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### EPAREGISTRATION DIVISION COMPANY NOTICE OFFILINGFOR PESTICIDE PETITIONS PUBLISHED IN THE FEDERAL REGISTER EPARegistration Division contact: Linda Hollis, Biochemical Pesticides Branch Chief, 703-308-8733 CPBio, Inc. EPAReg. # 4F8287 Choline Chloride
EPAhas received a pesticide petition 

### Choline Chloride from 

### CPBio Inc. P. 

### 0.Box 3367 Ontario, CA91761, proposing, pursuant to section 408(d) ofthe Federal Food. Drug, and Cosmetic Act (FFDCA), 21 U.S.C. 346a (d), to amend 40 CFRpart 180.
The addition ofan exemption from the requirement ofa tolerance for 

### Choline Chloride in or on the raw agricultural commodity 

### growing crops under40 CFR. EPA has determined that the petition contains data or information regarding the elements set forth in section 408 (d) (2) ofFDDCA; however, EPA has not fully evaluated the sufficiency ofthe submitted data at this time or whether the data supports granting ofthe petition. Additional data may be needed before EPArules on the 

### r--- petition. A. Residue Chemistry 

### The petitioner has provided EPA with sufficient toxicity data to supportthe issuance ofthis tolerance exemption for this inert ingredient. The data submitted will allow EPA to determine the sensitivities ofmajor identifiable subgroups of consumers, including infants and children.
1. Analyticalmethod. 

### The purpose ofthis study was to confirm a chromatographic method as the enforcement analytical method for the analysis of the active ingredient, in Choline Chloride 98o/o Technical. The Choline Chloride concentration is 98o/o) Technical was determined to be 99.03o/o Choline Chloride by high-performance liquid chromatographic (HPLC) analysis. Theidentity ofthe active ingredient, Choline Chloride, was confirmed bychromatographydata and spectral scans performed by GC/MS. 2. Magnitude of residues. 

### CP Bio, Inc. is petitioning the agency to establish an exemption from the requirement ofa tolerance for this natural class ofchemistry

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that is well known. Therefore information regarding the nature and magnitude of chemical re idue · re ultingfrom the usc ofthi product i. notr-equired. B. Toxicological Profile I. utt toxicity. Thetest substance, Choline Chloride 98 % Technical, wa. evaJuated for it. acute oral toxicity potential in female albino rats when administered as a gavage dose ata level of5000 mg/kg. ince the test substance failed the limit te t, the main testwas conducted following the up-and-down procedure (UDP) at 175, 550, 1750 and 5000 mglkg. The tudywas terminated following the stopping rule ofthi procedure. Mortality occurred only atthe 5000 

### mg/kg level. Therewere no clinical 

### signs oftoxicity during the tudy. 

### AnimaJ su_rviving to termination exhibited weekly weight gain, with thee ception ofone that to t weight between Days 7 and 14. Abnormal necropsy findings that occurred, only m the animals dying on te
.t, pertained to contents ofthe tomach and intestines. The acute oral 

### LDSO was estimated to 

### be 3129 mglkg. Derma) ensitization (OPPT 870.2600) A primary dermal irritation ,tudywas conducted on three albino r-.tbbits u ing te. t sub t-ance Choline Chloride 98o/u Technical. Therewas one intactte t itc per animal. Each test ite wa treated with 500 mg ofte t substance moistened witb O.S mL ofDl water and covered with a semi-permeable dre. ing. Thete. t sub tance wa maintained m contact witb the skin for 4 hour •Ob ervation for dermal irritation and defects were made at 1, 24, 48 and 72 bours after removal ofthe dressing • Irritation scores derived from there ·pcctive erythema and edema scores through the 72-hourobservations for each animal arepresented below. Acute Inhalation Toxicity (OPPT 870.1300) Testsub tance Choline Chloride98% Technical wa. to bee • aluated for acute inhalation toxicity potential in albino rats. Two pre-study trial assays were conducted to determine ifaero olizing the test substance into the e. posure chamber wouJd produce an acceptable concentration ofatleast2 mgfL with mass median aerodynamic diameter (MMAD) between 1 and 4um. Prior to one aero olization, test substancewas heated and ground. An aerosol was attempted using a Venturi A pirator connected to a SOOL e. po ure chamber. ln both attempts, the powder test

ub ·tance began to liquefy during aerosolization; the delivery method could not be adju ted for that occurrence and eoncentratjon information could notbe taken. Due to the nature ofthe tc t sub

**tam~e,** it · con idcred that an inhalation exposure to provide meaningful acute to icity data could not be done. PrimaryEye Irritation (OPPT 870.2400) An acute e e irritation study a conducted on three albino rabbit using test sub tance Choline Chloride 98% Technical. The test substance, 100 mg, was placed into the conjunctiva sac ofthe right eye ofeach animal elected fortesting. All treated eyes werewa bed with room teblperature DI waterfor one minute immediately afterrecording the24-hour observation. Therewereno positive effects exhibited in any eyes after treatment. Thetest ubstance i. rated minimall3 irritating and assigned to Toxicity Category IV. PrimarySkin Irritation (OPPT 870.2500) A skin sensitization tudy was conducted on 15 male and 15 female

