are considered dried shelled peas and require a minimum of five tri als. Furthermore , two independently composited samples of treated commodity should be co ll ected and separately analyzed in each field trial. Although inadequate , the results of the pea field trials from the current submission indicate that the maximum residues of pronamide (expressed as pronamide 

### + its metabolites containing the MDCB moiety) were 0

. 98 ppm in/on fresh pea vines, 2.40 ppm in/on dry pea hay, and 0.08 ppm in/on dry peas following a single broadcast foliar application of pronamide at 1 .48 lb ail A (Ix). Because residues are above the tolerance currently estab li shed for field pea seed (0.05 ppm) it is recommended to establish an interim tolerance of 0. 10 ppm. Moreover , tolerances are not established for pea vines and dry pea hay because a restriction is currently included in the labels. This restriction is impractical and should be removed from the labels. An interim tolerance of 1.5 ppm and 3.0 ppm is recommended for pea vines and dry pea hay , respectively. Leaf Lettuce As recommended by the 2009 scoping document , the tolerance for l eaf lettuce was removed from the 40 CFR 180.317 and the use was removed from the l abels. However, magnitude of residue studies were submitted by Dow Agrosciences and the IR- 4 project to re-establish a tolerance for leaf lettuce (I. 

### Negron-Encarnacion , D424598, 01115 / 2015). A tolerance of2 ppm was recommended provided that tolerance recommendations and label amendments are followed.

Page 15of20Pronamid e S ummary of Analytical Ch emistry and Re si due Data Barcode 04 2708 1



### 5.3.2 Fie ld Rotational C rops (860.1900) Res idu e Chemistry Ch apt er for TRE D (J. Mor ales. 02/ 28 /2002; 0275536) A n acceptab le limi ted fie ld rotationa l crop s tud y (M RID 44661501) w ith pronamide is available. B ased on a r e - evaluation of this s tud y, the 02/ 2 8/2 002 Residue C he mi stry C h apte r recommended plant back inte rva ls of 30-day for leafy ve ge tab l es (excep t Bra ss ica vegetabl es ) (C rop Group 4) ; 90 -d ay fo r root and tuber vegetable s (Crop Group l ); 360-day fo r cerea l gra in s (C rop Group 1 5) and the forage, fodder , and s traw of cereal g r ai n s (C rop Group 1 6)

. The se are based on limited field rotat iona l crop studies performed after application of2 lb s a i/A of pro namide to bare so il and planting of lettuce , carro t s , w heat and sorg hum at plant back interval s of 30 , 90, 180 , and/or 360 days. The current rota ti o n res triction s are adeq ua te. Rotat ion to oth e r cro p s than l eafy vegetab les c rop gro up 4 , roo t and tub e r vege tab les crop group I , and cereal g rain s is all owe d by fo ll owi n g a 365-day PBI. 

### 5.3.3 Processed Food and Feed (86 0

. 1520) Residue C he mi s try Chapter for TRED (J

. Morales, 02/ 28 /2002; 02755 36) The re quir e m e nt for da ta d e p ic tin g ma g ni tud e of the re s idu e in the proc essed co mmoditi es of app les, g rapes , and plum s is fu l filled

. In 1 995 , H E D waiv e d th e require ment to conduct processi n g st udie s for apples , grapes , and p l ums based o n th e resu lts of fie l d trials conducted at exagge ra t ed rates equiva le nt t o or greater than th e maximum th eoret ical concentration factors from processing (S

. Funk , 0 2 1 3037, 3/ 29/ 95). Fie ld trial s at exaggerat ed ra t es s howe d that residu es in processed commod i ties would be a nti c ipated to be < 0.1 ppm , the reassesse d tolerance s for the RA Cs. No other proc ess in g s tudie s hav e been s ubmitted or required. 

### 5

. 3.4 Meat, M ilk, Poultry and Eggs (860.1480) Residue Chemistry Chapter for T R ED ( J. Morales. 02/ 2 8/2002; 0 275536) No addi ti ona l data we re req uired fo r re s idu es of pronamid e o n meat , milk , poul try and eggs in the 20 0 2 TRE D. Toleran ces fo r kidney and liv e r of cattle , goats, hog s, hor ses, an d s heep were recomm e nd e d to be in c re a se d to 0.4 ppm du e to recent improv e ment 

### in th e effic ie n cy of th e revi se d e nforc e ment meth od fo r g oat li ve r an d the leve l s of transfer found in the cattl e feeding stud y . Th ese tole rances are c urre ntl y estab li s hed in the 40 CF R 180.317

. 

### 5.3.4.1 Dietary Burden The li vestock dietary b urd e n of pronamid e was estimated in th e 20 02 TRED to consi der re assessed tolerances for li vestock feed ite m s. The maxi mum th eore tical d ietary burde n s of pronamide to beef cattle , dairy catt le, swi n e, and poultry we re re-ca lc ulated to be 10. 7 ppm , l l.2 ppm , 1 .0 ppm , and 1.0 ppm , respect ive l y . A reassessm e nt of the dietary burde n was performed to co n s id e r r es idues in pea feedstuff commodities , includ e maximum re s idu es ins t e ad of toleranc e leve l residue s, and to u se the mo s t c urrent ve rs i o n of Tab l e 1 ( OPP TS Test Guidelin es 860.1000 ) r e lease d in Jun e 2 008 (Tabl e 5.3.4. 1).

