UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON,

D.C. 20460 OFFICE OF CHEMICAL SAFETY AND POLLUTION PREVENTION MEMORANDUM Date: August 30, 2012 TXR No. 0056440 SUBJECT: Propoxur: Review
of Acute Comparative Cholinesterase Assay (CCA) Studies, Benchmark Dose Analysis and Derivation

of Acute Point of Departure (POD) DP Barcode: D401491/ D404194 Registration No.: NA PC Code: 047802 Decision No.: 464412 Petition No.: NA Risk Assessment Type: NA TXR No.: 0056440 Regulatory Action: Generic Data Call In Case No.: NA MRID No.: 48784801,48784802,48784803, 48784804

CAS No.: NA 40CFR:NA FROM: Deborah Smegal, MPH, Toxicologist
C\ J.YI... J-- (__ **c:J(__** Registration Action Branch

6 (RAB6) (})' Health Effects Division (7509P) THROUGH: Felicia Fort, Branch Chief

. f/1

(l --1_
_ Registration Action Branch

6 **(RAB~C1 ~­**  Health Effects Division (7509P) TO: Kaitlin Keller, Chemical Review Manager (CRM) Risk Management
and Implementation Branch

3 Pesticide Reevaluation Division (PRD) (7508P) I. CONCLUSIONS The series of non-guideline cholinesterase (ChE) inhibition studies are acceptable and satisfy the generic data call-in (DCI) requirement

for propoxur
for

a comparative cholinesterase assay (CCA) study in adult rats versus postnatal day (PND)

11 pups. Page

1 of2

\OPP'sreviewindicatesthatthedatashowlifestagesensitivity(i.e,pupsaremoresensitivethanadults),andthatRBCChEismorethanbrainAChEinhibition.Inrecentyears,itistypicalOPPpracticetoperformB,MDanalysisonCCAstudies.OPPperformedbenchmarkdose(BMD)analysesoftheCCAonpropoxurinordertoestablishapointofdeparture(POD)forthesingle-chemicalriskassessmentusingtheagency'sBenchmarkDoseSoftware(BMDS).OPPanalyzedtheregistrant-supplieddataofredbloodcell(RBC)acetylcholinesteraseinhibition(AChEI)inmaleandfemalePND11pups(combinedsexdata)andcalculatedaBMD10(BenchmarkDosefor10%inhibition)as0.0427mg/kg/dayandtheBMDL10(i.e.95%LowerBoundConfidenceLimitsontheBMDlO)wasestimatedas0.0285mg/kg/day.OPPhasconcludedthattheuseofaBMDL10of0.0285mg/kg/daywillprovideahealth-protectiveandscientificallysupportableapproachforanacutePOD.BasedonacomparisonofBMD1Os,pupswereapproximately11foldmoresensitivethantheadultratsforbrainandRBCcholinesteraseinhibition(combinedsexdata).II.ACTIONREQUESTEDReviewtheseriesofnon-guideline comparativecholinesterase(ChE)assaystudies(MRID48784801-48784803)thatweresubmittedasrequiredbytheregistrationreviewDataCallIn(DCI).III.BACKGROUNDAseriesofnon-guidelinecholinesterase(ChE)inhibitionstudies(MRID48784801-48784803)wereundertakentoevaluateanydifferencesbetweenpostnatalday11(PND11)pupsandadultrats(approximately7-8weeksofage)withregardtocholinesteraseinhibitionforpropoxur.ThesestudiesareacceptableandsatisfythegenericDCIrequirementforpropoxurforaCCAstudyinadultratsversuspostnatalday(PND)11pups.Consistentwiththen-methylcarbamatesmodeofaction(MOA),thesestudiesevaluatedcholinesteraseinhibitionfollowingasingledoseatthetimeofpeakeffect.Boththerangefindinganddefinitivestudiesprovidegooddoseresponsedata.Thedefinitivestudyuseddosesfrom0.1to1mg/kg,whiletherangefindingstudyusesdosesfrom1to10mg/kginadultsand0.3to3mg/kginpups.IV.RESULTS/DISCUSSION(orMRIDSummaryTable,etc.)MRIDSummarya eTblEIxamp1eStudyTypeMRIDCommentsNon-GuidelineAcuteTimeCourseStudy48784801NewDERNon-GuidelineAcuteDoseRange48784802NewDERFindingStudyNon-GuidelineAcuteDose-Response48784803NewDERStudyNon-GuidelineStudy,Estimationof48784804ConsideredinEPABenchmarkBenchmarkDoseforPropoxurDoseAnalysisPage2of2! Cholinesterase Inhibition

in Pups and Adult Rats (2012)

I Page

1 of27 PROPOXUR/

c CODE 047802

Non-Guideline EPA Reviewer:

' Deborah Smegal, MPH **Signature**

**11 .......,~.,........~~~~-**  Risk Assessment Branch VI, Health Effects Division (7509P) Date._---="""+..f.C9,........._---.-. EPA Secondary Reviewer: Elissa Reaves, PhD Signature: **~~~~~~~~-** Risk Assessment Branch IV, Health Effects Division (7509P) Date __ **~'----¥--6-F-'---** I TXR#: 0056440

,

! DATAEVALUATIONRECORD

**~** STUDY TYPE:

; Non-guideline; Effects on Brain and RBC Cholinesterase

in Adult and Juvenile ! Rats PC CODE: 047802

I

DP BARCODE: D401491 TEST MATERIAL (PURITY): Propoxur (98.6% a.i.) SYNONYMS: CITATION:

I

• Tqot, J.D. (2012).

A Dose-Response Study of Red Blood Cell and Bram Cl1.olinesterase

in Juvenile Rats following Oral (Gavage) Administration

of Prbpoxur WIL Research Laboratories, LLC, Ashland, OH. Laboratory Project ID: WIL-551 010, March

9, 2012. MRID 48784803. Unpublished. Toot, J.D. (2011).

A Time Course Study ofthe Effects ofPropoxur

on Red Blood Ceil and Brain Cholinesterase

in Adult and Juvenile Rats Following Oral **(G:~vage)** Administration.
WIL Research Laboratories, LLC, Ashland, OH. Laboratory Project ID: WIL-551008, November
21, 2011. MRID 48784801. un:published. Toot, J.D (2012).

A Dose-Range Finding Study of the Effects ofPropoxur

on Red Blood Cell and Brain Cholinesterase

in Adult and Juvenile Rats Following Oral (Gavage) Administration.
WIL Research Laboratories, LLC, Ashland, OH. I

• Laboratory ProJect ID: WIL-551009, March

5, 2012. MRID 48784802. Unpublished.

' I SPONSOR: Wellmark International, Schaumburg,

IL and Bayer HealthCare, Shawnee **~ission,** KS

. :i EXECUTIVE SUMMARY

- This series of non-guideline cholinesterase (ChE) inhiqition studies (MRID 48784801-48784803)

was undertaken

to evaluate any differences between postnatal day 11 (PND 11) pups and adult rats (approximately

7-8 weeks of age) with rega,d
to cholinesterase **bition.**

1 I

. Time-Course **Stu~y** (MRID 48784801): The study was divided into

3 phases (Phase

I, Phase

II and Phase III). Pmpoxur (98.6% a.i.; Lot#: 1104080407)

in com oil was administered once via gavage

(5 mL!kg

) adult male or PND
11 male pups to determine
the time of peak cholinesterase inhibition. **Dos~g** was as follows:
(1) Phase

I dosing was to 40 adult Crl: CD (SD) rats at a **:~**

J ·ICholinesteraseInhibitioninPupsandAdultRats(2012)IPage2of27PROPOXUR/pcCODE047802Non-Guidelinedoseof5mglkg;(2)PhaseIIconsistedof12malePND11Crl:CD(SD)ratsatadoseof3mglkgtoassesssignsofoverttoxicityandguidedoseselectionforPhaseIII;(3)PhaseIIIconsistedof40malePND11Crl:CD(SD)ratsatadoseof3mglkg.'RBCandbraincholinesteraseactivitiesweredeterminedat15,30,60,120,and240minutesafterdosingintheadult5mglkgandPND11pup3mglkggroups(controlsevaluatedat15or249minutes).Results:Allrats(bothage"groups)survivedtoscheduledsacrifice.Inphase!IIIPND11pups,tremorswerenotedfor12/40malesreceiving3mglkg,whilenotreatment-relatedclinicalfmdingswerenotedinadultmalesinPhaseIorPND11malepupsinPhaseII.MaximalRBC(63%)andbrain(46%)inhibitionwasobservedat15minutesat5mg/kginmales.InthePND11malepup,maximalRBC(74%)andbrain(64%)inhibitionwasobservedat30minutesat3mglkg.However,RBCandwholebraincholinesteraseactivityin3.0mglkggroupincreasedfrom0.5-to4-hourspost-dosing(106%and83%,respectively,at4hours),providingevidenceofpartialrecoveryinbrain.Overallresultsfromthedosesevaluated,inthetimecoursestudysuggestthatthetimeofpeakinhibitionoccurredslightlyfasterinadults:(15minutes)thaninPND11pups(30minutes),butthePND11pupstooklongertorecover.Basedontheresultsofthistime-coursestudy,15minutesand30minuteswereselectedasthetimeofpeakcholinesterasedeterminationinthedefmitivedose-responsestudyforadultsandPND11pups,respectively.Followingacuteoralexposuretopropoxur,brainweightswerecomparableamongthegroupsinbothagegroups.'DoseRange-FindingStudy(MRID48784802):Thisstudywasperformedtodeterminethedoselevelsforuseinthedefinitivedose-responsestudy(MRID48784803).Adult1Crl:CD(SD)rats8weeksofage(1,2,3,5,or10mglkg)andPND11pups(0.3,0.5,1,2,1or3mglkg)wereadministeredsingledosesofpropoxurincomoil5ml/kgviagavage(6/sexldoseforbothagegroups).At15and30minutespostdoseforadultsandpups,respectivelycholinesteraseactivitywasassessedintheRBCandbraincompartments.Results:Notestsubstance-relatedclinicalsignswereobservedintheadultratsattimeofdosing(clinicalsignswerenotrecordedinpupsduetotechnicianerror).Adose-relatedreductioninChEactivity(bothcompartments)wasobservedatalldoselevelsinbothsexesandagegroups.Inadultrats,meanRBCcholinesteraseinhibition(ChEI)rangedfrom15%to84%,whilemeanwholebrainChEIrangedfrom3%to53%.InPND11pups,meanRBCChEIrangedfrom36%to82%,whilemeanwholebrainChEIrangedfrom22%to64%.Atthecommondoseof1.0mg/kg,RBCcholinesterasewasreduced15-16%intheadultratscomparedto57-58%inthePND11pups.SincesignificantChEinhibitionwasobservedatthelowestdoseinPND11pups,dosagelevelsof0.1,0.3,and1.0mglkgwereselectedtofurtherelucidateChEinhibitionatlowdosesforbenchmarkanalysesinthedefinitivedose-responsestudy.Brainweightswerecomparableamongthegroupsinbothagegroupsandsexes.Dose-ResponseStudyinPups(MRID48784803):Inthedefmitivedoseresponsestudy,propoxur(98.6%a.i.;Lot#:1104080407)wasadministeredonceviagavageincomoil(5mllkg)to11PNDIIpups/sex/doseatdoselevelsofO,0.1,0.3,or1.0mg/kg.AdultswerenotevaluatedCholinesterase Inhibition

in Pups and Adult Rats (2012)

I Page

3 of27 PROPOXUR!pc CODE 047802

Non-Guideline the range-fmding study resulted

in adequate inhibition

of RBC and brain ChE. RBC and brain ChE were evaluated

at the pre-determined
time of peak effect of 30 minutes. Results: Clinical signs. There were no treatment-related effects on mortality

in either age group. Clinical signs were not recorded
for any of the treated pups
as per the protocol prior to euthanasia due to technician error. Brain weights were comparable among the male and female PND 11pups. RBC Cholinesterase Results. As suggested

in the dose range-finding study, PND
11 pups were more sensitive than the adult rats based on
a comparison

of RBC cholinesterase inhibition

. In the PND
11 pups, both sexes displayed

a dose-related reduction

in RBC cholinesterase activity, with females being more sensitive than males. The magnitude

of RBC inhibition being 33-49% at 0.3 mg/kg and **~4-66%**

at the highest dose tested (1.0 mglkg)
in both male and female PND11 pups. At 0.1 mg/kg (lowest dose tested), 31% RBC cholinesterase inhibition was observed

in the female and 12% in male PND 11 pups. Brain Cholinesterase.
PND
11 pups were more sensitive than the adult rats, based on
a comparison

of brain ChE inhibition. Brain ChE activity was decreased

at all doses in the PND 11 pups (both sexes). The magnitude

of the decrease was 7.7% (M) and 8% (F) at 0.1 mg/kg, 23% (M) and-31%
(F) at 0.3 mg/kg, and 55% (M) and 48%(F)

at 1.0 mg/kg
in the PND11 pups (both sexes). HED conducted BMD analyses
on the propoxur comparative cholinesterase
data (CCA) for both adults (MRID 48784802)
and PND
11 pups (MRID 48784803)

for both RBC and brain compartments.

In additiofl, HED considered
the registrant's submitted benchmark dose (BMD) analysis
of the cholinesterase findings (Mihlan and Sheets (2012). Estimation

of Benchmark Dose for Propoxur Based on RBC Cholinesterase Activity

in Post-Natal Day 11 and Adult Rats; MRID 48784804). HED was able to reproduce most of the registrant's BMD analyses for the male and females.

In addition, HED evaluated
the male and female pup RBC data from the dose response study to see if it was appropriate

to combine these data for BMD analysis. Although the statistical tests that show

a borderline significant difference

in these datasets (p=0.067), HED concluded that **th~se** datasets can be combined

to provide more robust ChE measures

as there
is no biological basis to support sex-differences

for PND
11 pups, and the observed difference

is likely due to **vari~bility**

in the RBC ChE measurements. Details of Agency's BMD analysis are presented

in a separate BMD memo (D404194 memo from

J. Liccione/ B.Sarkar August 2012 TXR 0056440

' These studies are classified

as acceptable/non-guideline.

These studies
do not satisfy

a guideline requirep1ent

for propoxur. They satisfy the generic data call-in requirement

for propoxur
for

a comparative cholinesterase study in adult rats versus postnatal day (PND)

11 pups.

1 COMPLIANCE

i - Signed and dated Data Confidentiality,
GLP Compliance, Flagging, and Quality **Assuranc~** statements were provided. ICholinesteraseInhibitioninPupsandAdultRats(2012)IPage4of27PROPOXUR/pcCODE047802.Non-GuidelineI.MATERIALSANDMETHODSA.MATERIALS1.Testmaterial:Description:Lot#:Purity(w/w):Stability:CAS#:Structure:2.Vehicle-comoil3.TestanimalsSpecies:Strain:PropoxurWhitecrystallinepowder110408040798.6-98.5%a.i.Expirationdate:April26,2012;WILID#1100A4114-26-1RatCrl:CD(SD)Adultage/weightatstudyinitiation:adultrats5-6weeksatreceipt;time-mateddamsreceivedonGD10,11or12(#551008:GD10;#551009:GD11or12);usedtosupplyPND11pups;selectedadults:::17-8weeksold;bodyweights:#551008PhaseI8weekmales244-303g;#5510097weekmales:206-267gandfemales:151-195g.PupAge/weightatdosing:Source:Housing:Diet:Water:Environmentalconditions:Temperature:Humidity:Airchanges:Photoperiod:Acclimationperiod:11daysold;males:18.8-28.6g;#551009PhaseII20.9-28.9gmalesand19-27.2gfemales;#551108PhaseIImales21-28g;PhaseIIImales19-27gCharlesRiverLaboratories,Inc.(Raleigh,NC)Eachdamwashousedwithherlitterinanestingboxduringthepost-natalperiod.Adultswereindividuallyhousedinstainlesssteel,wirebottomedcages.CertifiedRodentDiet#5002(PMINutritionInternational,LLC.),adlibitumReverseosmosistreated(on-site)drinkingwater,adlibitum22±3°C50±20%10/hr12hrsdark/12hrslightAdults,16-14days;pups0-1daysPROPOXUR!pc CODE 047802 B. STUDY DESIGNS Cholinesterase Inhibition

in Pups and Adult Rats (2012)

I Page

5 of 27 Non-Guideline This series of non-guideline cholinesterase inhibition studies (MRID 48784801-48784803)

was undertaken

to evaluate any differences between neonatal (postnatal day 11; PND 11) pups and adult rats with regard
to cholinesterase (ChE) inhibition

as a biomarker
for more general neurological effects. Preliminary studies (MRID 48784801; and MRID48784802)
were performed

to determine
the appropriate dose levels for the time to peak effect study and for the definitive dose response study in PND11 pups (MRID48784803)

with regard
to inhibition

of red blood cell (RBC) and brain ChE. The adult ChE data in the range-fmding study (n=6/sex/dose) were determined

to be adequate
by EPA (email from M. Mizens

to K. Keller summarizing conclusions

of Dose-Response Conference
call on 10.25

. 11)

, and thus only additional pup data were collected

in the definitive dose-response study (MRID 48784803). 2. In-life dates- MRID 48784801: WIL 551008: Phase

1 Start: May 10, 2011; End: May 24, 2011; Phase
II Start: May 24, 2011; End: June 16, 2011; Phase III Start

: June 7, 2011; End: June 30, 2011 MRID 48784802: WIL 551009: Phase

1 Start: August
16, 2011

; End: September
1-2, 2011; Phase

II Start: August 30, 2011; End: September
21, 2011 MRID 48784803: WIL 551010: Start: December

12, 2011 End: December

14, 2011 3. Animal assignment
and treatment -Dose Range Finding Study (MRID 48784802). Phase

I (adult rats): There were

5 propoxur-treated groups and one control group, each consisting

of 6 adult rats/sex/group (Table
1). After randomization (based on body weight stratification randomized

in a block design), these rats were randomized into
2 study replicates

to allow for the reasonable conduct
of cholinesterase assessments. Each dose group and sex was **~qually** represented within each study replicate. Phase
II (PND
11 pups): Pups were randomly assigned

to one of 5 treatment groups and one control group; such that no more than

1 pup/sex/litter
was assigned

to each treatment group; each group consisted

of 11 pups/sex/dose. Each dose group and sex was equally represented within each study replicate
for dose administration. Adults and pups (on PND 11) received

a single gavage dose at a volume
of 5 mL/kg body weight. Study design: Table

1 shows the objectives and treatment groups allocated
for the three studies.PROPOXUR/pc CODE 047802 CboUnesterase Inhibition

in Pups and Adult Rats (2012)

I Page

6 of 27 Non-Guideline TABLE

1. Study Design for Cholinesterase Inhibition Studies
on Propoxur MRID Dose(s) #rats/sex Objective/Treatment/Termination Study# (mglkg) 48784801 Phase

I (adult):

0, 5 Phase

I (adult Determine time to peak inhibition (RBC and brain cholinesterase). WIL-551008 Phase

II (PND
11

8 weeks of Single oral dose in adults (Phase

I) and PND
11 (Phase III) male rats

. pups):
0, 3 age):

8
An additional group ofPND11 male pups (Phase II) were Toot 2011 males/time administered propoxur prior to Phase III to assess for signs of overt Time Course Phase III (PND
11 point toxicity

in PND 11 pups; Study
pups):
0, 3 Controls

: 8 adult (Phase

I) and
9 PND
11 pups (Phase III) terminated Phase

II (PND

at 0.25 and

4 hrs, 11 ): 12 males

8 adults (Phase

1) and
8 PND
11 pups (Phase III) terminated

at 0.25, treated and
3 0.5,
1, 2, and
4 hrs post dosing. control males Phase III (PND 11):

8 males/ /time point and 17 control males 48784802 Phase

I (adult):

0, 1, 6/sex Determine dose levels (dose-range fmding) for the inhibition ofRBC WIL-551009

2, 3, 5, 10

and brain cholinesterase following

a single oral dose in adults (Phase Phase

II (PND
11

I) and PND 11 (Phase II) rats. Toot2012 pups):
0, 0.3, 0.5,
1, 2
Single oral dose; clinical observations

at approximately

15 and 30 Dose Range and3

min post-dose
for adults and PND 11 pups, respectively; Finding

cholinesterase activity (brain and RBC) evaluated within

1 hourpostStudy

dose (termination)
and sample collection. 48784803 PND 11 pups: 0.1, Determine
the dose-response

for inhibition ofRBC and brain WIL

0.3,

1 11pupsl cholinesterase

in PND 11 (Phase II) male and female rats. Single oral 551010

sex/dose
dose: PND
11 pups at 30 minutes post dose. Toot2012DoseResponse Study
in Pups PND

= postnatal day 4. Dose selection and sampling time rationale; The dose levels and sampling times selected for the dose-response
study in pups (MRID 48784803) were based on the results of the two studies, described below (a, b). Propoxur was administered once via gavage

(5 mllkg)

to 11 PND
11 pups/sex/dose

at dose levels of 0, 0.1, 0.3, 1.0 mg/kg
in the definitive dose-response study (MRID 48784803).Cholinesterase Inhibition

in Pups and Adult Rats (2012)

I Page

7 of 27 PROPOXUR/pc CODE 047802

Non-Guideline a) Time point selection rationale-

A time-course study (MRID48784801;
WIL 551008) was conducted

in both adults and PND 11 male rats in three phases.

