RTI-909; Project 0209189.000 Final Report



### i Table of Contents
Page

QUALITY ASSURANCE STATEMENT ........................................................................... iii

COMPLIANCE STATEMENT .......................................................................................... v

ABSTRACT...................................................................................................................... 1

INTRODUCTION ............................................................................................................. 4

MATERIALS AND METHODS ......................................................................................... 5 Designation of Responsibilities .................................................................................. 5

Test Material............................................................................................................... 5 Animals and Husbandry ............................................................................................. 6 Study Schedule and Design....................................................................................... 9 Table A. Study Schedule........................................................................................... 9 Table B. Number of Animals Assigned to Study Groups........................................... 10

Statistics.................................................................................................................... . 13 Storage of Records .................................................................................................... 15

Personnel ................................................................................................................... 1 5

RESULTS ........................................................................................................................ 16 Test Chamber Analyses ............................................................................................. 16 Maternal Findings....................................................................................................... 17 Uterine and Embryofetal Findings .............................................................................. 19

DISCUSSION................................................................................................................... 2 1

CONCLUSIONS............................................................................................................... 26

REFERENCES ................................................................................................................ 28

LIST OF PROTOCOL DEVIATIONS ............................................................................... 32

Appendices Appendix I: HLS Inhalation Report Appendix II: Individual Animal Data Tables Appendix III: Protocol and 2 AmendmentsRTI-909; Project 0209189.000 Final Report



### ii List of Tables
Page
Table 1 Analysis of Test Atmospheres........................................................................ 34

Table 2 Summary and Statistical Analysis of the Maternal Body Weights and Body Weight Changes............................................................................................. 35

Table 3 Summary of the Maternal Clinical Observations ............................................ 41

Table 4 Summary and Statistical Analysis of the Maternal Feed Consumption.......... 43

Table 5 Summary and Statistical Analysis of the Maternal Absolute and Relative Organ Weights ............................................................................................... 49

Table 6 Summary and Statistical Analysis of Ovarian Corpora Lutea, Uterine Contents, Live Fetal Sex and Live Fetal Body Weight ................................... 51

Table 7 Summary and Statistical Analysis of Fetal External Malformations and Variations ....................................................................................................... 55

Table 8 Summary of Morphological Abnormalities in CD-1 Mouse Fetuses: Listing by Defect Type.................................................................................... 58

Table 9 External Malformations in Control CD-1® Mouse Litters From Studies Performed for Governmental Clients at RTI From 1997 to 2002.................... 59RTI-909; Project 0209189.000 Final Report



### iv 

## THIS PAGE LEFT INTENTIONALLY BLANK**RTI-909; Project** **0209189.000 Final** **Report GLP** **COMPLIANCE STATEMENT** **This study** **was conducted** **in accordance** **with the** **U.S. EPA** **Good Laboratory** **Practice (GLP)** **standards for** **the 21** **l(b) program** **(40 CFR** **79; U.S.** **EPA, 1994),** **and performed** **according to** **the protocol** **(and two** **amendments) and** **following the** **SOPs of** **HLS and** **RTI. This** **study complied** **with all** **appropriate parts** **of the** **USD A** **Animal Welfare** **Act regulations:** **9 CFR** **Parts 1** **and 2 Final** **Rules, Federal** **Register, Vol.** **54, No.** **168, August** **31,1989, pp.** **36112-36163, effective** **October 30,** **1989, and** **9 CFR** **Part 3** **Animal Welfare** **Standards; Final** **Rule, Federal** **Register, Volume** **55, No.** **32, February** **15,1991, pp.** **6426-6505, effective** **March 18,1991.** **It was** **the Sponsor's** **responsibility to** **maintain the** **method of** **synthesis, fabrication,** **or derivation** **of the** **test fuel,** **and this** **was not** **completed at** **the time** **of the** **study conduct** **but has been** **completed since** **and is** **on file** **with the** **Sponsor.**

### Rbchelle**W. Tyl,** **Ph.D., DABT** **Date Study** **Director**

### /^**M. Gray,** **IVLS^DABT ^f** **' Date** **^/Sponsor's Representative** RTI-909; Project 0209189.000 Final Report



### 1 ABSTRACT 

### This study was conducted to provide a robust evaluation of the developmental toxicity
potential of an inhaled vapor c ondensate of gasoline/methyl tert iary butyl ether (G/MTBE) in

mice. It was conducted in accordance with the U.S. EPA Good Laboratory Practice (GLP)
standards for the 211(b) program (40 CFR 79; U.S. EPA, 1994). In the mouse study at
ExxonMobil Biomedical Sciences, Inc. (EMB SI, 2009b), several uncommon ventral closure
defects (malformations) were observed in a nonexposure responsive in cidence. The study
reported herein was conducted to confirm and/or extend the findings obser ved in that earlier
mouse study.
Twenty-three plug-positive female CD-1 mi ce each were distribut ed on gestational day
(gd) 0 into the 0, 2000, 10,000, and 20,000 mg/m 3 

### target concentration groups, and an additional
38 plug-positive CD-1 mice were dist ributed on gd 0 into the 30,000 mg/m 3 

### target concentration
group. Exposures were for 6 hours/day on gd 5 through 16 for the 0-20,000 mg/m 3 

### groups and
for 6 hours/day on gd 5 through 10 for the 30,000 mg/m 3 

### group. The females were weighed on
gd 0 and daily on gd 5 through 17; feed consumption and clinical observations were also
recorded daily. Clinical observations were reco rded individually before and after each exposure
period and recorded at least on ce, using general categories (e. g., few, some, most, all, etc.)
during each exposure period. At scheduled necropsy on gd 17, all dams were euthanized, with
body weight, gravid uterine weight , liver weight, paired adrena l gland weights, and paired
kidney weights recorded. Ovaria n corpora lutea were counted a nd uterine total implantations,
resorptions, late fetal deaths, and live fetuses re corded for each pregnant dam. Each live fetus
was euthanized by intraperitoneal injection of sodium pentobarbital, sexed, and examined
externally for gross malformations and variations (including examination for cleft palate). Each
fetus was then dissected by a ve ntral longitudinal cut; the thoracic and abdominal viscera were

removed and retained in buffered neutral 10% formalin for possible subsequent visceral

examination. The carcasses were skinned afte r blanching and retained in 70% ethanol for
possible subsequent staining and skeletal examination.
Mean analytical exposure concen trations were 0, 2074, 9925, 20,342 and 29,250 mg/m 3



### . No females died or were sacrificed moribund; 1 female was removed due to a pre-existing

condition. There were no differences across groups in matern al body weights or weight changes
before, during, or after the exposure period, exce pt for significant decreases in body weightRTI-909; Project 0209189.000 Final Report



### 2 change from gd 12 to 13 at 2000 and 20,000 mg/m 3 

### . These findings on gd 12-13 were

considered incidental and unrelated to treatmen t since they were observed on only 1 day and not
in a dose-response pattern. Clinical observations that appeared treatment related included
labored breathing in 1 female each at 20,000 and 30,000 mg/m 3 

### and lacrimation in 1 female at
20,000 mg/m 3 

### and in 3 females at 30,000 mg/m 3 

### . Absolute maternal feed consumption (g/day)
was uniformly decreased at 20,000 and 30,000 mg/m 3 

### and increased at 10,000 mg/m 3 

### (and at
30,000 mg/m 3 

### for gd 13-14) in the exposure period, with sporadic increases and decreases in the
postexposure period in all exposed groups. Rela tive maternal feed consumption (g/kg body
weight/day) was redu ced at 20,000 and 30,000 mg/m 3 

### during the exposure period (with
significant increases dur ing exposure at 10,000 mg/m 3 

### [gd 5-6 and 6-7] and at 30,000 mg/m 3 

### [gd
13-14]), variable during the postexposure pe riod, and unaffected across all groups for the
gestational period: gd 0-17. These decreased feed consumption effects at 20,000 and 30,000
mg/m 3 

### were considered related to exposure to the te st material at these exposure concentrations,
exacerbating effects from the procedures for inha lation exposures, per se (e.g., moving animals,
removal of feed during exposures).
There were no differences in maternal gravid uterine weight or in absolute or relative
paired adrenal gland weights across groups. Absolute maternal liver weight was significantly
increased at 10,000 mg/m 3 

### , and relative maternal liver wei ght was significantly increased at
2000, 10,000, and 20,000 mg/m 3 

### , likely due to induction of me tabolizing enzymes during gd 5-
16 exposures (Conney, 1967). There were no di fferences across groups for the number of
ovarian corpora lutea, uterine impl antation sites, resorptions, late fe tal deaths or live fetuses per
litter, or percent preimplantation loss. There were also no statistically significant differences in
the number (or %) of nonlive (resorp tions plus late fetal deaths) or adversely affected (nonlive
plus malformed) implantations/lit ter, although there were 3 (of 36) females with fully resorbed
litters at 30,000 mg/m 3 

### (with 0, 1, 0, and 2 fully resorbed litters at 0, 2000, 10,000, and 20,000
mg/m 3 

### , respectively).
For live litters, there were no differen ces across groups on the number of live
fetuses/litter, % male fetuses/litter, number of ma le and female fetuses/litter, or on average fetal
body weight per litter for all fetuses or by sexes separately. There were no statistically
significant differences across groups for incidences of external malformation or variations by
fetuses or by litter. External fetal malformati ons included encephalocoele in 1 fetus (in 1 litter)RTI-909; Project 0209189.000 Final Report



### 3 at 2000 mg/m 3 

### and cleft palate in 2 fetu ses (in 2 litters) at 0 mg/m 3 

### , in 1 fetus (in 1 litter) each at
2000, 10,000, and 20,000 mg/m 3 

### , and in 7 fetuses (in 4 litters) at 30,000 mg/m 3 

### . The increased
incidence of cleft palate at 30,000 mg/m 3 

### was not statistically significant; it is likely due to

increased maternal stress and other toxicity at this exposur e concentration during the 6-day
exposure period, during the time of initial palatal formation (gd 5-10). Gastroschisis was also

observed in 1 female fetu s (in 1 litter) at 30,000 mg/m 3 

### (this female also had cleft palate). Fetal

external variations included abnor mal rugae in the palatal midline in 1 fetus (in 1 litter) each at
10,000 and 20,000 mg/m 3 

### , and hematomas of the face, head, neck, and shoulder at 0-20,000
mg/m 3 

### (but not at 30,000 mg/m 3 

### ). The external variations are considered incidental and

unrelated to treatment.
In conclusion, the current study did not c onfirm the presence of ectopia cordis observed
in a previous EMBSI (2009b) study in any fetus in any litter of any group, a nd therefore this fetal
finding is considered unlikely to be related to ma ternal exposure to the test material. In addition,
the study did not confirm the presence of gastroschisis in fetuses at 10,000 mg/m 3 

### (observed in
the EMBSI study) or at 2000 or 20,000 mg/m 3 

### (not observed in the EMBSI study or in the
present study). Gastroschisis was observed in 1 female fetus in 1 litter at 30,000 mg/m 3 

### ; she also
exhibited severely reduced body we ight and cleft palate and was part of a litter with 2 other
fetuses with cleft palate. In total, gastroschi sis was observed in 1 fetus (out of 407 fetuses;
0.24%) in 1 litter (out of 33 litters; 3.03%) at 30,000 mg/m 3 

### , and there was an increased
incidence (not statistically significant) of cleft palate (7 fetuse s in 4 litters), likely from maternal
stress also at 30,000 mg/m 3 

### . Cleft palate is the most common external malformation observed in
mouse fetuses. Maternal treatment-related clinical signs of distress, consisting of labored
breathing observed at 20,000 and 30,000 mg/m 3 

### , likely produced an increased incidence of cleft
palate at 30,000 mg/m 3 

### . This increase in cleft palate is in terpreted as likely to be secondary to
maternal stress (during the gd 5-10 exposure peri od), which would likely result in increased
corticosteroid synthesis (and cleft palate, as note d in the published literature; e.g., Carmichael et
al., 2007; Senda et al., 2005; Pradat et al., 2003; Hemm et al., 1977). The increased
corticosteroid levels likely would have resolv ed in this group by scheduled necropsy on gd 17.
The results of this study indicate that the effects on fetuses at 30,000 mg/m 3 

### were most likely due
to or exacerbated by maternal toxicity. The ab sence of gastroschisis in any of the 3,641 control
CD-1 mouse fetuses, in 288 contro l litters, in RTI’s historical c ontrol database (Table 9) lendsRTI-909; Project 0209189.000 Final Report



### 4 support to the conclusion that ga stroschisis may have been treatment related, occurring in only 1
compromised fetus at 30,000 mg/m 3 

### , with concomitant other fetot oxicity and some indication of
maternal toxicity. Indication of slight fetotoxic ity (gastroschisis in 1 compromised fetus) only at
30,000 mg/m 3 

### and of maternal toxicity (la bored breathing) at 20,000 and 30,000 mg/m 3 

### in this
study results in the following dete rminations: the maternal toxi city No Observable Adverse
Effect Level (NOAEL) was 10,000 mg/m 3 

### , and the developmental toxicity NOAEL was 20,000
mg/m 3



### .

INTRODUCTION 

### A very early inhalation toxi city study of MTBE for 6 hours/day on gd 6-15 in SpragueDawley rats and CD-1 mice showed no matern al or embryofetal effects at any exposure
concentration in rats (Conaway et al., 1985). In mice, there were increases in fetal resorptions at
the low (250 ppm) and high (2500 ppm) concentra tions, attributed to 2 females in each group
with a high number of resorptions . There were no treatment-relate d fetal external, visceral, or
skeletal malformations or variations. Slight ly increased sternebral fusions in the high
concentration group were “attr ibuted to fetotoxicity”.
A previous developmental toxicity study (EMBSI, 2009b) of G/MTBE vapor condensate
by inhalation in mice was one of a series of test s required in accordance with the Alternative Tier
2 provisions of fuels and fuels additives health effects testing regula tions (U.S. EPA, 1994, 40
C.F.R. § 79; Oge 1998). That study provided suggestiv e evidence of an increa se (not statistically
significant or exposure related) in midline de fects among the offspring of dams exposed to
G/MTBE by inhalation at the low and mid ( but not high) vapor concentrations, reporting
gastroschisis and ectopia cordis (2 very rare external malfor mations) in offspring at these
exposure concentrations in the absence of dose-response patterns. That study involved wholebody inhalation exposure of timed-pregnant CD-1 mice for at least 6 hours/day, on gd 5 through
17, to baseline gasoline vapor condensate with 25.5% MTBE at target concentrations of 0, 2000,
10,000, and 20,000 mg/m 3 

### (the last is 50% of the lower explosive limit).
The purpose of the present study was to provide maternal and developmental toxicity data
relative to a 6- or 12-day exposure regimen of inhaled G/MTBE during the period of early or
major organogenesis in gravid mice in order to confirm and extend the findings observed in the
EMBSI mouse study (2009b). The present st udy was conducted with the same exposureRTI-909; Project 0209189.000 Final Report



### 5 concentrations used in the EMBSI study (2009b) for 6 hours/day on gd 5 through 16 (23
females/group) and an additional group of 38 timed-mated mice exposed to 30,000 mg/m 3 

### for 6
hours/day on gd 5 through 10. Fetal malformations of specific interest in this study (ventral wall
closure defects) are formed early in the embr yonic period of gestation; gd 7 through 9 in the
mouse (e.g., Rugh, 1968), hence the shortened expos ure period for this group. A range-finding
study was previously conducted to evaluate the top exposure leve l and exposure duration to be
used in this study (RTI, 2009 )



### .

MATERIALS AND METHODS Designation of Responsibilities 

### RTI International was responsible for study design, protocol ge neration, designed the
procedures and trained the Huntingdon Life Scie nces (HLS) staff for pa iring and detection of
successful mating, assignment of plug-positive study females to groups, necropsy of the maternal
and fetal animals on gd 17, generati on of summary and individual data tables, and draft and final
reports (with RTI QA oversight). RTI’s Qualit y Assurance Unit performed a prestudy on-site
inspection, reviewed the protocol and any amendm ents, and monitored all phases of the study in
which RTI personnel participated. HLS was responsib le for receipt of the test material, prestudy
and study generation and analyses of the test vapo rs, receipt, quarantine and housing of the test
females and breeder males, determining the su ccessful mating and a ssignment of the study
females, in-life observations, loading and unloadi ng study females into and out of chambers, and
submission of interim and final inhalation reports. The Quality Assurance Unit of HLS reviewed
the protocol and monitored the facilities, equipm ent, personnel, methods, practices, records, raw
data, draft and final inhalation report s, and controls used in this st udy to assure that they were in
conformance with company standard operating pr ocedures and the referenced GLP regulations.

Test Material


### The test material, G/MTBE (MRD- 00-713; “API 211BG with MTBE Vapor
Condensate”), was a colorless liquid and identified by the s upplier (Chevron Global Technology
Services Company (CRTC; Richmond, CA) as Lot/Batch Number API 00-02. Methods of
synthesis, fabrication, or derivation were doc umented by the Sponsor and located at API.
Information on identity, strength, purity, a nd composition of G/MTBE was provided by theRTI-909; Project 0209189.000 Final Report



### 6 Sponsor and documented in the raw data and in this final report (Appendix III, protocol
attachment).
Two separate types of chemical analysis we re performed on the test material. CRTC
conducted proprietary characteri zation of all 120 com ponents. EMBSI conducted a GLP-compliant
characterization of 18 representa tive hydrocarbons and MTBE. In the CRTC method, the weight
percent of each of the components was measured, whereas in the EMBSI method, the relative

amount of each of the re presentative components was measured on area -percent basis.
EMBSI developed their method for API to shar e with CRTC and the la boratories performing
toxicological studies on 2 11(b) testing program test substances. The objective of the method was to

monitor and docum ent the chemical and com positional stability of G/ MTBE from manufacture
through transporta tion, storage, and animal ex posure using a standard method that all users could
perform.

The test material was stable an d stored under ambient conditions in an outside solvent shed
except when in use in the inhala tion laboratory. The test substa nce was handled as a flammable
liquid. Detailed information on chemical handli ng is provided in the MSDS attached to the
protocol (Appendix III).

Animals and Husbandry



### The test animals were Caesarean-ori ginated Virus Antibody Free (VAF) Crl:CD-1 ®
(ICR) BR outbred albino mice s upplied by Charles River Laborato ries, Inc., Raleigh, NC. The
use of live animals was requested by the Spons or and required by U.S. EPA OPPTS Testing
Guidelines (U.S. EPA, 1998). Alternative test systems are not available for the assessment of
chemical effects on prenatal mammalian develo pment. The Charles River CD-1® mouse has
been the mouse strain of choice on developmenta l toxicology contracts at RTI since 1976. Large
historical databases for reproduc tive performance and prevalence of spontaneous malformations

in control mice are available from studies c onducted at RTI (currently based on 288 control
litters and over 3600 fetuses from 16 studies).
One hundred seventy (170) nulliparous fema le mice were ordered for this study and
arrived at HLS on December 23, 2004. One hundr ed (100) male mice, 9-11 weeks old upon
arrival at HLS (on August 31, 2004), of the same strain and from the same supplier, were
received for the previous range -finding study, and the remaining 99 males were used as a male
breeding colony for this study. The exact number of females ordered was received, so there wereRTI-909; Project 0209189.000 Final Report



### 7 no replacements available. However, there were no animals with clinical signs, injury, and/or
reduced feed consumption during quarantine. Th e 99 males were used to generate timed-mated
animals for this definitive developmental toxic ity study, which required the mating of 170 female
mice (1:1, with the subsequent addition of naïve females to males who inseminated their original
females) to generate 130 plug-positive females. Females were 7-9 weeks old at arrival and 9-11
weeks of age and ~20-35 g in weight on gd 0. One hundred seventy (170) females were required
to generate 130 plug-positive females in 11 consecu tive days (the protocol indicated that we
expected 130 plug-positive females in 4-5 da ys, but it took longer; Amendment 2); 130 plugpositive females (23/group for 4 groups and 38/group for the fifth group) were required to supply
the optimal number (based on EPA’s guidanc e; e.g., OPPTS 870.3600; U.S. EPA, 1996; for
inhalation developmental toxicity studies) of pregnant animals a nd litters to assess any maternal
and/or embryo/fetal toxicity to the test substance and to confir m and extend the fetal findings
from the previous EMBSI study (2009b).
During an approximately 14-day quarantine/acc limation period at the HLS testing facility,
animals were checked for viability twice daily. Prior to study assignment, all animals were

examined to ascertain suitability for study. Th e HLS veterinarian formally released these
animals for use by signature and date. Males and females were individually housed in stainless
steel suspended cages with wire mesh floors and fronts, except for the mating period when
1 male and 1 female were housed together. During cohabitation, male and female mice were
housed in polycarbonate “shoebox” cages with st ainless steel lids and Alpha-Dri® bedding
(Shepherd Specialty Papers, Watertown, TN). Each cage was fitted to secure a glass feeder jar

with a stainless steel lid. Clean feed jars and fresh feed were pr ovided at least weekly. After the
gd 14 exposure (for Groups 1-4) or on the af ternoon of gd 14 (Group 5), a stainless steel,
perforated insert was placed on the wire-mesh floor of the stainless steel suspended cage of each
female and 1 Nestlet® (Ancare, Bellmore, NY) added to each cage until scheduled sacrifice on
gd 17. Females not undergoing dail y exposures after gd 10 (Group 5) were removed from their
home cage and placed in another suspended cage wit hout feed to match as closely as possible the
conditions of Group 1-4 females for the 6-hour expos ure period. They were then returned to
their home cage at the same time as the exposed females for feed measurement overnight. Feed
(PMI 5002 Certified M eal) was available ad libitum , except during the daily 6-hour inhalation
periods. Analytical certifications of batches of feed provided by the manufacturer are maintainedRTI-909; Project 0209189.000 Final Report



### 8 on file at the HLS testing facility, and there were no known contaminants found in the feed.
Facility water (supplied by Elizabethtown Wa ter Company, Westfiel d, NJ) was available ad

libitum via the automatic watering system or wate r bottles (during mati ng), except during the
daily 6-hour inhalation periods. Water anal yses were conducted by Elizabethtown Water
Company to assure that water met standards specified under the EPA Federal Safe Drinking
Water Act Regulations (40 CFR Part 141). Water analyses provided by the supplier are
maintained on file at the HLS testing facility. There were no known contaminants that interfered
with the objectives of this study. At all times , animals were housed, handled, and used according
to the National Research Council Guide (NRC, 1996).
A 12-hour light/dark cycle was provided via au tomatic timer. Temperature and relative
humidity were monitored in accordance with Testi ng Facility SOPs to ensure that the desired
range of 18 to 26°C for temperature and 30 to 70 % relative humidity was maintained to the
maximum extent possible (NRC, 1996).
Each animal was assigned a temporary iden tification number (designated on each cage)
upon receipt. During the second week of the quarantine/acclimation period, the 170 females

received were tail tatt ooed with consecutive numbers 1 through 170. The 99 remaining males
had been tail tattooed during the range-finding study with c onsecutive numbers 1 through 100
(except for No. 87). After se lection for use on the study, mati ng, indication of copulation, and
assignment to 1 of the five groups, each female was ear tagged with a number assigned by the
HLS testing facility. This number, plus the study number, comprised the unique animal number
for each animal. Each cage was provided with a cage card that was color coded for exposure

level identification and contained the study and animal numbers.
It was anticipated that the concentrations employed would not re sult in irritation or
corrosion to the respiratory tract of the test an imals (based on previous studies with the test
material; e.g., Conaway et al ., 1985; Bevan et al., 1997a; EMBS I, 2008, 2009a,b). Animals were
not subjected to undue pain or distress. All proce dures used in this study were designed to avoid
discomfort, distress, and pain to the animals. The HLS IACUC (Institutional Animal Care and
Use Committee) Protocol Review Subcommittee and the RTI IACU C reviewed the protocol and
found it to be in compliance with appr opriate animal welfare regulations.
Immediately prior to pairing, each female was weighed and subjected to a clinical
examination. For breeding, 1 male with 1 fema le pairing was employed since other pairingRTI-909; Project 0209189.000 Final Report



### 9 patterns (e.g., 1 male with 2 females) may ha ve resulted in an unacceptable number of plugpositive, nonpregnant females and/or sire eff ects. Individual females were placed in
polycarbonate “shoebox” cages with stainle ss steel lids with singly-housed males. On the
following morning and each morning thereafter, the females were examined for the presence of a
vaginal copulation plug (Hafez, 1970). The da y on which copulation plugs were found was
designated as gd 0. Plug-positive females (dams) were individually housed until scheduled
sacrifice on gd 17. Plug-negative females were reta ined in the same male's cage and checked for
plugs on successive mornings until insemination occurred or the treatment groups were filled,

whichever came first. HLS staff evaluated fema les for vaginal copulation plugs until all groups
were filled and then completed the exposure sche dule. When all treatment groups were filled,
the remaining presumed plug- negative females were sacrif iced by asphyxiation with CO 2 and
examined for pregnancy status; many of the fe males were in fact pregnant (see Protocol
Deviation No. 7). The males were also euthan ized by HLS staff after the breeding period was
completed. The fate of all animals is fully documented in the study records.