Page 1 6 of20Pronamide Summary of Analytica l C hemistry a nd Residue D ata

Table 5.3.4. 1

. Ca lculation of Dietary Burden s of Pron amid e Res i dues to Livestock. % Dry Feedst uff Type ' Matter2 %D iet2
B eef Catt le (R = 15 %; CC = 80 %; P C= 5 % ) Alfa l fa, hay4 R 89 1 5 TOTAL BURDEN 

### -- Dairy Ca ttl e (R = 45 %; CC= 45%; P C= 10 %) Alfa l fa , h ay4 R 89 5 Alfa l fa, forage 4 R 35 20 A l falfa , s il age4 R 40 20 A lfalfa , mea ls PC 89 10 TOT AL BURDEN

-- Sw in e (CC= 85 %; PC = 15 % ) Alfalfa, mea1 5 PC NA 5 Pea, field, seed PC NA 10 TOTA L BURDEN


### -- Poultry (CC = 75%; PC= 25 %) Alfalfa , m ea ls PC NA 5 Pea , field , seed PC NA 20 TOTAL BURDEN


### -- 1 R: Roug hage; CC: Carboh yd rate concentrate; PC: Pr otem conce ntrate
. 2 Table I Feedstuffs (J ul y 2008)

. HAFT (pp m )

9.52

9.52 9.52 9.52 9.52

9.52 0

. 08

9.52 0.08 Barc ode D42708 l

D ietary Co ntr ibution ( ppm) 3

1.6 1.6
0.53 5.4 4.8 I. I 11.8
0.476 0.008 0.484
0.476 0.0 1 6 0 .492

3 Contributi on =( [tolerance / % DM] X % diet) fo r beef and da iry cattle ; contribut ion = ([to leran ce] X % di e t ) for

poultry and s win e. 4 Alfalfa hay , forage and s ila ge b e lon gs to the n o n grass animal feeds gro up for which the highe s t residue observed

that results from a u se pattern s imilar to the lab eled u se pattern is 9.52 ppm ( Pronam id e Reg istration Standa rd , C. Trichilo , I I / 03 / 1985). 5 Data are not available (or r e quired) for a l fa l fa meal; howev er, a l fa l fa meal s hould be inc lu ded in the diet. Th e tolerance for nongrass an imal feeds (hay) is u sed for a l falfa mea l. 

### Conclusion: The updated dietary burden is 1

. 6 ppm fo r beef cattl e , 11.8 ppm fo r dairy cattle, 0.484 ppm for sw in e, and 0.492 ppm for poultry. These val ues are s i g nificantl y low e r than th e dietary burden previous l y calc ulated to estab li s h the c urr ent to lerances in mi l k, me at po u ltry and eggs, i. e. 10. 7 , 1.0 and 1.0 for beef catt le , sw ine and poultry , respectively , and sl ig htl y hi g her to that of dai ry cattle , 11. 2 ppm . B ased o n th e low e r or s imil ar re-calculated d ie tary burden s the current to l erances fo r pronam id e in liv estock commodities are co n s id e re d ad e quate. 

### 5.3.5. Foo d Handling (860.1460) Pronamide is n ot registered fo r u se in food-ha ndlin g es tabli s hment s; therefore , no res idu e chemistry data are required

. 

### 5.3.6 Water, Fish , a nd Irrigated Crops (860.1400) Pronamide is not reg istered for aquatic uses; th erefore, no res idu e chem istry data are requi red.
Page 17of20Pronamide Summary of Analytical Chemistry and R esidue Data Barcode 0427081

5.4 Food Res i d u e Pr o fil e Tol e rances ar e es tabli s he d i n th e 40 CFR § 1 80.317 fo r the u se of pron a m id e o n alfalfa, apple , arti c hoke , berri es, e ndi ve, s to ne fruits, grape , head lett uce, pear, radicchio , fie ld p ea , and rhubarb. In a dditi on , to lerances are estab li s he d fo r livestock commodities , and several feed commoditi es. Generally , residues below 0.4 ppm are expected in most food commodi ti es with the exceptio n of head lettuce (1.0 ppm) , endive ( 1.0 ppm) and radicchio (2.0 ppm)

. A tolerance of 2. 0 ppm was recommended to support a proposed new use on leaf lettuce. T hi s toleran ce is s upp orted by adequate residue data. In terim to l e rances of 0.10 ppm , 1.5 ppm and 3

. 0 ppm are r ecommende d for field pea seed , v in es and hay , respective ly . Add iti o nal crop fi e ld trial data is r eq uire d t o su pp ort the u se on field pea. The li vestock dietary burden was r eca lcul ate d to d e te rmine if ne w to lera nces for li vestock com moditi es we r e needed

. The no n -grass animal feeds are the maj o r contribu to rs to the dietary burd e n , field pea fee d st uff co mm od iti es be i ng a minor contri bu tor. B ased o n the lower or s imil ar re-calculated dietary burdens the current tolerances fo r pronamide in li vestock commodi ti es are conside red adeq uate. The current labels for pronam id e in c lud e plan t back intervals for other c rops than those recommended in the 2002 TRED. These PBls need to be removed. Rotation to othe r crops than leafy vege tables crop group 4 , root and tuber vegetables crop group I , and cereal grains is a ll owed by fo ll owing a 365- day PBI.