In Phase

1, 8 adult rats/time point were treated with single (gavage) doses ofO or 5 mg/kg.
In Phases

II (12 pups) and III (8 pups/time point), PND11 pups were given

a single dose of3 mg/kg; Phase
II pups were assessed for signs of overt toxicity within
12 hours and were discarded, while Phase III pups were evaluated further. For adults (Phase

I) and pups (Phase III), RBC and brain cholinesterase activities were analyzed

at sacrifice

at approximately

15, 30, 60, 120, or 240 minutes post dose (Tables

2 and 3). All animals (both age groups) survived

to scheduled sacrifice.

In Phase III, tremors were noted in 12/40 males in the
3 mg/kg group. No substance-related clinical signs were noted in adults (Phase

I) or the pups in Phase II. In the adult male rats, maximal RBC and brain inhibition was present

at 15 minutes post dosing (63 and 46%, respectively)

to 5 mg/kg. PND 11 male pups displayed

a maximal RBC and brain inhibition response

at 30 minutes

at 3 mg!kg (74% and 64%, respectively). Regarding
the time to peak effect, time points of 30 minutes and 15 minutes were selected for the PND 11 pups and adult rats, respectively,

for the definitive dose-response study (MRID48784803;
WIL 551010). a) Dose selection rationale
for time to peak effect study

- Dose levels were selected by the Sponsor based on the results of previous studies (Toot,2011, WLL-551008 MRID 48784801 and Toot, 2012, WIL-551009 MRlD 48784802).

In the time course study (Toot, 2011, WLL551 008), inhibition

of RBC and whole brain cholinesterase activity

in PND
11 males was observed

in the 3.0 mg!kg group, with peak inhibition noted
at 0.5 hours following dose administration. However, RBC and whole brain cholinesterase activity
in the 3.0 mg/kg group increased (106.0% and 83.0%, respectively) from 0.5- to 4-hours post-dosing

, providing evidence of partial recovery.

In the dose range-finding study (Toot, 2012, WIL-551009), administration

of the test substance

at doses of 0.3, 0.5, 1.0, 2.0, and 3.0 mg/kg
to PND
II pups resulted

in a doserelated, statistically significant, reduction

in ChE activity

in both the brain and RBC compartments

at all dose levels in both sexes and age groups (Tables

4 and 5). The PND
11 pups showed

a greater reduction

in RBC cholinesterase activity

at 1 mg/kg (males/females
57% ChEI) than the adult rats (males 14% ChEI/females
16% ChEI)
at the same dose level. Also at 1 mg!kg, PND
11 pups (both sexes) displayed

a greater reduction

in brain cholinesterase activity

( 42-4 7% ChEI) than the adult rats (3-1 0% ChEI).
In pups, RBC inhibition ranged from 36% to 78% in males and from 38% to 82% in females, while whole brain cholinesterase ranged from 22% to 61% in males and from 25% to 64% in females

at 0.5 hours following dose administration.Cholinesterase Inhibition

in Pups and Adult Rats (2012)



### I Page

8 of27 PROPOXUR!pc CODE 047802

Non-Guideline Table
2. Time Course Study (MRID48784801;
WIL 551008)- Mean (±SD) Cholinesterase Activity

in Male PND 

### 11 Pups Administered Propoxur 

### via Gavage (acute)• Time post-dose Dose

Red Blood Cells (minutes) (mg/kg) cholinesterase (UIL)



### % inhibition 

### " Males

**l~** ...

**~**
**!/'~** 15

0

6327±920 (adjusted
for



# - outlier) 3 3288±2595

(a)
48* 30

0



# -

- 3 1639±279
(a)

. 74** 60

01,;



# -- 3 2527±1342

(a) 60** 120

0



# - 3

2251±1231 (adjusted) 

### i(b)

64** 240

0 4780±993



# - 3 3375±1246

29.4* a Data were obtained from Table

S 1 0 (pages 66-68) of the study report. n = 8, except where noted; (a) n=7, 

### (b) n=5 * **p~0.05;**

 p~0.01** 

Brain cholinesterase (UiL)



### % inhibition 

### .



# . 24230±1150



# - 14345±5489

(a) 40.8** 

# -

- 8762±1424
63.8** 

# --

- 9659±957

60** 

# -

- 13820±5098

43* 24097±1039.6



# - 16020±2426
33.8** Table
3. Time Course Study (MRID48784801; WIL551008). Mean (±SD) Cholinesterase Activity

in Male Adult Rats Administered Propoxur 

### via Gavage (acute). Time post-dose Dose

Red Blood Cells

Brain (minutes) (mg/kg) Cholinesterase (UIL)



### % inhibition Cholinesterase



### % inhibition (UIL) 

## /' .... **~~**



### -

.;:



### , 

## ..

.. -· Males

**·~** 

## .. 

# ,

**~ .~.r**  .c. 15

0 3150±530
49994±1008 

### 5 1175±471**
62.7** 27084±5859**
45.8** 30

0



# - 5 1901±419

40* 30966±2599

38* 60

0



# - 5 2572±403

18.3* 36666±2459

26.6* 120

0



# - 5 3093±635

2 43835±1989

12.3 240

0 3482±379
50118±1782 

### 5 3467±332

0.4 47978±2392

4.3 a Data were obtained from Table S5 (pages 58-60) of the study report. n = 8; * **p~0.05;**

** pS:O.OlPROPOXUR/pc CODE 047802 Cholinesterase Inhibition

in Pups and Adult Rats (2012)

I Page

9 of 27 Non-Guideline Table

4. Study_ Design and Results (RBC) of Range-Finding Study (MRID48784802;
WIL

5 51009) Dose #of
RBC Cholinesterase (U/L) Sample Time (mg/kg) rats/sex Males Females (minutes post- dosing) Adults 0

6 2928±1187.6 3220±894

15 1

6 2499±468!14.7% 2697±728

t 16%

15 2

6 2061±792 !29.6% 2158±630* !33%

15 3

6 1701±934* !41.9.7% 1515±645** !53%

15 5

6 1453±408* !50.4% 1246±427** !61%

15 10

6 1120±636** !61.7% 485±239** !85%

15 PND 11 Pups 0

6 7657±1187 7158±884

30 0

. 3

5 4902±683** !36% 4444±879** !37.9%

30 5 4459±1062** 4124±1001**

30 0.5
!41.8% !42.4% 1

6 3259±573** !57.4% 3029±900** !57.7%

30 2

4 2470±749** !67.7% 1302±294** !81.8%

30 3

5 1726±328** !77.5% 1547±604**

t 78.4%

30 a Data obtained from Table S9 and S10 (pages 52-53) and Table S15 and S16 (pages 62-63) of the study report. * **p~0.05;** mean± s.d.  p~0.01;**  mean± s.d. Table
5. Study Design and Results (Brain)
of Range-Finding Study (MRID48784802;
WIL 551009) Brain Cholinesterase (UIL) Sample Time Dose #of Males Females (minutes post- (mglkg) rats/sex

dosing) Adults 0

6 52184±1887.5 51558±1322

15 1

6 50600±2619!3% 46481±2540 !9.8%

15 6 43686±2300 42136±5139 U8.3%

15 2

* !16.3% 6 41315±3712* 34790±2268** !32%

15 3
!20.8%

\\PROPOXUR/pcCODE047802Dose#of(mglkg)rats/sex56106060.360.56162636CholinesteraseInhibitioninPupsandAdultRats(2012)IPage10of27Non-GuidelineBrainCholinesterase(UIL)SampleTimeMalesFemales(minutespost-dosing)34964±4661**L33%26276±6965**t49%1528107±8309**24324±13626**15t46.1%L52.8%PNDllPups23640±198324486±8123018456±2756**18378±2164**30L21.9%L24.9%16627±2221**15268±2395**30L29.7%L37.6%13719±3004**12856±1936**30L42%L47.5%10073±1541**10058±3756**30L57.4%L58.9%9149±2023**8683±1743**30L61.3%t64.5%aDataobtainedfromTableS9andS10(pages52-53)andTableSISandS16(pages62-63)ofthestudyreport.*mean±s.d.**mean±s.d.5.Testsubstancepreparationandanalysis-Thedoseformulationswerepreparedonceassingleformulationsforeachdosedividedintoaliquotsfordispensation,andstoredrefrigerated.The doseformulationswerestirredcontinuouslythroughoutuse.Results:Homogeneityanalysis:Thedosing formulationswereinspectedvisuallyandwerefoundtobevisiblyhomogeneousand acceptableforadministration.HomogeneityanalysesinMRID48784803showedmean%oftargettobe91.7%-99.4%forformulationsof0.02,and0.2mg/mL.HomogeneityanalysesinMRID48784802showedmean%oftargettobe95.7%-111%forformulationsof0.06,and0.2mg/mL.Stabilityanalysis:Inthedose-responsestudy(MRID48784803),formulationsof0.02,and0.2mg/mLwere101%,and97.1%,respectively,ofthemean%oftimezeroafterstorage(refrigeration)for7days.Concentrationanalysis:Inthedose-responsestudy(MRID48784803),thedosingformulationswerewithinthetestinglaboratoriesstandardoperatingproceduresrangeforsuspensions(85%to115%)andwerewithintheprotocolrequirementforconcentrationacceptability(90%to110%oftarget)exceptforinitial0.02mg/Lformulationthatwas157%oftargetconcentration,andwasnotadministeredtoanimals.The0.02mg/Ldoselevelwasreformulatedtobe95.6%ofthetargetconcentration.Intherangefmdingstudy(MRID48784802)concentrationanalyseshadmean%oftargetof97.5%-107%andwerewithinWILResearchStandardOperatingProcedurePROPOXUR!pc CODE 047802 (SOP) range for suspensions. Cholinesterase Inhibition

in Pups and Adult Rats (2012)

I Page 11 of 27 Non-Guideline Concentration

(%» of target): Dose Response Study (PND
11 Pups) (MRID 48784803): 95.6- 104%; Dose Range Finding Study (MRID 48784802): Phase

I (adults) 97.5- 103%; Phase

II
( pups): 101-107%. 6. Statistics Time to peak effect (MRID 48784801) Dose Range Finding Study (MRID 48784802): and dose response (MRID 48784803) studies

- All statistical tests were performed using appropriate computing devices
or programs. Analyses were conducted using two-tailed tests (except

as noted otherwise)
for minimum significance levels of 1% and 5%, comparing each test substancetreated group to the control group by sex. Each mean was presented with the standard deviation (S.D.), standard error (S.E.), and the number
of animals (N) used to **cal~ulate** the mean. Due to the use of significant figures and the different rounding conventions inherent

in the types of software used, the means, standard deviations, and/or standard errors on the summary and individual tables may differ slightly. RBC and whole-brain cholinesterase
data were subjected

to a parametric one-way ANOV

A (Snedecor and Cochran, 1980) to determine intergroup differences between the control and test substance-treated groups. Ifthe ANOVA revealed significant (p<0.05) intergroup variance, Dunnett's test (Dunnett, 1964) was used to compare the test substance-treated groups
to the control group. Intergroup comparative statistics were not conducted
for body weight and brain weight data. C. METHODS 1. Observations

- Dose-Response Study (MRID 48784803) (Pup data) and Dose Range Finding Study (MRID 48784802) (Adult data) a. Adults

- All adult rats, including dams of offspring, were observed

at least twice daily for mortality and moribundity. Clinical examinations were performed prior to dosing, ::::20 minutes following dose administration,

and immediately prior to sacrifice (::::5-10 and 20-25 minutes post dose for Phases

I and II, respectively). b. Pups -Pups were sexed on PND 0, 4, 7, and 10. Pretest body weights and detailed physical examinations

for each pup were recorded
on PND

1, 4, 7, and 10. To reduce variability, litters were culled to 10 pups/litter, 5/sex when possible,
on PND

4 using

a computerized randomization procedure. 2. Body weight

- Body weights
of all adult rats and PND
11 pups were recorded initially and used as the basis of randomization
into dose groups. Additionally,

all rats were weighed
on the day of dose administration

to determine individual doses. 3. Cholinesterase activity determination

- Blood samples and whole brains were collected from each pup at approximately

30 minutes (Dose-Response Study MRID 48784803),
and each adult
at approximately

15 minutes (Range-Finding Study MRID 48784802) following dose administration

for determination

of cholinesterase activities

as follows. PND 11 pups in the dose

) ']__ response study were anesthetized with isoflurane and euthanized

by exsanguination during the JPROPOXUR/pcCODE047802CholinesteraseInhibitioninPupsandAdultRats(2012)IPage12of27Non-Guidelinebloodcollectionprocedure.Intherangefindingstudy,adultsandPND11pupswereeuthanizedbycarbondioxideinhalation.Bloodsamples(atleast1mLforadultanimalsand0.5mLforPND11pups)werecollectedfromtheinferiorvenacavaintochilledtubescontainingsodiumheparinastheanticoagulantandcentrifugedfor10minutesat4°C.Theplasmawasdiscardedandthepackedredbloodcells(RBC)werediluted1:20(w:v)using1%TritonX100solution(buffered).Thecontentsofthetubeweremixed(vortexmixer)andtheRBCpreparationwasanalyzed.Brainswereexcisedfromtheskullandweighed.Wholebrainswerediluted1:10(w:v)using1%TritonX-100solution(unbuffered)andhomogenized.Thehomogenatewascentrifugedforminutes4°C,andthesupernatantwasanalyzed.RBCandwholebraincholinesteraseactivitiesweredeterminedusinganassaybasedonamodification(Hunteretal.,1997)oftheEllmanreaction(Ellmanetal.,1961),whichusesacetylthiocholineasasubstratetomeasuretotalcholinesterase(i.e.,theassayisnotspecifictoacetylcholinesterase)viaaphotometricperiod(WILSOPNo.T5-146-2,dated9/12/2006).Topreventreactivationoftheenzyme,allsamplesweremaintainedinanice-waterbathfromthepointofcollectionuntilanalysisforcholinesteraseactivity.Thetimeofeachbloodsampleandbraincollectionwasrecordedandsampleswereanalyzedwithin1hourofsamplecollection,basedontheresultsofapreviousvalidationstudyofcarbamateinhibitionofcholinesterase(Roegge,2009,WIL-99420).II.RESULTS-Pups:Def"mitiveDose-ResponseStudy(MRID48784803)Adults:DoseRange-FindingStudy(MRID48784802)A.OBSERVATIONS1.Clinicalsignsoftoxicity.!}Pups-Noclinicalsignswereobservedpriortodosing.Noclinicalobservationswererecordedpostdosingduetotechnicianerror,asdocumentedintheprotocoldeviations.b)Adults-Noclinicalsignsoftoxicitywerenotedintheadultspriortodosing,orjustpriortoeuthanasia(approximately5-10minutesfollowingdoseadministration)atanydoselevel.2.Mortalitya)Pups-Allpupssurvivedtoscheduledtermination.b)Adults-Alladultratssurviveduntilscheduledtermination.B.BODYWEIGHTS-PupsandAdults-BodyweightsofthetreatedPND11pupsandadultratswerecomparabletotheirrespectivecontrolsonthedayofdoseadministration.C.CHOLINESTERASEACTIVITY-ThecholinesterasedataforbothcompartmentsandagegroupsaresummarizedandpresentedbelowinTables6and7.1.RedBloodCell(RBC)Cholinesterase-Pups.RBCdatainpupswasevaluatedinthedefinitivedose-responsestudy.RBCcholinesteraseactivitywasdecreased(dose-related)inbothsexesat0.5hourfollowingacuteexposure(malesdecreased12-66%/femalesdec.reased31-65%))todoselevelsrangingfrom0.1to1.0mg/kg(Table7).Inmales,themeanRBCcholinesterasePROPOXUR!pc CODE 047802 Cholinesterase Inhibition

in Pups and Adult Rats (2012)

I Page 13 of27 Non-Guideline activity

in the 0.3 and 1.0 mg/kg groups was statistically significantly (p<0.01) lower (33% and 66%, respectively), than the control group.
In females, the mean RBC cholinesterase activity

in the 0.1, 0.3 and 1.0 mg/kg groups was statistically significantly (p<0.01) lower (31%, 49%, and 65% lower, respectively) than the control group. Adults. Adult ChE data was evaluated

in the range-finding study, not the definitive doseresponse study. RBC cholinesterase activity was decreased (dose-related)

in all dose groups
in the adult rats of both sexes
at 15 minutes following acute exposure (males decreased
15- 62%/females decreased 16-85%)

to dose levels ranging from 1.0 to 10 mg/kg (Table 8). With the sexes combined,
the reductions were 15-74%.
At lower doses, males and females had similar responses

(1 and
2 mg/kg), but at higher doses females had

a greater response than males (i.e., 42% vs. 53% at 3 mg/kg, 50% vs. 61% at 5 mg/kg and 62 vs. 85% at 10 mg/kg for males and females, respectively).

At the common dose of 1.0 mg/kg, RBC cholinesterase
was reduced 15- 16% in the adult rats compared

to 65% in the PND
11 pups. 2. Brain Cholinesterase- Pups- Brain cholinesterase activity was decreased

at all doses compared

to the control values
in the male (8-55%) and female (8-48%) PND
11 pups (Table 7). A BMD analysis will provide

a level
at which 10% inhibition was observed.

In males, mean whole brain activity was statistically significantly (p<0.01

or p<O.OS) lower
in all three dose groups relative

to the control group.

In females, whole brain cholinesterase activity was statistically significantly (p<0.01) lower than the control

in the 0.3 and

1 mglkg/day groups. Adults- Brain cholinesterase activity was decreased

at all doses in male (16-46%) and female (10-53%) adult rats except the lowest dose in the male rats (Table 8). At the common dose of 1.0 mg/kg, brain cholinesterase
was reduced 3-10%
in the adult rats compared

to 48-55%

in the PND 11 pups. Table
6. PND 11 pup cholinesterase results from definitive dose-response study (MRID 48784803) Sex omwk!

0.1 mwk! 0.3m2fk! t.omwk! RBC Cholinesterase (U/L_l PND 11 Males 6737±1121.1 %!
5916±986.3 4532±1319.4 2323±591.8 12.2

32.7 ..
65.5•• range

4796-7918 5416-8782

2182-6364 1474-3396 PND

11 6994±2057.7 4856±489.9 3544±1166 2460±900.1 Females %!

30.6••
49.3••
64.8•• range 4188-10758 4244-5530 2196-6124 1474-3848 Brain Cholinesterase (UIL) PND 11 Males 21887±1315.5 18250±1

. 639.2 10594±1190 %! 23708±1480.1

7.7•

23••
55.3•• range 21082-26230 19796-24692 14277-19866 9040-12595 PND

11 24263±1220.8 22320±1610.7 16844±2432,9 12592±2300 Females 8 /o!