**short~haircd** albino guinea pig to determine iftest substance Choline Chloride 98%) Technical produced a en itizing reaction. Males and female were assigned to each oftwo groups, de.ignated Nah• e (5/ ex) and Test (10/ ex). aive control group animals remained untreated during the induction phase ofthe study. Test group animals were treated with 400 mg oftest ub tance moistened with 

### DJ water (selected from previou creenjng). Tc t animal were treated once weeki for threeweeks for a total ofthree treatments. After a twoweek rest period, all animals (both group ) were challenged at a virgin test ite witb an application of400 mg oftest ub tanee moi tencd with Dl water. The te t substance producedno reaction m eithertheTc t animal. ortheNaive control animals after the challenge treatment. Therefore, Choline bloride 98% Technical is not a ensitizer in guinea pigs.

2. Geuotoxit'ly. The literaturewa extensively searched for studies on the genotoxicity ofcholine chloride. TOXNET, Pubmed, FDA website, TJ and Google cholar searche wereconducted. Tbi waiver request for the genoto icity studies i based on the results Ames abnonella a says and in vitro chromo omaJ aberration and si ter chromatid e, change a. ays conducted on choline chloride and Ame almonclla and mou.c micronuclcu a ay conducted on PC. All assayi\ were negative except for one outof

### mo i ·ter chromatid exchange as ays which was considered positive by TP and "ambiguous' by OE D. A weight ofthe evidence evaluation on the available data on choUne chl
. oride indicate it bas little genoto ic potential. The literaturewa extensively earchcd for tudies on tbe toxicity to plants ofcholine chloride. TOXNET, the internet in general and Google Scholar searches were conducted. o studie meeting the guideline requirement ofthe

tudie ; however, tudies were found on the effect ofcholine chloride on pea seedling , choline on rice eedlings, emergence and elongation ofradish root and algal tudie u ing lobjbition (%)ofPseudokircllerilla ubcpilata andScenedesmus subspicatus were located. 

### In aU cases, choline, choline chloride and acetyl choline bad beneficial effect or no effect on the parameters studied. A waiver is requested for the above tudie based on the natural occurrence ofcholine in pJant andits lack ofadver
.e effects described in the cited studies.

'3. /lr 

### ·,lfluc :ve anddeveloom 11/a/

### toxic:'v. The literaturewas extensi\ 'ely earched for studies on the developmental/reproductive toxicjty ofcholine chloride.4 

### TOXNET, Pubmed, the FDA website, TIS andGoogle cbolar earche were conducted. Awaiver is 

### requested forte ·ting choline chloride in a preliminary developmental to .icity 

### crecn based on the existence ofinformatio
. n available on the developmental toxicity ofcholine chloride in 

### mice and ofAPC in rat , on reproductive toxicity ofcholine in male rats and on the po itive influence ofcholine on learning and memory in young rats. Overall, the study authors concluded 

### thatthe re u!t indicate that prenatal choline

upplementation facilitate. cognitive function aero . thelifespan, whereas prenatal choline deficiency impairs divided attention and

**accelerate.~** 

### age-related·decline in temporal processing (4). In another tudy,the arne laboratory studie the effectof perinatal choline upptementation on timing and temporal memory u ing a similar treatment regime. Reaction to nicotine e. posur was determined. holine enhanced the temporal proce ing domain. The tudy authors related the effect to modification in cholinergic function (5).
4. Subchronic: toxicity
. 

### The literature was extensively searched for tudie on the subchronkinhalation toxicity ofcholine chloride. TO ET, Pubmed the FDA web itc, TI and Google cbolar earcbes were conducted. o repeated do e inhalation to. icity tudies were found; however, a 28-day intranasal tudy was located. A waiver is requested for 90-dayinhalation toxicity te ting on choline chloride based on its low to icity, 

### GRA tatu., its acute inhalation L SO 

### of>1.6 mg/L (no cUnical sign ofto icity reported) and the exi tence ofa 28-day intranasal toxicity tudy in 

### rat·. The result ofthe intrana
.al tudy are summarized below. 28-day lntr-lna at · tudy. In tbi tudy, Balb/c mice (10/se.x/group)were administered choline chloride at a dose of200 mg/kg in 50

# ,,L aline very 

### alternate day·for 28 day

**~** 

### The study wa conducted under OECD GLP regulation
. Body weight and food consumption were recorded daily. Mice were e amined daily for clinical signs ofto icity. Hematology (hemoglobin, RBC, WB count, monocytes, lymphoc · tes, neutrophil and eo inophoils) and clinical chemi try (urea, BUN, 

### HDL, total chole terol, AL T,A T and creatinine) parameter were evaluated at study termination. The pleenwas asepticaJJy removed and single ceJJ suspensions were made. Viable splenic cells were counted by 

### the trypan blue exclusion test. A detailed gross necropsy was performed on all animals and the lungs, heart, liver, spleen and kidney were weighed. Microscopic examination wa done on ' vital ' organs including tbe lung, heart, liver, spleen andkidney. No mortality occurred and no statistically significant effects on body weight gain orfood con umption were ob enred. o tatistically significant changes were observed in hematology and clinical cbemi. try parameters orspleen cell count. Organ weight were comparable5 

### between the control and treated group. otreatment-related hi topathology was ob erved in the examined organs. The NOAEL was con idered to be >200 mglkglday in mice (1).
5. Fmlr>crine disruptio
. 