Page 18 of20Pronamide Summary of Analytical Chemistry and Residue Data Barcode 0427081


### A pp e ndix A . Inte rn ational Residue Limits Tab l e 

### Pronam ide (aka Propyzamide; PC Code 101701) Summary of 

### US and Internationa l Tolerances and Max i mum Residue Limits Res idu e D e finition: 

# us Canada M ex ico ' Codex 40 C FR 180.31 7 : 3.5-dichloro-J\T-(I , 1- No ne (a) Genera l. (I) propyz amide residues dim e th y l-2-propy n- Ico nvertible to meth y l 3 , 5-dic hl orobenzoate, yl)benza mide , includin g ex pre ss ed as the stoich iometri c eq uivalent of metab o lit es co ntainin g propyzamide, 3,5-d i c hl oro-N-( I , l-d im eth y l -2- th e 3 , 5-d ich lorobe n zoate propyny l )benzam id e m o iety Co mmodit y Toleranc e (po m ) /

Maximum Residue Limit**(111}.{ k~**  

# us Ca nada Mexico' Codex A lfalfa , seed 10.0 Animal feed, nongrass, g roup 18 10.0 Aoole 0.1 0.1 Art ic hoke , globe 0.01 Blackberry 0

.05 Blueberry 0.05 0.05 lowbush blueberries Bo yse nberry 0.05 Cattle, fat 0.2 Cattle , kidne y 0.4 Cattl e, liver 0.4 Cattle, meat 0.02 Cattl e, meat byproducts. excep t 0.02 kidne y and liver Egg 0.02 End ive 1.0 Fruit, s tone , group 12 0.1 Goat , fat 0.2 Goat, kidney 0.4 Goat, liver 0.4 Goa t , meat 0.02 Goa t , meat byproducts, exce pt 0

.02 kidney and liver Grape 0.1 Hog , fa t 0.2 I l og, kidne y 0.4 Hog, liver 0.4 Ho g, meat 0.02 Ho g, meat byproducts, except 0.02 kidney and liver Horse. fat 0.2 Ho rse, kidne y 0.4 Ho rse, li ve r 0.4 Horse, meat 0.02 I lo rse, meat byproducts , except 0.0 2 kidn ey and li ver Lettuce , head 1.0 I lettuc e

P age 

### 1 9 of20Pronamide Summary of Analytical Chemistry and Residue Data Barcode 0427081 

### Summary of US an d In te rn ation al To l erances and Maximum Re sidue Limi ts Res i due Definition: 

# us Ca n a d a Mex ico' Co d ex Milk 0.02 Pear 0. 1 0. 1 Pou ltry, fat 0.02 Poultry , liver 0.2 Poultry, meat 0.02 Poultry. meat byproducts, except 0.02 li ver Radicchio 2.0 Raspberry 0.05 Shee p , fat 0.2 Sheep, kid ney 0.4 Sheep , liver 0.4 Sheep , meat 0.02 Sheep , meat byproducts , except 0.02 kidney and liver Completed: M. Negu ss ie; 05 /07 / 15

1 Mexico adopts US tolerances and/or Codex MRL s for it s export purposes. (c) Tolerances with r egional r egistrations. Tole r ances with regiona l registration , as defined in § 1 80. I( I ), are establis hed fo r res idues of t he herbic ide pro pyzam ide , in c l udi ng its me t ab o lites and degrada t es, in or on the commod ities in th e tab le in this paragrap h. Co m p lia nce with the to lerance leve ls spec ified in this paragraph is to be determined by measu r ing on ly those p ropyzamide residue s convertib le to me thy l 3 , 5-dichlo robenzoate , exp ressed as the stoichiometric equivalent of propyzamide , 3,5-dichloro-N-( I , l-d imethyl-2-propynyl)benzamide , in or on the commodity.



## - - C ommodity Parts p e r mill io n Pea , licld , seed 0.05 Rh ubarb 0. 1 (d) I ndir ect or inadv e rt ent residue s. To lera nces are estab li s hed for ind irect or inadvertent residues of the he rbicide propyzamide , including its metabo lites and degradate s, in or on the commodities in the table in this paragraph. Compliance with the tolerance levels specified in this paragraph is to be determined by mea s uring only those prop yzamide residues convertible to methyl 3.5-dichlorobenzoate , expressed as the stoichiometric equivalent of prop yzamide , 3 , 5-dichloro- N

- ( I , l-dimethyl-2-propyn y l) benzamide , in or on the commodit y .

G G G C omm o dity

rain , cerea l, fo rage, group 16 rain, ce r ea l, hay , group 16 rain , cereal , straw, group 16



### Page 20 of20 Pa r ts p e r milli on 0.6



## - 0.2 0.3



### -

# Volume 3 

# Annex B



# Propyzamide (Pronamide)

B.5 METHODS OF ANALYSIS

B .5.2 ANALYTICAL METHODS FOR THE DETERMINATION OF RESIDUES (ALL COMPONENTS INCLUDED IN THE R ES IDUE DEFINITION PROPOSED)Propyzamide [PC Code I0 1 70 I] / Dow AgroSciences LLC

Note: This Data Eva lu ation Record (DER) was originally prepared under contract by Versar. I nc. (6850 Versar Center

. Springfield, VA 22151: submillcd 5 / 2 1115). The DER has been reviewed by HED and revised to rcnect current Office of Pesticide Pro grams (O PP ) po licies.