8 30.6••

48.1 .. range 22959-27078 19117-24191 14305-21859 9641-17993PROPOXUR!pcCODE047802CholinesteraseInhibitioninPupsandAdultRats(2012)IPage14of27Non-GuidelineDataobtainedfromTableS7andS8(pages42-43)andTableA7/A8(pages187--194)ofthestudyreport;*Dunnett'sp-value#<0.05or**p<0.01;<IIIIDunnett'stestwasusedtomakepairwisecomparisonsofeachindividualtreatedgroupwiththecontrolgroup.Table7.Adultcholinesteraseresultsfromtherange-findingstudy(MRID48784802)Sex0ml!fk21ml!fk22ml!fk23ml!fk25ml!fk210ml!fk2RBCCholinesterase(UIL)Males2928±1187.62499±468.42061±792.51701±934.81453±408.51120±636.4153042*50*62**Range1892-5020 1920-31961058-3168520-2928874-1980794-2362Females3220±894.92697±728.82158±630.11515±645.91246±427.5485±239.51633*53**61**85**Range1962-42701652-35321270-3220606-2278818-1764252-902BrainCholinesterase(U/L)Males52184±1887.550600±2619.943686±2300.241315±3712.834964±466128107±8309.1316*21**33**46**Ranee50068-5479647460-5464741150-4671934861-4362730790-4097119611-39910Females51558±1322.946481±2540.1 42136±5139.134790±2268.726276±696524324±13626101833**49**53**Range49750-5339642747-4968636182-4812432210-3826717979-3704816975-52060DataobtainedfromTable4(pages48-49)andTablesA9/A10(pages215-270)ofthestudyreport;n=6rat/sex/group*statisticallysignificant*Dunnett'sp-value*p<0.05,or**p<0.01D.BRAINWEIGHT-Followingacuteoralexposuretopropoxur,brainweightswerecomparableamongthegroupsinbothagegroupsandsexesinallthreestudies(timecourse,doseresponseanddoserangefmding).E.BENCHMARKDOSECBMD)ANALYSIS:REDconductedBMDanalysesonthepropoxurcomparativecholinesterasedata(CCA)forbothadults(MRID48784803)andPND11pups(MRID48784802)forbothRBCandbraincompartments.TheresultsarepresentedinTable8.Inaddition,REDconsideredtheregistrant'ssubmittedbenchmarkdose(BMD)analysisofthecholinesterasefindings(MihlanandSheets(2012).EstimationofBenchmarkDoseforPropoxurBasedonRBCCholinesteraseActivityinPost-NatalDay11andAdultRats;MRID48784804).REDhasreviewedthesubmissiontoverifytheBMDanalysisofthecholinesterasedatapresentedinMRID48784804.REDconductedaconfirmatoryanalysisusingEPA'sBenchmarkDoseSoftware(BMDS)2.1.1,thesamesoftwareusedinMRID48,784804andwasabletoreproducetheregistrant'sBMDanalysesforthemaleandfemales.Inaddition,REDevaluatedthemaleandfemalepupRBCdatafromthedoseresponsestudytoseeifitwasappropriatetocombinethesedataforBMDanalysis.TheregistrantproposedthattheAgency usethecombinedBMDLlOforregulatorypurposes.Toevaluatethemeritofthisapproach,REDconductedaLikelihoodratiotest(LRT)forRBCpupdatabasedontheBMDoutput.TheresultoftheLRTisnotstatisticallysignificantbecausepvalueisgreaterthanconventionalalphalevelof0.05.Thusthelikelihood-basedevaluationsuggeststhatthemalelandfemalePNDllRBCdatacanbecombinedinthe dose-responsemodeling.EPAalsobelievesthatthereisnobiologicalbasisforgender-baseddifferencesinAChEinhibitioninPROPOXUR/pc CODE 047802 Cholinesterase Inhibition

in Pups and Adult Rats (2012)

I Page 15 of27 Non-Guideline PND11 rats with N-methyl carbamates,
like propoxur. Given the lack of statistical significance, and lack of biological basis for gender based difference

in RBC at PND11 HED has concluded that male and female data for PND11 pups can be combined
for deriving more robust POD for AChE inhibition

at this age. Details
of Agency's BMD analysis are presented

in a separate BMD memo (D404194 memo from
J. Liccione/ B.Sarkar August 30, 2012 TXR 0056440). Sex Male " Female I · Combined Sexes

Table

8 Summary

of OPPs Benchmark Dose (BMD) Analyses for RBC and Brain ChE inhibition Adult"
PND 11 PupsA PND 11 PupsA Target (MRID 48784802 (MRID 48784803 (MRID 48784802 Range Range Finding Study) Dose ReSj >Q_nse Study) Findinrz Study) Organ BMD1o BMDL1o BMD1o BMDL1o BMD1o BMDL1o Brain 1.26 1.09 0.13
0.12 0.14 0.11 - RBC 0.73 0.498 0.094 0.075

0.1 0.08 Brain 0.88 0.76 0.08
0.06

0.1 o.os ; RBC 0.55 0.46 0.02
0.01 0.088 O.Q67 Brain 1.29 1.04 0.122 0.0875

--

--- RBC 0.474 0.339
0.042.7 0.0285

--

-- A Adult BMDs based on data m MRID 48784802; PNDII pup BMDs based on data m MRID 48784802,
and MRID 48784803. Ill. DISCUSSION
and CONCLUSIONS A.INVESTIGATORS' CONCLUSIONS (from Report Summary)

- Based on the time to peak effect study (MRID 48784801)

in adult rats and PND
11 pups, the investigators concluded that 15 minutes post-dosing
was the appropriate time to sample blood and brain tissue for cholinesterase activity
in the adult phase and 30 minutes post-dosing was appropriate

in the PND 11 pup phase of the comparative dose-response study. Recovery ofChE activity was observed

at 4 hours post dosing for adults administered

5 mglkg, while complete recovery time was not identified
for pups given

3 mglkg. In the range finding study (MRID 48784802)

a dose-related reduction

in cholinesterase activity (both compartments)
was observed

at all dose levels in both sexes of adults and PND
11 pups (adults:

1, 2, 3, 5, or 10 mg/kg and PND
11 pups: 0.3, 0.5,
1, 2, or 3 mg/kg).

In adult rats, mean RBC cholinesterase inhibition (ChEI) ranged from 15% to 85%, while mean whole brain ChEI ranged from 3% to 53%. In pups, the lowest dose tested of 0.3 mglkg resulted

in 36-38% RBC ChE inhibition and 22-28% brain ChEI (both statistically significant).

\lCholinesteraseInhibitioninPupsandAdultRats(2012)IPage16of27PROPOXUR/pcCODE047802Non-GuidelineInthedoseresponsestudy(MRID48784803)withPND11pups,bothsexesdisplayedadoserelatedreductioninRBCcholinesteraseactivityat0.3mg!kgformalesand0.1mg!kgforfemales.TheinhibitionofbrainChEactivitywasobservedat0.3mg!kgformalesandfemales.InPND11males,meanRBCChEIrangedfrom33to66%andbrainChEIrangedfrom23-55%.InPND11females,meanRBCinhibitionrangedfrom31%to65%andbrainChEIrangedfrom31%to48%.Theinvestigatorsdidnotidentifyano-observed-effectlevel(NOEL)foradultsorpups,althoughat2mg!kgstatisticallysignificantRBCChEI(33%)wasnotedinadultfemalerats,andstatisticallysignificantbrainChEI(16%)wasnotedinadultmalerats.Inpups,thelowestdosetestedof0.1mg!kginthedefinitivedoseresponsestudy resultedin31%and12%RBCChEinhibitioninfemalepups(statisticallysignificant)andmalepups(notsignificant),respectively.B.REVIEWERCOMMENTS-Therewerenotreatment-relatedeffectsonmortalityfollowingacuteoralexposuretopropoxurineitheradultratsorPND11pups.Thetimetopeakeffectis15minutespost-do.singforadultsand30minutesforPND11pupsfollowingpropoxurdosesof5mg!kgand3mg!kg,respectively.Inthetimecoursestudy,tremorswerenotedfor12/40PND11pupsgiven3mg!kgpropoxurbetween0.25and1hourpost-dosing,whilenotreatment-relatedclinicalfindingswerenotedinadultmalesdosedwith5mg!kgpropoxur.PND11malepupswithtremorsdisplayedcorrespondingRBCandbraininhibitionof48-74%and41-64%,respectivelybetween15and60minutespost-dosingwith3mg!kg.Inadultmales,RBCandwhole-braincholinesteraselevelsweresimilartothoseinthecontrolgroupatapproximately4-hourspost-dosing,indicatingcompleterecovery.ForPND11malepups,RBCandwholebraincholinesteraseactivityinthe3.0mg!kgpupgroupincreasedfrom0.5-to4-hourspost-dosing(106%and83%,respectively,at4hours),providingevidenceofpartialrecoveryinbrainofpups.Overallresultsfromthedosesevaluatedinthetimecoursestudysuggestthatthetimeofpeakinhibitionoccurredslightlyfasterinadults(15minutes)thaninPND11pups(30minutes),butthePND11pupstooklongertorecover.Intherangefindingstudy(MRID48784802)therewasagreateramountofinhibitioninpupsatasimilardoseinadultsandtherewassignificantinhibitionatamuchlowerdosethanadults.BothsexesofPND11pupsdisplayedsignificantbraincholinesteraseinhibition(22%-64%)atdoselevelsof0.3mg!kg-3mg!kgforbothsexes,whereasasimilarbutlowerlevelofbraininhibition(3%-53%)occurredinbothsexesoftheadultratsat1mg!kgto10mg!kg.SignificantRBCcholinesteraseinhibition(>36%)wasobservedinthePND11pupsat0.3mg!kgandabove,whereastheadultratdisplayedthislevelofRBCcholinesteraseinhibitionat2mg!kgandabove.Atthecommondoseof1.0mg!kg,RBCcholinesterasewasreduced15-16%intheadultratscomparedto57-58%inbothsexesofPND11pups.Inthedoseresponsestudy(MRID48784803)withonlyPND11pups,bothsexesdisplayedadose-relatedreductioninRBCcholinesteraseactivity,withfemalesbeingmoresensitivethanmales.Themagnitudeofinhibitionwas33%formalesand49%forfemalesat0.3mg!kgandapproximately65%(bothsexes)atthehighestdosetested(1.0mg!kg).At0.1mg!kg(lowestdosetested),31%and12%RBCcholinesteraseinhibitionwasobservedinthefemaleandmalePND11pups,respectively,whichwasstatisticallysignificantforfemalepups.-InPNDpups,braincholinesteraseactivitywasdecreasedatalldosesinbothsexes.ThemagnitudeofthePROPOXUR/pc CODE 047802 Cholinesterase Inhibition

in Pups and Adult Rats (2012)

I Page 17 of 27 Non-Guideline decrease was 7.7-8%

at 0.1 mg/kg, 23-31%

at 0.3 mg/kg, and 48-55%

at 1.0 mg/kg
in the PND 11 pups (both sexes). The studies

are classified

as acceptable/non-guideline.

These studies
do not satisfy

a guideline requirement

for propoxur. They satisfy the generic data call-in requirement

for propoxur
for
a comparative cholinesterase study in adult versus postnatal day (PND)

11 pups. D. STUDY DEFICIENCIES-
None that would impact study interpretation.
The description

of the cholinesterase assay procedures lacks sufficient details.PROPOXUR/pc CODE 047802 Cholinesterase Inhibition

in Pups and Adult Rats (2012)

I Page 18 of 27 Non-Guideline APPENDIX

A- SUMMARY TABLES Summary Table

1. PNDll Rat Control Values Across Studies Males PND11 RBC U/L MRID 48784802A range 7657±1187 finding study MRID 48784801(timecourse) 15 minutes
6327±920 240 minutes
4780±993 MRID 48784803A 6737±1121.1 (dose-response) Females PNDll RBC UIL MRID 48784802A
7158±884 MRID 48784801

Not tested MRID 48784803A 6994±2057.7 Males PNDll Brain U/L MRID 48784802A 23940±1983 MRID 48784801 15 minutes 24230±1150 240 minutes 24097+1039.6 MRID 48784803A 23708±1480.1 Females PNDll Brain U/L MRID 48784802A 24486±812 MRID 48784801

Not tested MRID 48784803A 24263±1220.8 30 mmutes post dose Summary Table
2. Adult Rat Control Values Across Studies Males- Adult RBC U/L MRID 48784802A 2928±1187.6 MRID 48784801 15 minutes
3150±530 240 minutes
3482±379 MRID 48784803A 2928±1187.6 Females- Adult RBC U/L

I MRID 48784802A
3220±894 MRID 48784801

Not tested MRID 48784803

A 3220±894.9 Males -Adult Brain U/L MRID 48784802A 52184±1887.5 MRID 48784801 15 minutes 49994±1008 240 minutes 50118±1782 MRID 48784803

A 52184±1887.5 Females -Adult Brain U/L MRID 48784802A 51558±1322 MRID 48784801

Not tested MRID 48784803

A 51558±1322

. 9 A 15 mmutes post dose, NT females not tested


# ~0PROPOXUR/pc CODE 047802 Cholinesterase Inhibition

in Pups and Adult Rats (2012)

I Page 19 ofl7 Non-Guideline Summary Table
3. PND 11 Pup Cholinesterase Values Across Studies
at 2 Common Dose Levels PND 11 Pup RBC Cholinesterase Values UIL Study Control

0

. 3 ml!!lu!

1.0 ml!!lu! Males MRID

7657

± 1187 4902±683 U36%)** 3259±573 U57.4%)** 48784802A range finding study MRID

6737

± 112l.l 4532±1319.4 U32.7%)** 2323±591.8 U65.5%)** 48784803A

- (dose-response)

Females MRID 7158±884 4444±879 (37.90/o)** 3029±900 U57.7%)** 48784802A range imding study MRID 6994±2057.7 3544±1166 (49.3%)** 2460±900

. 1 U64.8%)** 48784803A

- (dose-response)

PND 11 Pup Brain Cholinesterase Values UIL Males MRID 23640±1983 18456±2756 (21.90/o) 13719±3004 U42%)** 48784802A range fmding study MRID 23708±1480.1 18250±1639.2 (23%)** 10594±1190 U55.3%)** 48784803A (dose-response)

Females MRID 24486±812 18378±2164 U24

. 9%)** 12856±1936 U47.5%)** 48784802A range finding study MRID 24263±1220.8 16844±2432.9 (J.30.6%)** 12592±2300 U48%)** 48784803A (dose-response) (!%inhibition

at 30 minutes;* p<0.05;

** p<O.Ol; mean± s.d.UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON,

D.C. 20460 MEMORANDUM Date: August 30, 2012

OFFICE OF CHEMICAL SAFETY AND POLLUTION PREVENTION SUBJECT: Propoxur: **Ben~hmark** Dose Analysis

of **A~ute** Oral Studies and Derivation of a Point of Departure. PC Code: 047802 Decision No.: 464412 Petition No.: NA Risk Assessment Type: NA TXR No.: 0056440 MRIDNo.:NA FROM: John Liccione Toxicologist/Ris RABV

DP **Bar~ode:** D404194 Registration No.: NA Regulatory **A~tion:** NA Case No.: NA CASNo.:NA 40CFR: NA Health Effects Division (7905P)

r And

C'L

.. n **~ s~~~Nt)LJ.!L~**  Bayazid Sarkar **V.J~ilNt{**

..S: Mathematical **Statisti~**

0. Chemistry and Exposure Branch (CEB) Health Effects Division (7905P) THROUGH: Anna Lowit, Ph.D. **~~**

' n Senior Scientist

**~ ~**  Health Effects Division 7905P) **~!hael** Metzger

n n **~** K:

· Acting Branch Chie **~** RABV Health Effects Division

· 7905P) and

Page

1 of84TO:DavidMillerf);jlA0'{6</.f--;/41'I.-BranchCEBHealthEffectsDivision(7905P)ShaluShelatRiskAssessorRABVIHealthEffectsDivision(7905P)I.CONCLUSIONSOPPperformedbenchmarkdose(BMD)analysesofthecomparativecholinesteraseassay(CCA)onpropoxurinordertoestablishapointofdeparture(POD)forthesingle-chemicalriskassessmentusingtheagency'sBenchmarkDoseSoftware(BMDS).OPPanalyzedtheregistrant-supplieddataofredbloodcell(RBC)acetylcholinesteraseinhibition(AChEI)inmaleandfemalePND11pups(combinedsexdata)andcalculatedaBMD10(BenchmarkDosefor10%inhibition)as0.0427mg/kg/dayandtheBMDL10(i.e.95%LowerBoundConfidenceLimitsontheBMD10)wasestimatedas0.0285mg/kg/day.OPPhasconcludedthattheuseofaBMDL10of0.0285mg/kg/daywillprovideahealth-protectiveandscientificallysupportableapproachforanacutePOD.BasedonacomparisonofBMD10s,pupswereapproximately11foldmoresensitivethantheadultratsforbrainandRBCcholinesteraseinhibition(combinedsexdata).However,malepupswerebetween7and10foldmoresensitivethanmaleadults,whilefemalepupswerebetween6and28foldmoresensitivethanfemaleadultsforbrainandRBCcholinesteraseinhibition.ThismemosummarizestheapproachandpresentstheresultsofBMDanalysesandPODderivation.II.BACKGROUNDAseriesofnon-guidelinecholinesterase(ChE)inhibitionstudies(MRID48784801-48784803)wereundertakentoevaluateanydifferencesbetweenpostnatalday11(PND11)pupsandadultrats(approximately7-8weeksofage)withregardtocholinesteraseinhibitionforpropoxur.TheseCCAstudiesconsistofaTime-CourseStudy(MRID48784801),aDoseRange-FindingStudyinbothadultsandpups(MRID48784802)andaDose-ResponseStudyinPups(MRID48784803).Consistentwiththen-methylcarbamatesmodeofaction(MOA),thesestudiesevaluatedcholinesteraseinhibitionfollowingasingledoseatthetimeofpeakeffect.Thesestudiesareclassifiedasacceptable/non-guidelineandsatisfythegenericdatacall-inrequirementforpropoxurforacomparativecholinesterasestudyinadultratsversuspostnatalday(PND)11pups.Boththerangefindinganddefinitivestudiesprovidegooddose-responsedata.Thedefinitivestudyuseddosesfrom0.1to1mg/kg,whiletherangefmdingstudyusesdosesfrom1to10mg/kginadultsand0.3to3mg/kginpups.Page2of84OPP's review indicates that the data show lifestage sensitivity (i.e, pups are more sensitive than adults), and that RBC ChE is more than brain AChE inhibition.

In recent years,

it is typical OPP practice

to perform BMD analysis
on CCA studies. The Registrant conducted

an analysis and submitted

in findings (Mihlan and Sheets (2012). Estimation

of Benchmark Dose for Propoxur Based on RBC Cholinesterase Activity

in PostNatal Day 11 and Adult Rats; MRID 48784804). OPP has conducted

an independent analysis. Both are discussed

in this memo. III. METHODS OPP's BMD analyses were performed with EPA's Benchmark Dose Software (Version 2.1.1). The data selected for evaluation consisted

of brain and red blood cell (RBC) acetylcholinesterase (AChE) activities from PND11 and adult rat OPP has previously used the exponential model for modeling ChE activity, except when the data fit another model better.
In the case of the propoxur CCA study, OPP has used an exponential model, except for female and combined PND 11 pups where the Hill model provided

a better fit of the RBC cholinesterase
data. Model runs for ChE activity were conducted with the benchmark response level of 10%

, and statistical (e.g., goodness
of fit values, AIC criteria, scaled residual, etc.) and graphical results were used in model evaluation (USEP
A 2000, 2002). The registrant proposed that the Agency use the combined BMDL10
for regulatory purposes.