Study Schedule and Design 

### The actual dates of all major phases of the study are presented in Table A.

Table A. Study Schedule

Event Dates

Females arrived at HLS: December 23, 2004 Quarantine (14 days): December 23, 2004 – January 5, 2005
Animals paired: January 6-11, 2005 Dates of gd 0: January 7-17, 2005 TSCA experimental start date: January 12, 2005
Exposure dates: gd 5 through 10 January 12 – January 22, 2005 gd 5 through 16 January 12 – February 2, 2005 Terminal necropsy (gd 17) January 24 – February 3, 2005 TSCA experimental termination date: February 3, 2005

Submission of draft data on test atmospheres to Sponsor: February 9, 2005 (within 1 week after the last exposure date, February 2, 2005)RTI-909; Project 0209189.000 Final Report



### 10 This study was conducted with 4 treatment groups and 1 vehicle control group. Groups
1-4 were each comprised of 23 plug-positive female mice, and Group 5 was comprised of 38
plug-positive female mice (Table B).

Table B. Number of Animals Assigned to Study Groups

Group No. No. Animals Exposed No. Days Exposed Exposure Period (gd) Target Exposure Concentration (mg/m 3 )

1 23 12 5 through 16 0

2 23 12 5 through 16 2000
3 23 12 5 through 16 10,000

4 23 12 5 through 16 20,000

5 38 6 5 through 10 30,000



### The exposure period for Group 5, at 30,000 mg/m 3 

### G/MTBE on gd 5 through 10, was
selected to reduce the number of days of genera tion of test atmosphere at a concentration that
was 75% of the lower explosive limit. In addition, the fetal malformations of interest are formed
early in the embryonic period of gestation; gd 7-9 in the mouse (e.g., Rugh, 1968), so extending
the exposure period to gd 16 was considered unnecessary.
The test substance was administered as a vapor in the breathing air of the animals. The test
atmosphere was generated by an appropriate proc edure determined during prestudy trials. The
prestudy trials were performed (at least two 6-hour periods) to evaluate the optimal set of
conditions and equipment to genera te a stable atmosphere at th e target exposure levels and
maintain uniform conditions throughout the exposure chambers. Th e whole-body exposure

chambers each had a vo lume of approximately 1000 liters. The chambers were operated at a

minimum flow rate of 200 liters per minute. The final airflow was set to provide at least 1 air
change in 5 minutes (12 air changes/hour) and a T 99 

### equilibrium time of at most 23 minutes. This

chamber size and airflow rate wa s considered adequate to mainta in the oxygen level at least 19%
and the animal loading factor be low 5%. At the end of each daily 6-hour exposure, all animals

remained in the chamber for a minimum of the T 99 

### equilibrium time. During this time, the chamber
was operated at approximately the same flow rate usin g clean air only.
A nominal exposure concentra tion of G/MTBE was calculated. The flow of air through the

chamber was monitored us ing appropriate calibrated equipment. The test substance consumedRTI-909; Project 0209189.000 Final Report



### 11 during the exposure was divided by the total volume of air passing through the chamber (volumetric

flow rate times total ex posure time) to give the nominal concentration.
During each 6-hour exposure, measurements of airborne concentrations were performed in
the animals’ breathing zone at least 4 times using an appropriate sampling procedure and IR
analytical procedure. Specified airborne test ma terial concentrations were within +/- 10% of the
target concentrations. One sample per chamber du ring the trials period and the treatment period
was analyzed by gas chromatography to characteri ze at least 10 major com ponents (comprising at
least 80% by weight of the test substance) to show test substance stab ility and comparison between
the neat liquid test substance and the vaporized te st atmospheres. During the treatment period,
particle size determinations were performed on ce per chamber using a TSI Aerodynamic Particle
Sizer to confirm the absence of pa rticulate test substance condensate in the exposure atmosphere.
Chamber temperature, humidity, airflow ra te, and static pressu re were monitored
continuously and recorded every 30 minutes duri ng exposure. Chamber temperature and relative
humidity were maintained, to the maximum exte nt possible, between 20 to 24°C and 40 to 60%,
respectively. Chamber oxygen levels (maintained at least 19%) were measured pretest and at the
beginning, middle, and end of the exposure period for the study. Air samples were taken in the
vapor generation area pretest and at the beginning, middle, and end of the exposure period for the
study. Light (maintained approximately 30 foot-candles at 1.0 meter above the floor) and noise
levels (maintained below 85 decibels) in the expo sure room were measured pretest and at the
beginning, middle, and end of the exposure peri od for the study. The minimum frequency of
chamber activity during the treatment period is summarized below:

Activity Frequency/Chamber

Measured test substance concentration 4X/day
Measured test substance characterization 1X
Particle size 1X Temperature 13X/day Relative humidity 13X/day
Airflow rate 13X/day Static pressure 13X/day Nominal test substance concentration (exc luding the air control chamber) 1X/day Rotation pattern of exposure cages 1X/day Loading/unloading verification 1X/dayRTI-909; Project 0209189.000 Final Report



### 12 Plug-positive female mice (dams) were as signed to treatment groups by a stratified
randomization method designed to provide uniform mean body wei ghts and equal distribution of
females mated to the same male among dose groups using data from gd 0. A total of 23 plugpositive dams were assigned on gd 0 by stratified randomization ( by body weight) to each of 4
groups (0 [Group 1], 2000 [Group 2], 10,000 [Group 3], and 20,000 [Group 4] mg/m 3 

### ), and 38
plug-positive dams were simila rly assigned to Group 5 (30,000 mg/m 3 

### ). Because of the
unexpectedly prolonged mating period, the decision was made at HLS (see Protocol Deviation
No. 2) to use 130 plug-positive females rather than wait for 140 plug-positive females, and use
23/group (versus 25/group) for Group 4 and 38/group (versus 40/group) for Group 5. Plugpositive females were exposed to G/MTBE or air 6 hours per day from gd 5 through 16 for
Groups 1-4 and for gd 5 through 10 for Group 5, 30,000 mg/m 3 

### , since the malformations of
interest (gastroschesis and ectopia cordis) result fr om ventral midline closure defects early in the
embryologic period, and the shorter exposure period could possibly reduce the risk of prenatal
deaths, which could limit the ability of the study to detect these early malformations. For each
daily exposure, females were tran sferred to inhalation cages, and the cages were moved into the
appropriate chambers for exposur e. Following each daily exposur e, females were transferred
back to home caging for feed consumption measurements overnight.

Clinical observations of all animals were ma de once daily on gd 0 through 4 (prior to the
start of the exposure period), twi ce daily (prior to and immediat ely after each daily exposure)
throughout the exposure period (gd 5 through 10 or gd 5 thro ugh 16), and once daily on gd 11
through 17 or on gd 17 (after exposure period ended). In addition, during each daily exposure
period, animals were observed at least once during each exposure. This was routinely performed
near the middle of each exposure.
Dams were weighed in the mornings (prior to exposures for those days that exposures
occurred) on gd 0 and 5-17. Maternal weight ga ins were calculated fo r gd 0-5 (pre-exposure
period), gd 5-6, 6-7, 7-8, 8- 9, 9-10, 10-11, 11-12, 12-13, 13-14, 14-15,15-16, gd 5-10 or 5-16
(exposure period), gd 10-17 or 16-17 (postexpos ure period), and gd 0-17 (gestational period).
Maternal feed consumption was evaluated in the mornings from gd 0-5 (pre-exposure
period), gd 5-6, 6-7, 7-8, 8-9, 9-10, 10-11, 11-12, 12-13, 13- 14, 14-15, 15-16, gd 5-10 or 5-16
(exposure period), 10-17 or 16-17 (postexposur e period), and gd 0- 17 (gestation period).RTI-909; Project 0209189.000 Final Report



### 13 On gd 17, approximately 1 to 11⁄2 days before expected parturition, al l surviving maternal
animals from all groups were killed by CO 2 asphyxiation at HLS by RTI staff. The thoracic and

abdominal cavities and organs were examined, a nd pregnancy status was confirmed by uterine
examination. Uteri that presented no visible implantation sites were stained with ammonium
sulfide (10%) in order to visu alize any implantation sites that may have undergone very early
resorption (Salewski, 1964). At sacrifice, the body, liver, uterus, paired adrenal glands, and
paired kidneys of each plug-positive female were weighed. Ovarian corpora lutea were counted
and uterine contents (i.e., number of implantation sites, early and late resorptions, dead fetuses,
live fetuses) recorded.

Live and dead fetuses were removed from the uterus, counted, weighed, sexed externally,
and examined externally for gross malformations (including cleft palate) and variations by RTI
staff. Each fetus was killed by intraperitoneal in jection of sodium pentobarbital. Live and dead
fetuses were dissected longitudinally, and the th oracic and abdominal vis cera removed intact and
retained individually in labeled scintillation vial s in buffered neutral 10% formalin for possible
subsequent visceral examination. The fetal carc asses were blanched, skinned, and retained in
individually labeled scintilla tion vials in 70% ethanol for po ssible subsequent double staining
(alizarin Red S and alcian blue) and skeletal evaluation. All mate rnal organs and carcasses were
destroyed by incineration.

Statistics 

### The unit of comparison was the pregnant female or litter. Quantitative continuous data
(e.g., maternal body weights, feed consumption, fetal body weights, etc.) were compared among
the 4 treatment groups and 1 vehicle control group using either parametric ANOVA under the
standard assumptions or robus t regression methods (Zeger an d Liang, 1986; Royall, 1986;
Huber, 1967), which do not assume homogeneity of variance or normality. If the ANOVA test
was statistically significant (i.e., if one or more of the pairwise comparisons, not necessarily to
the vehicle control group, were st atistically significant), then st atistical pairwise comparisons
were made (see below). The homogeneity of variance assumption was examined via Levene’s
Test (Levene, 1960), which is more robust to th e underlying distribution of the data than the

traditional Bartlett’s Test. If Levene’s Test in dicated lack of homogene ity of variance (p<0.05),
robust regression methods were used to test all treatment effects. Th e heterogeneous varianceRTI-909; Project 0209189.000 Final Report



### 14 models (also known as robust regression met hods) use variance estimators that make no
assumptions regarding homogeneity of variance or nor mality of the data. They were used to test
for overall treatment group differences (via Wa ld Chi-Square Tests), followed by individual t

tests for exposed vs. control group comparisons when the overall treatment effect was
significant. At the time these methods were impl emented in this study, the software did not have
p-value adjustments for multiple pairwise compar isons. This problem was recognized, and to
protect against spurious findings, the significance of individual pair wise comparisons to controls
(repeated t-tests) were not reported for a given parameter unless the overall treatment effect was
statistically significant at p<0.05 (Wald chi-square Test). The presence of linear trends was
analyzed by robust regression methods for nonhom ogenous data. Robust regression methods are

available in the REGRE SS procedure of SUDAAN ® Release 8. (RTI, 2001).

If Levene’s Test did not re ject the hypothesis of hom ogeneous variances, standard
ANOVA techniques were applied for comparing th e treatment groups. The GLM procedure in

SAS ® Release 8 was used to evaluate the overall effect of treatment and, when a significant
treatment effect was present, to compare each exposed group to cont rol via Dunnett’s Test
(Dunnett, 1955, 1964). Prior to GLM analysis, an arcsine-square root transformation was
performed on all litter-derived percentage data (Snedecor and Cochran, 1967) to allow use of
parametric methods. For the litter-derived pe rcentage data, the ANOVA was weighted according
to litter size. The pr esence of linear trends was analyzed by GLM procedures for homogenous
data (SAS Institute Inc., 1999a, b, c, d, e; 2000; 2001). A one-tailed test (i.e., Dunnett’s Test)
was used for all pairwise comparisons to the ve hicle control group, except that a two-tailed test
was used for maternal body and organ weight pa rameters, maternal feed consumption, fetal body
weight, and percent males per litt er. Standard ANOVA methods, as well as Levene’s Test, are

available in the GLM procedure of SAS ® Release 8 (SAS Institute Inc., 1999a, b, c, d, e; 2000;
2001).
Nominal scale measures were analyzed by Chi-Square Test for Independence for
differences among treatment groups (Snedeco r and Cochran, 1967) and by the CochranArmitage Test for Linear Trend on Proportions (Cochran, 1954; Armitage, 1955; Agresti, 1990).
When Chi-Square revealed significant (p<0.05) di fferences among groups, then a Fisher’s Exact
Probability Test, with appropriate adjustments for multiple comparisons, was used for pairwise
comparisons between each treatment group and the control group.RTI-909; Project 0209189.000 Final Report



### 15 A test for statistical outliers (SAS Institu te, Inc., 1999b) was performed on female body
weights, feed consumption (in g/day), and se lected organ weights. Per RTI’s SOPs, if
examination of pertinent study data did not provide a plausible, biologically sound reason for
inclusion of the data flagged as “outlier,” then the data were excluded from su mmarization and
analysis and designated as outliers. If there was a plausible, bi ologically sound reason to retain
the flagged data, the data were included in the summarization and analysis. Unless otherwise
specified, the level of significance us ed for the various tests was p<0.05.

Storage of Records



### All data documenting experimental detail s and study procedures and observations were

recorded and maintained as raw data. At th e completion of the study, all reports, raw data,
preserved specimens, and retained samples will be maintained in RTI’s secure archives for a
period of 1 year after submission of the signed final report. The Sponsor will be contacted in
order to determine the final disposition of these materials.

Personnel 

### This study was conducted by RTI under contract to the API (Mr. T.M. Gray, Sponsor’s
Representative) at HLS (Mr. G.M. Hoffman, Prin cipal Investigator; Animal Research Facility
Veterinarian, Dr. Teresa S. Kusznir; Animal Research Facility Dir ector, Mr. I. Vanterpool;
Necropsy Laboratory Supervisor, Ms. G.E. Baxt er; Inhalation Laboratory Supervisor, Mr. S.
Cracknell; Formulation Chemistry Services, Ms. K. Saladdin; Reproductive Consultant, Mr. K.P.
Hazelden; and Quality Assurance, Ms. N.S. Iacono). Dr. R.W. Tyl of RTI served as Study
Director. RTI Reproductive and Developmenta l Toxicology personnel included Ms. M.C. Marr
(Laboratory Supervisor), Ms. C.B. Myers (Rep roductive Toxicity Study Supervisor and Data
Analyst), Mr. W.P. Ross, Mr. C.G. Leach, Ms. L.L. Macdonald, Ms. N.M. Kuney, and Ms. A.J.
Parham. RTI Quality Assurance personnel were Ms. D.A. Drissel (Manager), Ms. C.A. Ingalls,
Ms. M.M. Oh, and Ms. S.C. Wade. Mr. W.P. Ro ss, Ms. N.M. Kuney, Mr. C.G. Leach, Ms. L.L.
Macdonald, and Ms. M.C. Marr were present at HLS to perform the necropsy and external fetal
evaluations. Ms. S.C. Wade was also presen t to observe the procedures and evaluations.
The final report was prepared by Dr. R.W. Tyl and Ms. M.C. Marr, with assistance from
Ms. C.B. Myers for statistical analyses and gene ration of tables, and by Mr. T.W. Wiley for dataRTI-909; Project 0209189.000 Final Report



### 16 entry. Ms. M.C. Marr was responsible for all tr ansfer of custody proced ures for transfer of

records and tissues from HLS to RTI, and for archiving th e study records at RTI.

RESULTS
Test Chamber Analyses
Table 1, Appendix I 

### Prestudy chamber distribution analyses show ed that the test substance was evenly
distributed within the chamber. Prestudy a nd in-study chamber monitoring showed that the
chamber oxygen levels were 20%.
The analytically measured exposure levels of the airborne test substance were reasonably
close to the targeted exposure levels. The measured and nominal concentrations varied
somewhat (less than 10%), but r easonably, from the expected 1:1 ratio for this type of vapor
exposure. Chamber environmental conditions averaged 20.8oC temperature and 29.6% (based on
RTI calculations; 29.2% from HLS calculations ) relative humidity. Mean particle size

distribution measurements for th e exposures indicated that the atmospheres were essentially
vapor only, as expected, since th ere was no substantial differen ce between the test substance

chamber and the air control chambe r for particle size distribution.
Analysis of the major components in the neat test substance and the test atmospheres
showed a reasonably close concordance between th e neat test substance and the vaporized test

substance. These data demonstrated that the test animals were exposed, as expected, to all of the
major components of the test subs tance in their proper proportions . The data were consistent
between the prestudy and in-study analyses, indicating stability of the test substance and the
atmosphere generation techniques.
The test atmospheres were generated to w ithin 97.5 to 103.7% of the target (grand mean
of daily means/chamber). There was no test materi al detected in the control chamber, with an
estimated limit of quantif ication (LOQ) of 433 mg/m 3 

### (see HLS Study File Note in Appendix I;
last page). The relative content of MTBE was 21.3%, as provided by the supplier (see Appendix
III, attachments to protocol). The analytical profile of G/MTBE at HLS indicated 26-27%
MTBE, confounded by coelution with 2,3-dimethylbut ane, which could not be separated using
gas chromatography with a flame ionization det ector and a previously used Supelco Petrocol TMRTI-909; Project 0209189.000 Final Report



### 17 column for the range-finding study (range-finding final study report, Appendix I, HLS report),
and ~23-25% MTBE (confounded by coelution with 3-methylpentane) using a new column for
this endpoint-specific developmental toxicity study (see Appendix I, HLS report). Net MTBE

concentrations were 21.66- 22.72% (Appendix I, Table I).

Maternal Findings
Tables 2-5, Appendix II 

### One female (No. 3814) at 10,000 mg/m 3 

### was removed from study due to a pre-existing
condition (right side undescended testis, seminal vesicle and pros tate, left side ovary, oviduct,
uterus, cervix and vagina). No females died or were euthanized moribund. The numbers of
confirmed nonpregnant females (at scheduled sacr ifice) were 0, 1, 3, 1, and 2 and fully resorbed
litters were 0, 1, 0, 2, and 3 at 0, 2000, 10,000, 20,000, and 30,000 mg/m 3 

### , respectively. The
number (and %) pregnant were 23 (100.0), 22 (9 5.7), 19 (86.4), 22 (95.7), and 36 (94.7) at 0,
2000, 10,000, 20,000, and 30,000 mg/m 3 

### , respectively (Table 2). There were no statistically or
biologically significant differences between groups for % pregnant.
There were no effects of exposure across al l groups on maternal body weights for gd 0, 5,
6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 (in-life), and gd 17 (at sacrifice). Ma ternal body weight
changes were also unaffected across all groups fo r the following intervals: gd 0-5 (pre-exposure
period for all groups), gd 5-10 (exposure peri od for Group 5), gd 5-16 (exposure period for
Groups 1-4), gd 10-17 (postexposure period fo r Group 5), gd 16-17 (postexposure period for
Groups 1-4), and gd 0-17 (gestati on period), except for decreased maternal body weight changes
for gd 12-13 at 2000 (p<0.01) and 20,000 (p<0.05) mg/m 3 

### (with exposures on gd 5-16). These
findings were only for 1 day at the lowest and next to the highest exposure concentrations
(during the exposure period for Groups 1-4) and are considered incidental since they only
occurred once and did not display a dose-response pattern. Matern al gestational weight change
(gestational body weight gain minus gravid ut erine weight) was unaff ected across all groups
(Table 2).

Maternal clinical observati ons for gd 0-4 (pre-exposure period), prior to and after each
daily exposure period (gd 5-16 for Groups 1-4 or gd 5-10 for Group 5) and postexposure (gd 17
for Groups 1-4 or gd 10-17 for Group 5), are pres ented in Table 3. There were no clinical
observations of interest recorded for any dam in any group during the daily exposures. ModerateRTI-909; Project 0209189.000 Final Report



### 18 alopecia on extremities/snout was observed starting on gd 5 in 1 female (No. 4807) at 20,000
mg/m 3 

### . Enophthalmos (eyeball sunk into orbital cavity), left, was observed in 1 female (No.
5829) at 30,000 mg/m 3 

### . Labored breathing was observed on gd 9 postexposure for 1 female (No.
4823) at 20,000 mg/m 3 

### and on gd 10 postexposure for 1 female (No. 5838) at 30,000 mg/m 3



### . Lacrimation, either unilateral or bilateral, was observed for a total of 1 female (No. 4803) at
20,000 mg/m 3 

### and 3 females (Nos. 5802, 5805, and 5824) at 30,000 mg/m 3 

### . Unilateral moderate
lacrimation was observed in 1 female (No. 5824) at 30,000 mg/m 3 

### postexposure on gd 5, and
2 females (1 each at 20,000 [No. 4803] and at 30,000 [No. 5802] mg/m 3 

### ) postexposure on gd 6,
possibly treatment and dose related. Bilateral m oderate lacrimation was observed in 1 female
(No. 5805) at 30,000 mg/m 3 

### postexposure on gd 6. Lacrimation and labored breathing, observed
in more than one female at 20,000 and 30,000 mg/m 3 

### , appeared to likely be treatment related.
Also, red exudates were observed from the anogenital area of 2 females with resorptions: 1
female each at 10,000 mg/m 3 

### (No. 3808 with 2 mid resorptions) on gd 12 pre-exposure and at
20,000 mg/m 3 

### (No. 4814 with 1 mid resorption) on gd 11 postexposure (Table 3).
Maternal feed consumption (in g/day) was significantly reduced at 20,000 mg/m 3 

### for
gd 0-5 (pre-exposure period), signi ficantly increased at 10,000 mg/m 3 

### for gd 5-6, significantly
increased at 2000 and 10,000 mg/m 3 

### for gd 6-7, significantly reduced at 20,000 mg/m 3 

### for gd
7-8, and significantly reduced at 20,000 and 30,000 mg/m 3 

### for gd 8-9. Feed consumption (in
g/day) was also significantly reduced at 30,000 mg/m 3 

### for gd 10-11, significantly increased at
10,000 mg/m 3 

### for gd 12-13, and significan tly increased at 30,000 mg/m 3 

### for gd 13-14. Feed
consumption in g/day was equivalent acros s all groups for gd 9-10, 11-12, 14-15, 15-16, 16-17
(postexposure period, Groups 1-4), gd 5-10 (e xposure period only for Group 5, 30,000 mg/m 3 

### ),
gd 5-16 (exposure period for Groups 1-4), gd 10-17 (postexposure period for Group 5), and gd
0-17 (gestational period) (Table 4). None of these feed cons umption changes are considered
adverse or treatment related; all are likely incidental.
Maternal feed consumption (in g/kg body weight/day) was significantly reduced at
20,000 mg/m 3 

### for gd 0-5 (pre-exposure period), significantly increased at 10,000 mg/m 3 

### for
gd 5-6, significantly increased at 2000 and 10,000 mg/m 3 

### for gd 6-7, significantly reduced at
20,000 mg/m 3 

### for gd 7-8, 8-9, 9-10, and 11-12, significantly reduced at 30,000 mg/m 3 

### for gd 8-9,
10-11, and 5-10 (exposure period for Group 5), and significantly increased at 30,000 mg/m 3 

### for
gd 13-14. There were no differences across gro ups for feed consumption (in g/kg/day) for gdRTI-909; Project 0209189.000 Final Report



### 19 12-13, 14-15, 15-16, 16-17 (postexposure period for Groups 1-4), gd 5-16 (exposure period,
Groups 1-4), gd 10-17 (postexposure period for Gr oup 5), and gd 0-17 (gestation period) (Table
4). Changes in relative feed consumption at 20,000 and 30,000 mg/m 3 

### during the exposure
period were likely due to the processes for inha lation exposure; after th e exposure period, there
were essentially no effects on feed consumption.
At scheduled necropsy on gd 17, maternal abso lute gravid uterine weight, paired adrenal
gland weight, and paired kidney we ight were unaffected across a ll groups. Maternal absolute
liver weight was equivalent across 0, 2000, 20,000, and 30,000 mg/m 3 

### and was significantly
increased (11%; p<0.05) at 10,000 mg/m 3 

### . Maternal paired adrena l gland and paired kidney
weights (relative to terminal body weights) were equivalent across all groups. Relative (to
terminal body weight) maternal liver weight was significantly increased in a concentrationrelated manner at 2000 (6%; p<0.01), 10,000 (11%; p<0.001), and 20,000 (11%; p<0.001)
mg/m 3 

### , likely due to induction of metabolizing enzymes detected in dams exposed through gd 16
(observed in Groups 1-4, with exposures ending on gd 16); relativ e liver weight was unaffected
at 30,000 mg/m 3 

### , with exposures ending on gd 10, most likely due to down regulation of
metabolizing enzymes postexposure (Conney, 1967) (Table 5).