Tab l e B .5 . 2- 1. Overv iew of th e A n a l y tica l Met ho ds for t he D ete rmin a ti o n of Pro na mid e R esi du es. Data Matrix Analytcs Method Type Limit of Quantitation Reference Requirement (ppm) Enforcement Lettuce Propyzamide and GLC/E CD 0

. 2 PAM Vol. II. Method

- Plant its metabolites Method I Commodities convertible to Residue Chemistry methyl 3.5- Chapter for diehlorobenzoate Pronamide TR.ED. (MDCB) DP#275536. J. Morale s. 2 /28 / 02 Data-Gathering Ac i d i

c crons: Propyzamide and GC! MS 0.0 1 Study No. 04 10 I 0 Method

- Plant app l e; lem on: metabolite s (common moiety) MRID 49 1 26806 Commod itie s orange: orange containing the 3.5- peel and pulp dichlorobenzoyl Orv crons : moiety. including: alfalfa forag e. RH-24580. RHhay. and s eed: 246M. RI 1

- 24848. barle y forage. RH-25891. and grain. and s t raw: RH-26521 corn forage, g rain. and stove r : wheal forage, grain. and s traw**Oil~ 1**  croes: canola s eed: olive oil : soybean s eed: sunno"er s eed Wet crons: broccoli: grape: green bean; lettuce: potato ILV of Not required for the Enfo rcement P/\M, Vo lum e II

. Method- Plant Method I. for plant s Commodit i es because the primary enforcement method is the PAM MRM

. Radiovalidation Alfalfa Pr opyzamide and GLC !ECD 0

. 2 PAM Vol. II. of Methods

- Lelluce its metabolite s Method I Plant convertible to Residue Chemi s try Commodities methyl 3 , 5- Chapter for di ch lorobenzoate Pronamidc TRED. (MDCB) DP#275536. J. Morales , 2 / 28 / 02 Enforcement PAM Vol. II. Method- Method I Livestock (T R 34

- 9 1-68) Commodities Residue Chemi s try Chapter for Pronamide TRED. DP # 275536. J

. Morales. 2/28/ 02

EPA MR ID #: 46413406 Page 2 of 10Propyzamide (PC Code I 0170 I]/ Dow AgroSciences LL C

T a ble B.5.2-1. Overview of the Ana lyti ca l Method s for th e Determination of Pronamid e Residue s . Data Matrix Analytcs Method Type Limit ofQuantitation R eference Requirement (ppm) Dara-Gathering 51 31

85 Residu e Method- Chemistry Chapter L ivestock of the Re g istration Commodi ti es S tandard (MRIDs 00 I 07959 and 00 107967) lLVof Chicken fat Propyzamide and GC/MS 0.01 Study o.020063 Enfo rcement Oovine liver metabolites l GRM 02.2 1] MRJD 46414306 Method- containing the 3.5- (common moiety) DP # 427081 Livestock dichlorobenzoyl Co mmodities moiety Radiovalidation No t avai lable of Methods

- Livestock Commodities Multiresidue f-DJ\-Pest i eide Resid ue Chemistry Method Testing Analytical Chapter for Methods Multi- Pronamide TRED. Residue testing DP#275536. J. program Morales. 2128/ 02 MRID 43493203

Table B.5.2-2. C h e mi ca l Structures of Ana ly te s Addre ss ed b y Methods for the Analy s is of Propyzamide. Met hod ID C he mic a l Name C hemical S tru ctu re GRM 02.21 Co mmon Name :

# av~ Propyzamidc ( Pron amide) C h emica l Na me: 3.5- 

## I 0 

### NCit,

### ~II dichloro-N-( I

. I-dimethyl2-prop) ny l)bcnzamidc Cl GRM 02.21 Com mon Na me: 3 , 5- 

# CP-fl
D CB/\ (common moiety) C h e m ical Na me : 3,5

- C-OH di c h l orobc nzoic acid Cl GRM 02.2 1 Co mmon Name: 2,3

- 

# cO-R DCBA (common moiety) C hemi ca l Na me : 2.3

- C-OH dichlorobcnzoic acid

B.5.2.J 

### Ana lytica l Meth ods for P l ant Matrices (Annex llA 4.3, Annex IH A 5.3) Not ap plicabl e.

EPA MRID #: 46413406 P age 3of10Propyzam idc [PC Code I 0170 I) / Dow AgroSciences LLC B.5.2.2

B.5.2.2.1 Document 

### ID : Report:

Guid e lin es:

GLP C ompliance : Acce ptabili ty:

Eva lu ator : A nal ytic al Method s for Foodst uff of A nimal Ori g in (Livestock Matrices ; Annex IIA 4 .3, Annex lll A 5

. 3) Post-Registr ati on Method (Enforcement) MRID No. 46413406 Reed , R

. (2002) Ind epe ndent Laboratory Validation of Dow AgroSciences LL C M ethod G RM 02.21

- Determinati o n of R es idu es of Propy zami d e in Animal T iss ues by Gas Chromatography with Negative- Ion Che mic a l Ion ization Mass Spec trometry. Study Nos. 020063 and ML02

- I029- DOW. Unpubl i shed s tud y prepared by Dow AgroSciences LLC. 82 p. EPA OCSPP Hannonized Test Guideline 860.1340 Re s idu e Analytical Method (A ug u st 19 96) PMRA Re g ulator y Direc ti ve D IR98-02