To evaluate the merit of this approach, HED conducted

a Likelihood ratio test (LRT) for RBC pup data based on the BMD output. This analysis

is provided

in Appendix

A. The result of the LRT is not statistically significant because

p value
is greater than conventional alpha level of 0.05. Thus the likelihood-based evaluation suggests that the male and female PND11 RBC data can be combined

in the dose-response modeling. EPA also believes that there
is no biological basis for gender-based differences

in AChE inhibition

in PND11 rats with N-methyl carbamates, like propoxur. Given the lack of statistical significance,
and lack of biological basis for gender based difference

in RBC at PND 11 HED has concluded that male and female data for PND 11 pups can be combined
for deriving more robust POD for AChE inhibition

at this age. IV. Comparison

to Registrant BMD analysis The registrant used

a slightly different version
of BMD (although there
is no important difference between the versions), and only conducted BMDs for RBC ChE.
In addition, the registrant utilized

a heterogeneous variance model for both male and female RBC datasets; however, HED applied

a homogeneous variance model
to the male RBC dataset since
it provided a better fit. Also, for adult RBC ChE, the registrant dropped the high dose in their analyses for both males and females. HED only dropped the high dose for the males because HED could not obtain an adequate

fit for male data. However, the female adult data analyses resulted

in an adequate

fit (without dropping the high dose group).

It is HED policy
to utilize the entire dose Page

3 of84responsewheneverpossible.TheHEDBMD10andBMDL10resultsforadultmaleRBCChEarecomparabletotheregistrant'sresults,whiletheBMDlOandBMDLlOresultsforadultfemaleRBCChEwereslightlyhigherthantheregistrant'sestimates(likelybecauseHEDdidnotdropthehighdosegroupintheanalysis).FormaleandfemalepupRBCChE,HEDobtainedcomparableresultsastheregistrant(BMDLlOapproximately=0.08mg/kgformalesand0.01mg/kgforfemales).HEDagreeswiththeregistrantthattheHillmodelisthebestfitforthefemaleandcombinedpupRBCChEdata.Inaddition,HEDcalculatedBMDsusingtheexponentialmodelforcomparisonpurposes,althoughtheHillmodelprovidesabetterfitofthedata(asshowninTable2).TheregistrantalsoincludedLikelihoodRatioTest(LRntoevaluatetheappropriatenessofcombiningsexesstatistically.HEDreviewedtheanalysisofLRTtestandwasabletoreplicatetheresult(i.e.pvalue)ofLRTtestforPNDllRBC.SeeAppendixA.V.RESULTSTheresultsoftheBMDanalysesaresummarizedinTable1below.DetailsareincludedinAppendixA.HEDwasabletoreproducemostoftheregistrant'sBMDanalysesforthemaleandfemales(MihlanandSheets2012).Inaddition,HEDevaluatedthemaleandfemalepupRBCdatafromthedoseresponsestudytoseeifitwasappropriatetocombinethesedataforBMDanalysis.HEDconcludedthatthesedatasetscanbecombinedtoprovidemorerobustChEmeasuresasthereisnobiologicalbasistosupportsex-differencesforPND11pups.Inaddition,thestatisticaltestalsodidnotfmdsignificantdifferenceinthesedatasets(p=0.067)atconventionalalphalevel0.05.HEDalsoconductedBMDanalysisofthePND11pupdatafromtherangefmdingstudythatusedasmallersamplesize(n=6sex/dose)forcomparisonwiththeBMDestimatesfromthedoseresponsestudy(n=11/sex/dose)andtheseBMDsareshowninTable1.DetailsofAgency'sBMDanalysisarepresentedinAppendixB.Page4of84Sex Male Female Combined Sexes

Table

1 Summary

of OPPs Benchmark Dose (BMD) Analyses for RBC and Brain ChE inhibition **Adul~**

PND 11 PupsA PND 11 PupsA Target (MRID 48784802 (MRID 48784803 (MRID 48784802 Range Range Finding Study) Dose Response Study) Findin Study) Organ BMDto BMDLto BMDto BMDLto BMDto BMDLto Brain 1.26 1.09 0.13
0.12 0.14 0.11 RBC 0.73 0.498 0.094 0.075

0.1 0.08 Brain 0.88 0.76 0.08 0.06

0.1 0.08 RBC 0.55 0.46 0.02
0.01 0.088 0.067 Brain 1.29 1.04 0.12Q 0.0875

- - RBC 0.474 0.339 0.0427 0.0285

--

--- A Adult BMDs based on data m MRID 48784802; PNDll pup BMDs based on data m MRID 48784802, and MRID 48784803. Table

2 Summary Statistics

of PND 11 Pup Cholinesterase Data as Reported
by the Registrant (MRID 48784803)

(a) Time

Mean post Time ChE ChE sex
age dosing unit dose Activity unit

sd

n AcuteRBC PND11

0.5 hr

0 6737 UIL 1121.1

9 PND11

0

. 5 hr
0.1 5916 UIL 986.3

9 Males PND11

0.5 hr 0.3 4532 UIL 1319.4

9 PND11

0

. 5 hr

1 2323 UIL 591.8

8 PND11

0.5 hr

0 6994 UIL 2057.7

9 PND11

0.5 hr
0.1 4856 UIL 489.9

8 PND11

0.5 hr 0.3 3544 UIL 1166
10 Females PND11

0

. 5 hr

1 2460 UIL 900.1
10 PND11

0.5 hr

0 6865.67 UIL 1612.92

18 PND11

0.5 hr
0.1 5416.94 UIL 942.63

17 PND11

0.5 hr 0.3 4012.21 UIL 1307.87

19 M+F PND11

0.5 hr

1 2398.9 UIL 760.28

18 Acute Brain Males PND11

0.5 hr

0 23708 UIL 1480.1

11 PND11

0.5 hr
0.1 21887 UIL 1315.5

11 **~** Page

5 of84sexFemalesM+FSexAcuteRBCMMMMMMFFFFFFM+FM+FM+FM+FM+FTimeMeanpostTimeChEChEagedosingunitdoseActivityunitsdnPND110.5hr0.318250UIL1639.211PND110.5hr110594UIL119011PND110.5hr024263UIL1220.811PND110.5hr0.122320UIL1610.711PND110.5hr0.316844UIL2432.911PND110.5hr112592UIL230011PND110.5hr023985.82UIL1354.1322PND110.5hr0.122103.86UIL1452.1422PND110.5hr0.317547.18UIL2148.5122PND110.5hr111593.18UIL2058.8522(a)CombinedmaleandfemalestatisticscalculatedbyEPA.Table3SummaryStatisticsofAcuteAdultCholinesteraseDatainRangeFindingStudy(MRID48784802)TimepostTimeDoseChEChEdosin2unitm2/k2Activityunitsdn-.0.25hr02928UIL1187.660.25hr12499UIL468.460.25hr22061UIL792.560.25hr31701UIL934.860.25hr51453UIL408.560.25hr101120UIL636.460.25hr03220UIL894.960.25hr12697UIL728.860.25hr22158UIL630.160.25hr31515UIL645.960.25hr51246UIL427.560.25hr10485UIL239.560.25hr03074UIL1014120.25hr12598UIL593120.25hr22109.5UIL684120.25hr31607UIL772120.25hr51349UIL41312Page6of84Time post Time Dose ChE ChE Sex dosing unit mg/kg Activity unit

sd

n M+F

0.25 hr

10 802.8 UIL 565.8
12 **A:cute** Brain

rr **~~~,'If~~**
**~'11**

f M

0.25 hr

0 52184 UIL 1887.5

6 M

0.25 hr

1 50600 UIL 2619.9

6 M

0.25 hr

2 43686 UIL 2300.2

6 M

0.25 hr

3 41315 UIL 3712.8

6 M

0.25
hr

5 34964 UIL 4661.7

6 M

0.25 hr

10 28107 UIL 8309.1

6 F

0.25 hr

0 51558 UIL 1322.9

6 F

0.25 hr

1 46481 UIL 2540.1

6 F

0.25 hr

2 42136 UIL 5139.1

6 F

0.25 hr

3 34790 UIL 2268.7

6 F

0.25 hr

5 26276 UIL 6965.7

6 F

0.25 hr

10 24324 UIL 13626.8

6 M+F

0.25 hr

0 51870 UIL 1587.9

12 M+F

0.25 hr

1 48540 UIL 3268
12 M+F

0.25 hr

2 42911 UIL 3881
12 M+F

0.25 hr

3 37755 UIL 4438
12 M+F

0.25 hr

5 30620 UIL 7247
12 M+F

0.25 hr

10 26215 UIL 10940

12 VI. Recommendation

for POD for Single Chemical Risk Assessment. Based on the analyses,

it is concluded that the use of a BMDL10
of 0.0285 mg/kg/day

for red blood cell (RBC) cholinesterase inhibition (ChEI)
in male and female PND11 pups (combined sex data) will provide

a health-protective

and scientifically supportable approach
for an acute POD. VII. References U.S. EPA 2000. Benchmark Dose Guidance Document. October 2000. U.S EPA 2002. "Organophosphate pesticides: revised cumulative risk assessment." June 2002. Page

7 of84AppendixA.LRTcalculationforPNDllRBCChEdata:ItshouldbenotedthatthebestfittedmodelintermsofAIC,scaledresidualandvisualinspectionforPND11maleRBCisExponentialmodel(2);ontheotherhand,thebestfittedmodelforfemalePND11RBCandcombinedPND11RBCdatasetisHillmodel(lowestAICvalue).-However,acommonmodelisneededthatprovidesatleastamarginalfittoallthreedatasets(male,femaleandcombinedsexes)toconductalikelihoodratiotest(LRT).TheExponentialmodel4meetsthiscriterion.TheLikelihoodRatiotestcanbeconductedusingthefollowingasymptoticresult:Chi-Square(''NumberofParameters")=-2x{LogarithmLikelihoodModelFittoCombinedMales&Females-(LogarithmLikelihoodModelFittoMales+LogarithmLikelihoodModelFittoFemales)};Where''NumberofParameters"isthedegreesoffreedomandisequaltothenumberofparametersinthemodel.ThedistributionofteststatisticfollowsaChi-Squaredistributionwith''NumberofParameters"degreesoffreedom.TheLog-LikelihoodvalueforPND11maleforexponentialmodel(4)withheterogeneousvariance=-257.6347-32.16=-289.7947TheLog-LikelihoodvalueforPND11femaleforexponentialmodel(4)withheterogeneousvariance=-278.8974-34=-312.8974TheLog-LikelihoodforPND11combineddatasetforexponentialmodel(4)withheterogeneousvariance=-541.693-66.16=-607.853Thechi-squareteststatistic=-2*(-607.853-(-289.7947-312.8974))=-2*(-5.1609)=10.3218DF=5pvalue=0.067.Noteregistrantalsoderivedthesamepvalue.TheresultoftheLRTisnotstatisticallysignificantatconventionalalpha=0.05level.Thusthelikelihood-basedevaluationsuggeststhatthemaleandfemalePND11RBCdatacanbecombinedinthedose-responsemodelingPage8of84APPENDIXB (EPA Analyses) Table B-1 Results
of EPA BMD Modeling
for Brain

& RBC ChE in PNDll M I dF I T d .

hP a es an emaes reate w1t ropoxur Sex Compartment

BMD Results

8 (mg/kg/day) BMDto BMDLto Dose Response Study (48784802 (n=11 sex/dose)
(b) 0.13

0.12 Male Exponential

2 Exponential

2 0.08

0.06 Female

Exponential

4 Brain Exponential

4 Combined

0.122 0.0875 Male and Female Exponential

5 Exponential

5 Male

0.094

0.075 Exponential

2 Exponential

2 0.021
0.010 Female

Hill Model

Hill model (Best Fit of data) (Best Fit of data) RBC
0.032
0.019 Exponential

2 Exponential

2 Combined

0.0427
0.0285 Male and

Hill Model

Hill Model Female Range Finding Study_(48784802 (n=4-6 sex/dose}_ Male Brain
0.145 0.1095 Female

0.1
0.082 Male
RBC

0.1
0.080 Female

0.088

0.067 8 Exponential Model with BMR

= 10%, unless noted. EPA BMDS Software (b) source: MRID 48784803 Page9 of84TableB-2ResultsofEPABMDModelingforBrain&RBCChEinADULTM I d F I T t d'thPaaesanemaesreaeWIropoxurSexCompartmentBMDResults8(mg/kg/day)BMD10BMDL101.26°1.09°MaleExponential2Exponential20.880.76FemaleBrainExponential2Exponential2Combined1.291.04MaleandFemaleExponential5Exponential5Male0.7309°0.4982°Exponential2Exponential2Female0.550.46RBCExponential2Exponential2Combined0.4740.339MaleandFemaleExponential4Exponential4Source:MRID487848028ExponentialModelwithBMR=10%.EPABMDSSoftwarebHighdosedropped.Page10of84PND 11 Female Pup BMD Analyses for RBC ChE Inhibition Using Hill Model PROPOXUR- PNDll Female RBC BMDS 2.1.2 -Hill Model (model variance) BMR= 10°/o Data set: MRID 48784803 acute RBC Females PND11 time

dose ChE

SD 0.5

hr

0 6994
UIL 2057.7

9 0.5

hr 0.1 4856
UIL 489.9

8 0.5

hr 0.3 3544
UIL 1166

10 0.5

hr
2460
UIL 900.1

10 ==================================================================== Hill Model. {Version: 2.15; Date: 10/28/2009) Input Data File: C:/Usepa/BMDS212/Data/hil Testrunscontinuous

Opt. {d) Gnuplot Plotting File: C:/Usepa/BMDS212/Data/hil_Testrunscontinuous_Opt.plt

Men Jun 18 11:24:15 2012 ==================================================================== BMDS Model Run The form of the response function
is: Dependent variable

= Mean Independent variable

= Dose Power parameter restricted

to be greater than

1 The variance

is to be modeled

as Var{i)

= exp{lalpha +rho* ln{mean{i))) Total number
of dose groups

= 4 Total number
of records with missing values

= 0 Maximum number
of iterations

= 250 Relative Function Convergence
has been set to: 1e-008 Parameter Convergence
has been set to: 1e-008 Default Initial Parameter Values lalpha 14.3278 rho

0 intercept

v n k 6994 -4534 0.94713 0.119665 Asymptotic Correlation Matrix
of Parameter Estimates Page 11 of84( 

### *** The model parameter(s)

-n have been estimated

at

a boundary point,
or have been specified

by the user,

and do not appear
in the correlation matrix lalpha

rho intercept lalpha

1

- 1
0.1 rho

-1

1 -0.11 intercept

0.1 -0.11

1 v -0.23

0

. 23

-0

. 71 k 0.086 -0.086 -0.51 Parameter Estimates Interval Variable Estimate

Std. Err. Limit lalpha 2.76865 4.68109 11.9434
rho

1. 35729 0.562229 2.45924 intercept 7056.86 534.476 8104.41

v -5125

. 22

640

. 528 3869.81

n

1

NA k 0.128476 0.0582209 0.242587 NA

- Indicates that this parameter
has hit

a bound implied
by some inequality constraint
and thus has no standard error. Table of Data and Estimated Values
of Interest Dose

N Obs Mean Est Mean Obs Std Dev 

## -------- -------- 

# ----------- 0

9

6

. 99e+003

7

. 06e+003

2

. 06e+003 0

. 1 8 4.86e+003 4.81e+003

490 0.3
10 3.54e+003 3.47e+003 1.17e+003 1 10 2.46e+003 2.52e+003

900 Model Descriptions

for likelihoods calculated Model A1:

Yij Var{e(ijl} Model A2:

Yij Mu(i)

+ e(ij) Sigma

... 2 Mu(i)

+ e(ij) 

### Page 12 o f 8 4

v

k -0.23 0.086 0.23 -0.086 -0.71 -0.51 1 -0.16 -0.16

1 95.0% Wald Confidence Lower Conf. Limit Upper -6.40613 0.255342 6009.3 -6380

. 63 0.0143652 Est Std Dev Scaled Res

. 

# -----------

- - -------- 1

. 63e+003

-0

. 115 1.26e+003

0

. 0951 1.01e+003
0.237 811 -0.215 Conf.var{e(ij)} Sigma(i)

... 2 Model A3:
Yij Mu(i)

+ e(ij) var{e(ij)} exp(lalpha

+ rho*ln(Mu(i))) Model A3 uses any fixed variance parameters that were specified

by the user Model

R:

Yi Test
1: Test 2: Test 3: Test 4: (Note: Test Test Test Test Test

1 2 3 4 Var{e(il} Likelihoods

of Interest Model A1 A2 A3
Log (likelihood)

# Param•s

AIC fitted

R -281.447743 -273.238355 -278.207838 -278.338748 -301.433091 Explanation

of Tests

5 572.895487 8 562.476711 6 568.415675 5 566.677496 2 606.866183 Do responses and/or variances differ among Dose levels? (A2 vs. R) Are Variances Homogeneous?

(A1 vs. A2) Are variances adequately modeled?
(A2 vs. A3) Does the Model for the Mean Fit? (A3 vs. fitted) When rho=O the results

of Test

3 and Test

2 will be the same.) Tests of Interest -2*log(Likelihood Ratio) Test df p-value 56.3895

6 <.0001 16.4188

3 0.0009304 9.93896

2 0.006947 0.261821

1 0.6089 The p-value for Test

1 is less than .OS. There appears

to be

a difference between response and/or variances among the dose levels It seems appropriate

to model the data The p-value for Test

2 is less than .1.
A non-homogeneous variance model appears

to be appropriate The p-value for Test

3 is less than .1. You may want to consider

a different variance model The p-value for Test

4 is greater than .1. The model chosen seems to adequately describe the data Benchmark Dose Computation Specified effect

0.1 Risk Type Relative risk Confidence level

0.95 BMD 0.0205143
Page 13 of84BMDL

0

. 0103939

Hill Model with 0.95 Confidence Level 9000

Hill 8000 7000 Q) 6000 Ul c: 0 c. Ul Q) 0::: 5000 c: I'll Q) ::!: 4000 3000 2000

MD 0

0.2

0.4

0

. 6

0

. 8

1 dose 11:24 06/18 2012

Page 14 of84PND 11 Female Pup BMD Analysis for Brain ChE Inhibition Using Exponential Model PROPOXUR- PNDll Female Brain BMDS 2.1.2

- Exponential Model (homogeneous variance model) BMR=lO% Data Set: 48784803 acute brain Females PND11 Time

dose ChE

SD

n 0.5 hr

0 24263 U/L 1220.8

11 0.5 hr

0.1 22320 U/L 1610.7

11 0.5 hr

0.3 16844 U/L 2432.9

11 0.5 hr

12592 U/L
2300

11 ==================================-========-======================== Exponential Model. (Version: 1.7; Date: 12/10/2009) Input Data File: C:/Usepa/BMDS212/Data/exp_Testrunscontinuous_Setting.