Uterine and Embryofetal Findings
Tables 6-8, Appendix II 

### For all pregnant females, there were no effects across groups for any reproductive
parameter, including no statistically significan t effects on the number of ovarian corpora
lutea/dam, number of uterine impl antation sites/litter, percent pr eimplantation loss/litter, number
(and %) of resorptions/litter, number (and %) of l itters with resorptions, number (and %) of late
fetal deaths/litter, number (and %) of litters with late fetal deaths, number (and %) of nonlive
(late fetal deaths plus resorptions) implants/l itter, number (and %) of litters with nonlive
implants, number (and %) of litters with 100% nonlive implants (f ully resorbed), number (or %)
of adversely affected (nonlive plus malformed) implants/litter, and number (and %) of litters
with adversely affected implants. There were 0, 1, 0, 2, and 3 fully resorbed litters (100%
resorptions) at 0, 2000, 10,000, 20,000, and 30,000 mg/m 3 

### , respectively. These findings did not
differ statistically across groups but might indicate matern al stress at 20,000 and 30,000 mg/m 3



### ,RTI-909; Project 0209189.000 Final Report



### 20 with slightly increased numbers of fully resorbed litters (full litter losses are almost always

ascribed to effects on the ma ternal animal) (Table 6).
For live litters, there were no effects across groups for the number of live fetuses/litter,
percent male fetuses/litter, numbe r of male or female fetuses /litter, or for average fetal body
weight/litter for all fetuses or separately by sex (Table 6).
Summary and statistical analysis of fetal external malformations and variations are
presented in Table 7. Presentation of fetal extern al malformations and variations by defect type
is in Table 8. The number of fetuses (litte rs) examined were 276 (23), 236 (21), 225 (19),
252 (20), and 407 (33) at 0, 2000, 10,000, 20,000, and 30,000 mg/m 3 

### , respectively. There were
no statistically significant differences across groups for any of the parameters evaluated. These
parameters included the number a nd percentage of fetuses with ex ternal malformations per litter
(total and separately by sex), the number and percentage of fetuses (pooled by group) with
external malformations, and of litters with at l east 1 fetus with external malformations. Also,
there were no differences across groups for the same parameters as above for fetal external

variations. There were fetal external malformati ons and variations observed at low incidence in
all 5 groups (Table 7).
The fetal external malformations included encep halocele in 1 fetus in 1 litter (Dam No.
2819, Implant No. 5) at 2000 mg/m 3 

### , cleft palate in 2 fetuses (2 li tters), 1(1), 1(1), 1(1), and 7(4)
at 0, 2000, 10,000, 20,000, and 30,000 mg/m 3 

### respectively, and gastroschi sis in 1 fetus in 1 litter
(Dam No. 5810, Implant No. 7) at 30,000 mg/m 3 

### . This apparent (nonstatistically significant)
increase in the fetal (and litter) in cidence of cleft palate at 30,000 mg/m 3 

### may be indirect
evidence of maternal stress during exposures at this high concentration (see Discussion below).

The fetal external variations included abnor mal rugae in the midlin e of the palate in
1 fetus in 1 litter each at 10,000 (Dam No. 3822, Implant No. 15) and 20,000 mg/m 3 

### (Dam No.
4822, Implant No. 3), not cleft palate, and hematoma s at various locations (face, head, neck, and
shoulder) in 4 fetuses (in 4 litters) at 0 and 2000 mg/m 3 

### , in 1 fetus (in 1 litter) at 10,000 mg/m 3



### , in 2 fetuses (in 2 litters) at 20,000 mg/m 3 

### , and in no fetuses at 30,000 mg/m 3 

### (Table 8). None of

these external variations are considered related to treatment or exposure concentrations.

The historical control data for 288 litter s of CD-1 mice at RT I International for
governmental clients from 1997-2002 are presented in Table 9.RTI-909; Project 0209189.000 Final Report



### 21 DISCUSSION
This study was designed and performed:

1. To confirm or refute the fetal malforma tion finding of ectopia cordis observed in
1 fetus at 2000 mg/m 3 

### and in 2 fetuses (in th e same litter) at 10,000 mg/m 3 

### in the
previous developmental toxicity st udy on this test material in CD-1 ® mice
(EMBSI, 2009b);

2. To confirm or refute the fetal malforma tion finding of gastroschisis observed in 1
fetus at 10,000 mg/m 3 

### (but not at 2000 or 20,000 mg/m 3 

### ) in the previous
developmental toxicity study at EMBSI ( 2009b) on this test material in CD-1®
mice;
3. To extend the test atmospheric c oncentration range from 0, 2000, 10,000, and
20,000 mg/m 3 

### on gd 5 through 16 employed previously (EMBSI, 2009b), to 0,
2000, 10,000, 20,000, and 30,000 mg/m 3 

### (the last concentration at 75% of the
lower explosive limit), with daily e xposures on gd 5 through 16 for the 0-20,000
mg/m 3 

### groups and on gd 5 through 10 for the 30,000 mg/m 3 

### group. This last
gestation interval was in cluded to encompass the time of embryonic ventral wall
closure, the failure of which is likely responsible for both ectopia cordis and
gastroschisis. There were 23 pl ug-positive females/group at 0-20,000 mg/m 3 

### and
38 plug-positive females at 30,000 mg/m 3 

### to improve the possibility of detection

of these rare fetal malformations.

There were no apparent treatment-related effects on maternal body weights or weight
gains and no consistent treatment- or con centration-related effects on maternal feed
consumption. The treatment-related increases in absolute (at 10,000 mg/m 3 

### ) and relative (at
2000, 10,000, and 20,000 mg/m 3 

### ) maternal liver weights are most likely due to the induction of
hepatic metabolizing enzymes, with the concom itant increase in liver weight (Conney, 1967).

This is not considered maternal toxicity, per se, but an adapti ve metabolic response to exposure

to a xenobiotic. The absence of an increased liver weight at 30,000 mg/m 3 

### might reflect the 7-
day interval between the end of treatment and necropsy of animals in this group. It seems likely,
given the increases seen in the other test-substance-treate d groups, that liver weights were
initially increased in all treatment groups.RTI-909; Project 0209189.000 Final Report



### 22 Terminal maternal adrenal gland weights were not cha nged across groups, although the
current thinking is that there ma y be increased maternal producti on of corticosterone (shown to
cause fetal cleft palate; Hemm et al., 1977; Pradat et al., 2003; Se nda et al., 2005; Carmichael et
al., 2007) in response to the stre ss of moving the animals in and out of chambers in all groups
and in the high “dose” group also from the stress of the narcotic/letha rgic effect of MTBE at this
concentration (see Bevan et al., 199 7a,b). It is possible (if not pr obable) that the adrenal glands
of the dams exposed to 30,000 mg/m 3 

### on gd 5-10 were producing incr eased corticosterone (with
temporary glandular enlargement) during the ex posure period, with resolution of increased
hormone output and glandular changes by sche duled termination on gd 17. Interestingly,
lethargy was observed in the females at 30,000 mg/m 3 

### in the range-finding study but was not
documented in this study during th e daily exposures; it is likely the admittedly subjective effect
was present in this study since it was present in the range-finding study at the same exposure

concentration and duration. Clinical observation s of the dams did indicate treatment-related
findings, i.e., labored breath ing only at 20,000 and 30,000 mg/m 3 

### postexposure on gd 9 and 10,
respectively, in 1 female in each group, and lacrimation in 1 female on gd 6 at 20,000 mg/m 3 

### and
in 3 females on gd 5 and 6 at 30,000 mg/m 3 

### , early in the exposure period.

The CD-1 ® (Swiss) mice used in the previous study (EMBSI, 2009b) were from the
Charles River, Portage, MI, facility; the CD-1 ® (Swiss) mice used in the current study were
from the Charles River, Raleigh, NC, facility, be cause RTI International ha s a historical control
database for developmental toxicity studies on this mouse strain from this source, and to
preclude the possibility that the fetal findings from the EMBSI study were due to a different
spontaneous rate of these two fe tal malformations in the Portage colony (due to founder effects,
genetic drift, etc.). Females like the pseudohermaphroditic adult female at 10,000 mg/m 3



### (removed from study) have been observed at very low incidence in other studies with this mouse
strain at RTI International from the Charles River, Raleigh, NC, facility.
1. This study did not confirm the presence of ectopia cordis in any mouse fetus at
any exposure concentration out of a tota l 122 litters and 1396 fetuses. In the
absence of a clear dose-response pattern to this finding in the EMBSI (2009b)
study and the total absence of this findi ng in the present st udy, it is the Study
Director’s opinion that it is appropriate (a nd prudent) to conclude that this fetal
finding is likely not related to maternal exposure to the test material.RTI-909; Project 0209189.000 Final Report



### 23 2. This study did not confirm the presen ce of gastroschisis in fetuses at 2000,
10,000, or 20,000 mg/m 3 

### ; it was also not found at 2000 or 20,000 mg/m 3 

### at
EMBSI (2009b). One fetus (out of 407) did exhibit gastroschisis at 30,000 mg/m 3



### in the present study. This fetus (No. 6 female) was from Female No. 5810; her
litter included 15 implants and 14 live fetuse s. In her litter, fetus No. 5 female
and fetus No. 12 male exhibited cleft pala tes, and fetus No. 6 female had cleft
palate as well as gastroschisi s. In this group at 30,000 mg/m 3 

### , there were 7

fetuses in 4 litters with cl eft palate (greater inciden ce relative to other 4 groups,
but not statistically significantly different), wi th 3 of them in this index litter. The
body weight of the single female fetus w ith gastroschisis and cleft palate was
much lower (0.6057 g) than the body weights of the remaining fetuses in the
litter: females 0.8034-0.9768 g; males 0.8406-0.8893 g (Table A-4 in Appendix
II). Her body weight was also much lower than the mean female fetal body
weight/litter for this group (1.0141±0.0239 [S.E .M.] g); i.e., she was classified as
a “runt,” with a body weight below 3 standard deviations (and 3 standard errors)
from the litter mean by sex in this gr oup (Table 6). Although there is a known
relationship between fetal body weight and cleft palate (i.e., malformed fetuses
tend to be lighter at term than normal fetu ses; Ryan et al., 1991), there are not yet
any data on the biological relationship between reduced fetal body weight and
other malformations (i.e., is it cause and/or effect?). This 30,000 mg/m 3 

### group also contained 3 fully resorbed litter s (out of 36 pregnant). At 20,000 mg/m 3 

### , 2
litters were fully resorbed (out of 22 pr egnant), and 1 fetus in 1 litter exhibited
cleft palate (and no incidence of gastrosc hisis). There were no fully resorbed
litters at 0 or 10,000 mg/m 3 

### and only 1 fully resorbed litter at 2000 mg/m 3 

### (Table
2). Cleft palate was present in 2 fetuses in 2 litters at 0 mg/m 3 

### and in 1 fetus in 1
litter each at 2000, 10,000, and 20,000 mg/m 3 

### (Table 8). The increase in fetal and
litter incidence of cleft palate and the in crease in fully resorbed litters at 30,000
mg/m 3 

### provide circumstantial evidence for ma ternal toxicity (i.e., stress from the
exposure procedures, per se, and from a very high level of G/MTBE, resulting in

additional stress and excessive corticoste roid production known to cause cleft
palate; Hemm et al., 1977; Pr adat et al., 2003; Senda et al., 2005; Carmichael etRTI-909; Project 0209189.000 Final Report



### 24 al., 2007). Maternal toxicity (stress) is also known to cause full litter resorptions;
it is ascribed to the dam, not the conceptuses, and exposure on gd 5-10
corresponded to periods of early implan tation and palatal sensitivities. The
presence of gastroschisis in 1 fetus at 30,000 mg/m 3 

### may also indicate a possible

association with maternal stress and G/MT BE only at this very high atmospheric

concentration.
In the present study, gastroschisis was obs erved in only 1 fetus, only at 30,000 mg/m 3 

### and
only in the presence of profound developmental t oxicity for that fetus (very low body weight and
cleft palate). There was also ge neral evidence of toxic ity in this group (e.g., lethargy observed in
dams in the range-finding study at 30,000 mg/m 3 

### ) and lower individual maternal body weight
gain (1.9 g) for Dam No. 5810 with the fetus with gastroschisis, versus the 30,000 mg/m 3 

### mean (3.0 g) and the vehicle control group mean (3.3 g) for weight gain dur ing gd 5-10, the exposure
period. Historical control data from govern mental studies with the Charles River CD-1 ®
(Swiss) mouse at RTI (Table 9), with 288 litte rs and 3641 fetuses, indicates no fetuses with
gastroschisis or ectopia cordis. There were 18 fetuses in 11 control litters (in 6 studies) with cleft
palate (1 to 4 litters affected/affected study) an d 2 fetuses in 2 control litters (1/l itter) with
exencephaly in 2 studies, 1 litter affected/affected study. The absence of gastroschisis in any of
the 3,641 control CD-1 mouse fetu ses in 288 control litters (Tab le 9) lends support to the
conclusion that gastroschisis in this study may be treatment rela ted, occurring in a compromised
fetus at 30,000 mg/m 3 

### with likely concomitant maternal to xicity (see above a nd conclusion No. 4
below). No other historical control data on ma ternal and fetal findings in the Charles River

CD-1 ® mouse could be found in the open literature.
Neither gastroschisis nor ectopia cordis wa s observed in CD-1® mouse fetuses from
mothers exposed to 0, 1000, 4000, or 8000 pp m MTBE by whole-body inhalation (in the
presence of maternal and embryofetal toxicity at 4000 and 8000 ppm MTBE; Bevan et al.,
1997a), nor was either of these malformations observed in mice exposed to a vapor condensate
of gasoline as part of this te sting program (EMBSI, 2009a). It does not appear that exposure to
either MTBE or gasoline vapor, at atmospheric concentrations < 8000 ppm, causes ectopia cordis
or gastroschisis in mice. Maternal ataxia , hypoactivity, prostrati on, labored breathing, and
lacrimation were observed at 4000 and 8000 ppm MTBE, and the resultant stress was considered
most likely responsible for (or at least exacerbated) the increased incidence of fetal cleft palate atRTI-909; Project 0209189.000 Final Report



### 25 8000 ppm. Reduced fetal body weights and concomita nt reduced fetal skeletal ossification were
also observed at 4000 and 8000 ppm MTBE vapor, w ith 4 fully resorbed litters at 8000 ppm
MTBE (Bevan et al., 1997a). Neither gastroschi sis nor ectopia cordis was observed in CD® rat
offspring in a 2-generation study of inhaled MTBE at 400, 3000, or 8000 ppm (Bevan et al.,
1997b) or in rabbit fetuses from does exposed to 1000, 4000, or 8000 ppm MTBE in a
developmental toxicity study (Bevan et al., 1997 a). Ventral closure de fects were also not
observed in rat reproductive and developmental t oxicity studies conducted with gasoline vapor
condensate or G/MTBE (EMBSI, 2009a,b; Hunti ngdon Life Sciences, Inc., 2009a,b), nor with
light ends of gasoline (Roberts et. al., 2001), nor in the mouse developmental toxicity study
conducted with gasoline vapor condensate (EMBSI, 2009a).

The incidence of fetal cleft palate in the EMBSI study was only 1 fetus (in 1 litter), and
only at 20,000 mg/m 3 

### (that study’s highest concentration). In the present st udy, cleft palate was
observed in all 5 groups, includ ing the air control group (2 fe tuses in 2 litters), at 2000-
20,000 mg/m 3 

### (1 fetus in 1 litter in each group) and at 30,000 mg/m 3 

### (7 fetuses in 4 litters). The

increased incidence of cleft palate observed at 30,000 mg/m 3 

### , although not statistically
significantly different from the control value, was considered to be biolog ically relevant. Cleft
palate in fetal mice is inducible by increased co rticosterone levels in the dam (and presumably
transported to the fetuses; Carmichael et al., 2007; Senda et al., 2005; Pr adat et al., 2003; Hemm
et al., 1997). Maternal increased corticosterone levels may be attributed to increased maternal
stress from inhalation exposures, per se (moving dams into and out of chambers, exposure to
dynamic air flows, no feed or water during exposu re periods, no solid floo ring in exposure cages,
etc.), and from test materials at anesthetic concentrations. In f act, maternal inhalation of MTBE
has been shown to produce cleft palates in fe tuses from CD-1 mouse dams which exhibited
lethargy and apparent unconsciousness (Bevan et al., 1997a). Mate rnal lethargy during
exposures was also observed by HLS staff duri ng the daily exposure periods of G/MTBE at
30,000 mg/m 3 

### in the range-finding study at HLS (it wa s not noted by HLS staff during the daily
exposure periods at any concentration in this defi nitive study). Therefore, the presence of fetal
cleft palate in all groups (inc luding the control group) was not unexpected, and the increased
incidence at 30,000 mg/m 3 

### (from both inhalation procedures, per se, and the anesthetic qualities
of the MTBE in the G/MTBE at this atmospheric concentration) was also anticipated. The
increased cleft palate incidence at 30,000 mg/m 3 

### is interpreted as mostlikely secondary toRTI-909; Project 0209189.000 Final Report



### 26 maternal stress effects. Maternal stress may have also played a role in the single case of
gastroschisis in a vulnerable, compromised fetu s developing in a dam exposed to anesthetic
levels of MTBE. Since the maternal exposures to 30,000 mg/m 3 

### were from gd 5-10, it is highly
likely that any indications of maternal stress or other toxic ity (e.g., increased adrenal weights)
would have resolved during the postexposure period (gd 10-17) and were therefore not present at
scheduled necropsy on gd 17.

CONCLUSIONS
In conclusion, this study has demonstrated the following:
1. No confirmation of fetal ectopia cordis at any test atmospheric concentration
employed;

2. No confirmation of fetal gastroschisis at 0-20,000 mg/m 3 

### ; 3. One fetus (out of 407 fetuses, 0.24%) in one litter (out of 33 litters with live
fetuses, 3.03%) exhibited ga stroschisis at 30,000 mg/m 3 

### ; this female fetus had
very low body weight (designated a “runt”) , also exhibited cl eft palate, and was
clearly compromised. The single inciden ce of gastroschisis and the increased
incidence of cleft palate in this group may both be related to fetal toxicity,
secondary to maternal stress, only at 30,000 mg/m 3



### . 4. Fetal cleft palate was present at a lo w incidence (1-2 fetuses/group) at 0-20,000
mg/m 3 

### , with increased incidences (7 fetuses in 4 litters) at 30,000 mg/m 3 

### , likely
due to greater maternal stress from the an esthetic qualities of the test atmosphere

at this concentration. See above for discussion on maternal stress causing
elevated corticosteroids, which in turn cause offspring cleft palate. Since the
dams at 30,000 mg/m 3 

### were exposed only on gd 5-10, the offspring palates would
have been affected (Hemm et al., 1977; Pradat et al., 2003; Senda et al., 2005;
Carmichael et al., 2007), but any elevated corticosteroid levels (and any effects on
adrenal gland weights) w ould likely have resolved by gd 17 at the time of
necropsy. Relatively minor maternal trea tment-related clinical signs of distress
were observed at 20,000 and 30,000 mg/m 3 

### in this study, with greater maternal
clinical signs observed at 30,000 mg/m 3 

### in the range-finding study. The increased
incidence of fetal cleft palate and the pres ence of gastroschisis in 1 fetus in thisRTI-909; Project 0209189.000 Final Report



### 27 group, in this study, may indicate effects on compromised fetuses in the presence
of maternal stress (toxicity). Th e presence of gastroschisis at 30,000 mg/m 3 

### and

its absence from all available CD-1 historical control databases lends some
support to the possible conclu sion that the presence of 1 fetus with gastroschisis
may have been treatment related, secondary to maternal stress.
5. Therefore, in the Study Director’s opini on, maternal exposure to the test chemical
at 30,000 mg/m 3 

### , an extremely high atmospheric concentration, in the presence of

fetal and demonstrable maternal toxici ty during the embryonic period of ventral
body wall closure, may have resulted in a ve ry low incidence of gastroschisis (1
out of 407 fetuses, 0.24%; 1 out of 33 litters with live fetuses, 3.03%) in
vulnerable mouse fetuses in the highest concentration group. The one affected
fetus also exhibited reduced body weight a nd cleft palate. With lower fetal and
maternal toxicity at 20,000 mg/m 3 

### and below, there was no incidence of
gastroschisis. The absence of any m ouse fetus with gastroschisis in the
performing laboratory’s historical control database lends some support to the
possible conclusion that the single case of gastroschisis at a very high atmosphere
of G/MTBE (30,000 mg/m 3 

### ) in this study may have been treatment related. Since
the one affected female fetus at 30,000 mg/m 3 

### was also very small (a “runt”) and
also exhibited cleft palate, her malformation findings may have been caused by
(or exacerbated by) maternal exposure to G/MTBE. Alternatively, the fetal
effects may have been caused by (or exacerba ted by) maternal toxicity (stress) or
may have been a spontaneous fetal malformation.

Under the conditions of this study, th e NOAELs for maternal and developmental
toxicity, based on the maternal effects observed at 20,000 and 30,000 mg/m 3 

### and the
developmental effects observed at 30,000 mg/m 3 

### , were determined by the Study Director
to be 10,000 mg/m 3 

### for maternal toxicity and 20,000 mg/m 3 

### for developmental toxicity.RTI-909; Project 0209189.000 Final Report



### 28 REFERENCES

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Bevan, C., T.L. Neeper-Bradley, R.W. Tyl, L.C. Fisher, R.D. Panson, J.J. Kneiss, and L.S. Andrews (1997b). Two-generation reproductive toxicity study of methyl tertiary-butyl ether (MTBE) in rats. J. Appl. Toxicol. 17(1), 513-519.

Carmichael, S.L., G.M. Shaw, C. Ma, M.M. Werler, S.A. Rasmussen, and E.J. Lammer (2007). Maternal corticosteroid us e and orofacial clefts. Am. J. Obstet. Gynecol. 197(6), 585, e1-7; discussion 683-4, e1-7.

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Dunnett, C.W. (1964). New tables for multiple comparisons with a control. Biometrics 20 , 482- 491.

ExxonMobil Biomedical Sciences, Inc. (E MBSI) (2009a). Whole-Body Inhalation Developmental Toxicity Study in Mice with Baseline Gasoline Vapor Condensate (MRD-00- 695). Study Director, Mr. G.W. Trimmer, EMBSI. Performed for the American Petroleum Institute, Washington, DC. Final Report, Project No. 169534M, by EMBSI, Annandale, NJ.

ExxonMobil Biomedical Sciences, Inc. (EMBSI) (2009b). Whole-Body Inhalation Developmental Toxicity Study in Mice With Gasoline With MTBE Vapor Condensate (MRD00-713). Study Director, Mr. G.W. Trimmer, EMBSI. Performe d for the American Petroleum Institute, Washington, DC. Final report, Project No. 171334M, by EMBSI, Annandale, NJ.

ExxonMobil Biomedical Sciences, Inc. (EMBSI) (2008). Whole-Body Inhalation Developmental Toxicity Study in Rats with Baseline Gasoline Va por Condensate (MRD-00-RTI-909; Project 0209189.000 Final Report



### 29 695). Study Director, Mr. G.W. Trimmer, EMBSI. Performed for the American Petroleum Institute, Washington, DC. Final Report, Project No. 169534, by EMBSI, Annandale, NJ.

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Huntingdon Life Sciences, Inc. (2009b). Gaso line MTBE Vapor Condensate: A Two-Generation Whole-Body Inhalation Reproductio n Toxicity Study in Rats. Study Director, Mr. G. M. Hoffman, HLS. Performed for the American Petroleum Institute, Washington, DC. Draft Report, Project No. 00-4208, by HLS, East Millstone, NJ.

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Pradat, P., E. Robert-Gnansia, G.L. Di Tanna, A. Rosano, A. Lisi, and P. Mastroiacovo (2003). First trimester exposure to cor ticosteroids and oral clefts. Birth Defects Res. A Clin. Mol. Teratol. 67(12), 968-70.

Roberts, L., R. White, Q. Bui, W. Daughtrey, FR . Koschier, S. Rodney, C. Schreiner, ED. Steup, R. Breglia, R. Rhoden, R. Schroeder, and P. Ne wton (2001). Developmenta l toxicity evaluation of unleaded gasoline vapor in the rat. Reprod. Toxicol. 15(5), 487-494.

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### 30 RTI (Research Triangle Institute) (2001). SUDAAN User's Manual, Release 8.0. Research Triangle Park, NC.