- Res idue C he mi stry Guide l ines , Section 3

- Residue Analytical Method EU SANCO 825 / 00/ rev. 7 (17 / 3 / 04) O ECD Guidance Document on Pesticide Residue Analytical Methods No deviations from reg ulatory requirements were repo rt ed which would have a n impact on the va lidi ty of th e study. T he s tudy is conside r e d sc ientific a ll y acceptab l e. Th e acceptability of this stud y fo r r egu l atory purposes is a ddr essed in the forthcoming U.S

. EPA Re s idue Che m istry Summary**D~;nent ,**  DP # 42708 l. ld e li z Negr6n

- Encarnaci6 n 

# ry

EXECUT I VE SUMMARY
D ow AgroSciences LLC ha s s ubmi tted th e results of an independent laboratory valid ation ( ILV) of a gas chromatography method w ith negative-ion chemical io ni zation ma ss spec trometry detection (GC/ MS) , Method GRM 02

. 2 1 , for the determination of residues of propyzam ide (pron a mid e) and metabol ites contai n ing the 3,5-dic hlorob e nz oy l moi e ty as 3,5-d ic hlorob enzo ic acid (3,5-DC BA ) in poultry fat and bovine liv e r. The ILV was co nducted by Morse L abo rat ories (Sac ramento , CA). In addition to the IL V report , the submission included a complete method descript io n of the original method , along with supporting method validation data for milk , bovin e liver , kidney, mu scle, and fat , and eggs and poultry muscle and fat.
For the method as written , fat samp l es we r e ex tracte d with et hy l acetate , evaporated to dryness , and re di sso l ve d in 1 , 4

- dioxane. Th e samp les we re the n hydro lyze d by add i ng 9 N s ulfuri c ac id and heatin g at 100 ° C fo r at least 12 hours. During hydrol ys is . bound re s idu es of propyzamide and metabo lites were so lubilized and hydro l yzed to 3,5 -D CBA. Samples of milk , eggs , and tissues other than fat were extracted and hydrolyzed by add in g 4 N sodium hydrox ide and heating at 80 ° C for at least l 2 hours . During this initial hydrolysis, bound re s idu es of propyzam ide and metabolites were so lu b ili zed and hydro l yzed to 3,5- D CBA. Fo ll owing alkaline hydroly s i s, the sodium hydroxide so lu t ion was acidified with 12 N H C l (p H <2) and extracted and hydrol yze d as described above for fat samples. During the second hydrolys is , the bound residue s of propyzamide and metabolite s we re further so lu bi li zed and hydrol yzed to 3 , 5 -D C BA. Followin g hydrolysis , all samples we re cleaned up using an NH 2 solid pha se extractio n (S P E) column. Res idues were e luted with dichl orome than e : formic ac id (98:2, v:v). Afte r elution , EPA MR.JO # : 464 13406 Page 4of10Propy zamide [P C Code I0170 I] / Dow AgroSciences LLC
aceto n e 

### with peanut oil 

### was added as a 

### keeper , the 

### e luate was 

### evaporated to dryness , and residues were redissolved in isooctane:aceto ne (80:2 0 , v:v). A n internal standard , 2,3- dichlorobenzoic acid -tert-butyldimethylsilate (2,3- DCB A-TBDMS) , and a derivatizing agent , Nmethyl-N-(tert-b uty ldim ethylsilyl) -triflu oroacetamide (MTBSTF A) , wer e added , and the sample was derivati z ed for 60 minutes at 60 °C. After cooling , residues were analyzed by GC/MS. Th e method monitors two ions for determination of 3,5-DCBA fo ll owing derivatization. The limit of quantitation (LOQ, determined as the lowest limit of method validation ; LLMV) is 0.01 ppm. The reported limit of detection i s 0.003 ppm. The method provides for quantitation of residues in parent equivalents by use of ca librati on standards prepared in propyzamide equ ivalen t s. The method as written was ade quately va lid ated using samples of milk, bovine kidney , liver , muscle , a nd fat , and eggs a nd poultry muscle a nd fat fortified with propyzamide at 0.01, 0.05 , 0.25, and 0.50 ppm. R ecove ri es were within the acceptable range of 70-120 % at all fort i fication levels. Overall mean recoveries (and relativ e standard deviations) for prop yzamide we re: 86% (4 .5 %) for milk , 92% (6.8%) for bovine kidney , 93% (7.3%) fo r bovine liver , 94% ( 19 %) for bovine muscle , 90% (8.6%) for bovine fat , 98% (3.0%) for egg , 92% (6.7%) for poultry muscle , and 91 % (7.7%) for poultry fat. An ILV st udy for GC/MS m ethod GRM 02.21 was conducted by Morse Laboratories, Inc. (Sacramento , CA) u s in g samp le s of untreat ed bovine liver and c hicke n fat obtained from Dow AgroSciences fortified with propyzamide at 0.0 1 , 0.10, and 0.50 ppm (LOQ , 1 Ox the LOQ , and 50x the LOQ , respectively). The ILV was s u ccessful on the first attempt with c hicken fat; recoveries were within the acce ptabl e range of 70-120%. One recovery (268%) at the LOQ fo1tification leve l was determined to be an outlying data point u s in g th e Grubb's Method and was exc lud ed from the statistical calcu lations. The ILV was also considered s ucc essful on the first attempt with bovine liver. Th e first GC run was suspect due to an unacceptable calibration curve generated from the derivatized analytical standards accompanyi n g the sample set. This was determined to be the result of a double addition of internal standard to two of the ca libration standards. The original GC-ready extracts were re-injected against a new se t of d e rivati zed ana l ytical standards , a nd recoveries were w i thin the acce ptabl e range of 70-120%. Apparen t residues of propyzamide were below the LOQ ( < 0.0 1 ppm) in/on two unfortified samples each of chicken fat and bovine li v er. The ILV laboratory provided a table detailing a ll communic ation s with th e method developer during the s tud y. The IL V labo ratory rep o rted that profilin g th e e lution pattern of th e N H 2 SPE cartridge prior to analysis of actual samples is imperative and id e ntifi e d this as a critical step. The IL V laboratory reported that excluding overnight incubation , one set of 16 chicken fat samples required approximately 14.5 person h ours**2** calendar days) and one set of 16 bovine liver samples required 13.5 person hour s (
- 2 ca l e ndar days) to prepare samp le extracts for GC analysis. Automate d GC analysis was conducted overnight. No confirmatory analysis procedures were included. The registrant indicated that the GC/MS method is specific for the determination of propyzamide and metabolites as 3 , 5-DCBA by virtue of the chromatographic separat ion and selective detection system u sed