(d) Gnuplot Plotting File: Man May

07 14:14:11 2012 BMDS Model Run the response function

by Model: The form of Model
2: Y[dose]
a * exp{sign * b * dose} Model
3: Model
4: Model
5: Y[dose]
a* exp{sign

* (b
* dose)Ad} Y[dose]
a* [c-(c-1)

* exp{-b *dose}] Y[dose]
a* [c-(c-1)

* exp{-(b

* dose)Ad}] Note: Y[dose]

is the median response for **e~osure**

= dose; sign
+1 for increasing trend in data; sign

= -1 for decreasing trend. Model

2 is nested within Models

3 and 4. Model

3 is nested within Model
5. Model

4 is nested within Model
5. Dependent variable

= Mean Independent variable

= Dose Data are assumed
to be distributed: normally Variance Model: exp(lnalpha +rho *ln(Y[dose])) rho is set to o. A constant variance model is fit. Total number
of dose groups

= 4 Total number
of records with missing values

= 0 Maximum number
of iterations

= 250 Relative Function Convergence
has been set to: 1e-008 Parameter Convergence
has been set to: 1e-008 MLE solution provided: Exact Initial Parameter values Page 15 of84Variable
Model

2 Model

3 Model

4 -------- ------- ------- ------- lnalpha 15.0613 15.0613 15.0613 rho(S)

0

0

0 a 14742
14742 25476.2 b

0. 634777 0.634777

3

. 13297 c

0.47073 0.47073

d

1 (S) Specified Parameter Estimates

by Model Variable lnalpha rho a b c d Table Dose

N 0
11 0.1

11 0.3

11 1
11 Model

2 3 4 5

of Model

2 15.4424

0 23437.7 0.700996 Stats From Input Obs Mean ---------- 2.426e+004 2.232e+004 1.684e+004 1.259e+004

Model

3 15.4424

0 23437.7 0.700996

1 Data Obs Std Dev ------------- 1221 1611 2433 2300 Estimated Values
of Interest Dose
Est Mean
Est Std ---------- --------- 0 2.344e+004

2256 0.1 2.185e+004

2256 0.3 1.899e+004

2256 1 1.163e+004

2256 0 2.344e+004

2256 0.1 2.185e+004

2256 0

. 3 1.899e+004

2256 1 1.163e+004

2256 0 2.465e+004

1933 0.1 2.153e+004

1933 0.3 1.735e+004

1933 1
1. 249e+004

1933 0 2.426e+004

1864 0.1 2.232e+004

1864 0.3 1.684e+004

1864 1 1. 259e+004

1864 Other models for which likelihoods

are calculated: Model A1:
Yij Var{e(ij)} Mu(i)

+ e(ij) Sigma

... 2 Page 16 of84 Model

4 15.134

0 24649.9 2. 75113 0.473097 Scaled Residual ---------------- 1.213 0.6896 -3.159 1.419 1.213 0.6896 -3.159 1.419 -0.6638 1. 362 -0.871 0.1729 -8.431e-008 2.676e-008 -1.391e-007 1.543e-008

Model

5 ------- 15.0613

0 25476.2 3.13297

1 Model

5 15.0613

0 24263 3.34891 0.518308 1.55833 

# ~lModel A2:
Yij Mu(i)

+ e(ij) Var{e(ijl} Sigma(i)

... 2 Model A3:
Yij Mu(i)

+ e (ij) var{e(ijl} exp(lalpha

+ log(mean(i))

* rho) Model

R:
Yij Mu
+ e(i) Var{e(ij)} Sigma

... 2 Likelihoods

of Interest Model
Log (likelihood) ------- ----------------- A1 -3S3.3496 A2 -3S0.2243 A3 -3S3.3496 R -396.3873 2 -361.7322 3 -361.7322 4 -3S4.9483 s -3S3.3496 Additive constant for all log-likelihoods

= above values gives the log-likelihood including depend
on the model parameters.

DF s 8 s 2 3 3 4 s -40.43. the term Explanation

of Tests

AIC ------------ 716.6993 716.448S 716.6993 796.774S 729.4644 729.4644 717.8966 716.6993 This constant that does not added to the Test
1: Does response and/or variances differ among Dose levels?
(A2 vs. R) Test 2: Are variances Homogeneous?

(A2 vs. A1) Test 3: Are variances adequately modeled?
(A2 vs. A3) Test 4: Does Model

2 fit the data? (A3 vs. 2) Test Sa: Does Model

3 fit the data

· ? (A3 vs. 3) Test Sb: Is Model

3 better than Model 2? (3 vs. 2) Test 6a: Does Model

4 fit the data? {A3 vs. 4) Test 6b: Is Model

4 better than Model
2? (4 vs. 2) Test 7a: Does Model

s fit the data? {A3 VB. S) Test 7b: Is Model

s better than Model 3? (S VB. 3) Test 7c: Is Model

s better than Model 4? (S vs. 4) Tests of Interest Test -2*log(Likelihood Ratio)

D. F. -------- ------------------------

------ Test

1

92.33

6 Test

2

6.2S1

3 Test

3

6.2S1

3 Test

4

16.77

2 Test Sa

16.77

2 Test Sb

0

0 Test 6a

3.197

1 Test 6b

13.S7

1 Test 7a

0

0 Test 7b

16.77

2 Test 7C

3.197

1 Page 17 of84 p-value < 0.0001 0.1 0.1 0.0002288 0.0002288 N/A 0.07376 0.0002301 N/A 0.0002288 0.07376Thep-valueforTest1islessthan.05.Thereappearstobeadifferencebetweenresponseand/orvariancesamongthedoselevels,itseemsappropriatetomodelthedata.Thep-valueforTest2isgreaterthan.1.Ahomogeneousvariancemodelappearstobeappropriatehere.Thep-valueforTest3isgreaterthan.1.Themodeledvarianceappearstobeappropriatehere.Thep-valueforTest4islessthan.1.Model2maynotadequatelydescribethedata;youmaywanttoconsideranothermodel.Thep-valueforTestSaislessthan.1.Model3maynotadequatelydescribethedata;youmaywanttoconsideranothermodel.DegreesoffreedomforTestSbarelessthanorequalto0.TheChi-Squaretestforfitisnotvalid.Thep-valueforTest6aislessthan.1.Model4maynotadequatelydescribethedata;youmaywanttoconsideranothermodel.Thep-valueforTest6bislessthan.05.Model4appearstofitthedatabetterthanModel2.DegreesoffreedomforTest7aarelessthanorequalto0.TheChi-Squaretestforfitisnotvalid.Thep-valueforTest7bislessthan.05.Model5appearstofitthedatabetterthanModel3.Thep-valueforTest7cisgreaterthan.05.Model5doesnotseemtofitthedatabetterthanModel4.BenchmarkDoseComputations:SpecifiedEffect0.100000RiskTypeRelativedeviationConfidenceLevel0.950000Model2345BMDandBMDLbyModelBMD0.1503010.1503010.07649950.117152BMDL0.1295080.1295080.06001840.0753261Page18of84•i7.Exponential Model

2 with 0.95 Confidence Level 26000 Exponential 24000 22000 G) 20000 Ill c 0 Q. Ill G) 18000 0:: c cu G) ::::! 16000

l 14000 12000 10000 BMD BMD 0

0.2

0.4

0.6

0.8

1 dose 14:14 05/07 2012

0 Page 19 of84ExponentialModel3with0.95ConfidenceLevel26000Exponential2400022000Q)20000Illc::0a.IllQ)18000a::c::caQ):!!:16000I140001200010000BMD BMD00.20.40.60.81dose14:1405/072012Page20of84Exponential Model

4 with

0

. 95 Confidence Level 26000 Exponential 24000 22000 Q) 20000 II) c: 0 c. II) Q) 18000 0:: c: I'll Q) **~** 16000 14000 12000 10000 BMD BMD 0

0

. 2

0

. 4

0

. 6

0

. 8 dose 14:14 05/07 2012

Page 21 of84ExponentialModel5with0.95ConfidenceLevel26000Exponential24000J22000CD20000tilc:0a.tilCD180000:::c:IllCD:::!16000140001200010000BMDMD00.20.40.60.8dose14:1405/072012Page22of84studytype acute time acute acute 4 acute
PND 11 Male Pup BMD Analysis for RBC ChE Inhibition Using Exponential Model PROPOXUR- PNDll Male RBC BMDS 2.1.2

- Exponential Model (homogeneous variance model) BMR=lO% Data Set: Males PND 11 RBC 48784803 timeunit dose chei chunit

sd

n 0.5 hr

0 6737 UIL 1121.1

9 0.5 hr

0.1 5916 UIL
986.3

9 0.5 hr

0.3 4532 UIL 1319.4

9 0.5 hr

2323 UIL
591.8

8 ========================z=========================================== Exponential Model. (Version: 1.7; Date: 12/10/2009) Input Data File: C:/Usepa/BMDS212/Data/exp_Testrunscontinuous_Setting.

(d) Gnuplot Plotting File:

Mon May 07 11:58:51 2012 ==================================================================== BMDS Model Run The form of the Model
2: Model
3: Model
4: Model
5: response function

by Model: Y[dose]
a * exp{sign * b * dose} Y[dose]
a * exp{sign

* (b
* dose)Ad} Y[dose]
a * [c- (c-1)

* exp{ -b
* dose}] Y[dosel
a* [c-(c-1)

* exp{-(b

* dose)Ad}l Note: Y[dose)

is the median response for exposure

= dose; sign

+1 for increasing trend in data; sign

= -1 for decreasing trend. I Model

2 is

' nested within Models

3 and 4. Model

3 is nested within Model
5. Model

4 is nested within Model
5. Dependent variable

= Mean Independent variable

= Dose Data are assumed

to be distributed: normally Variance Mode

' l: exp(lnalpha +rho *ln(Y[dose])) rho is set to

· o. A constant variance model is fit. Total number
of dose groups

= 4 Total number
of records with missing values

= 0 Maximum number
of iterations

= 250 Relative Function Convergence
has been set to: 1e-008 Parameter Convergence

has been set to: 1e-008 MLE solution provided: Exact Page23 of84InitialParameterValuesvariablelnalpha0.164196rho(S)abcd(S)SpecifiedModel213.792803256.411.05077Model313.792803256.411.050771Model413.792807073.851.652470.164196ParameterEstimatesbyModelVariableModel2Model3Model4-----------------------------lnalpha13.812813.812813.7932rho000a6625.926625.926753.37b1.117391.117391.77608c0.209792d1TableofStatsFromInputDataDoseNObsMeanObsStdDev-----------------------09673711210.195916986.30.39453213191 82323591.8EstimatedValuesofInterestModelDoseEstMeanEstStdScaledResidual------------------------------------------------206626998.60.33370.15925998.6-0.028280.34739998.6-0.621112168998.60.4403306626998.60.33370.15925998.6-0.028280.34739998.6-0.621112168998.60.4403406753988.9-0.049670.15885988.90.094170.34549988.9-0.0518112320988.90.007758506737988.71.987e-0070.15916988.7-4.962e-0070.34532988.72.309e-00712323988.79.523e-007Othermodelsforwhichlikelihoodsarecalculated:Page24of84Model513.792807073.851.652471Model5-------13.7928067371.985880.2514671.06876Model A1:
Yij Mu(i)

+ e(ij) Var{e(ij)} Sigma"2 Model A2:

Yij Mu(i)

+ e (ij) var{e(ij)} Sigma(i)"2 Model A3:

Yij Mu(i)

+ e(ij) var{e (ij)} exp(lalpha

+ log(mean(i))

* rho) Model

R:
Yij Mu
+ e(i) Var{e(ij)} Sigma"2 Likelihoods

of Interest Model
Log (likelihood) -----

- -

- - --------------- A1 -2S8.8746 A2 -2S6.3498 A3 -2S8.8746 R -281.9819 2 -2S9.224 3 -2S9.224 4

- 2S8

. 8817 s -2S8.8746 Additive constant for all log

- likelihoods

= above values gives the log-likelihood including depend
on the

' model parameters.

DF s 8 s 2 3 3 4 s -32

. 16. the term Explanation

of Tests

AIC -----------

- S27.7493 S28.6996 S27.7493 S67.9638 S24.4479 S24.4479 S2S.7634 S27.7493 This constant that does not added to the Test
1: Does response and/or variances differ among Dose levels?
(A2 vs. R) Test

2

: Are variances Homogeneous?

(A2 vs. A1) Test 3: Are variances adequately modeled?
(A2 vs

. A3) Test 4: Does Model

2 fit the data? (A3 vs. 2) Test Sa: Does Model

3 fit the data? (A3 vs. 3) Test Sb: Is Model

3 better than Model
2? (3 vs. 2) Test 6a: Does Model

4 fit the data? (A3 vs. 4) Test 6b: Is Model

4 better than Model
2? (4 vs. 2) Test 7a: Does Model

s fit the data? (A3 vs. S) Test 7b: Is Model

s better than Model
3? (S vs. 3) Test 7c

: Is Model

s better than Model
4? (S vs. 4) Tests of Interest Test -2*log(Likelihood Ratio)

D. -------- ------------------------ Test

1

S1.26

6 Test

2

s.os

3 Test

3

s.os

3 Test

4

0.6986

2 Test Sa

0.6986

2 Test Sb

0

0 Test 6a
0.01408

1 Test 6b

0.6846

1 Test 7a

1. 364e-012

0 Page25 of84

F. p-value --------------

< 0.0001 0.1682 0.1682 0.70S2 0.70S2 N/A 0.90SS 0

. 408 N/ATest7bTest7c0.69860.01408210.70S20.90SSThep-valueforTest1islessthan.OS.Thereappearstobeadifferencebetweenresponseand/orvariancesamongthedoselevels,itseemsappropriatetomodelthedata.Thep-valueforTest2isgreaterthan.1.Ahomogeneousvariancemodelappearstobeappropriatehere.Thep-valueforTest3isgreaterthan.1.Themodeledvarianceappearstobeappropriatehere.Thep-valueforTest4isgreaterthan.1.Model2seemstoadequatelydescribethedata.Thep-valueforTestSaisgreaterthan.1.Model3seemstoadequatelydescribethedata.DegreesoffreedomforTestSbarelessthanorequalto0.TheChi-Squaretestforfitisnotvalid.Thep-valueforTest6aisgreaterthan.1.Model4seemstoadequatelydescribethedata.Thep-valueforTest6bisgreaterthan.OS.Model4doesnotseemtofitthedatabetterthanModel2.DegreesoffreedomforTest7aarelessthanorequalto0.TheChi-Squaretestforfitisnotvalid.Thep-valueforTest7bisgreaterthan.OS.ModelSdoesnotseemtofitthedatabetterthanModel3.Thep-valueforTest7cisgreaterthan.OS.ModelSdoesnotseemtofitthedatabetterthanModel4.BenchmarkDoseComputations:SpecifiedEffect0.100000RiskTypeRelativedeviationConfidenceLevel0.9SOOOOModel234sBMDandBMDLbyModelBMD0.09429180.09429180.07618090.0818214BMDL0.07481610.0748161O.OS06134O.OS06648Page26of84PND 11 Male Pup BMD Analysis for Brain ChE Inhibition Using Exponential Model PROPOXUR- PNDll Male Brain BMDS 2.1.2

- Exponential Model (homogeneous variance model) BMR=lO% Data Set: 48784803 acute brain males PND11 Time

Dose ChE

SD

n 0.5 hr

0 23708 UIL 1480.1

11 0.5 hr

0.1 21887 UIL 1315.5

11 0.5 hr

0.3 18250 UIL 1639.2

11 0.5 hr

10594 UIL

1190

11 ==================================================================== Exponential Model. (Version: 1.7; Date: 12/10/2009) Input Data File: C:/Usepa/BMDS212/Data/exp_Testrunscontinuous_Setting.

(d) Gnuplot Plotting File:

Mon May 07 12:25:36 2012 ==================================================================== BMDS Model Run The form of the response function

by Model: Model
2:

Y [dose]

a * exp{sign

* b * dose} Model
3: Y[dose]
a * exp{sign

* (b
* dose)"d} Model
4:

Y [dose]

a * [c- (c-1)

* exp{-b

* dose}l Model
5:

Y [dose]

a * [c- (c-1)

* exp{-(b

* dose)"d}l Note: Y[dose]

is the median response for exposure

= dose; sign

+1 for increasing trend in data; sign

= -1 for decreasing trend. Model

2 is nested within Models

3 and 4. Model

3 is nested within Model
5. Model

4 is nested within Model
5. Dependent variable

= Mean Independent variable

= Dose Data are assumed
to be distributed: normally Variance Model: exp(lnalpha +rho *ln(Y[dose])) rho is set to 0. A constant variance model is fit. Total number
of dose groups

= 4 Total number
of records with missing values

= 0 Maximum number
of iterations

= 250 Relative Function Convergence
has been set to: 1e-008 Parameter Convergence
has been set to: 1e-008 MLE solution provided: Exact Initial Parameter Values Page27 of84VariableModel2Model3Model4---------------------- -------lnalpha14.416414.416414.4164rho(S)000a13431.313431.324893.4b0.8041460.8041461.31733c0.2127870.212787d1(S)SpecifiedParameterEstimatesbyModelVariablelnalpharhoabcdModel214.43023640.50.814242Model3-------14.43023640.50.8142421TableofStatsFromInputDataDoseNObsMeanObsStdDev-----------------------0112.371e+00414800.1112.189e+00413160.3111.825e+00416391111.059e+0041190EstimatedValuesofInterestModelDoseEstMeanEstStd-------------------------202.364e+00413600.12.179e+00413600.31.852e+004136011.047e+0041360302.364e+00413600.12.179e+00413600.31.852e+004136011.047e+0041360402.378e+00413530.12.176e+00413530.31.831e+004135311.059e+0041353502.371e+00413500.12.189e+00413500.31.825e+004135011.059e+0041350Othermodelsforwhichlikelihoodsarecalculated:ModelA1:Yij=Mu(i)+e(ij)Page28of84Model4-------14.4197023777.21.038830.14148ScaledResidual----------------0.16460.232-0.65130.29720.16460.232-0.65130.2972-0.16960.304-0.15030.01585-9.875e-0081.349e-0071.266e-007-1.187e-007Model5-------14.4164024893.41.317331Model5-------14.41640237081.444180.2898781.12009Var{e(ij)} Sigma"2 Model A2:
Yij Mu(i)

+ e(ij) var{e(ij)} Sigma(i)"2 Model A3:
Yij Mu(i)

+ e(ij) Var{e(ij)} exp(lalpha

+ log(mean(i))

* rho) Model

R:
Yij Mu
+ e(i) Var{e(ij)} Sigma"2 Likelihoods

of Interest Model
Log (likelihood) ------- ----------------- A1 -339.1604 A2 -338.S238 A3 -339.1604 R -398.S328 2 -339.4S92 3 -339.4S92 4 -339.232S s -339.1604 Additive constant for all log-likelihoods

= above values gives the log-likelihood including depend
on the model parameters.

DF s 8 s 2 3 3 4 s -40.43. the term Explanation

of Tests

AIC ------------ 688.3208 693.0476 688.3208 801.06S6 684.9183 684.9183 686.46S1 688.3208 This constant that does not added to the Test
1: Does response and/or variances differ among Dose levels?
(A2 vs. R) Test 2: Are Variances Homogeneous?