RTI (Research Triangle Inst itute) (2009). Range-Findi ng Tolerance Study for the Developmental Toxicity Evalua tion of Inhaled Gasoline Methyl Tertiary Butyl Ether Vapor Condensate (G/MTBE) in CD-1 ® Mice, RTI ID No. 0209189.000, for the American Petroleum Institute by RTI International ).



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SAS Institute Inc. (1999d). SAS® Procedures Guide , Version 8, Cary, NC: SAS Institute Inc. 1643 pp.

SAS Institute Inc. (1999e). SAS® Companion for the Microsoft Windows Environment , Version 8, Cary, NC: SAS Institute Inc. 562 pp.

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Senda, T., N. Natsume, J. Kuno, T. Toyoda, and K. Shimozato (2005). Rate of occurrence of dexamethasone-induced cleft palate affected by uterine environment in the mouse. Plast. Reconstr. Surg. 115(4), 1208-1210.

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U.S. EPA (1994). U.S. Environmental Protec tion Agency. Good laborat ory practices (GLP) standards for inhalation expos ure health effects testing. Federal Register 59 (122) , 33119-33124, June 27, 1994 (40 CFR, part 79, subpart F, §79.60).RTI-909; Project 0209189.000 Final Report



### 31 U.S. EPA (1996). U.S. Environmental Protec tion Agency; Prevention, Pesticides and Toxic Substances (OPPTS), Health Effects Test Guideline OPPTS 870.3600, Inhalation Developmental Toxicity Study (Public Draft, June 1996).

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### 32 LIST OF PROTOCOL DEVIATIONS

Study: Mi04-HLS2 Master Protocol No.: RTI-909 HLS Study Code: 04-4263

HLS

1. Due to technician error, food left weights were record ed for the Exposure Day 12-13 interval, but food fed weights for the Exposur e Day 13-14 interval were not entered into the computer system for Animal Nos. 1805, 1806, 1807, 1808, 1809, 1810, 1811, 1812, 1814, 1815, 1816, 1818, 1819, 1820, 1823, 2804, 2806, 2808, 2814, 2815, 2817, 2818, 2819, 2821, 2822, and 2823. Since the feeders for thes e animals were returned to the cages after obtaining the food left wei ghts and did not need to be ch anged, a series of edits was performed which provided a food fed weight equivalent to the food left weight.

2. Due to the unexpectedly extended mating peri od, only 130 mice were placed on test rather than 140. This resulted in the following group sizes: Group 1-4 ha d 23 mice and Group 5 had 38 mice.

3. The group means for MIRAN sampling were outside of the stated protocol range of ± 10% on Exposure Days 6, 9, 16 (Group 2), Exposure Days 16 and 19 (Group 3), and Exposure Day 2 and 16 (Group 4).

4. Individual MIRAN samples were outside the protocol specifi ed range of ± 10% for the following Sample Nos.: 2001, 2002, 2020, 2021, 2022, 2032, 2034, 2035, 2036, 2037, 2044, 2052, 2063, 2064, 3061, 3062, 3063, 3064, 3065, 3066, 3077, 3078, 4001, 4004, 4005, 4406, 4016, 4024, 4063, 4064, 4070, 5001, 5005 and 5025. Chamber concentration values were confirmed as needed in accordance with Testing Facility SOP.

5. Due to technician oversight, a Nestlet® was given to Animal No. 5838 during the morning of Exposure Day 16 (removed afte r a total of 37 minutes) and th en again in the afternoon. The protocol specifications were for afternoon only.

6. Due to technician oversight, animals in Gr oups 1-4 chambers and Group 5 chamber were not rotated on Exposure Days 11 and 13, respectively.

7. At RTI’s request, with the Sponsor’s approva l, nonpregnant females were sacrificed and examined macroscopically in or der to determine their actual state of pregnancy, although not required by protocol.

8. Due to a communication error with the Spons or’s Representative, Group 5 dams were removed from their cages with Nestlets® and inserts only from GD 14-16 and placed into cages without Nestlets® and inserts to simula te the Groups 1-4 e xposure regimen. Per intent of Sponsor, the Group 5 dams should ha ve been removed from their cages with Nestlets® and inserts from GD 10-16 and placed into cages without Nestlets® and insertsRTI-909; Project 0209189.000 Final Report



### 33 to simulate the cage environment of the fe males in Groups 1-4 during the remaining exposure on gd 16.

9. Due to the HLS Principal Investigator’s over sight, Protocol Amendment No. 2 was signed by one IACUC member instead of tw o, as designated by the protocol.

In the Study Director’s profe ssional opinion, these protocol devi ations did not affect the study design, performance, or interpretation and are presented for completeness.RTI-909; Project 0209189.000 Final Report



### 34 Table 1. Analysis of Test Atmospheres (page 1 of 1)

Target Concentrations (mg/m 3 )

0 2000 10,000 20,000 30,000
Mean analytical concentration ± SD (% of target) a 0 ± 0 b (NA) 2074 ± 248 (103.7) 9925 ± 688 (99.25) 20,342 ± 1815 (101.7) 29,250 ± 1480 (97.5)

Particle Size Determination: c
MMAD ( μ m) 2.179 5.699 9.319 3.845 1.144

GSD 1.676 2.117 2.071 1.955 2.910 TMC (mg/m 3 ) 2.56 x 10 -2 5.05 x 10 -3 3.41 x 10 -3 1.47 x 10 -3 1.54 x 10 -2

Mean temperature ( ° C ± SD) d 20.3 ± 0.9 20.8 ± 1.2 21.5 ± 0.9 20.7 ±0.9 20.7 ±0.8
Mean relative humidity (% ± SD) d 31.2 ± 4.9 32.0 ± 7.1 28.6 ± 4.2 28.4 ± 4.1 27.6 ± 4.7

a Mean of 4 assays/chamber/day (20 days for Group 1, 18 days for Group 2, 22 days for Group 3,

19 days for Group 4, and 12 days for Group 5) measured by infrared spectroscopy

b Estimated limit of quantification (LOQ) = 433 mg/m 3 (Appendix I)

c Measured 1 time/chamber

d Measured 13 times/chamber/day

SD = Standard deviation
MMAD = mass median aerodynamic diameter GSD = geometric standard deviation TMC = total mass concentration (measure of aerosol concentration)RTI-909; Project 0209189.000 Final Report



### 35 Table 2. Summary and Statistical Analysis of the Ma ternal Body Weights and Body Weight Changes (page 1 of 6)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000
Subjects (No. Dams) No. on Study 23 23 23 23 38 No. Removed 0 0 1 a 0 0
No. Dead or Euthanized 0 0 0 0 0 No. Nonpregnant 0 1 3 1 2 No. (%) Pregnant at Scheduled Sacrifice 23 (100.0) 22 (95.7) 19 (86.4) 22 (95.7) 36 (94.7)

No. (%) with 100% Resorptions 0 (0.0) 1 (4.5) 0 (0.0) 2 (9.1) 3 (8.3)

Maternal Body Weight (gd 0) (g) b
26.7 27.2 27.0 27.3 27.5

+ 0.4 + 0.3 + 0.3 + 0.3 + 0.2 N=23 N=22 N=19 N=22 N=36
Maternal Body Weight (gd 5) (g) b
27.6 28.4 27.8 28.5 28.3

+ 0.4 + 0.4 + 0.4 + 0.3 + 0.3 N=23 N=22 N=19 N=22 N=36
Maternal Body Weight (gd 6) (g) b
28.2 29.1 28.7 29.0 28.8

+ 0.4 + 0.4 + 0.4 + 0.3 + 0.3 N=23 N=22 N=19 N=22 N=36
Maternal Body Weight (gd 7) (g) b
28.7 29.7 29.1 29.6 29.3

+ 0.4 + 0.4 + 0.4 + 0.3 + 0.3 N=23 N=22 N=19 N=22 N=36
Maternal Body Weight (gd 8) (g) b
29.2 30.2 29.7 30.0 29.8

+ 0.4 + 0.4 + 0.4 + 0.3 + 0.3 N=23 N=22 N=19 N=22 N=36RTI-909; Project 0209189.000 Final Report



### 36 Table 2. Summary and Statistical Analysis of the Ma ternal Body Weights and Body Weight Changes (page 2 of 6)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000

Maternal Body Weight (gd 9) (g) b
29.7 30.6 30.3 30.6 30.4

+ 0.4 + 0.5 + 0.5 + 0.4 + 0.4 N=23 N=22 N=19 N=22 N=36
Maternal Body Weight (gd 10) (g) b
30.9 31.8 31.6 31.5 31.3

+ 0.5 + 0.6 + 0.5 + 0.4 + 0.4 N=23 N=22 N=19 N=22 N=36
Maternal Body Weight (gd 11) (g) b
32.8 33.9 33.6 33.2 33.0

+ 0.5 + 0.6 + 0.6 + 0.5 + 0.5 N=23 N=22 N=19 N=22 N=36
Maternal Body Weight (gd 12) (g) b
34.9 35.8 35.5 35.1 35.3

+ 0.5 + 0.8 + 0.5 + 0.6 + 0.5 N=23 N=22 N=19 N=22 N=36
Maternal Body Weight (gd 13) (g) b
36.9 37.2 37.3 36.7 37.4

+ 0.5 + 0.9 + 0.6 + 0.7 + 0.6 N=23 N=22 N=19 N=22 N=36
Maternal Body Weight (gd 14) (g) b
39.2 39.2 39.4 38.7 39.7

+ 0.6 + 1.0 + 0.6 + 0.9 + 0.7 N=23 N=22 N=19 N=22 N=36RTI-909; Project 0209189.000 Final Report



### 37 Table 2. Summary and Statistical Analysis of the Ma ternal Body Weights and Body Weight Changes (page 3 of 6)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000

Maternal Body Weight (gd 15) (g) b
42.0 42.0 42.2 41.2 42.4

+ 0.6 + 1.1 + 0.7 + 1.0 + 0.9 N=23 N=22 N=19 N=22 N=36
Maternal Body Weight (gd 16) (g) b
45.2 45.1 45.3 44.0 45.2

+ 0.7 + 1.4 + 0.7 + 1.2 + 1.1 N=23 N=22 N=19 N=22 N=36
Maternal Body Weight (gd 17) (g) b
48.4 48.0 48.3 46.7 48.3

+ 0.7 + 1.5 + 0.8 + 1.4 + 1.2 N=23 N=22 N=19 N=22 N=36 Maternal Body Weight (gd 17 at sacrifice) (g) b
47.14 46.71 47.52 45.91 47.44

+ 0.73 + 1.50 + 0.79 + 1.43 + 1.18 N=23 N=22 N=19 N=22 N=36

Maternal Body Weight Change (gd 0 to 5) (g) b
0.8 1.2 0.8 1.2 0.8

+ 0.2 + 0.3 + 0.3 + 0.2 + 0.2 N=23 N=22 N=19 N=22 N=36 Maternal Body Weight Change (gd 5 to 6) (g) b
0.6 0.7 0.9 0.5 0.5

+ 0.1 + 0.1 + 0.2 + 0.1 + 0.1 N=23 N=22 N=19 N=22 N=36RTI-909; Project 0209189.000 Final Report



### 38 Table 2. Summary and Statistical Analysis of the Ma ternal Body Weights and Body Weight Changes (page 4 of 6)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000
Maternal Body Weight Change (gd 6 to 7) (g) b
# 0.6 0.6 0.5 0.6 0.5

+ 0.1 + 0.1 + 0.1 + 0.1 + 0.1 N=23 N=22 N=19 N=22 N=36 Maternal Body Weight Change (gd 7 to 8) (g) b
0.5 0.5 0.6 0.4 0.5

+ 0.1 + 0.1 + 0.1 + 0.1 + 0.1 N=23 N=22 N=19 N=22 N=36 Maternal Body Weight Change (gd 8 to 9) (g) b
0.5 0.4 0.6 0.5 0.6

+ 0.1 + 0.1 + 0.1 + 0.1 + 0.1 N=23 N=22 N=19 N=22 N=36 Maternal Body Weight Change (gd 9 to 10) (g) b
1.2 1.2 1.3 0.9 0.9

+ 0.1 § + 0.1 + 0.2 + 0.1 + 0.1 N=23 N=22 N=19 N=22 N=36 Maternal Body Weight Change (gd 10 to 11) (g) b
1.9 2.0 2.0 1.7 1.7

+ 0.1 + 0.2 + 0.1 + 0.2 + 0.1 N=23 N=22 N=19 N=22 N=36 Maternal Body Weight Change (gd 11 to 12) (g) b
2.1 1.9 1.8 2.0 2.3

+ 0.1 § + 0.2 + 0.1 + 0.2 + 0.1 N=23 N=22 N=19 N=22 N=36RTI-909; Project 0209189.000 Final Report



### 39 Table 2. Summary and Statistical Analysis of the Ma ternal Body Weights and Body Weight Changes (page 5 of 6)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000
Maternal Body Weight Change (gd 12 to 13) (g) b
2.1 ‡‡ 1.4 ** 1.9 1.5 * 2.0

+ 0.1 + 0.2 + 0.1 + 0.2 + 0.1 N=23 N=22 N=19 N=22 N=36 Maternal Body Weight Change (gd 13 to 14) (g) b
2.3 2.0 2.1 2.0 2.3

+ 0.1 + 0.2 + 0.1 + 0.2 + 0.2 N=23 N=22 N=19 N=22 N=36 Maternal Body Weight Change (gd 14 to 15) (g) b
2.8 2.8 2.8 2.5 2.7

+ 0.1 + 0.2 + 0.1 + 0.2 + 0.3 N=23 N=22 N=19 N=22 N=36 Maternal Body Weight Change (gd 15 to 16) (g) b
3.2 3.1 3.0 2.8 2.9

+ 0.2 + 0.2 + 0.1 + 0.3 + 0.2 N=23 N=22 N=19 N=22 N=36 Maternal Body Weight Change (gd 16 to 17) (g) b
3.2 2.9 3.1 2.7 3.1

+ 0.2 + 0.2 + 0.1 + 0.2 + 0.1 N=23 N=22 N=19 N=22 N=36 Maternal Body Weight Change (gd 5 to 10) (g) b,c
3.3 3.0

+ 0.2 + 0.2 N=23 N=36 Maternal Body Weight Change (gd 5 to 16) (g) b,d
17.6 16.7 17.5 15.5

+ 0.4 + 1.2 + 0.5 + 1.2 N=23 N=22 N=19 N=22RTI-909; Project 0209189.000 Final Report



### 40 Table 2. Summary and Statistical Analysis of the Ma ternal Body Weights and Body Weight Changes (page 6 of 6)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000
Maternal Body Weight Change (gd 10 to 17) (g) b,c
17.5 17.0

+ 0.4 + 0.9 N=23 N=36
Maternal Body Weight Change (gestation) (g) b
20.4 19.6 20.5 18.6 20.0

+ 0.6 + 1.3 + 0.7 + 1.5 + 1.1 N=23 N=22 N=19 N=22 N=36
Maternal Body Weight Change (corrected) (g) b,e
3.14 3.98 3.95 2.88 3.20

+ 0.25 + 0.44 + 0.51 + 0.38 + 0.32 N=23 N=22 N=19 N=22 N=36

a Female 3814 was removed from study due to a pre- existing condition. At necropsy she was found to

have an undescended testis on the right and seminal vesicl es and prostate to the right of the vagina and cervix.

b Includes all pregnant dams until terminal sacrific e on gestational day 17. Reported as the mean +

S.E.M.; gd=gestational day.

c This endpoint was only calculated for the 0 and 30,000 mg/m 3 dose groups.

d This endpoint was only calculated for the 0, 2000, 10,000, and 20,000 mg/m 3 dose groups.

e Weight change during gestation (gestational day 17 sa crifice weight minus gestational day 0 weight)

minus gravid uterine weight.

# Levene’s Test for homogeneity of variances was significant (p<0.05); therefore, robust regression

methods were used to test all treatment effects.
‡‡ p<0.01; ANOVA Test.
§ p<0.05; Test for Linear Trend.

* p<0.05; Dunnett’s Test.

** p<0.01; Dunnett’s Test.RTI-909; Project 0209189.000 Final Report



### 41 Table 3. Summary of the Maternal Clinical Observations (page 1 of 2)

A. Clinical Observations Summarized by Group
Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled)

Dosed gd 5-16 Dosed gd 5-10 Observation 0 2000 10,000 20,000 30,000 Alopecia: extremities/snout, moderate 1
Eye: enophthalmos a , unilateral, left 1
Labored breathing 1 1
Lacrimation, bilateral, moderate 1
Lacrimation, unilateral, moderate 1 2 Red exudates from anogenital area 1 1

B. Clinical Observations Summarized by Group, Day, and Time

Gasoline MTBE Vapor Condensate (mg/m 3

, inhaled) Dosed gd 5-16 Dosed gd 5-10 Day b Time c Observation 0 2000 10,000 20,000 30,000

2 Prior Eye: enophthalmos, unilateral, left 1 3 Prior Eye: enophthalmos, unilateral, left 1 4 Prior Eye: enophthalmos, unilateral, left 1 5 Prior Eye: enophthalmos, unilateral, left 1 Post Alopecia: extremities/snout, moderate 1 Eye: enophthalmos, unilateral, left 1
Lacrimation, unilateral, moderate 1 6 Prior Alopecia: extremities/snout, moderate 1 Eye: enophthalmos, unilateral, left 1 Post Alopecia: extremities/snout, moderate 1 Eye: enophthalmos, unilateral, left 1
Lacrimation, bilateral, moderate 1 Lacrimation, unilateral, moderate 1 1 7 Prior Alopecia: extremities/snout, moderate 1 Eye: enophthalmos, unilateral, left 1 Post Alopecia: extremities/snout, moderate 1 Eye: enophthalmos, unilateral, left 1RTI-909; Project 0209189.000 Final Report



### 42 Table 3. Summary of the Maternal Clinical Observations (page 2 of 2)

B. Clinical Observations Summarized by Group, Day, and Time

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 Day b Time c Observation 0 2000 10,000 20,000 30,000

8 Prior Alopecia: extremities/snout, moderate 1 Eye: enophthalmos, unilateral, left 1 Post Alopecia: extremities/snout, moderate 1 Eye: enophthalmos, unilateral, left 1
9 Prior Alopecia: extremities/snout, moderate 1 Eye: enophthalmos, unilateral, left 1 Post Alopecia: extremities/snout, moderate 1 Eye: enophthalmos, unilateral, left 1 Labored breathing 1
10 Prior Alopecia: extremities/snout, moderate 1

Eye: enophthalmos, unilateral, left 1 Post Alopecia: extremities/snout, moderate 1 Eye: enophthalmos, unilateral, left 1 Labored breathing 1
11 Prior Alopecia: extremities/snout, moderate 1

Eye: enophthalmos, unilateral, left 1 Post Alopecia: extremities/snout, moderate 1 Red exudates from anogenital area 1
12 Prior Alopecia: extremities/snout, moderate 1

Eye: enophthalmos, unilateral, left 1 Red exudates from anogenital area 1 Post Alopecia: extremities/snout, moderate 1
13 Prior Eye: enophthalmos, unilateral, left 1
14 Prior Eye: enophthalmos, unilateral, left 1
15 Prior Eye: enophthalmos, unilateral, left 1
16 Prior Eye: enophthalmos, unilateral, left 1
17 Prior Eye: enophthalmos, unilateral, left 1

a A sinking of the eyeball into the orbital cavity.

b Gestational day.

c Time is prior to/post (after) exposures.RTI-909; Project 0209189.000 Final Report



### 43 Table 4. Summary and Statistical Analysis of t he Maternal Feed Consumption (page 1 of 6)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000

No. Dams 23 22 19 22 36

Maternal Feed Consumption (gd 0 to 5) (g/day) a # 6.1 ††† 6.5 6.2 5.3 ÞÞ 6.8

+ 0.2 + 0.4 + 0.4 + 0.2 + 0.4
N=17 b N=18 b,c N=11 b N=16 b,c N=25 b,c

Maternal Feed Consumption (gd 5 to 6) (g/day) a # 6.1 † 6.7 7.7 Þ 6.2 5.9

+ 0.3 Ÿ + 0.3 + 0.7 + 0.2 + 0.1
N=20 b,d N=21 b N=16 b N=21 d N=32 b,d

Maternal Feed Consumption (gd 6 to 7) (g/day) a # 6.1 †† 7.8 Þ 7.7 ÞÞÞ 6.2 6.3

+ 0.2 + 0.7 + 0.4 + 0.3 + 0.2
N=18 b,d N=19 b,c,d N=17 b N=17 b,d N=32 b,c

Maternal Feed Consumption (gd 7 to 8) (g/day) a # 7.0 ††† 7.4 8.2 6.2 Þ 6.4

+ 0.4 ŸŸ Ÿ + 0.3 + 0.7 + 0.2 + 0.2
N=21 d N=21 d N=19 N=18 b,d N=34 c,d
Maternal Feed Consumption (gd 8 to 9) (g/day) a # 7.6 ††† 6.8 7.5 6.1 ÞÞ 6.2 ÞÞ

+ 0.5 ŸŸ Ÿ + 0.3 + 0.4 + 0.2 + 0.1
N=21 b N=19 c,d N=16 b,c N=20 b,d N=33 b,c

Maternal Feed Consumption (gd 9 to 10) (g/day) a 6.7 ‡‡‡ 7.0 7.3 6.1 6.3

+ 0.2 §§ + 0.3 + 0.2 + 0.2 + 0.1 N=23 N=21 b N=18 b N=20 b,c N=35 bRTI-909; Project 0209189.000 Final Report



### 44 Table 4. Summary and Statistical Analysis of t he Maternal Feed Consumption (page 2 of 6)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000
Maternal Feed Consumption (gd 10 to 11) (g/day) a # 6.9 ††† 7.6 7.5 6.3 6.2 ÞÞ

+ 0.2 ŸŸŸ + 0.3 + 0.2 + 0.2 + 0.1
N=19 b N=22 N=18 b N=20 d N=33 c,d
Maternal Feed Consumption (gd 11 to 12) (g/day) a
7.7 ‡ 7.1 7.7 6.7 7.8

+ 0.4 + 0.2 + 0.3 + 0.2 + 0.3 N=23 N=21 b N=18 b N=20 b,d N=33 b,c,d
Maternal Feed Consumption (gd 12 to 13) (g/day) a 7.1 ‡‡ 7.3 8.0 * 6.9 7.8

+ 0.2 + 0.3 + 0.3 + 0.2 + 0.2 N=23 N=21 c N=19 N=22 N=34 c,d
Maternal Feed Consumption (gd 13 to 14) (g/day) a 7.3 ‡‡ 7.5 7.9 7.1 8.0 *

+ 0.1 § + 0.3 + 0.2 + 0.3 + 0.2 N=23 N=20 b,c N=18 b N=21 b N=36
Maternal Feed Consumption (gd 14 to 15) (g/day) a 7.4 7.4 7.8 7.1 7.6

+ 0.1 + 0.3 + 0.2 + 0.3 + 0.2 N=23 N=21 b N=19 N=22 N=36
Maternal Feed Consumption (gd 15 to 16) (g/day) a 7.5 ‡ 7.8 7.9 6.9 7.2

+ 0.2 § + 0.3 + 0.3 + 0.2 + 0.2 N=23 N=22 N=19 N=22 N=35 c
Maternal Feed Consumption (gd 16 to 17) (g/day) a 7.6 7.4 7.8 7.3 7.5

+ 0.2 + 0.3 + 0.2 + 0.2 + 0.2 N=23 N=22 N=19 N=22 N=35 cRTI-909; Project 0209189.000 Final Report



### 45 Table 4. Summary and Statistical Analysis of t he Maternal Feed Consumption (page 3 of 6)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000
Maternal Feed Consumption (gd 5 to 10) (g/day) a,e # 6.9 6.2

+ 0.4 + 0.1
N=20 f N=28 f
Maternal Feed Consumption (gd 5 to 16) (g/day) a,g 6.9 7.0 7.7 6.8

+ 0.2 + 0.3 + 0.3 + 0.3
N=17 f N=14 f N=12 f N=17 f
Maternal Feed Consumption (gd 10 to 17) (g/day) a,e 7.3 7.6

+ 0.2 + 0.2
N=19 f N=31 f

Maternal Feed Consumption (gd 0 to 17) (g/day) a 6.5 ‡ 6.8 7.0 6.1 6.9

+ 0.1 + 0.3 + 0.3 + 0.2 + 0.2
N=14 f N=13 f N=8 f N=14 f N=20 f

Relative Maternal Feed Consumption (gd 0 to 5) (g/kg/day) a # 224.8 ††† 230.4 223.1 189.4 ÞÞ 241.5