### EPA i 

### required under the FFDCA, a amended by 

### FQPA, to develop a creening program to determi
. ne whether certain ubstances (including all pesticide active and other ingredients) "mayha\ • e an effect in human that is 

### similarto 

### an effect produced by a naturally occurring estrogen, or other such endocrine effect as the dministrator may de
.ignate." Following recommendations ofit EndocrineDisruptor and Testing Advi ory Committe (ED TAC), EPA determined that there wa a cientitic basi for includin , a partofthe program, the androgen and thyroid hormone sy 

### terns, in addition to the c trogen hormone system. Then EPA also adopted ED TAC' recommendation thatthe Program include evaluations ofpotentialeffect in wildlife. orpe ticidc cbemital , EPA 

### will use FIFRA and, to the extent thateffects in wildlife ma help determine whether a sub tance may have an effectin human , FFD A authority to requirethe wildlife evaluations. As 

### the science del·e)ops and resource allow, creening ofadditional hormone sy terns may he added to the Endocrine Di ruptor creening Program (EDSP). 

### When additional appropriate screeningand/or testing protocols being con idered under the Agency's ED P have been developed, Choline Chloride may be objected to further creening and/ortesting to better charaderize effect related to endocrine disruption.
C. Aggregate Exposure 1. c:tary 

### exposure. ln 

### evaluating dietary exposure to Choline Chloride based on its u e in food, feed, co metics, drug and in plants CP Bio belie ve there is no ri k form 

### any increase in

### expo urefrom the use as a PGR in food orfeed crops. EPA e tabUshe exemption from the requirement ofa tolerance onl in those ca es where it can be demon trated thatthe ri ks 

### from aggregate e ·posure to pesticide chemical residues underrea onably foreseeable circumstance 

### will po.e no appreciable ri ks 

### to human health. In order to determine the ri. ks 

### from aggregate exposure to pesticide products, the Agency considers the toxicity ofthe product in conjunction with possible e po ure to re iducs ofthe productthrough food, drinkingwater, and through otherexpo ure that occur a a re ult ofpesticide u e 

### in residential settings. If

### EPA is able to determine that a finite tolerance is not necessary to ensure thatthere is a reltsonable certainty thatno harm will 

### result from aggregate expo ure to the product, an exemption from the requirement ofa tolerance may be established. Section 408(b)(2)(C) ofFFDCA provides thatEPA halJ apply an additional tenfold (1

### OX) margin of afety for infants and children in the case ofthreshold effect to6

account for prenatal and postnatal toxicity and the completene ofthe database on toxicity and exposure unle s EPA determines based on reliable data that a different margjn ofsafety will be safe for infants and chiJdren. This additional margin 

### of safety is commonly referred to as the 

### FQP A safety factor ( F). In applying 

### this provision, EPA eitherretains the default value oflOX, or use a different additional

afety factor when reliable data available to EPA support the choice ofa different factor. CP Bio believe that the 

### lOX standard safet y factor is unneces aryfor Choline Chloride a this sugar is used as an additiYe in baby formula a a food supplement.
D. ( 'umulative t.D.'ecrs ufficient data 

### are provided on the chemicalidentity ofCholine Chloride and

**~**ufficient data are available on the metabolism and environmental degradation of this compound. CPBio relied collectively on information provided on the representative chemical structure , the ubmitted phy ical chemical data, structure similarity information, a 

### wcU a information from
.the 

### GRA a es ment and the use in food and feed for thi natural sugar. CP Bio ha concluded that a risk as essment ba ed on toxicity data for the parent compounds is not likel to underestimaterisk.
E. Stifely Determination

. L. \' popu/ntic'l. Ba ed on these

**ri~;k** as essments, P Bio, Inc. conclude that there is a reasonable certainty that no harm will result to the general popuJation or to infant and children from aggregate exposure to Choline Chloride. 

### :!. /nf(pzts an I · 'ldrt
. Based on these risk as e sment , 

### P Bio, Inc. concludes that there

### is a rea onable certainty that no harm will re ultto the general population or to infants and children from aggregate expo ore to Choline Chloride. F. lnternatwnal Tolerances CP Bio, Inc. is not aware ofany country requiring 'a tolerancefor .Choline Chloride nor have any ODEXMaximum Re idue Level been est.ablished for any food crops at this time.