(A2 vs. A1) Test 3: Are variances adequately modeled?
(A2 vs. A3) Test 4: Does Model

2 fit the data? (A3 vs. 2) Test Sa: Does Model

3 fit the data? (A3 vs. 3) Test Sb: Is Model

3 better than Model
2? (3 vs. 2) Test 6a: Does Model

4 fit the data? (A3 vs. 4) Test 6b: Is Model

4 better than Model
2? (4 vs. 2) Test 7a: Does Model

s fit the data? (A3 vs. S) Test 7b: Is Model

s better than Model
3? (S vs. 3) Test 7c: Is Model

s better than Model 4? (S vs. 4) Tests of Interest Test -2*log(Likelihood Ratio)

D. F. -------- ------------------------ Test

1

120

6 Test

2

1. 273

3 Test

3

1.273

3 Test

4

O.S97S

2 Test sa

O.S97S

2 Test Sb 1.137e-013

0 Test 6a

0.1443

1 Test 6b

0.4S32

1 Test 7a

0

0 Test 7b

O.S97S

2 Page29 of84 p-value --------------

< 0.0001 0.73SS 0.73SS 0.7417 0.7417 N/A 0.7041 O.S008 N/A 0.7417Test7c0.144310.7041Thep-valueforTest1islessthan.05.Thereappearstobeadifferencebetweenresponseand/orvariancesamongthedoselevels,itseemsappropriatetomodelthedata.Thep-valueforTest2isgreaterthan.1.Ahomogeneousvariancemodelappearstobeappropriatehere.Thep-valueforTest3isgreaterthan.1.Themodeledvarianceappearstobeappropriatehere.Thep-valueforTest4isgreaterthan.1.Model2seemstoadequatelydescribethedata.Thep-valueforTestSaisgreaterthan.1.Model3seemstoadequatelydescribethedata.DegreesoffreedomforTestSbarelessthanorequalto0.TheChi-Squaretestforfitisnotvalid.Thep-valueforTest6aisgreaterthan.1.Model4seemstoadequatelydescribethedata.Thep-valueforTest6bisgreaterthan.05.Model4doesnotseemtofitthedatabetterthanModel2.DegreesoffreedomforTest7aarelessthanorequalto0.TheChi-Squaretestforfitisnotvalid.Thep-valueforTest7bisgreaterthan.05.Model5doesnotseemtofitthedatabetterthanModel3.Thep-valueforTest7cisgreaterthan.05.Model5doesnotseemtofitthedatabetterthanModel4.BenchmarkDoseComputations:SpecifiedEffect0.100000RiskTypeRelativedeviationConfidenceLevel0.950000BMDandBMDLbyModelModel2345BMD0.1293970.1293970.1192110.128639BMDL0.1191120.1191120.09741440.0979343Page30of8451Exponential Model

2 with

0

. 95 Confidence level 26000 Exponential 24000 22000 20000 5l c: 0 18000 a. II) CD a:: c: 111 16000 CD ::!: 14000 12000 10000 BMD BMD 0

0.2

0.4

0.6

0

. 8

1 dose 12:25 05/07 2012

Page 31 of84ExponentialModel3with0.95ConfidenceLevel26000Exponential240002200020000GlIllr::::018000c.IllGl0:::r::::Ill16000Gl::!140001200010000BMDMD00.20.40.60.8dose12:2505/072012Page32of84Exponential Model4 with 0.95 Confidence Level 26000 Exponential 24000 22000 20000 CD VI 1: 0 18000 c. VI CD 0::: 1: cu 16000 CD **~** 14000 12000 10000 BMD BMD 0

0.2

0.4

0.6

0.8

1 dose 12

: 25 05/07 2012

Page 33 of84ExponentialModel5with0.95ConfidenceLevel26000Exponential240002200020000Q)II)c:018000c.II)Q)0:::c:Cll16000Q):::!:140001200010000BMDMD00.20.40.60.81dose12:2505/072012Page34of84Adult Female BMD Analysis for RBC ChE Inhibition Using Exponential Model PROPOXUR- Adult Female RBC BMDS 2.1.2

- Exponential Model BMR=lO% DataSet: adult female RBC at 0.25 hr post dosing DOSE
Che

SD

n 0 3220 UIL 894.9

6 I 2697 UIL 728.8

6 2 2158 UIL 630.1

6 3 1515 UIL 645.9

6 1 5 1246 U/L 427.5

6 10 485 UIL 239.5

6 Exponential Model. (Version: 1.7; Date: 12/10/2009) Input Data File: C:/Usepa/BMDS212/Data/exp_Testrunscontinuous_Setting.

(d) Gnuplot Plotting File:

Thu Jun 07 09:03:13 2012 ==================================================================== BMDS Mode

1 Run The form of the Model
2: Model
3: Model
4: Model
5: response function

by Model: Y[dose]
a * exp{sign * b * dose} Y[dose]
a* exp{sign

* (b
* dose)Ad} Y[dose]
a* [c-(c-1)

* exp{-b *dose}] Y[dose]
a* [c-(c-1)

* exp{-(b

* dose)Ad}l Note: Y[dose]

is the median response for exposure

= dose; sign

= +1 for increasing trend in data; sign

= -1 for decreasing trend. Model

2 is nested within Models

3 and 4. Model

3 is nested within Model
5. Model

4 is nested within Model
5. Dependent variable

= Mean Independent

v

. ariable

= Dose Data are assumed
to be distributed: normally Variance **Mode~:** exp(lnalpha +rho *ln(Y[dose])) The variance

1 is to be modeled

as Var (i)
= exp (lalpha

+ log (mean {i))

* rho) Total number
of dose groups

= 6 Total number
of records with missing values

= 0 Maximum **numbe~**
of iterations

= 250 Relative Function Convergence
has been set to: 1e-008 Parameter Convergence
has been set to: 1e-008 MLE solution provided: Exact
Page 35 of84Initial Parameter Values Variable
Model

2 Model

3 Model

4 Model

5 -------- ------- ------- ------- ------- lnalpha 2.68472 2.68472 2.68472

2. 68472 rho 1.34512 1.34512 1.34512

1. 34512 a 831.979

831. 979

3381

3381 b 0.187911 0.187911 0.253582 0.253582 c

0. 0717243 0. 0717243

d

1

1 Parameter Estimates

by Model Variable
Model

2 Model

3 Model

4 Model

5 -------- ------- ------- -------

------- lnalpha 2.47268 2.47268 2.32367 2.32367 rho 1.35616 1.35616

1. 37493 1.37493 a 3145.43 3145.43 3224.88 3224.88 b 0.192137 0.192137 0.224761 0.224761 c

0.0507246 0.0507246 d

1

1 Table of Stats From Input Data Dose

N
Obs Mean Obs Std Dev ---------- ------------- 0

6 3220 894.9 1 6

2697 728.8 2 6

2158 630.1 3 6

1515 645.9 5 6

1246 427.5 10

6
485 239.5 Estimated Values
of Interest Model Dose
Est Mean
Est Std Scaled Residual ---------- --------- ---------------- 2

0 3145 810.3 0.2254 1 2596 711.3 0.3492 2 2142 624.4 0.06329 3 1767 548.2 -1.128 5 1204 422.4 0.2462 10 460.5 220.2 0.2724 3

0 3145 810.3 0.2254 1 2596 711.3 0.3492 2 2142 624.4 0.06329 3 1767 548.2 -1.128 5 1204 422.4 0.2462 10 460.5 220.2 0.2724 4

0 3225 825.2 -0.01447 1 2609 713.3 0.3034 2 2116 617.8 0.1646 3 1723 536.4 -0.9516 5 1159 408.3 0.5242 10

487

225 -0.0219 5

0 3225 825.2 -0.01447 1 2609 713.3 0.3034 Page 36 of842 3 5 10 2116 1723 1159 487 617.8 536.4 408.3 225 Other models for which likelihoods

are calculated: Model A1:

Yij Mu(i)

+ e(ij) var{e(ij)} Sigma"'2 Model A2:

Yij Mu(i)

+ e(ij) Var{e(ij)} Sigma(i)"'2 Model A3:

Yij Mu(i)

+ e(ij) Var{e(ij)} exp(lalpha

+ log(mean(i))

* Model

R:
Yij Mu
+ e(i) Var{e(ij)} Sigma"'2 Likelihoods

of Interest Model
Log (likelihood)

DF ------- ----------------- A1 -246.8013

7 A2 -241.7651

12 A3 -242.095

8 R -269.4793

2 2 -242.6912

4 3 -242.6912

4 4 -242.5467

5 5 -242.5467

5 0.1646 -0.9516 0.5242 -0.0219 rho) AIC ------------ 507.6026 507.5301 500.1899 542.9587 493.3823 493.3823 495.0933 495.0933 Additive constant for all log-likelihoods

= above values gives the log-likelihood including depend
on the model parameters.

-33.08. This constant the term that does not Explanation

of Tests

added to the Test
1: Does response and/or variances differ among Dose levels?
(A2 vs. R) Test 2: Are variances Homogeneous?

(A2 vs. A1) Test 3: Are variances adequately modeled?

(A2 vs. A3) Test 4: Does Model

2 fit the data? (A3 vs

. 2) Test Sa: Does Model

3 fit the data? (A3 vs

. 3) Test Sb: Is Model

3 better than Model
2? (3 vs. 2) Test 6a: Does Model

4 fit the data? (A3 vs. 4) Test 6b: Is Model

4 better than Model
2? (4 vs. 2) Test 7a: Does Model

5 fit the data? (A3 vs. 5) Test 7b: Is Model

5 better than Model
3? (5 vs. 3) Test 7c: Is Model

5 better than Model 4? (5 vs. 4) Tests of Interest Test -2*log(Likelihood Ratio)

D. F. Page 37 of84 p-valueTest1SS.4310<0.0001Test210.07s0.07321Test30.6S9840.9S62Test41.19240.8793Testsa1.19240.8793TestSb1.70Se-0130N/ATest6a0.903430.8246Test6b0.2891O.S909Test7a0.903430.8246Test7b0.2891O.S909Test7c-1.137e-0130N/AThep-valueforTest1islessthan.OS.Thereappearstobeadifferencebetweenresponseand/orvariancesamongthedoselevels,itseemsappropriatetomodelthedata.Thep-valueforTest2islessthan.1.Anon-homogeneousvariancemodelappearstobeappropriate.Thep-valueforTest3isgreaterthan.1.Themodeledvarianceappearstobeappropriatehere.Thep-valueforTest4isgreaterthan.1.Model2seemstoadequatelydescribethedata.Thep-valueforTestSaisgreaterthan.1.Model3seemstoadequatelydescribethedata.DegreesoffreedomforTestSbarelessthanorequalto0.TheChi-Squaretestforfitisnotvalid.Thep-valueforTest6aisgreaterthan.1.Model4seemstoadequatelydescribethedata.Thep-valueforTest6bisgreaterthan.OS.Model4doesnotseemtofitthedatabetterthanModel2.Thep-valueforTest7aisgreaterthan.1.Modelsseemstoadequatelydescribethedata.Thep-valueforTest7bisgreaterthan.OS.ModelsdoesnotseemtofitthedatabetterthanModel3.DegreesoffreedomforTest7carelessthanorequalto0.TheChi-Squaretestforfitisnotvalid.BenchmarkDoseComputations:SpecifiedEffect0.100000RiskTypeRelativedeviationConfidenceLevel0.9SOOOOBMDandBMDLbyModelModel23BMDO.S48362O.S48362BMDL0.4SS4290.4SS429Page38of84Q) Ul c: 0 a. Ul Q) 0:: c: IU Q) **~**

4 5 4500 4000 3500 3000 2500 2000 1500 1000 500 0.495262 0.495262 0.360857 0.360857 Exponential Model2 with 0.95 Confidence Level Exponential 0 BMD BMD 0

2

4

6 dose 09:03 06/07 2012

Page 39 of84

8

10 {)ExponentialModel3with0.95ConfidenceLevel4500Exponential400035003000Cl)Illc:25000Q.IllCl)0:::c:2000IllCl):E150010005000BMDMD0246810dose09:0306/072012Page40of84Exponential Model4 with 0.95 Confidence Level 4500 Exponential 4000 3500 3000 CD II) 2500 c: 0 a. II) CD **~** c: 2000 co CD :::::!: 1500 1000 500 0 BMD BMD 0

2

4

6

8

10 dose 09:03 06/07 2012

Page 41 of84ExponentialModel5with0.95ConfidenceLevel4500Exponential400035003000CD1/)2500c:0a.1/)CD0:::c:2000tUCD::::!150010005000BMDMD0246810dose09:0306/072012Page42of84Adult Female BMD Analysis for Brain ChE Inhibition Using Exponential Model BMDS 2.1.2

- Exponential Model (nonhomogeneous variance- best fit) BMR=lO% Dataset: acute brain female adult at 0.25 hrs post dosing Dose ChE

SD

n 0 51558 U/L 1322.9

6 1

4
, 6481 U/L 2540.1

6 2 42136 U/L 5139.1

6 3 34790 U/L 2268.7

6 5 26276 U/L 6965.7

6 10 24324 U/L 13626.8

6 Exponential Model. (Version: 1.7; Date: 12/10/2009) Input Data File: C:/Usepa/BMDS212/Data/exp_Testrunscontinuous_Setting.

(d) Gnuplot Plotting File:

Thu Jun 07 12:33:27 2012 BMDS Mode

1 Run The form of the Model
2: response function
by Model: Y[dose]
a * exp{sign * b * dose} Model
3: Model
4: Model
5: Y[dose]
a* exp{sign

* (b
* dose)Ad} Y[dose]
a* [c-(c-1)

* exp{-b *dose}] Y[dose]
a* [c-(c-1)

* exp{-(b

* dose)Ad}l Note: Y[dose]

is the median response for exposure

= dose; sign

+1 for increasing trend in data; sign

= -1 for decreasing trend. Model

2 is nested within Models

3 and 4. Model

3 is

. nested within Model
5. Model

4 is nested within Model
5. Dependent variable

= Mean Independent variable

= Dose Data are assumed
to be distributed: normally variance Model: exp(lnalpha +rho *ln(Y[dose])) The variance

is to be modeled

as Var(i)

= exp(lalpha

+ log(mean(i)) *rho) Total number
of dose groups

= 6 Total number
of records with missing values

= 0 Maximum number
of iterations

= 250 Relative Function Convergence
has been set to: 1e-008 Parameter Convergence
has been set to: 1e-008 MLE solution provided: Exact Initial Parameter Values variable
Model

2 Model

3 Page43 of84 Model

4 Model

5lnalpha65.798565.798565.798565:7985rho-4.69136 -4.69136-4.69136-4.69136a27502.127502.154135.954135.9b0.07840620.07840620.3493270.349327c0.4279180.427918d11ParameterEstimatesbyModelVariableModel2Model3Model4Model5--------------- ---------------------lnalpha53.180151.895663.382264.1543rho-3.48949-3.36982-4.44667-4.52536a51893.751832.151979.751579.9b0.1201470.1242560.1843540.273658c0.2884450.421209d1.033171.34443TableofStatsFromInputDataDoseNObsMeanObsStdDev-----------------------065.156e+0041323164.648e+0042540264.214e+00451393 63.479e+00422695 62.628e+00469661062.432e+0041.363e+004EstimatedValuesofInterestModelDoseEstMeanEstStdScaledResidual-----------------------------------205.189e+0042095-0.392514.602e+00425840.438124.081e+00431871.0233.619e+0043930-0.872152.846e+0045976-0.8947101.561e+0041.705e+0041.253305.183e+0042115-0.317414.616e+00425710.308324.088e+00431550.97333.613e+0043884-0.847852.812e+0045927-0.7619101.483e+0041.742e+0041.335405.198e+0041898-0.544114.575e+00425210.707624.057e+00432931.16233.627e+0044226-0.856352.971e+0046586-1.276102.085e+0041.447e+0040.5885505.158e+0041854-0.0288714.678e+0042312-0.320724.086e+00431400.993333.559e+0044293-0.456252.823e+0047253-0.6589102.235e+0041.23e+0040.3934Page44of84Other models for which likelihoods
are calculated: Model A1:

Yij Mu(i)

+ e(ij) Var{e(ij)}

= SigmaA2 Model A2:
Yij Var{e (ij)} Model A3:
Yij Mu(i)

+ e(ij) Sigma(i)A2 Mu(i)

+ e(ij) Var{e(ij)} exp(lalpha

+ log(mean(i)) *rho) · Model

R:
Yij var{e(ij)}

Mu

+ e(i) SigmaA2 Likelihoods

of Interest Model Log(likelihood) ------- ----------------- A1 -332.1801 A2 -312.7243 A3 -317.7667 R -3SS.S20S 2 -319. 7208 3 -319.6983 4 -319.2167 s -317.9674 Additive constant for all log-likelihoods

= above values gives the log-likelihood including depend
on the model parameters.

DF 7 12 8 2 4 s s 6 -33.08. the term Explanation

of Tests

AIC ------------ 678.3602 649.4486 6S1.S333 71S.041 647.4416 649.3966 648.4333 647.9348 This constant that does not added to the Test
1: Does response and/or variances differ among Dose levels?
(A2 vs. R) Test 2: Are Variances Homogeneous?

(A2 vs. A1) Test 3: Are variances adequately modeled?
(A2 vs. A3) Test 4: Does Model

2 fit the data? (A3 vs. 2) Test Sa: Does Model

3 fit the data? (A3 vs. 3) Test Sb: Is Model

3 better than Model
2? (3 vs. 2) Test 6a: Does Model

4 fit the data? (A3 vs. 4) Test 6b: Is Model

4 better than Model
2? (4 vs. 2) Test 7a: Does Model

s fit the data? (A3 vs. S) Test 7b: Is Model

S better than Model
3? (S vs. 3) Test 7C: Is Model

s better than Model
4? (S vs. 4) Tests of Interest I Test -2*log(Likelihood Ratio)

D. -------- -----------------------

- Test

1

8S.S9

10 Test

2

38.91

s Test

3

10.08

4 Test

4

3.908

4 Test sa

3.863

3 Page45 of84

F. p-value < 0.0001 < 0.0001 0.03902 0.4186 0.2766TestSb0.0450210.832Test6a2.930.4073Test6b1.00810.3153Test7a0.401520.8181Test7b3.46210.0628Test7c2.49910.114Thep-valueforTest1islessthan.05.Thereappearstobeadifferencebetweenresponseand/orvariancesamongthedoselevels,itseemsappropriatetomodelthedata.Thep-valueforTest2islessthan.1.Anon-homogeneousvariancemodelappearstobeappropriate.Thep-valueforTest3islessthan.1.Youmaywanttoconsideradifferentvariancemodel.Thep-valueforTest4isgreaterthan.1.Model2seemstoadequatelydescribethedata.Thep-valueforTestSaisgreaterthan.1.Model3seemstoadequatelydescribethedata.Thep-valueforTestSbisgreaterthan.05.Model3doesnotseemtofitthedatabetterthanModel2.Thep-valueforTest6aisgreaterthan.1.Model4seemstoadequatelydescribethedata.Thep-valueforTest6bisgreaterthan.05.Model4doesnotseemtofitthedatabetterthanModel2.Thep-valueforTest7aisgreaterthan.1.Model5seemstoadequatelydescribethedata.Thep-valueforTest7bisgreaterthan.05.Model5doesnotseemtofitthedatabetterthanModel3.Thep-valueforTest7cisgreaterthan.05.Model5doesnotseemtofitthedatabetterthanModel4.BenchmarkDoseComputations:SpecifiedEffect0.100000RiskTypeRelativedeviationConfidenceLevel0.950000BMDandBMDLbyModelModel2345BMD0.8769320.9114540.8215071.06114BMDL0.763350.7642250.6819060.774727Page46of8401Exponential Model2 with 0.95 Confidence Level Exponential 50000 40000 Q) tn c 0 c. tn Q) 0:: 30000 c 1'0 Q) :E 20000 10000 BMD BMD 0

2

4

6

8

10 dose 12:33 06/07 2012

Page47 of84ExponentialModel3with0.95ConfidenceLevelExponential5000040000Cl)1/)c:0c.1/)Cl)0:::30000c:<UCl):22000010000BMDMD0246810dose12:3306/072012Page48of84Exponential Model

4 with
0

. 95 Confidence Level Exponential 50000 40000 Q) U) r:::: 0 c. U) Q) 0::: 30000 r:::: «< Q) :::2!: 20000 10000 BMD BMD 0

2

4

6

8

10 dose 12:33 06/07 2012

Page49 of84

0Exponential Model

5 with 0.95 Confidence Level Exponential 50000 40000 Q) t/) c: 0 c.. t/) Q) 0:::: 30000 c: as Q) ::lE 20000 10000 BMD MD 0

2

4

6

8

1 0 dose 12:33 06/07 2012

Page 50 of84PROPOXUR- Adult Male RBC Note: high dose (10 mg/kg/day) dropped since there was no acceptable BMD fit BMDS 2.12

- Exponential Model BMR=lO% dose chei chunit 0 2928 U/L 1 2499 U/L 2 2061 U/L 3 1701 5 1453

sd

n 1187.6

6 468.4

6 792.5

6 U/L
934.8 U/L
408.5 ==================================================================== Exponential Model. (Version: 1.7; Date: 12/10/2009)

6 6 Input Data File: C:/Usepa/BMDS212/Data/exp_Testrunscontinuous_setting.