+ 10.3 + 11.3 + 13.5 + 5.5 + 14.3
N=17 b N=18 b,c N=11 b N=16 b,c N=25 b,c
Relative Maternal Feed Consumption (gd 5 to 6) (g/kg/day) a # 216.1 † 233.1 270.7 Þ 217.3 207.5

+ 8.3 Ÿ + 9.7 + 24.6 + 8.9 + 4.6
N=20 b,d N=21 b N=16 b N=21 d N=32 b,dRTI-909; Project 0209189.000 Final Report



### 46 Table 4. Summary and Statistical Analysis of t he Maternal Feed Consumption (page 4 of 6)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000
Relative Maternal Feed Consumption (gd 6 to 7) (g/kg/day) a # 214.6 †† 266.0 Þ 264.7 ÞÞ 213.3 218.1

+ 7.6 Ÿ + 22.5 + 14.5 + 8.4 + 7.2
N=18 b,d N=19 b,c,d N=17 b N=17 b,d N=32 b,c
Relative Maternal Feed Consumption (gd 7 to 8) (g/kg/day) a # 242.4 ††† 247.6 275.4 208.1 Þ 216.5

+ 14.8 ŸŸŸ + 10.4 + 21.8 + 5.9 + 5.3
N=21 d N=21 d N=19 N=18 b,d N=34 c,d
Relative Maternal Feed Consumption (gd 8 to 9) (g/kg/day) a # 255.3 ††† 224.2 249.3 202.6 ÞÞ 204.2 ÞÞ

+ 17.1 ŸŸŸ + 10.3 + 14.0 + 5.9 + 4.0
N=21 b N=19 c,d N=16 b,c N=20 b,d N=33 b,c
Relative Maternal Feed Consumption (gd 9 to 10) (g/kg/day) a 222.9 ‡‡‡ 225.2 234.9 198.8 * 204.8

+ 8.4 §§ + 8.7 + 7.6 + 4.7 + 3.4 N=23 N=21 b N=18 b N=20 b,c N=35 b
Relative Maternal Feed Consumption (gd 10 to 11) (g/kg/day) a 218.8 ‡‡‡ 229.9 230.3 197.4 193.4 *

+ 10.0 §§§ + 8.3 + 6.9 + 6.4 + 2.8
N=19 b N=22 N=18 b N=20 d N=33 c,d
Relative Maternal Feed Consumption (gd 11 to 12) (g/kg/day) a 228.1 ‡ 203.7 222.9 197.0 * 230.6

+ 11.6 + 5.1 + 9.0 + 4.6 + 8.0 N=23 N=21 b N=18 b N=20 b,d N=33 b,c,d
Relative Maternal Feed Consumption (gd 12 to 13) (g/kg/day) a 198.3 ‡‡ 198.8 221.2 192.0 213.4

+ 5.9 + 6.0 + 7.5 + 5.9 + 5.6 N=23 N=21 c N=19 N=22 N=34 c,dRTI-909; Project 0209189.000 Final Report



### 47 Table 4. Summary and Statistical Analysis of t he Maternal Feed Consumption (page 5 of 6)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000
Relative Maternal Feed Consumption (gd 13 to 14) (g/kg/day) a 191.7 ‡ 195.8 205.9 189.8 209.4 *

+ 4.2 § + 4.9 + 4.9 + 6.6 + 4.1 N=23 N=20 b,c N=18 b N=21 b N=36
Relative Maternal Feed Consumption (gd 14 to 15) (g/kg/day) a 182.2 182.8 190.7 177.8 187.3

+ 4.1 + 4.9 + 5.8 + 4.6 + 5.0 N=23 N=21 b N=19 N=22 N=36
Relative Maternal Feed Consumption (gd 15 to 16) (g/kg/day) a 171.5 ‡‡ 179.0 181.5 162.0 163.6

+ 3.4 §§ + 4.5 + 6.1 + 3.5 + 2.6 N=23 N=22 N=19 N=22 N=35 c
Relative Maternal Feed Consumption (gd 16 to 17) (g/kg/day) a 162.9 160.3 166.1 161.2 159.9

+ 4.7 + 3.4 + 3.4 + 4.6 + 2.4 N=23 N=22 N=19 N=22 N=35 c
Relative Maternal Feed Consumption (gd 5

to 10) (g/kg/day) a,e # 235.6 † 209.5 Þ

+ 12.1 Ÿ + 3.7
N=20 f N=28 f
Relative Maternal Feed Consumption (gd 5

to 16) (g/kg/day) a,g 202.6 206.9 222.0 199.3

+ 5.7 + 5.3 + 6.7 + 7.0
N=17 f N=14 f N=12 f N=17 f
Relative Maternal Feed Consumption (gd 10 to 17) (g/kg/day) a,e 189.6 194.5

+ 4.8 + 3.4
N=19 f N=31 fRTI-909; Project 0209189.000 Final Report



### 48 Table 4. Summary and Statistical Analysis of t he Maternal Feed Consumption (page 6 of 6)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000
Relative Maternal Feed Consumption (gd 0

to 17) (g/kg/day) a 187.1 197.0 198.2 178.6 194.3

+ 4.9 + 5.8 + 7.0 + 3.8 + 5.1
N=14 f N=13 f N=8 f N=14 f N=20 f

a Includes all pregnant dams until terminal sacrific e on gestational day 17. Reported as the mean +

S.E.M.; gd = gestational day. b Decrease in N is due to one or more feeders sp illing, and therefore the f eed weight was excluded.

c Decrease in N is due to the feed being contami nated for one or more animals, and therefore the feed

weight was excluded. d Decrease in N is due to the feed consumption value for one or more animals being a statistical outlier,

and therefore they were excluded.

e This endpoint was only calculated for the 0 and 30,000 mg/m 3 dose groups. f Decrease in N is due to interim feed consumption value(s) for one or more dams being missing, and

therefore the overall feed consumpt ion value could not be calculated. g This endpoint was only calculated for the 0, 2000, 10,000, and 20,000 mg/m 3 dose groups. # Levene’s Test for homogeneity of variances was significant (p<0.05); therefore, robust regression

methods were used to test all treatment effects. † p<0.05; Wald Chi-square Test for overall tr eatment effect in robust regression model. †† p<0.01; Wald Chi-square Test for overall tr eatment effect in robust regression model. ††† p<0.001; Wald Chi-square Test for overall treatment effect in robust regression model. Ϋ p<0.05; Linear trend test in robust regression model. ΫΫΫ p<0.001; Linear trend test in robust regression model. Þ p<0.05; Individual t-test for pairwise compar isons to control in robust regression model. ÞÞ p<0.01; Individual t-test for pairwise comparisons to control in robust regression model. ÞÞÞ p<0.001; Individual t-test for pairwise comparis ons to control in robust regression model. ‡ p<0.05; ANOVA Test. ‡‡ p<0.01; ANOVA Test. ‡‡‡ p<0.001; ANOVA Test. § p<0.05; Test for Linear Trend. §§ p<0.01; Test for Linear Trend. §§§ p<0.001; Test for Linear Trend.

* p<0.05; Dunnett’s Test.RTI-909; Project 0209189.000 Final Report



### 49 Table 5. Summary and Statistical Analysis of the Ma ternal Absolute and Relative Organ Weights (page 1 of 2)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000

Absolute Gravid Uterine Weight (g) a 17.2900 15.5703 16.5732 15.7215 16.7807

+ 0.4737 + 1.1217 + 0.3737 + 1.2039 + 0.9127 N=23 N=22 N=19 N=22 N=36

Absolute Maternal Liver Weight (g) a 2.4511 ‡ 2.5766 2.7247 * 2.6308 2.4253

+ 0.0498 + 0.0889 + 0.0538 + 0.0715 + 0.0604 N=23 N=22 N=19 N=22 N=36

Absolute Maternal Paired Adrenal Gland Weight (g) a 0.0136 0.0144 0.0132 0.0137 0.0135

+ 0.0006 + 0.0007 + 0.0004 + 0.0005 + 0.0003
N=22 b N=22 N=18 b N=22 N=36

Absolute Maternal Paired Kidney Weight (g) a 0.4277 0.4454 0.4394 0.4376 0.4311

+ 0.0089 + 0.0107 + 0.0054 + 0.0089 + 0.0073 N=23 N=22 N=19 N=22 N=36 Relative Maternal Liver Weight (% sacrifice weight) a # 5.1961 ††† 5.5269 ÞÞ 5.7418 ÞÞÞ 5.7610 ÞÞÞ 5.1550

+ 0.0569 + 0.0865 + 0.0927 + 0.0754 + 0.0939 N=23 N=22 N=19 N=22 N=36 Relative Maternal Paired Adrenal Gland Weight (% sacrifice weight) a 0.0290 0.0314 0.0279 0.0306 0.0297

+ 0.0014 + 0.0016 + 0.0009 + 0.0017 + 0.0015
N=22 b N=22 N=18 b N=22 N=36

Relative Maternal Paired Kidney Weight (% sacrifice weight) a 0.9067 0.9783 0.9281 0.9817 0.9345

+ 0.0109 + 0.0451 + 0.0156 + 0.0496 + 0.0337 N=23 N=22 N=19 N=22 N=36RTI-909; Project 0209189.000 Final Report



### 50 Table 5. Summary and Statistical Analysis of the Maternal Absolute and Relative Organ Weights (page 2 of 2)

a Includes all pregnant dams until terminal sacrific e on gestational day 17. Reported as the mean +

S.E.M.; gd=gestational day.

b Decrease in N is due to the paired adrenal weight fo r one animal being a statistical outlier and therefore

it was excluded.

# Levene’s Test for homogeneity of variances was significant (p<0.05); therefore, robust regression

methods were used to test all treatment effects.
‡ p<0.05; ANOVA Test.

* p<0.05; Dunnett’s Test.
††† p<0.001; Wald Chi-square Test for overall treatment effect in robust regression model.

ÞÞ p<0.01; Individual t-test for pairwise comparisons to control in robust regression model.

ÞÞÞ p<0.001; Individual t-test for pairwise comparis ons to control in robust regression model.RTI-909; Project 0209189.000 Final Report



### 51 Table 6. Summary and Statistical Analysis of Ovarian Corpora Lutea, Uterine Contents, Live Fetal Sex and Live Fetal Body Weight (page 1 of 4)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000



### A LL 

### L ITTERS 

### a : 23 22 19 22 36

No. Corpora Lutea per Dam b
12.96 12.18 12.78 13.18 13.19

+ 0.38 + 0.57 + 0.36 + 0.56 + 0.47 N=23 N=22 N=18 c N=22 N=36

No. Implantation Sites per
Litter b
12.74 12.00 12.68 13.23 12.89

+ 0.35 + 0.61 + 0.33 + 0.46 + 0.37 N=23 N=22 N=19 N=22 N=36
%Preimplantation Loss per
Litter b
2.06 5.99 3.93 2.61 4.81

+ 0.93 + 3.74 + 1.52 + 0.99 + 1.24 N=23 N=22 N=18 c N=22 N=36

No. Resorptions per Litter b
0.61 1.27 0.74 1.68 1.56

+ 0.16 + 0.52 + 0.18 + 0.86 + 0.48 N=23 N=22 N=19 N=22 N=36

% Resorptions per Litter b
4.88 11.03 5.57 12.91 13.75

+ 1.31 + 4.90 + 1.40 + 6.21 + 4.52 N=23 N=22 N=19 N=22 N=36
No. Litters with Resorptions 11 9 10 10 20
% Litters with Resorptions 47.83 40.91 52.63 45.45 55.56RTI-909; Project 0209189.000 Final Report



### 52 Table 6. Summary and Statistical Analysis of Ovarian Corpora Lutea, Uterine Contents, Live Fetal Sex and Live Fetal Body Weight (page 2 of 4)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000
No. Late Fetal Deaths per
Litter b
0.13 0.00 0.11 0.09 0.03

+ 0.07 + 0.00 + 0.07 + 0.06 + 0.03 N=23 N=22 N=19 N=22 N=36
%Late Fetal Deaths per
Litter b
1.02 0.00 0.70 0.65 0.20

+ 0.56 + 0.00 + 0.49 + 0.45 + 0.20 N=23 N=22 N=19 N=22 N=36

No. Litters with Late Fetal Deaths 3 0 2 2 1

% Litters with Late Fetal Deaths 13.04 0.00 10.53 9.09 2.78
No. Nonlive Implants per
Litter b,d
0.74 1.27 0.84 1.77 1.58

+ 0.19 + 0.52 + 0.22 + 0.86 + 0.48 N=23 N=22 N=19 N=22 N=36
% Nonlive Implants per
Litter b,d
5.90 11.03 6.28 13.56 13.95

+ 1.52 + 4.90 + 1.59 + 6.16 + 4.52 N=23 N=22 N=19 N=22 N=36

No. Litters with Nonlive Implants d
11 9 10 12 20

% Litters with Nonlive Implants d
47.83 40.91 52.63 54.55 55.56

No. Litters with 100% Nonlive Implants d
0 1 0 2 3

% Litters with 100% Nonlive Implants d
0.00 4.55 0.00 9.09 8.33RTI-909; Project 0209189.000 Final Report



### 53 Table 6. Summary and Statistical Analysis of Ovarian Corpora Lutea, Uterine Contents, Live Fetal Sex and Live Fetal Body Weight (page 3 of 4)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000
No. Adversely Affected Implants per Litter b,e
0.83 1.36 0.89 1.82 1.78

+ 0.20 + 0.52 + 0.23 + 0.85 + 0.48 N=23 N=22 N=19 N=22 N=36 % Adversely Affected Implants per Litter b,e
6.69 11.66 6.63 13.87 15.38

+ 1.64 + 4.85 + 1.63 + 6.15 + 4.50 N=23 N=22 N=19 N=22 N=36
No. Litters with Adversely Affected Implants e 12 11 10 12 21
% Litters with Adversely Affected Implants e 52.17 50.00 52.63 54.55 58.33

L IVE L ITTERS f : 23 21 19 20 33

No. Live Fetuses per Litter b
12.00 11.24 11.84 12.60 12.33

+ 0.39 + 0.68 + 0.27 + 0.49 + 0.26 N=23 N=21 N=19 N=20 N=33

% Male Fetuses per Litter b
50.03 46.46 52.95 44.67 45.58

+ 3.11 + 3.82 + 3.70 + 3.14 + 2.99 N=23 N=21 N=19 N=20 N=33

No. Male Fetuses per
Litter b
6.09 5.38 6.21 5.65 5.61

+ 0.47 + 0.45 + 0.41 + 0.48 + 0.36 N=23 N=21 N=19 N=20 N=33RTI-909; Project 0209189.000 Final Report



### 54 Table 6. Summary and Statistical Analysis of Ovarian Corpora Lutea, Uterine Contents, Live Fetal Sex and Live Fetal Body Weight (page 4 of 4)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000

No. Female Fetuses per Litter b
5.91 5.86 5.63 6.95 6.73

+ 0.37 + 0.52 + 0.48 + 0.47 + 0.39 N=23 N=21 N=19 N=20 N=33
Average Fetal Body Weight (g) per Litter b
0.9994 1.0114 0.9471 0.9492 1.0248

+ 0.0265 + 0.0212 + 0.0241 + 0.0288 + 0.0249 N=23 N=21 N=19 N=20 N=33
Average Male Fetal Body Weight (g) per Litter b
1.0114 1.0088 0.9605 0.9722 1.0367

+ 0.0309 + 0.0166 + 0.0254 + 0.0311 + 0.0261 N=23 N=20 g N=19 N=20 N=33

Average Female Fetal Body Weight (g) per Litter b
0.9961 0.9937 0.9344 0.9343 1.0141

+ 0.0246 + 0.0227 + 0.0231 + 0.0281 + 0.0239 N=23 N=21 N=19 N=20 N=33

a Includes all dams pregnant at terminal sacrifice on ge stational day 17; litter size = no. implantation sites

per dam.

b Reported as the mean + S.E.M.

c Decrease in N is due to the right ovary for one female inadvertently being lost prior to the corpora lutea

being counted.

d Nonlive = late fetal deaths plus resorptions.

e Adversely affected = nonlive plus malformed.

f Includes only dams with live fetuses; lit ter size = no. live fetuses per dam.

g Decrease in N is due to one litter having female fetuses only.RTI-909; Project 0209189.000 Final Report



### 55 Table 7. Summary and Statistical Analysis of Fetal Exter nal Malformations and Variations (page 1 of 3)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000

No. Fetuses Examined a 276 236 225 252 407

No. Litters Examined b
23 21 19 20 33

No. Fetuses with External Malformations per Litter c,d
0.09 0.10 0.05 0.05 0.21

+ 0.06 + 0.07 + 0.05 + 0.05 + 0.11 N=23 N=21 N=19 N=20 N=33

No. Male Fetuses with External Malformations per Litter c,d
0.04 0.05 0.05 0.00 0.09

+ 0.04 + 0.05 + 0.05 + 0.00 + 0.05 N=23 N=20 N=19 N=20 N=33

No. Female Fetuses with External Malformations per Litter c,d
0.04 0.05 0.00 0.05 0.12

+ 0.04 + 0.05 + 0.00 + 0.05 + 0.07 N=23 N=21 N=19 N=20 N=33
% Fetuses with External Malformations per
Litter c,d
0.85 0.66 0.38 0.36 1.64

+ 0.61 + 0.46 + 0.38 + 0.36 + 0.85 N=23 N=21 N=19 N=20 N=33

% Male Fetuses with External Malformations per Litter c,d
0.62 0.83 0.88 0.00 1.89

+ 0.62 + 0.83 + 0.88 + 0.00 + 1.16 N=23 N=20 N=19 N=20 N=33

% Female Fetuses with External Malformations per
Litter c,d
0.72 0.95 0.00 1.25 1.29

+ 0.72 + 0.95 + 0.00 + 1.25 + 0.74 N=23 N=21 N=19 N=20 N=33RTI-909; Project 0209189.000 Final Report



### 56 Table 7. Summary and Statistical Analysis of Fetal Exter nal Malformations and Variations (page 2 of 3)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000

No. Fetuses with External
Malformations d
2 2 1 1 7

% Fetuses with External
Malformations d
0.72 0.85 0.44 0.40 1.72

No. Litters with External
Malformations e 2 2 1 1 4

% Litters with External
Malformations e 8.70 9.52 5.26 5.00 12.12

No. Fetuses with External Variations per Litter c,d
0.13 0.14 0.11 0.15 0.00

+ 0.07 + 0.10 + 0.07 + 0.08 + 0.00 N=23 N=21 N=19 N=20 N=33

No. Male Fetuses with External Variations per Litter c,d
0.09 0.05 0.05 0.05 0.00

+ 0.06 + 0.05 + 0.05 + 0.05 + 0.00 N=23 N=20 N=19 N=20 N=33

No. Female Fetuses with External Variations per Litter c,d
0.04 0.10 0.05 0.10 0.00

+ 0.04 + 0.10 + 0.05 + 0.07 + 0.00 N=23 N=21 N=19 N=20 N=33
% Fetuses with External Variations per
Litter c,d
1.34 1.27 0.81 1.26 0.00

+ 0.76 + 0.99 + 0.56 + 0.69 + 0.00 N=23 N=21 N=19 N=20 N=33

% Male Fetuses with External Variations per Litter c,d
1.45 0.56 0.66 1.00 0.00

+ 1.00 + 0.56 + 0.66 + 1.00 + 0.00 N=23 N=20 N=19 N=20 N=33RTI-909; Project 0209189.000 Final Report



### 57 Table 7. Summary and Statistical Analysis of Fetal Exter nal Malformations and Variations (page 3 of 3)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000

% Female Fetuses with External Variations per Litter c,d
0.72 1.59 1.05 1.39 0.00

+ 0.72 + 1.59 + 1.05 + 0.98 + 0.00 N=23 N=21 N=19 N=20 N=33

No. Fetuses with External
Variations d
3 3 2 3 0

% Fetuses with External
Variations d
1.09 1.27 0.89 1.19 0.00

No. Litters with External
Variations e 3 2 2 3 0

% Litters with External
Variations e 13.04 9.52 10.53 15.00 0.00

a Only live fetuses were examined for malformations and variations.

b Includes only litters with live fetuses.

c Reported as the mean + S.E.M.

d Fetuses with one or more malformations or variations.

e Litters with one or more fetuses with malformations or variations.RTI-909; Project 0209189.000 Final Report



### 58 Table 8. Summary of Morphological Abnormalities in CD-1 Mouse Fetuses: Listing by Defect Type a (page 1 of 1)

Gasoline MTBE Vapor Condensate (mg/m 3 , inhaled) Dosed gd 5-16 Dosed gd 5-10 0 2000 10,000 20,000 30,000

EXTERNAL MALFORMATIONS

Total No. of Fetuses Examined for
External Malformations b 276 236 225 252 407

No. of Fetuses with External
Malformations c 2 2 1 1 7

% Fetuses with External Malformations 0.7% 0.8% 0.4% 0.4% 1.7%

Total No. of Litters Examined for
External Malformations d 23 21 19 20 33

No. of Litters with External
Malformations e 2 2 1 1 4

% Litters with External Malformations 8.7% 9.5% 5.3% 5.0% 12.1%

Encephalocele 1(1) Cleft Palate 2(2) 1(1) 1(1) 1(1) 7(4) Gastroschisis 1(1)

EXTERNAL VARIATIONS

Total No. of Fetuses Examined for
External Variations b 276 236 225 252 407

No. of Fetuses with External
Variations c 3 3 2 3 0

% Fetuses with External Variatio ns 1.1% 1.3% 0.9% 1.2% 0.0% Total No. of Litters Examined for
External Variations d 23 21 19 20 33

No. of Litters with External
Variations e 3 2 2 3 0

% Litters with External Variati ons 13.0% 9.5% 10. 5% 15.0% 0.0%
Abnormal Rugae in Midline of Palate 1(1) 1(1) Hematoma: Face 2(1) Hematoma: Head 2(2) 1(1) Hematoma: Neck 1(1) 1(1) 1(1) 2(2) Hematoma: Shoulder 1(1)

a A single fetus may be represented more than once in listing individual defects. Data are presented as

the number of fetuses (number of litters). b Only live fetuses were included.

c Fetuses with one or more malformations/variations. d Includes only litters with live fetuses.

e Litters with one or more malformed/variant fetuses.RTI-909; Project 0209189.000 Final Report



### 59 Table 9. External Malformations in Control CD-1® Mouse Litters From Studies Performed for Governmental Clients at RTI From 1997 to 2002 (page 1 of 1)

Control Group With Gastroschisis b Study Code Year Study Type a No. Dams No. Fetuses No. Fetuses No. Litters Incidence of External Malformations No. Fetuses (No. Litters)

A 2002 RF 10 138 0 0 1 (1) cleft palate

B 2002 RF 9 123 0 0 1 (1) exencephaly

C 1999 RF 15 184 0 0 0

D 1999 RF 15 200 0 0 0

E 2000 RF 14 172 0 0 0

F 1999 RF 15 187 0 0 0

G 2000 RF 12 165 0 0 0

H 1999 D 24 302 0 0 0

I 2000 D 24 326 0 0 2 (1) cleft palate

J 1999 D 23 291 0 0 0

K 1997 D 22 271 0 0 1 (1) cleft palate

L 2000 D 23 283 0 0 1 (1) exencephaly

M 2001 D 24 270 0 0 1 (1) cleft palate

N 2002 D 20 254 0 0 6 (4) cleft palate

O 2002 D 22 279 0 0 7 (3) cleft palate

P 2000 D 16 196 0 0 0

TOTAL 288 3641 0 0 18 (11) cleft palate 2 (2) exencephaly

a RF = range-finding study D = definitive study

b There were also no fetuses with ectopia cordis

# Appendix I:

Inhalation Report

### STUDY**FILE NOTE** **Study Number:** **04-4262/4263 Effective** **Date: 9DecO8** **cc: Subject:** **Ml RAN** **LOQ The** **control chamber** **limit of** **quantification was** **estimated at** **433 mg/m3** **based on** **the extrapolated** **value for** **an absorbance** **reading of 0.01** **which Is** **considered the** **lowest practical** **reading above** **the baseline.** **Since the** **nominal to** **measured ratio** **for even** **the low** **exposure level** **for the** **main study** **was close** **to 1:1,** **we did** **not see** **need to** **change the** **calibration curve.** **Prepared by** **and Principal** **Investigator Approval:** **• Date:** 

# Appendix II:

Individual Animal Data Tables

### Table of Contents

A-1 Individual Maternal Body Weights and Organ Weights (g) ................................. 1