. Confirmation i s by

EP A MRJD # : 46413406 Page 5of10Propyzamide [PC Code I 01701 ] / Dow AgroSciences LLC 

### comparison of the retention time (GC) as well as the peak area ratio resulting from se l ected ion monitoring following MS analysis in the negative chemical ionization mode. No radiovalidation data were s ubm itted. Ana lyti cal methods should be radiovalidated to determine whether total toxic residues (TTR) are extracted from weath ered residues. This issue is addressed in 04 27801

. I. 

### Princ ipl e of the Metho d : GRM 02.21

Tab le B.5.2.2.1-1. Sum m ary Parameters fo r the Post-Reg i stration Ana l yt ical Method for the A n a lys i s of Propyzam id e Re s idues in A n ima l Ti ss ue s. Method ID GRM 02.2 1 Ana l yte(s) Propyzamide and metabolites containing the 3.5-dichlorobenzoyl (DCBA) moiety Extraction so l vent and Fat samp l es were extracted with ethyl acetate and centrifuged. An aliquot of th e extract was technique evapo rated to dryness , redissolved in 1.4-dioxanc , and hydro l yzed by add in g 9 N sulfuric acid and heating at I 00 °C for at l east 12 hours. During hydrolysis , bound residues of propyzamide and metabolites were solubi li zed and hydrolyzed to 3 , 5-DCBA

. Samp l es of milk. eggs. and tissue s o th er than fat were ex tracted and hydrolyzed by adding 4 N sodium hydroxide and heating at 80 ° C for at l east 12 hours. During this initial hydrolysis , bound residues of propyzamide and metabo lites were solubi li zed and hydrolyzed to 3,5-DCBA. Following alk a lin e hydrolysis, the sodium hydroxide solution was acidified with 12 N HCI (pH <2), sodium chloride was added, and the mixture was partitioned with ethyl acetate and centrifuged. An aliquot of the ethyl acetate phase was evaporated to dryne ss, redis so lved in 1, 4- dioxane. and hydrolyzed by add in g 9 N s ulfuri c acid a nd heating at I 00 ° C for at least 12 hours. During the second hydrolysis , bound residues of propyzamide and metabolites were further solubi li zcd and hydrolyzed to 3.5-DCBA. Clean -up strategics For all samples following hydrolysis, the sulfu ric acid hydrolysalc was partitioned with ethyl acetate , and the ethy l acetate phase was evaporated to dryness and redissolved in ethyl acetate. The s ample was c l eaned up using an Nl-12 S P E column. Residues were eluted with dichloromethane:formic acid (98:2, v:v). Aller e luti o n , ace tone with peanut oil was added as a keeper, the eluate was evaporated to dryness, and residues were redissolved in isooctane:acetone (80:20 , v:v). An int ernal s tand ard , 2 ,3 -D CBA -TBDMS , and a derivatizing agent, MTBSTFA, were added and the sample was dcrivatizcd for 60 minutes at 60 °C. After cooling. residues were a nal yzed by GC/MS. Instrument and Detector GC/MS using a Restek capillary column. Rtx

- 50 liquid phase MS de t ec tion operated in the negative-ion chemical ionization mode with selected ion monitorin g. The following ions were monitored: 3.5-DCBA-TBDMS: m l = 304 (quan tit a tion) and m / z 306 (confirmation) 2 , 3-DCBA

-TBDMS (interna l standard): m l z 155 (quantitat ion ) and m lz 304 (confirmation) Standardizatio n method External standardization using a 8-point calibration curve (linear regression. least squares fit). Ca libra ti on s tandards of 3 , 5-DCBJ\ arc prepared in propyzamidc equ i va l ents and derivatized prio r to ana lysis. Internal s tand a rdi zati o n using an internal sta ndard of2.3-DCBA-TBDMS Stab ilit y of std solutions Stab ilit y of th e s ta ndard so lutions was not reported. Retention times 3,5-DCBA: 9.51 minutes 2,3-DCBA (interna l standard): I0.29-10.30 minutes 

### The method provides for quantitation of residues in parent equivalents by use of calibration standards prepared in propyzamide equivalents. 