(d) Gnuplot Plotting File

:

Mon Jun 11 12

: 38:17 2012 ==================================================================== BMDS Model Run The form of the response function

by Model: Model
2: Y[dose]

a * exp{sign

* b * dose} Model
3:

Y [dose]

a * exp{sign

* (b
* dose)"d} Model
4: Y[dose]

a * [c- (c-1)

* exp{-b

* dose}] Model
5: Y[dose]

a * [c- (c-1)

* exp{-(b

* dose)"d}J Note: Y[dose]

is the median response for exposure

= dose; sign

+1 for increasing trend in data; sign

= -1 for decreasing trend

. Model

2 is nested within Models

3 and 4. Model

3 is nested within Model
5. Model

4 is nested within Model
5. Dependent variable

= Mean Independent variable

= Dose Data are assumed

to be distributed: normally Variance Model: exp(lnalpha +rho *ln(Y[dose])) The variance

is to be modeled

as Var(i)

= exp(lalpha

+ log(mean(i)) *rho) Total number
of dose groups

= 5 Total number
of records with missing values

= 0 Maximum number
of iterations

= 250 Relative Function Convergence
has been set to: 1e-008 Parameter Convergence
has been set to: 1e-008 MLE solution provided: Exact Initial Parameter Values Variable
Model

2 Model

3 Page 51 of84 Model

4 Model

5lnalpha

1. 59226 1.59226 1.59226

1. 59226 rho 1.50831 1.50831

1. 50831

1. 50831 a 1500.99 1500.99
3074.4 3074.4 b 0.144329 0.144329 0.254213 0.254213 c

0.236306 0.236306

d

1

1 Parameter Estimates

by Model variable
Model

2 Model

3 Model

4 Model

5 

## -------- ------- -------

------- 

### ------- lnalpha -1.94991 117.185 -0.925051 -0.925052 rho

1. 97589 -13.5129
1.8413 1.8413 a 2834.45 2128.4 2913.48 2913.48 b 0.144133 0.0603954

0. 251355 0.251355 c

0.275837 0.275837 d
17.4896

1 Table of Stats From Input Data Dose

N
Obs Mean Obs Std Dev 

# ---------- ------------- 0

6 2928
1188 1

6 2499 468.4 2

6 2061 792.5 3

6 1701 934.8 5 6

1453 408.5 Estimated Values
of Interest Model Dose
Est Mean
Est Std Scaled Residual 

## ------- 

# ---------- 

# --------- ---------------- 2

0 2834 971.5 0.2359 1 2454 842.5 0.1308 2 2125 730.7 -0.2132 3 1839 633.7 -0.535 5 1379 476.7 0.3815 3

0 2128 913.1
2.145 1 2128 913.1 0.9942 2 2128 913.1 -0.1808 3 2128 913.1 -1.147 5 2128 913.1 -1.812 4

0 2913 974.2 0.0365 1 2445 828.9 0.1609 2 2080 714.3 -0.06469 3 1796 624.1 -0.3737 5 1404 497.5

0. 2411 5

0 2913 974.2 0.0365 1 2445 828.9 0.1609 2 2080 714.3 -0.06469 3 1796 624.1 -0.3737 5 1404 497.5

0. 2411 Other models for which likelihoods

are calculated: Model A1:
Yij

= Mu(i)

+ e(ij) 

### Page 52 of84var{e(ij)} Sigma"2 Model A2:
Yij Mu(i)

+ e(ij) var{e(ij)} Sigma(i)"2 Model A3:

Yij Mu(i)

+ e(ij) Var{e(ij)} exp(lalpha

+ log(mean(i))

* rho) Model

R:
Yij Mu
+ e(i) var{e(ij)} Sigma"2 Likelihoods

of Interest Model Log(likelihood) ------- ----------------- A1 -213.2S86 A2 -208.8074 A3 -211.7294 R -219.SOS8 2 -211.9663 3 -219.SOS8 4 -211.9129 s -211.9129 Additive constant for all log-likelihoods

= above values gives the log-likelihood including depend
on the model parameters.

DF 6 10 7 2 4 s s s - 27.S7. the term Explanation

of Tests

AIC ------------ 438.S172 437.6147 437.4S88 443.0117 431.9327 449.0117 433.82S9 433.82S9 This constant that does not added to the Test 1: Does response and/or variances differ among Dose levels?
(A2 vs. R) Test 2: Are variances Homogeneous?

(A2 vs. A1) Test 3: Are variances adequately modeled?

(A2 vs. A3) Test 4: Does Model

2 fit the data? (A3 vs. 2) Test Sa: Does Model

3 fit the data? (A3 vs. 3) Test Sb

: Is Model

3 better than Model
2? (3 vs. 2) Test 6a

: Does Model

4 fit the data? (A3 vs

. 4) Test 6b: Is Model

4 better than Model
2? (4 vs. 2) Test 7a: Does Model

s fit the data? (A3 vs. S) Test 7b

: Is Model

s better than Model
3? (S vs. 3) Test 7c

: Is Model

s better than Model 4? (S vs. 4) Tests of Interest Test

- 2*log(Likelihood Ratio)

D. -------- ------------------------ Test

1

21.4

8 Test

2

8.903

4 Test

3

S.844

3 Test

4

0.4739

3 Test sa

1S.SS

2 Test Sb

-1S.08

1 Test 6a

0.367

2 Test 6b

0.1068

1 Test 7a

0.367

2 Test 7b

1S

. 19

0 Page 53 of84

F. p-value 0.006164 0.063S8 0.1194 0.9246 0.000419S N/A 0

. 8323 0

. 7438 0.8323 N/A

## Test 7c -S.684e-014

0 

## The p-value for Test

1 is less than .OS. There appears

to be

a difference between response and/or variances among the dose levels,

it seems appropriate

to model the data. The p-value for Test

2 is less than .1.
A non-homogeneous variance model appears

to be appropriate. The p-value for Test

3 is greater than .1. The modeled variance appears

to be appropriate here. The p-value for Test

4 is greater than .1. Model

2 seems to adequately describe the data.

N/A 

## The p-value for Test sa is less than .1. Model

3 may not adequately describe the data; you may want to consider another model. The p-value for Test Sb is less than .OS. Model

3 appears to fit the data better than Model
2. The p-value for Test 6a is greater than .1. Model

4 seems to adequately describe the data. The p-value for Test 6b is greater than .OS. Model

4 does not seem to fit the data better than Model
2. The p-value for Test 7a is greater than .1. 

## to adequately describe the data. Degrees

of freedom for Test 7b are less than The Chi-Square test for fit is not valid. Degrees

of freedom for Test 7c are less than The Chi-Square test for fit is not valid. Benchmark Dose Computations: Specified Effect 0.100000 Risk Type Relative deviation Confidence Level 0.9SOOOO BMD and BMDL by Model Model 2 3 

### 4 

# s

BMD 0.73099S 14

. SS8S O.S9121S O.S9121S

BMDL 0.498177 0.27478S 0.2263S 0.2263S 

# Page 54 of84 Model

s seems or equal to
0
. or equal to 0.Exponential Model

2 with 0.95 Confidence Level 4500 Exponential 4000 3500 3000 Q) fl) c: 0 c.. fl) 2500 Q) 0::: c: Ill Q) :::!: 2000 1500 1000 500 BMD
MD 0

2

3

4

5 dose 12:38 06111 2012

Page 55 of84Exponential Model

3 with
0

. 95 Confidence Level 4500 Exponential 

# -- 4000 3500 

### 3000 Cl)



# -- U) r:::: 0

-rc. U) 2500 Cl) 0::: r:::: 111 Cl) ::!: 2000

_._



## ~ -r- 

## - 1500 1000

_._ 500 BMqL

**~Mi** 0

2

4

6

8

10

12

14 dose 12:38 06/11 2012



# 11 Page 56 of84Exponential Model

4 with 0.95 Confidence Level 4500 Exponential 4000 3500 3000 (J) "' c:: 0 Q. "' 2500 (J) a::: c:: 1'0 (J) **~** 2000 1500 1000 500 BMD 0

2

3

4

5 dose 12:3806/112012

Page 57 of84ExponentialModel5with0.95ConfidenceLevel4500Exponential400035003000Q)t/)c:0Q.t/)2500Q)0::c:coQ):!E200015001000500BMDMD02345dose12:3806/112012Page58of84PROPOXUR- Adult Male Brain

- high dose dropped (10 mg!kg/day)

[to improve

fit and convergence] BMDS 2.1.2

- Exponential Model (homogeneous variance

- best fit) BMR=lO% Dataset: acute male adult brain at 0.25 hrs postdosing Dose

0 2 3 5 ChE

sd

N 52184 U/L 1887.5

6 50600 U/L 2619.9

6 43686 U/L 2300.2

6 41315 U/L 3712.8

6 34964 U/L 4661.7

6 Exponential Model. (Version: 1.7; Date: 12/10/2009) Input Data File: C:/Usepa/BMDS212/Data/exp_Testrunscontinuous_Setting.

(d) Gnuplot Plotting File:

Mon Jun 11 13:01:24 2012 BMDS Model Run The form of the response function

by Model: Model
2:

Y [dose] a * exp{sign * b * dose} Model
3:

Y [dose] a * exp{sign

* (b
* dose)

... d} Model
4: Y[dose]
a * [c- (c-1)

* exp{-b

* dose}] Model
5: Y[dose]
a * [c- (c-1)

* exp{-(b

* dose)

... d}l Note: Y[dose]

is the median response for exposure

= dose; sign

+1 for increasing trend in data; sign

= -1 for decreasing trend

.

. Model

2 is nested within Models

3 and 4. Model

3 is nested within Model
5. Model

4 is nested within Model
5. Dependent variable

= Mean Independent variable

= Dose Data are assumed

to be distributed: normally variance Model: exp(lnalpha +rho *ln(Y[dose])) rho is set to 0. A constant variance model is fit. Total number
of dose groups

= 5 Total number
of records with missing values

= 0 Maximum number
**o~** iterations

= 250 Relative Function Convergence
has been set to: 1e-008 Parameter Convergence
has been set to: 1e-008 MLE solution provided: Exact Initial Parameter Values Page 59 of84VariableModel2Model3Model4-----------------------------lnalpha15.9615.9615.96rho(S)000a36693.236693.254793.2b0.08348520.08348520.152853c0.3190540.319054d1(S)SpecifiedParameterEstimatesbyModelVariableModel2Model3----------------------lnalpha16.079816.072rho00a5297952657.9b0.08333430.0897811cd1.08599TableofStatsFromInputDataDoseNObsMeanObsStdDev-----------------------065.218e+00418881 65.06e+0042620264.369e+00423003 64.132e+00437135 63.496e+0044661EstimatedValuesofInterestModelDoseEstMeanEstStd-------------------------205.298e+004310214.874e+004310224.485e+004310234.126e+004310253.493e+0043102305.266e+004309014.895e+004309024.51e+004309034.14e+004309053.463e+0043090405.298e+004310214.874e+004310224.485e+004310234.126e+004310253.493e+0043102505.249e+004303114.967e+004303124.496e+004303134.052e+004303153.511e+0043031Page60of84·Model416.07980529790.08333430ScaledResidual-----------------0.62771.466-0.91570.043460.03021-0.37571.306-1.122-0.064820.2629-0.62771.466-0.91570.043460.03021-0.24620.7494-1.0280.6459-0.1209Model5-------15.96054793.20.1528531Model516.033052488.60.3226790.6275721.64519Other models for which likelihoods

are calculated: Model A1:
Yij Mu(i)

+ e(ij) Var{e(ij)} Sigma"2 Model A2:
Yij Mu(i)

+ e(ij) Var{e(ij)} Sigma(i)"2 Model A3:
Yij Mu(i)

+ e(ij) var{e(ijl} exp(lalpha

+ log(mean(i))

* rho) Model

R:
Yij Mu
+ e(i) var{e(ij)} Sigma"2 Likelihoods

of Interest Model
Log (likelihood)

DF
AIC ------- -----------------

---

- -------- A1 -2S4.4003

6 S20.8007 A2 -2S1.1922

10 S22.384S A3 -2S4.4003

6 S20.8007 R -280.3427

2 S64.68S3 2 -2S6.1963

3 S18.3926 3 -2S6.0803

4 S20.1606 4 -2S6.1963

3 S18.3926 s -2SS.4949

s S20.9898 I Additive constant for all log-likelihoods

= -27.S7. This constant added to the above values **giv~s** the log-likelihood including
the term that does not depend
on the model parameters. Explanation

of Tests Test
1: Does response and/or variances differ among Dose levels?
(A2 vs. R) Test 2: Are vartances Homogeneous?

(A2 vs. A1) Test 3: Are varfances adequately modeled?

(A2 vs. A3) Test 4: Does Model

2 fit the data? (A3 vs. 2) Test Sa: Does Model

3 fit the data? (A3 vs. 3) I Test Sb: Is Model

3 better than Model
2? (3 vs. 2) ' Test 6a: Does Model

4 fit the data? (A3 vs. 4) Test 6b: Is Model

4 better than Model
2? (4 vs. 2) Test 7a: Does Model

s fit the data? (A3 vs. S) Test 7b: Is Modet

s better than Model
3? (S vs. 3) Test 7c: Is Model

s better than Model 4? (S vs. 4) Tests of Interest Test -2*log(Likelihood Ratio)

D. F. --------

---

- --------

- ----------- Test

1

S8.3

8 Test

2

6.416

4 Test

3

6.416

4 Test

4

3.S92

3 Test sa

3.36

2 Page 61 of84 p-value < 0.0001 0.1701 0.1701 0.309 **0.1864 ~** Testsb0.232Test6a3.S92Test6b0Test7a2.189Test7b1.171Test7c1.4031301120.630.309N/A0.1390.27920.49S9Thep-valueforTest1islessthan.OS.Thereappearstobeadifferencebetweenresponseand/orvariancesamongthedoselevels,itseemsappropriatetomodelthedata.Thep-valueforTest2isgreaterthan.1.Ahomogeneousvariancemodelappearstobeappropriatehere.Thep-valueforTest3isgreaterthan.1.Themodeledvarianceappearstobeappropriatehere.Thep-valueforTest4isgreaterthan.1.Model2seemstoadequatelydescribethedata.Thep-valueforTestSaisgreaterthan.1.Model3seemstoadequatelydescribethedata.Thep-valueforTestSbisgreaterthan.OS.Model3doesnotseemtofitthedatabetterthanModel2.Thep-valueforTest6aisgreaterthan.1.Model4seemstoadequatelydescribethedata.DegreesoffreedomforTest6barelessthanorequaltoo.TheChi-Squaretestforfitisnotvalid.Thep-valueforTest7aisgreaterthan.1.ModelSseemstoadequatelydescribethedata.Thep-valueforTest7bisgreaterthan.OS.ModelSdoesnotseemtofitthedatabetterthanModel3.Thep-valueforTest7cisgreaterthan.OS.ModelSdoesnotseemtofitthedatabetterthanModel4.BenchmarkDosecomputations:SpecifiedEffect0.100000RiskTypeRelativedeviationConfidenceLevel0.9SOOOOBMDandBMDLbyModelModel234sBMD1.264311.402411.264311.S2871BMDL1.089071.096070.9294S81.02944Page62of84Exponential Model2 with 0.95 Confidence Level 55000 Exponential 50000 Q) 45000 Ill c: 0 c.. Ill Q) 0::: c: (U 40000 Q) :::::!!: 35000 30000

BMD BMD 0

1

2

3

4

5 dose 13

: 01 06/11 2012

Page 63 of84Exponential Model3 with 0.95 Confidence Level 55000 Exponential 50000 Q) 45000 1/) c: 0 c.. 1/) Q) 

### a: c: «< 40000 Q) :::!: 35000 30000

0

2



### 3

4

5 dose 

### 13

: 01 06/11 2012

Page 64 of84Exponential Model

4 with 0.95 Confidence Level 55000 Exponential 50000 CD 45000 U) c: 0 c.. U) CD 0::: c: IV CD 40000 :E 35000 30000

0

2

3

4

5 dose 13

: 01 06111 2012

Page 65 of84

## Exponential Model5 with 0.95 Confidence Level 55000

Exponential 50000 Cl) 

## 45000 rn c::: 0 0.. 

## rn Cl) 

# a::: c::: CQ 

## 40000 Cl) 

### ~ 

## 35000 30000



## 0

2



## 3

4



# 5 dose 13:01 06/11 2012 

# Selected BMDS output for Likelihood Ratio Test: Exponential model (heterogeneous variance) Female PNDll RBC: Likelihoods

of Interest Model
Log (likelihood)



## DF
AIC 

# ------- 

# ----------

- ------

- ---

- ------- A1 -281.4468

5 572.8935 A2 -273.2373

8 562.4745 A3 -278.2065

6 568.4131 R 

## -301.4351

2 606.8702 2 -284.6608

4 577.3215 3 -284.6608

4 577.3215 4 -278.8974

5 567.7947 5 -278.8974

5 567.7947 Additive constant for all log-likelihoods



# = -34. This constant added to the above values gives the log-likelihood including
the term that does not depend
on the model parameters.



## Page 66 of84Exponential Model (heterogeneous variance) Male PND11 RBC: Likelihoods

of Interest Model
Log (likelihood)

DF
AIC ------- ----------------- ------------ A1 -258.8745

5 527.7491 A2 -256.3494

8 528.6987 A3 -257.6339

6 527.2678 R -281.9828

2 567.9655 2 -257.8321

4 523.6641 3 -499.1267

4 1006

. 253 4 -257.6347

5 525.2694 5 -257.6347

5 525.2694 Additive constant for all log-likelihoods

= -32.16. This constant added to the above values gives the log-likelihood including
the term that does not depend
on the model parameters

. Exponential Model (heterogeneous variance

) combined PND 11 RBC

: Likelihoods

of Interest Model
Log (likelihood) ------- ----------------- A1 -544.8439 A2 -539.0674 A3 -541.3211 R -584.2958 2 -545.789 3 -890.5382 4 -541.693 5 -541.693 Additive constant for all log-likelihoods

= above values gives the log-likelihood including depend
on the model parameters.

Page 67 of84

DF 5 8 6 2 4 4 5 5 -66.16. the term AIC ------------ 1099.688 1094.135 1094.642 1172.592 1099.578 1789.076 1093.386 1093.386 This constant that does not added to the

# Combined Male and Female PNDll RBC ChE Data for Dose- Response Study 

# ==================================================================== Hill Model. (Version: 2.14; Date: 06/26/2008) Input Data File: C:\USEPA\BMDS21\hi1Dax Setting.
(d) Gnuplot Plotting File: C:\USEPA\BMDS21\hi1Dax_Setting.plt

Tue Aug 14 11:07:55 2012 

# ==================================================================== BMDS Model Run The form of the response function
is: Dependent variable



# = mean Independent variable

= 

# dose Power parameter restricted

to be greater than

1 The variance

is to be modeled

as Var(i)



# = exp(lalpha +rho* ln(mean(i))) Total number
of dose groups



# = 4 Total number
of records with missing values

= 

# 0 Maximum number
of iterations



# = 250 Relative Function Convergence
has been set to: 1e-008 Parameter Convergence
has been set to: 1e-008 Default Initial Parameter Values lalpha 14.1917 rho

0 intercept

v 

# n 

# k 

# 6865.67 -4466.78 8.77777 0.211717 Asymptotic Correlation Matrix
of Parameter Estimates ( *** The model parameter(s)

-n have been estimated

at

a boundary point,
or have been specified

by the user,

and do not appear
in the correlation matrix lalpha

rho intercept

v

k 

# lalpha

1

-1 0.14 -0.13 -0.056 rho

-1

1 -0.15
0.13 0.056 intercept

0.14 -0.15

1 -0.2 -0.61 v -0.13
0.13
-0.2

1 -0.59 k 

# -0.056 0.056 -0.61 -0.59

1 

# Page 68 of84Parameter Estimates 95.0% Wald Confidence Interval variable Estimate

Std. Err. Lower Conf. Limit Upper Limit lalpha 4.19024 3.76642 11.5723
rho 1.17634 0.449021 2.05641 intercept 6839.55 330.275 7486.87

v -5892.8 572.828 4770.08

n

1

NA k 0.325016 0.115836 0.552051 NA

- Indicates that this parameter
has hit

a bound implied
by some inequality constraint
and thus has no standard error. Table of Data and Estimated Values
of Interest -3.19182 0.296275 6192.22 -7015.53 0.0979805 Dose

N Obs Mean Est Mean Obs Std Dev Est Std Dev Scaled Res. -------- -------- ----------- 0 18 6.87e+003 6.84e+003 1.61e+003 0.1
17 5.42e+003 5.45e+003

943 0.3
19 4.01e+003 4.01e+003 1.31e+003 1 18 2.4e+003 2.39e+003

760 Model Descriptipns

for likelihoods calculated Model A1:
Yij Var{e

· (ij}} I Model A2:
Yij Var{e

J (ij}} Mu(i}

+ e(ij} Sigma"2 Mu(i}

+ e(ij} Sigma(i}"2 Model A3:
Yij Mu(i}

+ e(ij} ----------- 1.46e+003 1. 28e+003 1.07e+003 789 Var{e(ij}} exp(lalpha

+ rho*ln(Mu(i}}} Model A3 uses any fixed variance parameters that were specified

by the user Model

R:

Yi var{e(i}} Model A1 A2 A3 fitted

R

Mu
+ e(i} Sigma"2 Likelihoods

of Interest Log (likelihood}

# Param•s -544.843935

5 -539.067392

8 -541.321057

6 -541.453167

5 -584.295799

2 Page 69 of84
AIC 1099.687870 1094.134785 1094.642113 1092.906334 1172.591599 ---------- 0.0757 -0.116 0.00464 0.036 Conf. qo

I

# Test
1: Test 2: Test 3: Test 4: (Note: Test Test Test Test Test

1 2 3 4 

# Explanation

of Tests Do responses and/or variances differ among Dose levels? (A2 vs. R) Are Variances Homogeneous?