A-2 Individual Maternal Clinical Observations........................................................... 5 A-3 Individual Maternal Feed Consumption (g/day) .................................................. 6 A-4 Individual Embryo/Fetal Data.............................................................................. 11Appendix II 1Table A-1. Individual Maternal Body Weights and Organ Weights (g) (page 1 of 4)Gestational DayDoseaDam ID056789101112131415161717bGravidUterineWt.LiverWt.PairedAdrenalGlandWt.PairedKidneyWt.0 1801 23.7 25.3 25.6 26.9 27.8 29.5 29.4 31.0 33.4 35.7 37.4 40.2 42.7 45.7 44.04 14.4811 2.3692 0.0136 0.44601802 27.4 28.6 29.3 30.1 30.7 31.0 32.3 34.7 37.2 40.0 41.5 44.1 49.0 52.3 51.46 19.7607 2.6311 0.0158 0.48191803 27.5 27.3 28.6 28.6 29.3 29.6 29.9 31.9 33.1 34.5 36.4 39.0 41.8 44.2 41.65 13.6798 2.1015 0.0115 0.37751804 24.0 24.5 24.9 25.6 26.2 26.3 27.4 29.6 31.8 34.3 36.3 40.2 43.3 46.1 45.15 17.7023 2.3577 0.0165 0.41121805 27.2 27.3 27.6 28.8 30.0 29.9 30.7 32.9 34.9 36.3 38.2 41.8 44.4 47.7 46.02 16.6375 2.2176 0.0115 0.39081806 27.2 27.5 28.3 29.2 29.5 30.0 31.0 32.9 35.6 37.4 39.6 43.5 46.2 49.3 48.23 18.2375 2.4033 0.0164 0.43951807 23.9 26.2 25.7 26.9 27.3 27.9 29.9 31.7 33.1 35.9 37.2 39.9 43.8 48.4 47.32 18.3623 2.4555 .c0.40201808 26.4 26.7 26.9 27.6 27.7 27.6 28.7 30.3 32.8 35.1 38.3 40.1 43.6 47.8 46.10 17.9075 2.3041 0.0141 0.42911809 26.5 28.4 29.2 29.0 29.6 30.2 31.1 32.4 34.6 36.2 38.0 39.4 41.1 44.4 43.49 14.7802 2.2250 0.0127 0.33791810 27.8 28.4 29.6 30.3 29.7 30.5 31.6 33.7 34.3 36.0 39.0 41.1 43.4 46.2 44.21 13.6742 2.3782 0.0128 0.39501811 27.9 27.6 28.4 28.6 29.5 29.8 31.3 34.0 37.3 40.1 41.2 43.7 48.6 53.1 52.05 20.7503 2.9608 0.0135 0.50551812 25.0 25.0 25.5 25.9 26.7 26.5 28.1 29.3 31.5 33.5 36.2 39.2 42.4 45.3 44.26 17.3355 2.2345 0.0135 0.40181813 27.2 28.3 29.2 30.2 30.7 30.9 32.5 33.5 35.0 36.8 39.1 42.3 44.8 48.3 46.90 15.3242 2.6188 0.0211 0.44321814 27.2 27.7 28.5 29.3 30.1 30.6 32.2 33.3 35.8 37.9 40.9 43.6 46.8 50.3 48.78 17.9133 2.7351 0.0106 0.42111815 28.3 28.6 29.3 30.3 30.9 31.2 32.9 35.3 36.6 38.4 40.6 42.8 45.9 49.2 47.46 16.4148 2.7521 0.0135 0.40911816 28.7 30.0 31.9 32.3 32.5 33.1 34.4 36.8 38.9 40.5 43.5 46.3 48.2 52.4 51.34 19.6942 2.6059 0.0132 0.49681817 23.9 24.6 24.5 24.6 25.1 25.4 26.3 27.8 29.6 31.5 33.6 36.0 38.9 40.5 40.07 13.4026 2.0213 0.0130 0.37581818 28.5 29.9 31.0 30.5 31.1 32.2 33.9 36.8 38.7 40.8 43.2 46.8 49.9 52.5 51.75 20.6578 2.5824 0.0112 0.48051819 28.8 30.0 31.2 30.9 31.2 32.5 34.0 36.1 38.1 40.4 43.0 46.1 49.4 51.8 51.28 20.1298 2.5947 0.0192 0.47351820 25.2 26.8 27.5 28.1 28.8 29.7 30.5 33.2 35.0 36.6 40.0 43.0 46.2 49.7 48.71 18.7321 2.3504 0.0088 0.41191821 24.3 25.4 25.7 26.5 26.6 27.2 28.0 29.3 31.6 33.6 35.5 38.0 41.5 43.9 43.40 15.6224 2.1432 0.0149 0.40961822 28.9 30.7 30.9 31.5 31.5 32.1 33.1 34.3 36.9 39.2 41.1 44.5 48.9 52.4 51.15 18.5153 2.6422 0.0099 0.47611823 28.8 29.0 28.9 29.3 29.6 29.7 31.5 34.0 36.3 38.7 41.6 44.9 48.9 51.8 49.47 17.9535 2.6913 0.0119 0.42042000 2801 25.0 27.6 27.9 28.4 28.5 28.9 28.9 29.7 30.2 30.1 30.5 32.0 32.4 33.5 31.58 2.3762 1.8423 0.0139 0.46692802 27.0 29.1 29.9 31.1 31.7 32.3 33.2 34.5 35.8 36.9 38.6 41.0 44.6 46.7 45.48 13.0643 2.4958 0.0162 0.45332803 27.5 27.1 27.3 28.6 28.2 28.5 30.1 34.4 36.3 37.8 40.6 45.8 49.6 52.9 50.08 21.7074 2.1868 0.0110 0.38652804 24.3 25.3 25.4 26.5 27.0 26.8 27.9 29.7 32.3 33.9 36.5 39.5 42.6 44.8 43.98 15.4744 2.4670 0.0114 0.40412805 26.7 26.0 26.2 27.0 28.4 28.3 29.1 30.7 33.1 34.8 36.8 40.2 43.7 45.5 44.36 16.1582 2.2875 0.0159 0.36872806 27.4 27.6 28.8 29.1 29.6 30.2 31.4 34.1 36.1 37.5 40.3 43.9 46.2 50.2 49.83 17.2433 2.7615 0.0162 0.44522807 24.6 24.8 26.4 27.7 27.5 28.0 29.3 31.2 32.9 34.8 36.0 38.4 41.6 44.6 43.51 14.5031 2.6123 0.0154 0.40262808 26.4 28.8 30.3 30.7 30.7 31.2 32.5 34.4 35.9 37.7 39.3 40.8 42.5 45.0 44.13 12.2719 2.5176 0.0146 0.49442809 26.8 30.4 31.0 31.5 32.6 32.9 34.7 36.5 38.8 40.0 40.9 44.1 47.6 49.5 48.54 15.9050 2.9708 0.0161 0.49682810 27.7 30.5 30.1 30.3 31.7 32.0 32.6 35.1 36.4 37.6 38.9 42.0 45.1 48.1 46.70 14.7170 2.5925 0.0142 0.40622811 28.0 28.6 29.0 29.6 30.0 30.7 31.8 33.7 35.4 37.6 39.8 42.1 45.3 47.8 46.65 17.7486 2.3583 0.0141 0.38722812 29.8 30.7 31.9 32.0 32.3 33.6 34.8 37.4 40.7 42.5 45.5 48.7 52.7 56.9 55.09 20.6185 2.9510 0.0177 0.45902813 25.2 25.3 26.0 26.0 26.7 26.6 26.3 26.1 25.9 24.9 24.4 25.1 24.7 26.5 24.98 0.2012 1.4584 0.0094 0.41662814 27.0 26.7 27.6 28.0 28.5 29.0 30.2 32.7 34.6 36.6 37.7 40.1 43.3 46.5 45.18 14.7611 2.5448 0.0227 0.4334Appendix II 2Table A-1. Individual Maternal Body Weights and Organ Weights (g) (page 2 of 4)Gestational DayDoseaDam ID056789101112131415161717bGravidUterineWt.LiverWt.PairedAdrenalGlandWt.PairedKidneyWt.20000 2815 27.5 31.4 32.4 32.7 33.4 33.9 36.0 38.0 40.3 41.5 44.1 47.4 51.5 54.2 53.35 18.9077 3.1841 0.0163 0.47822816 28.3 27.8 29.1 30.4 30.6 30.5 32.1 34.6 37.8 39.0 41.8 43.9 48.0 51.4 49.86 18.2901 2.6670 0.0133 0.42952817 28.8 29.8 29.7 30.0 30.6 31.0 32.2 34.0 35.4 36.7 38.7 41.1 43.8 46.8 46.53 17.2023 2.2908 0.0141 0.41202818 25.6 27.7 29.1 30.1 30.1 30.9 32.9 34.9 37.6 39.7 42.4 45.6 49.4 52.8 52.35 21.5024 2.8345 0.0151 0.50622819 28.2 30.7 31.5 31.9 32.5 33.3 34.7 37.2 39.5 42.0 44.1 47.2 50.7 55.0 53.58 18.9426 3.0684 0.0129 0.56202820 29.1 30.3 31.4 31.6 32.7 33.3 35.3 38.4 39.9 41.2 44.5 47.5 50.9 54.7 53.60 18.8395 2.9602 0.0172 0.53502821 .d2822 27.3 27.0 27.7 28.6 28.7 28.8 30.1 31.9 33.9 35.1 38.5 41.2 44.7 47.7 46.32 15.2662 2.5073 0.0069 0.41862823 29.3 30.8 31.5 32.4 32.7 33.3 34.3 36.1 38.3 39.9 42.3 46.2 50.4 54.0 51.95 16.8460 3.1252 0.0127 0.437410000 3801 25.4 26.5 26.8 27.3 27.5 28.4 29.6 31.8 33.7 35.5 37.1 39.8 42.3 44.4 44.62 14.7301 2.7557 0.0115 0.45763802 .d3803 27.5 28.0 28.7 29.2 30.3 30.4 31.3 33.4 35.5 37.2 39.3 41.7 44.1 47.1 47.34 16.9253 2.4204 0.0121 0.41913804 25.4 27.5 28.4 29.0 30.2 30.7 33.5 35.1 36.7 38.3 40.7 43.8 46.9 49.7 48.87 16.1002 2.6020 0.0127 0.43183805 26.5 28.2 28.7 29.0 30.1 31.1 32.5 35.6 37.3 38.9 41.5 44.7 46.9 49.3 48.51 17.6251 2.5656 0.0140 0.41623806 27.6 29.4 30.1 31.0 32.0 33.0 34.9 37.6 39.7 42.2 45.0 48.0 51.9 55.2 53.77 19.7811 2.9632 0.0135 0.49453807 25.0 24.8 25.3 25.7 26.7 27.3 28.4 30.5 32.1 33.7 35.2 37.4 41.0 44.3 43.27 14.5264 2.8788 0.0106 0.47093808 26.2 26.3 26.3 27.0 27.3 27.3 28.1 30.1 32.7 34.3 35.8 38.1 40.0 42.5 41.96 13.4726 2.4245 0.0157 0.42363809 26.9 27.4 27.6 28.5 28.8 29.2 30.4 32.1 34.0 35.1 37.0 39.6 42.7 45.2 44.35 16.7280 2.4484 0.0138 0.43453810 27.5 28.9 29.8 30.1 30.5 30.9 31.5 34.2 36.0 38.3 41.1 42.6 45.7 48.9 47.29 16.8350 2.5713 0.0152 0.44033811 28.1 26.1 27.2 27.2 27.9 27.7 28.8 30.3 32.2 34.4 36.5 39.3 41.7 44.3 43.58 14.9477 2.5122 0.0122 0.40603812 29.1 29.1 29.5 30.3 30.7 31.1 32.4 34.3 36.3 38.1 40.7 43.9 47.2 50.3 48.94 18.1387 2.7584 0.0129 0.43663813 25.5 24.2 27.3 27.1 27.1 27.6 29.3 30.5 32.0 34.4 35.7 39.4 41.8 44.8 43.89 15.3821 2.6385 0.0116 0.44573814 .e3815 27.7 29.0 30.5 31.2 31.8 32.7 35.1 37.0 38.4 40.5 42.1 45.7 48.9 52.8 52.65 17.5735 2.904f0.0141 0.45503816 28.2 28.0 29.0 29.6 30.2 30.3 32.0 34.5 36.3 38.7 41.3 43.9 48.0 51.5 50.10 18.6991 2.9257 0.0135 0.45363817 .d3818 25.8 27.3 28.8 28.7 28.5 30.5 30.3 32.4 34.1 35.4 37.2 40.5 43.4 46.1 45.33 14.8345 2.5348 0.0141 0.41113819 28.2 28.8 29.4 29.6 29.9 30.4 32.0 32.9 35.3 37.2 38.6 41.1 44.2 47.6 46.66 16.0488 2.9220 0.0151 0.42953820 26.9 30.3 32.1 32.4 33.6 34.7 35.4 37.9 39.0 41.0 43.2 45.6 48.7 53.2 52.04 16.8824 3.1119 0.0104 0.42743821 27.3 29.7 30.1 30.8 30.6 31.4 33.5 34.6 37.2 38.3 41.2 45.0 48.1 51.3 50.73 18.0837 3.2004 0.0141 0.47443822 28.1 29.1 29.4 29.9 30.8 31.3 31.4 33.7 35.1 37.7 39.8 42.5 46.7 50.1 48.90 17.5758 2.6313 .c0.42133823 .d20000 4801 25.7 26.3 26.7 27.7 28.0 28.1 28.6 30.4 32.5 33.8 36.7 39.0 42.6 45.0 44.82 16.7765 2.6705 0.0084 0.41494802 26.4 29.1 29.1 30.3 30.3 31.3 32.0 34.0 36.8 38.8 40.6 43.6 46.3 48.6 48.83 17.0931 2.8105 0.0141 0.52664803 28.0 30.0 30.4 31.3 31.4 31.4 32.2 34.7 36.9 38.6 41.4 43.9 47.3 50.8 50.04 18.8684 2.4162 0.0127 0.4472Appendix II 3Table A-1. Individual Maternal Body Weights and Organ Weights (g) (page 3 of 4)Gestational DayDoseaDam ID056789101112131415161717bGravidUterineWt.LiverWt.PairedAdrenalGlandWt.PairedKidneyWt.20000 4804 29.7 29.8 30.5 31.0 31.3 31.7 32.6 34.6 37.6 39.0 41.3 43.5 47.0 49.3 49.19 17.6044 2.6289 0.0172 0.42664805 25.7 26.8 27.6 28.3 28.7 28.7 29.3 30.9 33.5 35.4 37.1 40.3 42.7 46.6 45.87 16.9954 2.6732 0.0150 0.43204806 26.4 28.8 29.2 29.8 30.9 31.0 32.4 33.5 35.6 38.2 39.2 42.5 44.9 47.4 47.48 17.0330 2.8556 0.0132 0.41884807 27.7 29.0 28.9 30.1 30.8 30.7 31.6 34.3 36.2 38.3 41.0 44.2 46.9 50.4 49.18 17.5032 2.8624 0.0165 0.41164808 25.1 26.7 27.6 28.3 28.5 28.9 30.2 32.1 34.3 36.2 38.8 41.8 45.9 49.4 48.53 18.8523 2.8895 0.0168 0.41534809 26.0 29.3 29.6 30.8 31.6 32.3 34.0 36.7 39.6 41.8 44.9 47.5 52.5 54.8 55.12 22.6107 2.9528 0.0137 0.44314810 27.0 26.6 27.5 28.2 28.3 28.8 30.0 31.6 33.5 34.2 37.0 39.7 43.2 46.2 45.12 16.4734 2.7402 0.0161 0.39324811 27.5 28.2 29.9 30.9 31.3 32.5 34.3 36.0 38.2 39.1 42.4 43.6 46.4 51.0 49.15 19.6011 2.8417 0.0131 0.39394812 28.2 27.7 28.2 28.2 28.5 29.1 29.8 30.9 33.2 34.3 36.5 38.7 41.0 43.5 42.80 15.8511 2.2713 0.0131 0.37024813 29.0 29.1 29.5 30.5 30.4 31.8 32.9 36.5 39.2 41.1 44.2 47.4 51.8 55.3 53.28 21.9836 2.9297 0.0106 0.47954814 25.6 27.3 27.7 27.8 28.1 28.4 29.7 31.1 32.1 33.6 34.7 37.5 38.2 40.0 38.65 10.0827 2.2418 0.0152 0.40314815 .d4816 27.7 29.2 30.3 30.2 31.2 31.3 32.0 34.0 35.3 37.4 39.6 43.2 45.7 49.0 47.99 17.4621 2.6378 0.0105 0.47264817 28.2 28.0 29.0 29.5 30.3 30.9 32.0 33.6 35.7 37.5 39.6 42.5 45.9 49.2 48.70 17.1257 2.8046 0.0149 0.46534818 30.1 30.6 30.9 31.3 31.3 32.3 31.9 32.0 31.3 31.9 30.6 30.5 30.9 31.4 29.96 0.1257 1.8489 0.0149 0.49314819 26.0 26.5 26.2 26.3 27.2 27.4 27.3 27.6 27.7 27.0 27.7 27.3 27.3 27.0 26.27 0.1142 1.7097 0.0124 0.43254820 27.9 28.6 29.1 29.0 29.4 30.5 31.7 33.2 35.7 36.7 39.3 41.6 44.4 47.7 46.95 15.0383 2.7616 0.0165 0.47974821 27.3 29.3 29.8 30.3 31.2 31.8 32.4 34.6 36.6 38.9 40.8 44.3 47.7 50.5 50.09 18.7231 2.8499 0.0104 0.36624822 27.4 28.7 29.2 29.7 30.1 30.1 31.0 31.5 34.1 35.9 37.4 40.8 44.4 47.4 45.92 15.4136 2.7219 0.0139 0.48554823 28.1 31.5 31.4 32.2 32.1 33.5 34.0 35.6 37.3 39.3 40.7 43.3 44.7 46.7 46.09 14.5411 2.7599 0.0112 0.456030000 5801 .d5802 26.2 27.5 26.8 28.1 28.6 29.5 30.4 32.0 35.1 37.6 40.6 43.6 47.1 50.0 49.36 18.8132 2.2720 0.0147 0.49195803 26.3 27.4 28.0 28.5 29.5 30.0 30.3 33.3 35.3 37.1 39.9 43.5 45.3 48.6 46.62 18.7458 1.9859 0.0092 0.39405804 28.2 29.5 29.5 30.3 31.2 32.0 32.7 34.5 36.5 39.6 42.3 45.8 48.7 50.6 51.20 18.2262 2.4269 0.0115 0.50635805 28.2 28.4 29.3 29.5 30.3 30.9 30.9 32.7 34.5 36.8 39.1 36.0 42.6 46.5 45.64 18.1812 2.0216 0.0131 0.40785806 29.6 30.7 31.0 32.7 33.0 34.2 34.8 36.7 38.5 40.9 44.7 48.6 51.8 55.1 54.57 22.0056 2.3285 0.0131 0.40115807 25.9 26.4 27.1 27.8 29.0 29.6 30.0 31.9 34.7 36.8 40.3 42.8 45.4 48.6 47.78 18.3769 2.3540 0.0126 0.34565808 25.9 26.6 26.7 27.9 29.0 28.9 29.7 32.1 34.1 36.7 39.5 42.0 45.3 47.7 47.72 17.6901 2.3910 0.0138 0.40825809 26.3 26.6 27.2 28.3 29.1 29.3 30.8 32.7 35.4 37.1 40.8 43.6 46.0 49.8 48.81 18.6972 2.2420 0.0139 0.43985810 29.2 29.9 29.9 30.1 31.1 30.7 31.8 33.8 36.7 38.7 41.9 44.6 48.4 51.0 49.73 19.1084 2.5462 0.0119 0.40365811 31.3 31.9 32.1 32.6 33.6 33.8 35.1 37.3 40.3 43.0 47.0 50.6 53.8 57.0 55.58 21.4731 2.8532 0.0148 0.47615812 25.6 27.5 27.3 28.0 28.4 29.4 30.3 31.7 34.2 36.9 38.9 42.0 44.8 47.6 46.86 16.2375 2.5839 0.0155 0.50045813 25.7 26.7 27.0 27.8 27.3 26.5 26.2 26.5 27.1 27.6 27.3 26.7 25.9 27.3 26.21 0.1443 1.7424 0.0151 0.41385814 25.8 28.4 28.4 29.1 29.6 30.1 31.1 32.7 35.7 38.4 40.8 43.5 47.4 50.9 50.04 17.4432 2.8111 0.0134 0.48145815 25.9 26.4 27.3 28.5 28.5 28.8 29.7 31.8 34.2 36.5 38.2 41.2 44.9 47.7 46.86 17.2691 2.5748 0.0122 0.37225816 27.1 28.3 29.4 31.0 31.2 32.3 34.1 35.6 38.4 40.9 43.4 46.8 49.3 51.7 51.43 20.6765 2.6542 0.0140 0.43465817 .dAppendix II 4Table A-1. Individual Maternal Body Weights and Organ Weights (g) (page 4 of 4)Gestational DayDoseaDam ID056789101112131415161717bGravidUterineWt.LiverWt.PairedAdrenalGlandWt.PairedKidneyWt.30000 5818 27.3 27.0 28.1 27.6 27.7 28.7 30.5 32.0 34.6 37.5 39.7 41.4 44.7 48.9 47.49 19.4554 2.1213 0.0136 0.47445819 27.4 29.0 29.6 29.6 30.1 31.2 32.5 34.3 36.4 39.2 42.1 44.9 48.2 51.8 49.80 18.6413 2.5928 0.0122 0.46145820 28.3 28.6 28.8 29.5 29.8 29.9 31.3 32.3 35.3 37.1 39.3 42.6 45.4 48.9 47.34 16.7165 2.3715 0.0106 0.40885821 28.3 29.5 30.5 32.0 32.1 33.3 34.2 35.8 38.2 40.5 41.6 44.5 48.7 51.5 50.76 17.7549 2.8188 0.0123 0.47315822 28.4 28.4 28.3 29.0 28.2 28.5 29.3 30.5 32.8 34.2 36.8 39.3 41.4 44.9 43.85 16.2522 2.0635 0.0116 0.37505823 28.7 29.1 30.0 30.7 31.6 33.1 34.1 36.2 39.2 41.3 44.6 49.0 51.5 55.1 55.01 22.3565 2.3108 0.0144 0.34645824 25.8 24.9 25.5 24.2 23.4 24.0 23.8 24.0 25.2 25.4 25.5 25.2 25.0 25.7 25.39 0.1097 1.4946 0.0172 0.36395825 26.2 26.8 28.5 28.6 29.0 29.8 30.1 31.8 34.4 35.3 36.9 40.3 43.0 46.1 45.25 14.9805 2.3879 0.0143 0.41995826 26.6 27.1 28.3 28.5 28.7 29.2 30.3 32.3 35.8 38.1 41.0 44.1 47.2 50.7 50.20 19.2251 2.7888 0.0131 0.46475827 27.7 26.4 27.2 27.1 27.8 28.1 29.0 31.1 33.7 35.4 37.0 39.9 42.9 46.6 45.81 15.3833 2.3213 0.0143 0.37845828 27.8 28.5 29.3 28.9 29.1 29.9 30.9 32.9 36.5 37.6 39.1 42.7 44.2 47.7 46.68 15.6981 2.2852 0.0151 0.43365829 28.1 28.7 29.1 29.0 29.5 30.1 31.1 33.0 35.4 36.2 38.2 41.1 44.1 47.0 45.62 14.9195 2.4199 0.0111 0.48035830 30.3 33.4 33.1 33.0 33.5 34.3 35.0 37.2 40.1 42.8 45.4 50.5 53.5 58.0 56.48 20.2425 3.0487 0.0150 0.43655831 26.3 28.0 28.2 28.7 29.0 29.5 30.1 33.4 35.4 37.1 39.8 42.9 46.4 49.5 48.76 18.3770 2.6777 0.0132 0.46565832 26.9 30.0 30.2 31.4 32.2 33.2 35.0 37.4 39.3 41.4 44.0 47.3 51.1 54.7 54.13 20.3232 3.2612 0.0151 0.47215833 27.0 27.5 27.6 28.4 28.7 29.2 30.4 32.1 34.5 36.0 38.5 41.3 44.6 47.8 47.02 16.3016 2.7776 0.0161 0.46445834 27.9 27.3 28.0 28.2 29.5 29.1 31.2 32.5 33.6 36.2 38.0 39.7 42.6 45.9 44.84 14.7283 2.3049 0.0131 0.42995835 27.7 30.2 30.4 31.3 31.9 33.4 35.7 37.9 39.7 42.8 45.4 49.3 51.9 56.8 56.05 22.1501 2.7876 0.0131 0.40395836 28.1 28.4 29.3 30.1 30.5 31.6 33.1 34.4 36.6 38.5 41.1 44.9 48.9 52.4 51.13 19.7858 2.3708 0.0158 0.44205837 28.3 29.1 30.0 30.0 30.2 29.9 30.1 29.7 29.9 30.3 30.2 29.7 30.1 31.2 29.40 0.1544 1.8195 0.0131 0.45125838 28.0 28.1 28.7 28.9 30.0 31.3 31.5 32.8 35.1 37.7 39.7 43.1 46.7 49.2 48.33 17.4607 2.5002 0.0145 0.3965aMg/m3of gasoline MTBE vapor condensate.bBody weight at sacrifice.cPaired adrenal gland weight was a statistical outlier and therefore it was excluded.dFemale was not pregnant.eFemale was removed due to a preexisting condition. At necropsy she was found to have an undescended testis on the right and seminal vesicles and prostate tothe right of the vagina and cervix.fLiver weight was inadvertently recorded to only 3 decimal places.Appendix II 5