### II. Specificity The GC/MS method is specific for the determination of propyzam id e and metabolites as 3,5

- DCBA by virtue of the chromatog raphi c separation and selective detection system used. Confirmation is by comparison of the rete ntion time (GC) as well as the peak area ratio resulting
E PA MRIO # : 46413406 Page 6of10Propy zamide [PC Code I01 70 I) / Dow AgroSciences LLC from selected ion monitoring following MS ana lysis in the negative c hemical ionization mode. Ana lys is of control sa mpl es resulted in no significant sig nals at the ex pected retention times of the analytes. U nambi g uo u s identification of the co mmon moiety is ensured by monitoring two ions during MS determination as follows: 3 , 5-DCBA (derivatized) 

### m l z 304 ( quantitation ) mlz 306 (confirmat io n) Confirmation of the presence of the propyzamide and metabolites (as 3 , 5-DCBA) in the recovery s amples was indicated when the retention times of the analytes in the san1ples matched the retention times of the analytes in the calibration standards and the confirmation ratios (m/z 306 / 304) of the analytes in the samples were ± 20% of the average confirmation ratios of the standards. The confirmation ratios for each analyte were determined by calculat in g the ratio of the confirmation peak area to the quantitation p ea k area. The confirmation rat ios for eac h analyte in repre se ntati ve sample matrice s of bovine muscle and fat were within ± 20% of the average confirmation ratios of the standards , indicating that the method i s se lec ti ve for the determination of propy z amide and metabolites (as 3.5- D C BA ) in an imal tissues and fat. III . Linearity For propyzamide (as 3 , 5-DCBA), the linearity of detector response was eva luated using so lvent standard so luti ons. Ca li bration curves w e re ca lcu lated by linear regre ssio n ana l ysis. For the least-squares equation , which desc ribes the detecto r re spo nse as a function of the standard concentrat ion, a representative calibration curve reflect ing injection of e i g ht standards over the concentration ran ge of 0.075 -1 2.50 ng/mL 3 , 5-DCBA (equ ivale nt to 0.003-0.5 μgig propyzamide) demon s trated linearity with a coefficient of determination 

### (r2) o f 0.99983. IV. Accuracy (Recovery) a nd Precision ( Repeatability) Radiovalidation. No radiovalidation data were included in the submission. Ana lyt ical methods sho uld be radiova li dated to determine whether total toxic residues (TTR) arc extracted from weathered matric es. This i ss ue is addressed in 04 2780 1

. Method validation. Th e method as written was adequate ly validated u s in g samp le s of milk , bovine kidney, li ver, muscle , and fat, eggs , and p oultry muscle and fat fortified with propyzarnide at 0.01 , 0.05 , 0.25 , and 0.50 ppm. R ecove ri es were w ithin the acceptab le range of 70-120% at all forti fi cation leve ls. Overall me an recoveries (and relative standard deviations) for propyzamide were: 86% (4.5%) for milk , 92% (6.8%) for bovine kidney , 93% (7.3%) for bovine liver , 94% (19%) for bovine muscle , 90% (8 .6 %) for bovine fat , 98% (3.0%) for egg , 92% (6.7%) for pou ltry mu sc le, and 91 % (7.7%) fo r poultry fat. Repeatability data we re ge nerate d from at least two sa mpl es fortified at the LOQ and one or two s amples each fortified at 5x LOQ , 25x LOQ , and 50x LOQ for each matri x. The standard deviations (RSDs) obtai ned were less than 19 %. Recovery and repeatabi li ty data are presented in Tables B.5.2.2.1-3 , below.

EPA MRID # : 46413406 Page 7of10Prop y zamide [PC C ode I01 7 0 I] / Dow AgroScicnce s LL C
T a bl e B

. 5 .2.2. 1 -3. Acc u racy a n d Pre ci s io n D a t a fo r th e Va li d at io n o f GRM 02.2 1. An a lyte M a tri x Fortification Validation R eco ve ry(%) RS D L

e vel (ppm) Indi vidual Mea n Range (%) Propy zamid e Milk 0.01 83 , 84 86 83

- 92 4

. 5 111/

z 304 for 3 , 5- 0.05 92 D C BA 0

. 25 84 ( qu a ntitation ) 0

. 50 89 13 ov in c 0. 01 89.99 92 84-9 9 6.8 k i d ney 0.0 5 91 0. 25 98 0

. 5 0 84 B ov in e li ve r 0 .0 1 9 6 , 1 00 93 82-100 7.3 0.05 93 0.25 82 0.50 95 B o vine 0.01 97. 110 94 74-113 19 mu s cl e 0

. 05 74 0. 25 7 8 0 .5 0 113 B ov in e fat 0.0 1 79. 9 5. 1 0 1 90 79 -1 0 1 8.6 0

. 0 5 85 .9 6 0.25 89 , 95 0.50 79 , 88 Egg 0.01 99. 103 98 95- 103 3

. 0 0.05 95 0.25 97 0.50 9 8 Po ultry 0

. 01 9 7, 92 92 82-9 7 6.7 mu sc l e 0.05 82 0. 25 9 7 0 .50 94 Po ultry f a t 0.01 9 4. 81. I0 I. 99 9 1 81-101 7.7 0.05 82 , 86 0.25 93,98 0.50 87.87 Independent Laboratory Validat ion CILV). An lLV study for GC/MS method GRM 02.21 was conducted by Mor s e Laboratories , Inc. ( Sacramento , CA) using sample s of untreated bovine liver and chicken fat , obtained from Dow AgroSciences , fortified with prop y zan1ide at 0.01 , 0.10 , and 0.50 ppm and analyzed accord ing to the method procedures described in Table B.5.2.1.1