(A1 vs A2) Are variances adequately modeled?
(A2 vs. A3) Does the Model for the Mean Fit? (A3 vs. fitted) When rho=O the results

of Test

3 and Test

2 will be the same.) Tests of Interest -2*log(Likelihood Ratio) Test df

p - value 90.4568

6 <.0001 11.5531 3 0

. 009082 4.50733

2 0.105 0.264221

1

0. 6072 The p-value for Test

1 is less than .05. There appears

to be

a difference between response and/or variances among the dose levels It seems appropriate

to model the data The p-value for Test

2 is less than .1.
A non-homogeneous variance model appears
to be appropriate The p-value for Test

3 is greater than .1. The modeled variance appears to be appropriate here The p-value for Test

4 is greater than .1. The model chosen seems to adequately describe the data Benchmark Dose Computation Specified effect

0.1 Risk Type Relative risk Confidence level

0

. 95 BMD 

# 0.0426766 BMDL 0.0285396



# Page 70 of84Hill Model with 0.95 Confidence Level 8000

Hill 7000 6000 Q) Ul r::: 0 Q. 5000 Ul Q) 0::: r::: IV Q) ::::! 4000 3000 2000 BMD BMD 0

0.2

0

. 4

0.6

0.8 dose 11:07 08/14 2012

Page 71 of84

### Propoxur CCA Dose range study- combined sexes: adult male and female RBC combined EPA BMDS v2.12 

### Exponential model- 

## ==================================================================== Exponential Model. (Version: 1.7; Date: 12/10/2009) Input Data File: C:/Usepa/BMDS212/Data/exp_UntitledData_Setting.

(d) Gnuplot Plotting File:

Thu Aug 16 11:12:56 2012 BMDS Model Run The form of the response function

by Model: Model
2:

Y [dose] a * exp{sign * b * dose} Model
3:

Y [dose] a * exp{sign

* (b
* dose)Ad} Model
4: Y[dose]
a * [c- (c-1)

* exp{-b

* dose}] Model
5: Y[dose]
a * [c- (c-1)

* exp{-(b

* dose)Ad}l Note: Y[dose]

is the median response for exposure



## = dose; sign

+1 for increasing trend in data; sign



## = -1 for decreasing trend. Model

2 is nested within Models

3 and 4. Model

3 is nested within Model
5. Model

4 is nested within Model
5. Dependent variable

= mean Independent variable



### = Dose Data are assumed
to be distributed: normally Variance Model: exp(lnalpha +rho *ln(Y[dose])) The variance

is to be modeled

as Var(i)



### = exp(lalpha

+ log(mean(i)) *rho) Total number
of dose groups



### = 6 Total number
of records with missing values



## = 0 Maximum number
of iterations



## = 250 Relative Function Convergence

has been set to: 1e-008 Parameter Convergence
has been set to: 1e-008 MLE solution provided: Exact Initial Parameter Values Variable
Model

2 Model

3 

### -------- ------- 

## ------- lnalpha 7.48339 7.48339 rho 0.731572

0. 731572 a 1106.16 1106.16 b 0.132095 0.132095 c 0.236888

d

1 Parameter Estimates

by Model 

### Page 72 of84 Model

4 

### ------- 7.48339 0.731572 3227.7 0.385352 0.236888 Model

5 

### ------- 7.48339 0.731572 3227.7 0.385352

1Variable
Model

2 Model

3 -------- ------- ------- lnalpha 8.49327 8.49326 rho 0.605466 0.605468 a 2929.5 2929.5 b 0.149886 0.149886 c d

1 Table of Stats From Input Data Dose

N
Obs Mean Obs Std Dev ---------- ------------- 0
12 3074
1014 1
12 2598 593.1 2
12 2110 684.4 3
12 1608 772.2 5
12 1350 413.1 10
12 802.8 565.8 Estimated Values
of Interest Model Dose
Est Mean
Est Std ------- ---------- --------- 2

p 2929 783.1 1 2522 748.3 2 2171 715.1 B 1869 683.4 5 1385 624.1 10 654.4 497.4 3

0 2929 783.1 1 2522 748.3 2 2171 715.1 3 1869 683.4 5 1385 624.1 10 654.4 497.4 4

0 3134 824.7 1 2520 753.9 2 2061 694.1 3 1719

644 5 1271 568.8 10 836.8 478.9 5

0 3119 822.7 1 2537 755.8 2 2070 695.1 3 1716 643.6 5 1258 566.4 10 841.4 480.1 Other models for which likelihoods

are calculated: Model A1:
Yij = Mu(i)

+ e(ij) Var{e(ij)}

= SigmaA2 Model A2:
Yij Var{e(ij)} Mu(i)

+ e(ij) Sigma(i)A2 Model A3:

Yij = Mu(i)

+ e(ij)

Model

4 ------- 6.8027 0.823266 3134.12 0. 291311 0.224916 scaled Residual ---------------- 0.6392 0.3538 -0.2965 -1.322 -0.1938 1.034 0.6392 0.3538 -0.2965 -1.322 -0.1938 1.034 -0.2525 0.3582 0.2398 -0.596 0.479 -0.2459 -0.19 0.2788 0.1962 -0.5837 0.5599 -0.278 Var{e(ij)}

= exp(lalpha

+ log(mean(i)) *rho) Page 73 of84

Model

5 ------- 6.80955 0.82231 3119.12 0.298776 0.237499 1.05216

## Model

R:
Yij Var{e(ij)} Model 

# ------- A1 A2 A3 R 2 3 4 

# s Mu
+ 

## e(i) Sigma"2 Likelihoods

of Interest Log (likelihood)

DF 

# --------------

- -- -S04.4921

7 -499.0298

12 -S02.2S09

8 -S34.4764

2 -S04.3179

4 -S04.3179

4 -S02.3881



# s -S02.3774

6 AIC 

# -----------

- 1022.984 1022.06 1020.S02 1072.9S3 1016.636 1016.636 1014.776 1016.7SS Additive constant for all log-likelihoods



# = -66.16. This constant added to the above values gives the log-likelihood including
the term that does not depend
on the model parameters. Explanation

of Tests Test
1: Does response and/or variances differ among Dose levels?
(A2 vs. R) Test 2: Are Variances Homogeneous?

(A2 vs. A1) Test 3: Are variances adequately modeled?

(A2 vs. A3) Test 4: Does Model

2 fit the data? (A3 vs. 2) Test Sa: Does Model

3 fit the data? (A3 vs 3) Test Sb: Is Model

3 better than Model
2? (3 vs. 2) Test 6a: Does Model

4 fit the data? (A3 vs 4) Test 6b: Is Model

4 better than Model
2? (4 vs. 2) Test 7a: Does Model



# s fit the data? (A3 vs S) Test 7b: Is Model



# s better than Model
3? (S vs. 3) Test 7c: Is Model



# s better than Model 4? (S vs. 4) Tests of Interest Test -2*log(Likelihood Ratio)

D.
F

. 

# -------- 

# ------------------------ Test

1

70.89

10 Test

2

10.92



# s Test

3

6.442

4 Test

4

4.134

4 Test Sa

4.134

4 Test Sb -9.3e-011

0 Test 6a

0.2744

3 Test 6b

3.86

1 Test 7a

0.2S31

2 Test 7b

3.881

2 Test 7c
0.02129

1 p-value 

# --------------

< 

## 0.0001 O.OS29 0.168S 0.3882 0.3882 N/A 0.9648 0.04946 0.8811 0.1436 0

. 884 The p-value for Test

1 is less than .OS. There appears

to be

a difference between response and/or variances among the dose levels,

it seems appropriate

to model the data. 

# Page 74 of84The p-value for Test

2 is less than .1.
A non-homogeneous variance model appears

to be appropriate. The p-value for Test

3 is greater than .1. The modeled variance appears

to be appropriate here. The p-value for Test

4 is greater than .1. Model

2 seems to adequately describe the data. The p-value for Test Sa is greater than .1. Model

3 seems to adequately describe the data. Degrees
of freedom for Test Sb are less than or equal to 0. The **Chi~square** test for fit is not valid. The p-value for Test 6a is greater than .1. Model

4 seems to adequately describe the data. The p-value for Test 6b is less than .05. Model

4 appears to fit the data better than Model
2. The p-value for Test ?a is greater than .1. Model

5 seems to adequately describe the data. The p-value for

' Test 7b is greater than .05. Model

5 does not seem to fit

1 the data better than Model
3. The p-value for Test 7c is greater than .05. Model

5 does not seem to fit the data better than Model
4. Benchmark Dose Computations: Specified Effect 0.100000 Risk Type Relative deviation Confidence Level 0.950000 Model 2 3 4 5 BMD and BMDL by Model BMD I - ----------- 0.702937 0.702937 . 0. 474181 0.51859

BMDL 0.557075 0.557075 0.339178 0.339621 Page 75 of84

### Exponential Model

2 with

0

. 95 Confidence Level 4000 Exponential 3500 3000 

# 5l

2500 c: 

# &. 

# .., **~** 2000 c: Ill CD 

## ::!: 1500 1000 500 

### BMD MD 

## 0

2

4



## 6

8

10 dose 11

: 12 08116 2012



# Page 76 of843! c: 0 D. & c: :l **~** 4000 3500 3000 2500 2000 1500 1000 500 11:12 08116 2012 4000 3500 3000 2500 2000 1500 1000 500 11

: 12 08/16 2012 BMD MD 0 BMD MD 0 Exponential Model

3 with

0

. 95 Confidence Level Exponential 2

4

6

8

1 0 dose Exponential Model

4 with 0.95 Confidence Level Exponential 2

4

6

8

1 0 dose Page 77 of84

# Exponential Model

5 with

0

. 95 Confidence Level 4000 Exponential 3500 3000 

# 5l

2500 c 0 

# a. 

# XI 

# a: 2000 c al Q) 

## ::::!! 

# 1500 1000 500 BMD MD 0



# 2

4

6

8

10 dose 11:12 08/16 2012



# Page 78 of84Propoxur CCA Dose-Range study- combined sexes adult male and female brain EPA BMDS v2.12 Exponential model Exponential Model. (Version: 1.7; Date: 12/10/2009) Input Data File: C:/Usepa/BMDS212/Data/exp_UntitledData_Setting.

(d) Gnuplot Plotting File:

Thu Aug 16 12:06:49 2012 ==================================================================== BMDS Model Run The form of the Model
2: Model
3: Model
4: response function
by Model: Y[dose]
a * exp{sign * b * dose} Y[dose]
a * exp{sign

* (b
* dose)Ad} Y[dose]
a* [c-(c-1)

* exp{-b *dose}] Model
5:

Y [dose] a * [c- (c-1)

* exp{-
(b
* dose) Ad}] Note: Y[dose]

is the median response for exposure

= dose; sign
+1 for increasing trend in data; sign

= -1 for decreasing trend. Model

2 is nested within Models

3 and 4. Model

3 is nested within Model
5. Model

4 is nested within Model
5. Dependent variable

= mean Independent variable

= Dose Data are assumed

to be distributed: normally Variance Model: exp(lnalpha +rho *ln(Y[dose])) The variance

is to be modeled

as Var(i)

= exp(lalpha

+ log(mean(i)) *rho) Total number
of dose groups

= 6 Total number
of records with missing values

= 0 Maximum number
of iterations

= 250 Relative Function Convergence
has been set to: 1e-008 Parameter Convergence
has been set to: 1e-008 MLE solution provided: Exact Variable lnalpha 0.458415 rho a b c d Variable
Model

2 67.4078 -4.79547 30110 0.0704899 Model

2 Initial Parameter Values Model

3 67

. 4078 -4.79547 30110 0.0704899

1 Parameter Estimates

by Model Model

3 Page 79 of84 Model

4 67.4078 -4.79547 54464.4 0.313747 0.458415 Model

4 Model

5 67.4078 -4.79547 54464.4 0.313747

1 Model

5 }00

# -------- ------- ------- lnalpha 54.5481 54.5481 rho -3.58247 -3.58247 a 52289.6 52289.6 b 0.0990209 0.0990209 c d

1 Table of Stats From Input Data Dose

N


## Obs Mean Obs Std Dev 

# ---------- ------------- 0
12 5.187e+004

1588 1
12 4.854e+004

3268 2
12 4.291e+004

3881 3
12 3.776e+004

4438 5
12 3.062e+004

7247 10
12 2.622e+004 1.094e+004 Estimated Values
of Interest Model Dose
Est Mean
Est Std 

# ------- 

# ---------- --------- 2

0 5.229e+004

2473 1 4.736e+004

2953 2 4.289e+004

3526 3 3.885e+004

421.0 5 3.187e+004

6003 10 1.943e+004 1.457e+004 3

0 5.229e+004

2473 1 4.736e+004

2953 2 4.289e+004

3526 3 3.885e+004

4210 5 3.187e+004

6003 10 1.943e+004 1.457e+004 4

0 5.25e+004

2280 1 4.706e+004

2901 2 4.25e+004

3631 3 3.868e+004

4469 5 3.279e+004

6430 10 2.464e+004 1.207e+004 5

0 5.199e+004

2171 1 4.831e+004

2571 2 4.291e+004

3379 3 3.783e+004

4518 5 3.083e+004

7241 10 2.629e+004 1.045e+004 Other models for which likelihoods

are calculated: Model A1:

Yij Var{e(ij)} Model A2:
Yij Var{e(ij)} Model A3:
Yij Mu(i)

+ e(ij) Sigma

... 2 Mu(i)

+ e(ij) Sigma(i)"2 Mu(i)

+ e(ij)


# ------- 63.3663 -4.40747 52499.6 0.176593 0.359934 Scaled Residual 

# ---------------- -0.5866 1. 385 0.01581 -0.9012 -0.7219 1.614 -0.5866 1. 385 0.01581 -0.9012 -0.7219 1. 614 -0.9555 1. 768 0. 3913 -0.7163 -1.171 0.4512 -0.1973 0.3049 -0.002005 -0.05872 -0.1 -0.02404 var{e(ij)} exp(lalpha

+ log(mean(i)) *rho) 

# Page 80 of84



# ------- 65.4174 -4.60927 51994.5 0.284735 0.501383 1. 49384Model

R:
Yij Var{e(ij)}

Mu
+ e(i) Sigma"2 Likelihoods

of Interest Model
Log (likelihood) ------- ---------

- ------- A1 -6S9.9218 A2 -636

. 619S A3 -638

. 6038 R -70S

. 0244 2 -643.2422 3 -643.2422 4 -641.6889 s -638.619S Additive constant for all log-likelihoods

= above val

l es gives the log-likelihood including depend
on the model parameters.

DF 7 12 8 2 4 4 s 6 -66.16. the term Explanation

of Tests I

AIC ---

- -------- 1333.844 1297.239 1293.208 1414.049 1294.484 1294.484 1293.378 1289.239 This constant **adde~** that does not

I Test
1: Does response and/or variances differ among Dose levels?
(A2 vs. R)

' I Test

2
: Are Variances Homogeneous?

(A2 vs. A1) Test

3
: Are variances adequately modeled?

(A2 vs. A3) Test 4: Does Model

2 fit the data? (A3 vs. 2) I I Test Sa: koes Model

3 fit the data? (A3 VS 3) Test Sb:

s Model

3 better than Model
2? (3 vs. 2) I I

fit the data? (A3 VS 4) Test 6a: poes Model

4 Test 6b: Is Model

4 better than Model
2? (4 vs. 2) i I Test 7a: boes Model

s fit the data? (A3 VS S) Test 7b: Is Model

s better than Model
3? (S vs. 3) Test 7c: Is Model

s better than Model 4? (S vs. 4) Tests of Interest Test -2*log(Likelihood Ratio)

D

. F. p-value -------- ------------------------ -------------- Test

1

136.8

10

< 0.0001 Test

2

46.6

s

< 0.0001 Test

3

3.969

4 0.4103 Test

4

9.277

4 O.OS4S4 Test Sa

9

. 277

4 O.OS4S4 Test Sb

I 2.274e-013

0

N/A Test 6a

6.17

3 0.1036 Test 6b

3

. 107

1 0.07798 Test 7a
0.03139

2 0.9844 Test 7b

9.24S

2 0.009826 Test 7c

6

. 139

1 0.01322 The p-value for Test

1 is less than .OS. There appears

to be

a differe

h ce between response and/or variances among the dose levels,

J , it seems appropriate

to model the data. I ' The p-value for Test

2 is less than

.1

. A non-homogeneous Page 81 of84

to theI f ' 

### I { 

## I 

# I I i I 

# ! 

# i .I I 

## ! 

# I variance model appears

to be appropriate. The p-value for Test

3 is greater than .1. The modeled variance appears

to be appropriate here. The p-value for Test

4 is less than .1. Model

2 may not adequately describe the data; you may want to consider another model. The p-value for Test sa is less than

.1

. Model

3 may not adequately describe the data; you may want to consider another model. Degrees

of freedom for Test Sb are less than or equal to 0. The Chi-Square test for fit is not valid. The p-value for Test 6a is greater than .1. Model

4 seems to adequately describe the data. The p-value for Test 6b is greater than .OS. Model

4 does not seem to fit the data better than Model
2. The p-value for Test 7a is greater than .1. Model

S seems to adequately describe the data. The p-value for Test 7b is less than .OS. Model

S appears to fit the data better than Model
3. The p-value for Test 7c is less than .OS

. Model

s appears to fit the data better than Model
4. Benchmark Dose Computations: Specif

" ied Effect 0.100000 Risk Type Relative deviation Confidence Level 0.9SOOOO BMD and BMDL by Model Model 2 3 4 

### s

BMD 1.06402 1.06402 0

. 961987 1. 28942

BMDL 0.944464 0.944464 0.822887 1. 03778 

### Page 82 of84Exponential Model

2 with
0

. 95 Confidence Level 55000 Exponential 50000 45000 CD 40000 .., r:: 0 c. .., CD a:: 35000 r:: Ill Q) :::.! 30000 25000 20000 BMD MD 0

2

4

6

8

1 0 dose 12:06 08/16 2012 Exponential Model

3 with
0

. 95 Confidence Level 55000 Exponential 50000 45000 Q) 4oobo .., r:: 0 c. .., Q) a:: 35000 r:: Ill CD :::.! 30000 25000 20000 BMD BMD 0

2

4

6

8

1 0 dose 12

: 06 08/16 2012

)0 Page 83 of84ExponentialModel4with0.95ConfidenceLevel55000Exponential5000045000CD40000Illc:0c.Ill(!}_35000c:IllCD::!!300002500020000BMDMD0246810dose12:0608/162012ExponentialModel5with0.95ConfidenceLevel55000Exponential5000045000CD40000Illc:0c.Ill(!}_35000c:IllCD::!!300002500020000BMDMD0246810sdoset-12:0608/1620120Page84of84