Table A-2. Individual Maternal Clinical Observations (page 1 of 1)

Dose a Dam ID Day b Time c Observation
10000 3808 12 Prior Red exudate from anogenital area

20000 4803 6 Post Lacrimation, unilateral, moderate 4807 5 Post Alopecia: extremities/snout, moderate 6 Prior Alopecia: extremities/snout, moderate Post Alopecia: extremities/snout, moderate 7 Prior Alopecia: extremities/snout, moderate Post Alopecia: extremities/snout, moderate 8 Prior Alopecia: extremities/snout, moderate Post Alopecia: extremities/snout, moderate 9 Prior Alopecia: extremities/snout, moderate Post Alopecia: extremities/snout, moderate 10 Prior Alopecia: extremities/snout, moderate Post Alopecia: extremities/snout, moderate 11 Prior Alopecia: extremities/snout, moderate Post Alopecia: extremities/snout, moderate 12 Prior Alopecia: extremities/snout, moderate Post Alopecia: extremities/snout, moderate 4814 11 Post Red exudate from anogenital area 4823 9 Post Labored breathing

30000 5802 6 Post Lacrimation, unilateral, moderate 5805 6 Post Lacrimation, bilateral, moderate 5824 5 Post Lacrimation, unilateral, moderate 5829 2 Prior Eye: enophthalmos, unilateral, left 3 Prior Eye: enophthalmos, unilateral, left 4 Prior Eye: enophthalmos, unilateral, left 5 Prior Eye: enophthalmos, unilateral, left Post Eye: enophthalmos, unilateral, left 6 Prior Eye: enophthalmos, unilateral, left Post Eye: enophthalmos, unilateral, left 7 Prior Eye: enophthalmos, unilateral, left Post Eye: enophthalmos, unilateral, left 8 Prior Eye: enophthalmos, unilateral, left Post Eye: enophthalmos, unilateral, left 9 Prior Eye: enophthalmos, unilateral, left Post Eye: enophthalmos, unilateral, left 10 Prior Eye: enophthalmos, unilateral, left Post Eye: enophthalmos, unilateral, left 11 Prior Eye: enophthalmos, unilateral, left 12 Prior Eye: enophthalmos, unilateral, left 13 Prior Eye: enophthalmos, unilateral, left 14 Prior Eye: enophthalmos, unilateral, left 15 Prior Eye: enophthalmos, unilateral, left 16 Prior Eye: enophthalmos, unilateral, left 17 Prior Eye: enophthalmos, unilateral, left 5838 10 Post Labored breathing

a Mg/m 3 of gasoline MTBE vapor condensate. b Gestational day.
c Time is prior to dosing (Pri or) or after dosing (Post).Appendix II 6Table A-3. Individual Maternal Feed Consumption (g/day)a(page 1 of 5)Gestational DaysDosebDam ID 0-5 5-6 6-7 7-8 8-9 9-1010-11 11-12 12-13 13-14 14-15 15-16 16-17 5-10c5-16d10-17c0-170 1801 .e5.6 6.4 6.8 7.4 6.88.8 8.6 7.3 7.8 8.0 8.0 8.8 6.6 7.4 8.2 .f1802 5.5 6.0 6.2 6.2 6.86.9 7.2 7.6 7.5 7.3 7.4 8.7 8.56.4 7.1 7.7 6.71803 .e.e.e.g10.4 8.5 6.8 6.6 6.7 8.6 7.4 7.4 9.4 .f.f7.6 .f1804 5.6 5.1 5.8 5.8 6.85.8 9.4 8.2 7.0 6.8 7.2 7.9 7.95.9 6.9 7.8 6.61805 7.3 6.2 6.4 6.5 6.5 6.5 .e9.8 6.9 6.8 7.1 7.3 7.4 6.4 .f.f.f1806 4.9 4.9 5.9 5.7 5.75.6 6.1 6.0 6.5 6.2 7.5 7.1 7.15.6 6.1 6.6 5.81807 6.0 5.8 7.9 7.3 9.86.3 6.7 6.7 7.0 7.4 7.1 7.0 6.87.4 7.2 7.0 6.81808 5.4 5.2 5.4 5.5 5.65.9 5.2 5.5 5.7 6.6 6.2 6.1 6.65.5 5.7 6.0 5.71809 5.0 5.4 5.5 6.0 5.46.1 5.7 6.2 6.1 6.2 6.4 6.2 6.45.7 5.9 6.2 5.71810 6.5 8.7 7.1 6.1 6.66.7 7.7 6.6 6.4 7.9 7.3 7.0 7.97.0 7.1 7.3 7.01811 .e.g.g6.9 14.5 8.0 7.3 11.3 7.6 7.0 7.3 8.28.5 10.4 9.1 8.2 .f1812 .e4.1 5.1 .g.e6.7 .e5.6 5.8 7.58.2 6.6 7.1 .f.f.f.f1813 5.3 6.6 6.1 8.8 8.36.0 6.3 5.8 6.8 8.4 6.9 8.2 8.07.2 7.1 7.2 6.61814 .e8.4 .g9.0 9.7 9.7 .e8.3 8.2 8.1 7.9 7.9 8.1 10.6 .f.f.f1815 .e6.0 6.8 7.2 8.2 7.17.3 6.9 7.1 7.9 6.9 7.5 10.2 7.1 7.2 7.7 .f1816 7.4 8.1 .g9.7 10.1 8.6 .e10.2 7.3 7.6 8.2 6.5 7.9 10.8 .f.f.f1817 7.5 .e.e12.2 .e8.3 8.1 11.0 9.26.5 7.9 7.6 5.6 .f.f8.0 .f1818 7.4 7.1 5.4 6.0 5.65.8 6.4 10.8 9.57.9 7.1 8.4 7.86.0 7.3 8.3 7.31819 5.0 6.1 5.5 6.7 5.26.0 6.6 7.2 8.1 7.1 8.2 8.9 7.25.9 6.9 7.6 6.31820 5.2 6.0 6.3 5.8 6.55.7 6.3 7.4 7.1 7.1 7.1 7.6 6.96.1 6.6 7.1 6.21821 7.7 4.4 6.5 7.3 7.86.1 6.5 7.9 6.2 6.3 8.0 7.2 6.66.4 6.7 7.0 7.01822 5.7 5.5 5.2 5.8 5.46.0 6.5 5.8 6.3 7.3 7.3 6.9 7.35.6 6.2 6.8 6.11823 6.2 6.3 5.7 5.7 6.95.6 6.1 7.0 7.0 7.0 6.8 7.5 7.06.0 6.5 6.9 6.52000 2801 5.8 6.1 5.36.4 7.6 6.3 6.2 6.2 6.3 6.1 6.2 5.6 5.96.26.12802 6.3 6.9 9.5 8.0 6.5 8.3 7.0 6.9 7.1 7.0 6.0 7.8 8.07.47.12803 5.3 5.8 6.2 9.54.5 6.0 7.5 6.7 6.2 .e.e7.0 7.4.f.f2804 .e5.5 11.2 6.6 7.77.5 6.5 8.4 7.6 7.4 6.9 6.3 7.27.4.f2805 4.7 4.8 5.5 6.2 6.5 6.2 6.1 6.7 6.9 6.6 6.9 7.3 6.76.35.92806 .h7.7 16.0 8.2 .h6.8 7.5 8.0 6.1 .h6.8 7.9 7.3.f.f2807 .h8.1 .g10.9 .g10.6 10.6 .e7.5 7.1 6.3 8.2 6.9.f.f2808 7.9 9.5 8.2 7.5 6.9 7.2 9.1 8.1 7.7 8.1 9.0 8.5 7.98.2 8.12809 7.4 5.9 5.6 6.7 5.7 7.2 8.4 7.3 6.9 6.1 8.8 6.9 7.7 6.9 7.12810 7.5 6.0 7.1 7.7 6.8 6.9 7.6 7.2 8.0 7.1 9.3 8.1 7.6 7.4 7.5Appendix II 7Table A-3. Individual Maternal Feed Consumption (g/day)a(page 2 of 5)Gestational DaysDosebDam ID 0-5 5-6 6-7 7-8 8-9 9-1010-11 11-12 12-13 13-14 14-15 15-16 16-17 5-10c5-16d10-17c0-172000 2811 5.2 5.5 6.3 5.86.3 6.0 6.0 6.2 6.3 6.3 7.1 9.86.0 6.5 6.12812 8.2 .e12.8 9.8 11.4 8.08.1 7.8 11.4 11.28.8 9.0 9.3 .f.f2813 4.3 5.2 5.7 5.5 4.3 5.2 4.6 4.8 4.3 4.4 4.4 4.9 4.3 4.8 4.62814 6.6 8.2 6.0 6.2 6.7 6.8 7.5 7.9 7.4 7.3 7.5 8.1 8.6 7.2 7.12815 9.5 9.0 .e8.2 8.3 7.5 9.3 8.8 7.0 9.0 8.0 8.4 7.3 .f.f2816 7.7 7.2 7.6 7.7 7.9 7.0 7.3 7.6 7.5 9.0 7.7 9.0 8.8 7.8 7.82817 4.7 5.3 5.2 5.9 5.5 5.5 5.4 5.9 5.7 6.8 5.5 5.9 6.3 5.7 5.42818 4.9 6.5 6.2 7.2 6.3 .e8.6 6.9 7.3 7.87.5 7.5 7.3 .f.f2819 5.5 7.0 6.9 7.1 7.2 6.9 10.2 7.3 8.7 8.4 8.3 8.5 8.4 7.9 7.22820 8.2 8.2 9.5 7.4 7.4 7.9 8.4 7.7 8.6 8.2 8.5 8.7 8.7 8.2 8.32821 .i2822 .h5.0 7.0 6.5 6.5 5.5 7.3 5.9 .h7.9 6.6 8.3 6.8 .f.f2823 7.8 6.7 .h.g.g7.9 7.4 7.1 8.6 7.8 9.3 9.1 9.0 .f.f10000 3801 5.5 5.6 5.8 4.96.9 5.7 6.8 6.8 7.2 7.6 7.3 6.98.8 6.5 6.43802 .i3803 .e.e8.4 10.8 .h6.1 6.7 .e12.0 8.0 7.9 7.3 8.1 .f.f3804 .e5.9 9.4 6.8 6.6 8.1 7.4 7.3 8.3 10.3 8.2 8.2 8.4 7.9 .f3805 .e.e10.0 8.1 7.7 7.38.9 8.1 7.7 7.77.8 8.0 7.0 .f.f3806 6.7 6.8 11.4 8.2 8.0 7.2 8.1 7.7 8.3 8.9 7.4 10.2 8.3 8.4 7.93807 4.9 5.8 6.5 6.0 6.2 6.0 6.3 6.8 7.0 7.1 6.1 7.3 7.1 6.5 6.03808 .e6.0 8.1 6.9 9.3 8.4 6.3 10.7 6.9 6.9 7.2 6.6 6.9 7.6 .f3809 6.0 4.9 5.6 5.36.5 7.7 6.6 7.9 8.4 .e6.6 7.3 6.7 .f.f3810 8.9 13.0 8.8 7.4 7.2 7.0 7.2 7.6 8.3 7.7 8.6 7.6 8.7 8.2 8.53811 7.9 10.9 .e9.7 7.0 8.5 7.1 6.9 7.2 7.3 7.4 7.1 7.4 .f.f3812 5.4 6.0 5.7 6.0 5.9 6.4 6.4 6.5 7.0 7.4 8.1 7.5 7.2 6.6 6.33813 .e9.0 9.2 6.9 .e7.4 7.7 7.5 7.3 8.0 7.5 7.6 7.0 .f.f3814 .j3815 .e14.5 8.7 12.4 11.8 8.1 8.6 8.9 8.4 9.0 9.1 9.0 8.7 9.9 .f3816 5.8 6.6 6.0 6.4 10.4 6.2 9.5 7.3 6.7 8.3 6.8 7.4 7.3 7.4 6.93817 .i3818 .e.e.e11.9 .h.e9.5 9.1 8.4 7.610.5 11.7 7.9 .f.f3819 5.2 5.4 5.8 5.9 5.2 6.7 .e6.2 7.7 6.8 6.7 6.8 7.1 .f.f3820 .e9.3 8.1 17.8 8.9 8.9 7.7 9.1 8.7 7.7 7.9 8.9 9.2 9.4 .fAppendix II 8Table A-3. Individual Maternal Feed Consumption (g/day)a(page 3 of 5)Gestational DaysDosebDam ID 0-5 5-6 6-7 7-8 8-9 9-1010-11 11-12 12-13 13-14 14-15 15-16 16-17 5-10c5-16d10-17c0-1710000 3821 6.5 7.6 7.2 6.47.0 8.0 7.5 7.6 7.6 9.4 9.4 8.78.3 7.9 7.53822 5.1 5.3 5.8 7.3 5.8 7.0 6.8 6.0 9.6 6.9 7.2 6.8 7.4 6.8 6.33823 .i20000 4801 4.2 5.2 6.2 5.95.3 6.1 5.9 5.7 6.1 6.4 7.2 6.58.0 6.0 5.64802 5.7 6.1 6.5 6.0 6.6 6.5 7.1 7.5 7.8 7.7 8.4 6.8 8.3 7.0 6.74803 .e.g.g.g8.4 6.7 .g.g8.8 10.2 7.7 7.4 9.9 10.2 .f4804 5.1 5.0 6.1 5.6 5.5 5.5 5.5 6.4 6.2 6.6 6.9 7.1 8.9 6.0 5.94805 5.4 5.9 6.3 6.0 5.7 5.6 6.0 6.3 6.5 6.9 6.1 7.1 6.8 6.2 6.04806 .e6.0 .g6.1 6.4 6.8 6.2 .e7.1 7.2 6.4 7.7 6.2 .f.f4807 .e7.8 9.3 .g.g5.7 .g6.7 7.4 7.9 7.6 8.4 7.0 8.6 .f4808 4.8 5.6 6.0 5.7 5.6 5.5 5.7 6.7 6.7 6.5 7.2 7.0 7.0 6.2 5.84809 6.2 5.9 6.8 7.5 5.8 6.6 6.8 6.8 7.1 6.9 9.1 7.1 6.4 6.9 6.74810 .e7.3 5.0 7.1 5.2 7.3 6.3 7.1 7.3 6.8 8.4 6.3 6.9 6.7 .f4811 4.4 5.8 6.0 6.3 6.5 5.7 5.8 6.1 6.1 6.3 6.7 5.5 7.7 6.1 5.74812 4.0 4.9 4.7 5.1 5.0 5.1 5.1 5.0 5.2 5.3 7.7 5.9 6.5 5.4 5.04813 5.6 6.1 6.0 6.7 6.7 6.6 7.4 7.4 7.5 7.9 9.7 8.4 8.1 7.3 6.84814 5.3 9.8 .e5.8 6.7 6.2 6.3 6.8 6.1 7.7 6.1 6.3 6.8 .f.f4815 .i4816 5.4 7.1 6.7 6.2 6.5 6.4 6.9 7.2 6.9 8.8 7.6 7.8 8.0 7.1 6.74817 5.4 5.7 7.0 6.4 5.9 6.0 5.7 6.8 6.3 8.2 6.4 7.2 7.1 6.5 6.24818 5.8 6.7 6.7 5.1 6.5 5.2 6.4 6.1 6.2 7.6 4.2 5.7 6.4 6.0 6.04819 4.9 5.0 4.3 5.3 5.2 5.6 6.7 5.5 7.0 5.2 4.4 4.9 4.1 5.4 5.24820 .h6.2 .e.e.e.e8.3 8.6 8.3 .e8.1 7.4 7.1 .f.f4821 .e5.5 5.6 6.1 5.3 .h5.2 7.1 5.1 6.96.9 6.7 7.1 .f.f4822 6.1 6.5 6.8 8.0 7.8 6.2 7.7 6.9 7.6 6.9 7.2 7.8 8.7 7.2 7.04823 6.0 6.6 .g.e6.2 7.6 5.7 7.3 7.6 5.6 7.0 6.1 6.9 .f.f30000 5801 .i5802 5.4 4.7 5.5 5.4 5.6 6.7 6.5 11.9 7.3 7.7 6.9 7.1 7.7 5.6 7.9 6.55803 5.3 5.9 5.9 5.7 5.6 6.1 6.1 6.7 6.6 7.2 7.1 6.7 8.2 5.8 6.9 6.15804 5.2 5.4 6.5 5.8 5.6 6.9 6.1 7.2 7.9 7.4 7.6 7.7 7.4 6.0 7.3 6.35805 .h5.7 5.8 5.7 5.5 5.8 5.2 6.0 8.1 7.2 3.9 6.8 7.6 5.7 6.4 .fAppendix II 9Table A-3. Individual Maternal Feed Consumption (g/day)a(page 4 of 5)Gestational DaysDosebDam ID 0-5 5-6 6-7 7-8 8-9 9-1010-11 11-12 12-13 13-14 14-15 15-16 16-17 5-10c5-16d10-17c0-1730000 5806 .h5.9 7.0 6.2 6.2 6.6 .h.e8.3 7.2 6.4 7.0 8.0 6.4 .f.f5807 7.6 6.0 6.3 6.8 6.6 6.2 5.9 7.9 7.4 7.4 8.1 7.2 7.1 6.4 7.3 7.15808 8.3 5.3 6.4 .h5.0 6.8 6.2 7.1 6.2 7.4 8.1 6.5 .h.f.f.f5809 .h6.1 5.7 6.1 .h6.1 5.5 8.4 .h10.3 8.0 7.8 8.1 .f.f.f5810 5.4 6.5 5.0 5.3 5.7 6.9 6.7 6.5 7.6 7.3 7.3 6.8 7.9 5.9 7.2 6.35811 5.6 6.3 7.9 7.0 7.0 6.0 6.9 7.3 8.2 8.1 8.3 8.6 9.0 6.8 8.1 7.05812 9.4 5.7 5.6 6.4 7.0 6.4 6.4 9.7 7.8 8.5 10.0 8.8 8.8 6.2 8.6 8.15813 .e.g6.2 7.3 6.5 6.8 6.3 7.2 5.9 6.5 5.0 3.7 4.8 7.6 5.6 .f5814 5.3 5.7 5.2 6.5 6.4 5.8 6.5 7.0 9.6 8.0 7.4 8.2 8.2 5.9 7.8 6.55815 10.4 6.0 6.2 5.8 6.2 5.7 5.9 6.9 7.9 7.8 7.8 7.0 6.5 6.0 7.1 7.75816 6.2 6.5 8.9 7.1 6.1 6.5 6.1 7.4 6.8 8.2 8.3 7.0 7.1 7.0 7.3 6.95817 .i5818 4.7 6.3 4.6 6.3 5.85.7 6.7 7.2 6.6 7.7 5.6 .h6.8 5.7 .f.f5819 6.4 .e.e7.4 6.4 6.2 .g7.6 7.9 8.38.4 8.1 7.6 .f8.4 .f5820 9.0 5.1 5.4 6.9 5.3 5.6 5.6 6.3 11.4 8.5 7.2 7.2 7.1 5.7 7.6 7.45821 11.1 6.8 6.57.7 7.2 8.0 7.2 .g7.3 7.8 8.1 8.6 8.5 7.2 8.9 9.05822 5.2 5.4 4.6 7.0 6.5 6.5 4.9 5.4 5.3 6.9 6.7 6.2 6.6 6.0 6.0 5.85823 5.7 6.7 5.3 6.6 5.8 6.1 6.2 7.2 6.9 8.7 8.8 8.1 8.3 6.1 7.7 6.75824 .e4.7 4.7 3.9 5.0 5.2 4.7 6.8 6.2 6.3 6.6 3.7 4.2 4.7 5.5 .f5825 5.1 5.9 6.7 6.2 5.9 5.5 5.8 6.4 7.7 7.1 9.0 7.4 6.7 6.0 7.2 6.25826 .e6.2 7.4 5.8 6.2 6.6 6.3 8.7 8.6 9.4 9.1 7.7 7.5 6.4 8.2 .f5827 4.5 4.5 5.6 5.0 4.9 5.6 5.3 6.4 6.8 7.5 7.4 6.9 7.3 5.1 6.8 5.65828 5.3 6.0 6.8 6.2 6.4 7.1 6.3 8.0 8.3 8.4 8.3 7.0 7.5 6.5 7.7 6.65829 10.0 5.7 .e.g7.0 7.4 7.0 9.2 10.2 8.8 8.8 8.8 8.6 .f8.8 .f5830 7.2 6.6 .e9.3 6.6 6.5 6.6 13.8 9.1 9.0 8.5 8.8 8.3 .f9.2 .f5831 .e6.7 6.5 6.0 6.2 5.7 6.5 8.6 7.5 8.5 9.2 7.1 8.2 6.2 7.9 .f5832 7.5 7.7 8.4 7.7 6.9 6.8 6.8 8.8 8.5 8.1 9.2 8.8 8.9 7.5 8.4 7.95833 .e.g10.3 8.3 .e.e.g9.6 9.4 9.67.9 6.9 6.8 .f9.5 .f5834 .h4.5 .h7.7 .h6.0 6.0 .h.g10.1 6.0 6.2 8.5 .f.f.f5835 6.3 5.0 6.4 5.7 8.8 6.9 7.6 9.6 8.3 8.8 7.9 7.8 7.2 6.6 8.2 7.25836 .e8.1 6.8 5.8 5.9 6.4 6.6 8.2 7.9 8.4 8.7 7.2 7.1 6.6 7.7 .f5837 6.6 6.3 5.8 6.1 5.5 5.2 6.1 6.6 5.5 6.8 6.0 4.6 4.9 5.8 5.8 6.05838 .e.e6.1 5.3 5.7 6.4 5.7 6.9 8.5 7.7 7.4 7.5 7.9 .f7.4 .fAppendix II 10Table A-3. Individual Maternal Feed Consumption (g/day)a(page 5 of 5)aThese data represent the difference between the new feed jar weights (beginning weight; full feed jar) and the old feed jar (end weight; after theanimal has fed for the measurement interval) weights divided by the number of days inthe measurement interval.bMg/m3of gasoline MTBE vapor condensate.cThis endpoint was only calculated for the 0 and 30000 mg/m3dose groups.dThis endpoint was only calculated for the 0, 2000, 10000 and 20000 mg/m3dose groups.eFeed spilled therefore the feed weight was excluded.fInterim feed consumption value(s) are missing and thereforethe overall feed consumption value could not be calculated.gFeed consumption value was a statisticaloutlier and therefore it was excluded.hFeed was contaminated thereforethe feed weight was excluded.iFemale was not pregnant.jFemale was removed due to a preexisting condition. At necropsy she was found to have an undescended testis on the right and seminal vesiclesand prostate to the right of the vagina and cervix.Appendix II 11

Table A-4. Individual Embryo/Fetal Data (page 1 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

0 1801 11 1 A L 1 F 1.0230 2 A L 2 M 0.8182 3 A L 3 M 0.9328 4 D L 0.7402 5 E R 6 A R 4 M 0.9507 7 A R 5 M 0.9290 8 A R 6 F 0.9828 9 A R 7 F 1.0268 10 A R 8 M 0.9286 11 A R 9 M 0.9976 External Variation Hematoma: Head External Variation Hematoma: Neck 1802 13 1 A L 1 F 1.0746 2 A L 2 M 1.0913 3 A L 3 F 1.0557 4 A L 4 M 1.0646 5 A R 5 M 1.0013 6 A R 6 F 1.0405 7 A R 7 F 0.9931 8 A R 8 F 1.0460 9 A R 9 M 1.1404 10 A R 10 M 1.1569 11 A R 11 M 1.0598 12 A R 12 F 1.0618 13 A R 13 F 1.0505 1803 9 1 A L 1 F 1.2866 2 A L 2 F 1.4021 3 E L 4 A R 3 M 1.3324 5 A R 4 F 1.2360 6 A R 5 F 0.9690 External Malformation Cleft Palate 7 A R 6 F 1.1523 External Variation Hematoma: Head 8 A R 7 F 1.2401 9 A R 8 M 1.3432 1804 14 1 A L 1 F 1.0263 2 A L 2 M 1.0068 3 A L 3 M 0.9753 4 A L 4 F 0.8615 5 A L 5 F 0.9291 6 A R 6 M 1.0035 7 A R 7 M 0.9158 8 A R 8 M 0.9942 9 A R 9 F 0.9321 10 A R 10 F 0.9239 11 A R 11 M 0.9972 12 A R 12 F 0.9071 13 A R 13 F 0.9282 1805 13 1 A L 1 M 1.0623 2 A L 2 M 1.0939 3 A L 3 M 1.0992 4 A L 4 M 1.0797 5 A L 5 F 1.0217 6 A R 6 F 1.0826 7 A R 7 F 1.0288 8 A R 8 M 1.0531 9 A R 9 F 1.0119 10 A R 10 M 1.0624 11 A R 11 F 0.8523 12 A R 12 F 1.0232Appendix II 12