- 1. It was noted that minor modifications were necessa r y d ue to differe n t laboratory equip m en t (wate r bath , ce n tri fuge, evaporator , in cubator, oven , ul traso n ic cleaner , vac u um ma n ifo l d, vortex mi xe r , S P E cartridge , g lassware) ; h owever none of th ese adaptations h ad a n y i nfl u ence on the result s . The results of th e ILV are presented in Table B.5.2.2.1

- 4. The ILV was s uccessful on the first attempt with chicken 

### fat. One recovery 

### (268%) at the LOQ fortificat io n level was determined to b e an outlying data point using the Grubb ' s Method and excluded from the statistical calculations. Recov e r ie s were within the acceptable range of 

### 70-120%

. The ILV was also considered successful on the first attempt with bovine l iver. The first G C run was suspect due lo an unacceptable cal ibration curve generated from the derivatized ana lytical standards accompanying the samp le set w hi ch was the resu lt of a dou bl e addi t io n of in terna l standard to EPA MRJD # : 46413406 Page 8 of JOPropyzamide [PC Code I 0170 I]/ Dow AgroSciences LLC


### two of the calibration standard s. The original GC

- read y extracts we re re-injected against a new set of derivatized ana lytical standards and recoveries were within the acceptab l e range of 70- 120%. Apparent residues of propyzamide were below the LOQ (<0.01 ppm) in/on two unfortified samp le s each of chicken fat and bovine li ver. The IL V laboratory reported that profiling the e luti on pattern of the 

### N H2 S PE cartridge prior to analys is of actual samples is imperative and identified this as a critical ste p. There were several communi cations between th e Study Director at the confirmatory l aboratory and the method developers or others fami liar w ith the method

. These are listed below: • Availability of c lean control matrices • Approval of equivalent glassware and equ ipm ent • The Study Director asked for in struction on how to dilute sampl es with analyte responses falling outside the range of responses in the standard c urve. In struction s were to dilute the samples twofold w ith 20% acetone/ 80% isooctane so lution (con taining no interna l standard) prior to injection. • Discussion of typographical errors within the method write-up • Results of the first chicken fat ILV trial reported and discussed

. Di sc u ss ion regarding the high recovery at the LOQ and whether to treat it as an outl ier if shown to be so by the Grubb's test. • Discussion of the results from the first bovine liver trial and the unacceptable calibration curve that was produced. Th e Study Director suggested that a new set of derivatized calibration standards be prepared a nd to re-inject the original sample set with the new standards. This course of action was approved. • Results of the first bovine liver ILV trial repo11ed and approved

. The I L V laboratory reported that exc ludin g overnight in cubation, one set of 16 chicken fat samples required approximate ly 14.5 person hours (

- 2 calendar days) and one set of 16 bovine liver samp les required 1 3.5 person hours (

- 2 calendar days) to prepare samp l e extracts for GC analysis. Automated GC analysis was conducted overnight.

Tab le B.5.2.2.1-4. Accurac y and Prec is ion Data for t h e Independent Laboratory Va li datio n o f GRM 02.21 fo r Livestock Matri ces. Analyte Matrix Fortification Indep endent Laboratory Validation Recovery(%) RSD Level (ppm) Indi v idual Mean Ran ge (%) Propyzamide Chicken fat 0.01 268 1

, 106 , 1 02 , 114 , 9 1 103 91- 11 4 9.2 m

l = 304 for 3.5- 0.10 103 , 92 , 95. 106 , 84 96 84- 106 8.9 DCB/\ 0.50 79,88, 70 79 70-88 11.3 (quantitat i on) Bovine liver 0.01 I 05. I 08, I09, 98, 94 103 94-109 6.3 0.10 100, 120 , 1 20, 100, 1 10 110 100- 1 20 9.0 0.50 11 6, 111

. 111 113 111-11 6 2.6 1 Value determined to be an outlying data point using the Grubb 's Method. Not used for stat i st ical purposes. V. Lim i t of Q u antitation

Tab l e B.5.2.2.1-5. Sum m a r y of D etecti on and Qu a nt i tatio n L imits for GRM 02.21. Ana l )1e 

### I LOO (ppm)' 

### I LOQ (pp m) 2 Propyzamide. as 3.5-DCBA 

## I 0.003 

### I 0.01 1 LOD = limit of detection.
EP A MRID # : 46413406 Page 9of10Propyzamide [PC Code I0170 I] / Dow AgroSciences LLC

2 LOQ = limit of quantitation, defin ed as the lowest fortification level where acceptable precision and accuracy data were obtained. VI. 

### Co nclusion s The GC / MS Method GRM 02.21 was adequately va lidated for determining residues of propyzamide and metabolites containin g the 3,5-dichlorobenzoyl moiety as 3,5-DCBA following derivatization in milk , bovine tissues (kjdney, liver, muscle, and fat), eggs, and poultry tissues (m u scle and fa t). The val idated LOQ i s 0.01 ppm , and the LOO is 0.003 ppm. Addit ional information/data pertaining to radiovalidation are needed. The submitted ILV study is acceptable. Adequate recoveries were obtained fo r chicken fat and bovine li ver fort ifi ed with propyzamide at 0.0 I (LOQ), 0.10 ( 1 

### Ox LOQ), and 0.50 ppm (50x LOQ).

EPA MRJD # : 46413406 Page 10of10