Table A-4. Individual Embryo/Fetal Data (page 2 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

0 1806 14 1 A L 1 F 1.0370 2 A L 2 M 1.0681 3 A L 3 M 0.8841 4 A L 4 M 0.9099 5 A L 5 M 0.9751 6 A L 6 M 0.7932 7 A L 7 M 0.9739 8 A L 8 M 1.0688 9 A R 9 M 1.1047 10 A R 10 M 0.9852 11 A R 11 F 0.9760 12 A R 12 M 1.0491 13 A R 13 F 1.0917 1807 15 1 A L 1 M 0.8745 2 A L 2 F 0.9240 3 A L 3 M 0.8271 4 A L 4 F 0.9596 5 A L 5 M 0.8050 6 A L 6 M 0.9051 7 A L 7 M 0.8972 8 A L 8 M 0.9840 9 A R 9 M 0.9552 10 A R 10 M 0.9733 11 E R 12 A R 11 M 1.0288 13 A R 12 F 0.9371 14 A R 13 F 0.8945 15 A R 14 M 0.9640 1808 15 1 A L 1 F 0.8116 2 A L 2 M 0.8444 3 A L 3 M 0.8401 4 A L 4 M 0.7738 5 A L 5 F 0.7672 6 A L 6 F 0.8215 7 A R 7 F 0.8106 8 A R 8 M 0.7823 9 A R 9 M 0.7679 10 A R 10 M 0.6397 11 A R 11 F 0.7671 12 A R 12 M 0.8271 13 A R 13 M 0.8018 14 A R 14 M 0.7566 15 A R 15 M 0.8540 1809 11 1 A L 1 F 0.9641 2 A L 2 F 1.0304 3 A R 3 M 0.6548 4 A R 4 M 0.8226 5 A R 5 F 0.8699 6 A R 6 F 0.8221 7 A R 7 M 0.8814 8 A R 8 M 0.9828 9 A R 9 F 0.8574 10 A R 10 F 0.8405 11 A R 11 F 0.8094Appendix II 13

Table A-4. Individual Embryo/Fetal Data (page 3 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

0 1810 13 1 A L 1 M 1.1877 2 A L 2 M 0.9171 3 A L 3 M 1.1916 4 A L 4 F 1.1147 5 E R 6 E R 7 A R 5 F 1.1019 8 A R 6 F 1.2054 9 A R 7 F 1.0496 10 A R 8 F 1.1505 11 E R 12 A R 9 M 1.0525 1811 14 1 A L 1 M 1.1393 2 A L 2 F 1.0167 3 A L 3 M 1.0642 4 A L 4 M 1.0878 5 A L 5 F 1.0595 6 A L 6 M 1.0034 7 A L 7 M 0.9544 8 A L 8 M 0.9429 9 A L 9 M 1.0414 10 A R 10 M 1.1475 11 A R 11 M 1.0808 12 A R 12 F 1.0640 13 A R 13 F 1.0829 14 A R 14 M 1.0891 1812 13 1 A L 1 F 0.9006 2 A L 2 M 0.8712 3 A L 3 F 0.8832 4 A R 4 F 0.8264 5 A R 5 F 0.8169 6 A R 6 F 0.7940 7 A R 7 M 0.8558 8 A R 8 M 0.8438 9 A R 9 F 0.8537 10 A R 10 F 0.7710 11 A R 11 F 0.8807 12 A R 12 M 0.8740 13 A R 13 M 0.8937 1813 11 1 A L 1 F 1.0101 2 A L 2 M 1.1114 3 A L 3 M 1.1098 4 A L 4 M 1.0110 5 A L 5 M 0.9292 6 A L 6 F 1.0585 7 A L 7 M 1.0834 8 A R 8 M 1.1358 9 E R 10 A R 9 F 1.0958 11 A R 10 F 1.0459Appendix II 14

Table A-4. Individual Embryo/Fetal Data (page 4 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

0 1814 14 1 A L 1 F 0.9528 2 A L 2 F 0.7076 3 A L 3 F 0.9589 4 A L 4 F 1.0443 5 A L 5 M 1.0409 6 A L 6 F 1.0072 7 A R 7 M 0.9022 8 A R 8 F 0.9146 9 A R 9 F 0.8631 10 D R 0.6018 11 A R 10 F 0.9350 12 A R 11 M 0.9835 13 E R 14 A R 12 F 1.0240 1815 11 1 A L 1 M 1.0836 2 A L 2 F 0.9789 3 E L 4 A L 3 M 1.0703 5 A L 4 M 0.9151 6 A L 5 M 1.0118 7 A L 6 F 1.1307 8 A R 7 F 1.0776 9 A R 8 M 1.0655 10 A R 9 F 1.1202 11 A R 10 F 0.9812 12 A R 11 M 1.1028 1816 14 1 A L 1 M 0.9686 External Malformation Cleft Palate 2 A L 2 M 1.0881 3 A L 3 M 1.0086 4 A L 4 F 0.9104 5 A L 5 F 0.9647 6 A L 6 M 1.1311 7 A L 7 F 1.1062 8 A R 8 F 1.0862 9 A R 9 F 1.0008 10 A R 10 F 1.0492 11 A R 11 F 1.0072 12 A R 12 M 1.1214 13 A R 13 M 1.1400 14 A R 14 M 1.0859 1817 11 1 A L 1 M 0.8516 2 A L 2 M 0.8587 3 A L 3 M 0.8686 4 A L 4 F 0.8578 5 A L 5 F 0.9120 6 A L 6 F 0.8655 7 A R 7 F 0.8314 8 A R 8 F 0.9139 9 A R 9 F 1.0210 10 E R 11 A R 10 M 0.7935Appendix II 15

Table A-4. Individual Embryo/Fetal Data (page 5 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

0 1818 17 1 A L 1 M 1.3283 2 A L 2 M 1.4374 3 A L 3 F 1.2843 4 A L 4 F 1.3751 5 A L 5 M 1.4098 6 E R 7 A R 6 F 1.3769 8 A R 7 F 1.1389 9 D R 0.8652 10 A R 8 M 1.2370 11 A R 9 F 0.9656 12 A R 10 F 1.1105 13 A R 11 F 1.1893 14 A R 12 F 1.3430 1819 13 1 A L 1 F 1.2033 2 A L 2 F 1.2014 3 A L 3 M 1.2734 4 A L 4 M 1.2249 5 A L 5 M 1.2560 6 A L 6 F 1.2349 7 A L 7 M 1.3133 8 A L 8 M 1.2207 9 A R 9 M 1.1921 10 A R 10 M 1.2161 11 A R 11 M 1.2267 12 A R 12 F 1.1668 13 A R 13 F 1.2489 1820 15 1 A L 1 M 1.1172 2 A L 2 M 0.9227 3 A L 3 F 0.9747 4 E L 5 E L 6 A L 4 F 0.8975 7 A L 5 F 0.9357 8 A R 6 M 0.9353 External Variation Hematoma: Shoulder 9 A R 7 M 0.9363 10 A R 8 M 0.9574 11 A R 9 F 0.9115 12 A R 10 M 0.9384 13 A R 11 F 0.9389 14 A R 12 F 0.8823 15 A R 13 F 0.9283 16 A R 14 F 0.7267 1821 11 1 A L 1 M 0.8618 2 A L 2 M 0.9622 3 A L 3 F 0.9537 4 A R 4 M 0.7376 5 A R 5 M 0.8339 6 A R 6 F 0.8517 7 A R 7 M 0.8706 8 A R 8 M 0.9207 9 A R 9 F 0.8109 10 A R 10 M 0.7936 11 A R 11 F 0.9996Appendix II 16

Table A-4. Individual Embryo/Fetal Data (page 6 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

0 1822 12 1 A L 1 F 0.9341 2 A L 2 F 0.9481 3 A L 3 M 0.9897 4 A L 4 M 1.0243 5 A L 5 M 1.0367 6 A R 6 F 0.9537 7 A R 7 F 0.8929 8 A R 8 M 0.8955 9 A R 9 F 0.8876 10 A R 10 F 0.9153 11 A R 11 M 0.9270 12 A R 12 F 0.9892 1823 14 1 A L 1 F 0.7610 2 A L 2 M 0.9371 3 A L 3 F 1.0011 4 E L 5 A L 4 F 0.9726 6 A R 5 F 0.8709 7 A R 6 F 0.9728 8 A R 7 F 0.9841 9 A R 8 M 0.9371 10 A R 9 F 0.9246 11 A R 10 F 0.7944 12 A R 11 M 0.9476 13 A R 12 M 0.8361 14 A R 13 M 0.9707

2000 2801 5 1 A R 1 F 1.3441

2802 8 1 A L 1 M 1.2708 2 A L 2 F 1.0697 3 E L 4 E L 5 E L 6 A R 3 M 1.2216 7 A R 4 M 1.2408 8 E R 9 A R 5 F 1.0764 10 A R 6 M 1.1545 11 A R 7 F 1.0262 12 E R 13 A R 8 F 1.0773 2803 15 1 A L 1 M 0.9750 2 A L 2 M 1.0125 3 A L 3 F 1.0308 4 A L 4 M 1.0001 5 A L 5 M 1.0323 6 A L 6 F 0.9611 7 A R 7 M 1.0954 External Variation Hematoma: Head External Variation Hematoma: Neck 8 A R 8 M 1.1073 9 A R 9 F 0.9910 10 A R 10 M 0.9514 11 A R 11 M 0.9946 12 A R 12 M 1.0013 13 A R 13 F 1.0459 14 A R 14 F 0.9746 15 A R 15 F 1.0812Appendix II 17

Table A-4. Individual Embryo/Fetal Data (page 7 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

2000 2804 12 1 A L 1 F 0.9064

2 A L 2 M 0.9034 3 A R 3 M 0.9715 4 A R 4 F 0.8306 5 A R 5 F 0.7990 6 A R 6 F 0.8979 7 A R 7 M 0.7550 8 A R 8 M 0.8960 9 A R 9 F 0.7931 10 A R 10 F 0.7344 11 A R 11 F 0.8566 12 A R 12 M 0.9062 2805 12 1 A L 1 M 0.9858 2 A L 2 F 0.9488 3 A L 3 F 0.7516 4 A L 4 M 0.9525 5 A R 5 F 0.8988 6 A R 6 M 0.9142 7 A R 7 M 1.0188 8 A R 8 F 0.8397 9 A R 9 M 0.7913 10 A R 10 M 0.8964 11 A R 11 F 0.8909 12 A R 12 M 1.0129 2806 12 1 A L 1 M 1.0891 2 A L 2 M 1.0178 3 A L 3 M 1.0216 4 A R 4 F 0.9649 5 A R 5 M 0.9381 6 A R 6 M 0.9603 7 A R 7 F 0.8702 8 A R 8 M 1.0281 9 A R 9 F 0.9887 10 A R 10 M 1.0687 11 A R 11 M 1.0483 12 A R 12 M 0.9483 2807 12 1 A L 1 M 1.0771 2 A L 2 M 0.8771 3 E L 4 A L 3 M 1.1134 5 A L 4 M 1.0197 6 A R 5 M 1.0057 7 A R 6 F 0.8473 8 A R 7 F 0.8830 9 A R 8 M 1.0132 10 A R 9 F 0.8436 11 E R . 12 A R 10 M 1.0088 2808 13 1 A L 1 M 0.8984 2 M L 3 A L 2 M 0.9015 4 A L 3 F 0.9677 5 A L 4 M 0.9335 6 M L 7 A L 5 M 0.9938 8 A L 6 M 0.9303 9 M R 10 A R 7 M 1.1087 11 E R 12 E R 13 A R 8 F 1.0222Appendix II 18

Table A-4. Individual Embryo/Fetal Data (page 8 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

2000 2809 10 1 A L 1 M 1.0041

2 A L 2 F 1.0047 3 E L 4 A L 3 F 0.9698 5 A L 4 M 0.9814 6 A L 5 M 0.9031 7 A R 6 M 1.0715 8 E R 9 A R 7 F 1.0581 10 A R 8 F 1.0225 11 A R 9 F 0.8569 12 A R 10 F 0.9546 13 A R 11 M 0.9146 2810 10 1 A L 1 F 1.0928 2 A L 2 M 1.0999 3 A L 3 F 1.0128 4 A R 4 M 1.1283 5 A R 5 F 1.0145 6 A R 6 F 0.9308 7 A R 7 M 1.0088 8 A R 8 F 0.9268 9 A R 9 M 1.0164 10 A R 10 M 0.9720 2811 14 1 A L 1 F 1.0482 2 A L 2 F 0.9048 3 A L 3 F 0.9513 4 A L 4 F 0.9646 5 A L 5 M 0.9417 6 A L 6 M 0.7961 7 A L 7 M 0.9538 8 A L 8 F 0.8648 9 A R 9 F 0.8361 10 A R 10 F 0.9768 11 A R 11 M 0.9976 12 A R 12 M 0.9588 13 A R 13 M 0.9767 2812 14 1 A L 1 M 1.1579 2 A L 2 F 1.0919 3 A L 3 F 1.0422 4 A L 4 F 1.0483 5 A L 5 M 0.9578 6 A L 6 M 1.0596 7 A L 7 F 1.0260 8 A L 8 M 1.1331 9 A R 9 F 1.0548 10 A R 10 M 1.1496 11 A R 11 F 1.0310 12 A R 12 M 1.0993 13 A R 13 F 1.0189 14 A R 14 F 1.0590 2813 10 1 I L 2 I L 3 I L 4 I R 5 I R 6 I R 7 I R 8 I R 9 I R 10 I RAppendix II 19

Table A-4. Individual Embryo/Fetal Data (page 9 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

2000 2814 11 1 A L 1 M 1.0360

2 A L 2 M 0.9772 3 A L 3 M 0.8825 4 A L 4 F 0.9772 5 A L 5 M 0.9588 6 E L 7 A L 6 F 1.0073 8 A R 7 F 0.9526 9 A R 8 F 1.0928 10 A R 9 F 1.0102 11 A R 10 F 1.0398 2815 16 1 A L 1 F 1.0205 2 A L 2 F 1.0379 3 A L 3 F 0.9504 4 A L 4 M 1.0584 5 A L 5 F 1.1114 6 A L 6 F 0.8976 7 A L 7 F 1.0575 8 A L 8 F 1.0156 9 A L 9 M 1.0038 10 A R 10 F 0.9891 11 A R 11 M 1.1001 12 A R 12 M 1.0271 13 A R 13 M 1.1058 2816 12 1 A L 1 M 1.1100 2 A L 2 F 1.0718 3 A L 3 M 1.0676 4 A L 4 F 1.0803 5 A L 5 M 1.1129 6 A R 6 F 1.1096 7 A R 7 F 0.9815 8 A R 8 F 1.0332 9 A R 9 F 0.9981 10 A R 10 F 1.1400 11 A R 11 F 0.9800 12 A R 12 M 0.9989 2817 15 1 A L 1 F 1.0393 2 A L 2 F 0.9766 3 A L 3 M 1.1003 4 A L 4 F 1.0763 5 A R 5 F 1.0411 6 A R 6 F 0.9787 7 A R 7 F 0.9803 8 A R 8 M 1.0243 9 A R 9 M 1.0327 10 A R 10 F 0.9913 11 A R 11 F 1.0011 12 A R 12 F 0.9916Appendix II 20

Table A-4. Individual Embryo/Fetal Data (page 10 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

2000 2818 17 1 A L 1 F 0.9018

2 A L 2 M 0.9189 3 A L 3 M 1.0189 4 A L 4 F 0.9339 5 A L 5 F 0.7564 6 A L 6 F 0.9032 7 A L 7 M 0.8642 External Malformation Cleft Palate 8 A L 8 F 0.9189 9 A L 9 M 1.0005 10 A L 10 M 0.9558 11 A R 11 F 0.9258 12 A R 12 M 0.9915 13 A R 13 F 0.9614 14 A R 14 F 0.8635 15 A R 15 F 0.8936 16 A R 16 F 0.8928 2819 13 1 A L 1 F 1.0275 2 A L 2 F 1.1046 3 A L 3 M 1.0580 4 A R 4 M 1.0291 5 A R 5 F 0.7359 External Malformation Encephalocele 6 A R 6 M 0.8896 7 A R 7 M 0.9365 8 A R 8 M 0.9639 9 A R 9 F 0.9304 10 A R 10 M 1.0039 11 A R 11 M 1.0009 12 A R 12 F 0.9925 13 A R 13 M 1.0325 2820 14 1 A L 1 F 1.1862 2 A L 2 M 1.0684 3 A L 3 M 0.9071 4 A L 4 F 1.2382 5 A R 5 F 1.0283 6 A R 6 F 1.0491 7 A R 7 F 1.0592 8 E R 9 A R 8 F 0.9977 10 A R 9 F 1.0505 11 A R 10 F 0.8839 12 A R 11 M 0.9944 13 A R 12 M 0.7045 14 A R 13 F 0.9805 2821 . g 2822 11 1 A L 1 F 0.9312 2 A L 2 F 0.9493 External Variation Hematoma: Face 3 A L 3 F 1.0554 4 A L 4 F 1.0212 5 A R 5 M 1.0618 6 E R 7 A R 6 F 1.0410 External Variation Hematoma: Face 8 A R 7 M 1.0158 9 A R 8 F 1.0331 10 A R 9 M 1.0376 11 A R 10 M 0.9412Appendix II 21

Table A-4. Individual Embryo/Fetal Data (page 11 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

2000 2823 12 1 A L 1 F 0.9744

2 A L 2 F 1.0834 3 A L 3 M 1.0539 4 E L 5 A L 4 F 1.0137 6 A R 5 M 1.1048 7 A R 6 F 0.9936 8 A R 7 M 1.0680 9 A R 8 M 1.0025 10 A R 9 M 1.0029 11 A R 10 M 1.0112 12 A R 11 F 0.9788

10000 3801 13 1 A L 1 M 1.0190 2 A L 2 M 0.9014 3 A L 3 M 0.9821 4 A L 4 M 1.0398 5 A L 5 M 1.0434 6 A L 6 F 0.8964 7 A L 7 F 0.9460 8 A R 8 M 1.0947 9 A R 9 M 1.0239 10 A R 10 M 0.9811 11 E R 12 A R 11 M 0.9961 3802 . g 3803 10 1 A L 1 F 1.1517 2 A L 2 M 1.0631 3 A L 3 F 1.1021 4 E L 5 A L 4 F 1.1125 6 A L 5 M 1.2200 7 A R 6 F 1.0961 8 A R 7 F 1.0464 9 A R 8 F 1.0687 10 A R 9 M 1.2010 11 A R 10 F 1.1963 12 A R 11 M 1.2033 13 E R 3804 14 1 A L 1 M 0.9328 2 A L 2 M 0.9306 3 A L 3 F 0.8937 4 A L 4 F 0.9386 5 L R 6 A R 5 M 0.9326 7 A R 6 M 0.9338 8 A R 7 M 0.8966 9 A R 8 M 0.9382 10 A R 9 M 0.9291 11 A R 10 M 0.9202 12 A R 11 F 0.9163Appendix II 22

Table A-4. Individual Embryo/Fetal Data (page 12 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

10000 3805 16 1 A L 1 F 0.8832 2 A L 2 F 0.9752 3 A L 3 M 1.0583 4 A L 4 M 0.8836 5 E R 6 D R 0.7638 7 A R 5 F 0.9635 8 A R 6 M 0.8553 9 A R 7 F 0.8790 10 A R 8 F 0.8643 11 A R 9 M 0.9115 12 A R 10 M 0.9509 13 A R 11 F 0.8660 14 A R 12 F 0.8833 3806 13 1 A L 1 F 1.1007 2 A L 2 M 1.0647 3 A L 3 F 1.0466 4 A L 4 M 1.0508 5 A L 5 F 1.1125 6 A L 6 M 1.1215 7 A L 7 M 1.0254 8 A L 8 F 1.1635 9 A R 9 M 1.2134 10 A R 10 F 1.1405 11 A R 11 M 1.2279 12 A R 12 F 1.1768 13 A R 13 F 1.2030 3807 12 1 A L 1 F 0.8125 2 A L 2 F 0.8706 3 A L 3 F 0.7146 4 A L 4 F 0.7455 5 A L 5 F 0.6843 6 A R 6 M 0.7882 7 A R 7 F 0.7540 8 A R 8 F 0.7794 9 A R 9 F 0.7171 10 A R 10 M 0.7736 11 A R 11 M 0.7202 12 A R 12 M 0.7913 3808 11 1 A L 1 M 0.9736 2 A L 2 F 0.9106 3 A L 3 M 0.9582 4 A L 4 M 0.9339 5 A R 5 M 0.9355 6 A R 6 F 0.8832 7 A R 7 M 0.9395 8 A R 8 F 0.9138 9 M R 10 A R 9 M 0.8939 11 M RAppendix II 23

Table A-4. Individual Embryo/Fetal Data (page 13 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

10000 3809 13 1 A L 1 F 0.7510 2 A L 2 F 0.8011 3 A L 3 M 0.8512 4 A L 4 F 0.9113 5 A L 5 M 0.8740 6 A R 6 F 0.8957 7 A R 7 M 0.8702 8 A R 8 M 0.9312 External Variation Hematoma: Neck 9 A R 9 M 0.9074 10 A R 10 F 0.8759 11 A R 11 M 0.8169 12 A R 12 M 0.8717 13 A R 13 M 0.8925 3810 12 1 A L 1 F 0.9400 2 A L 2 F 0.8910 3 A L 3 M 1.0533 4 A L 4 M 0.9740 5 A L 5 F 0.9930 6 A L 6 F 0.9413 7 A R 7 F 0.9848 8 A R 8 M 1.0769 9 A R 9 F 0.9693 10 A R 10 M 1.0352 11 A R 11 M 1.0835 12 A R 12 F 0.9862 3811 11 1 A L 1 M 0.9645 2 A L 2 M 1.0063 3 A L 3 F 0.9305 4 A L 4 F 0.8759 5 A R 5 M 0.9009 6 A R 6 M 0.8406 7 A R 7 M 0.8920 8 A R 8 M 0.5587 9 A R 9 F 0.8932 10 A R 10 F 0.9297 11 A R 11 M 0.9312 3812 13 1 A L 1 M 1.0739 2 A L 2 M 0.9977 3 A L 3 F 1.0568 4 A L 4 F 0.9341 5 A L 5 F 0.8089 6 A L 6 F 1.0368 7 A L 7 M 1.0440 8 A R 8 F 1.0519 9 A R 9 F 0.9655 10 A R 10 F 1.0877 11 A R 11 F 1.0286 12 A R 12 F 1.0178 13 A R 13 M 0.9960 3813 12 1 A L 1 M 0.9396 2 A L 2 M 0.9545 3 E L 4 A L 3 M 0.9843 5 A L 4 F 0.9474 6 A L 5 M 0.9391 7 A R 6 M 0.9248 8 A R 7 F 0.9039 9 A R 8 F 0.9342 10 A R 9 F 0.9293 11 A R 10 F 0.9427 12 A R 11 F 0.9326Appendix II 24

Table A-4. Individual Embryo/Fetal Data (page 14 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

10000 3814 . h
3815 11 1 A L 1 F 1.1360 2 A L 2 M 1.1999 3 A L 3 F 1.1700 4 A L 4 M 1.1825 5 A L 5 M 1.1779 6 A L 6 M 1.1906 7 A R 7 M 1.1756 8 A R 8 M 1.1893 9 A R 9 M 1.2653 10 A R 10 M 1.1124 11 A R 11 M 1.1893 3816 15 1 A L 1 M 0.8593 2 A L 2 F 0.8340 3 A L 3 F 0.8706 4 A L 4 F 0.8719 5 A L 5 F 0.8671 6 A L 6 M 0.8532 7 A L 7 F 0.8176 8 A R 8 M 0.9453 9 A R 9 M 0.8392 External Malformation Cleft Palate 10 A R 10 M 0.8336 11 A R 11 F 0.8685 12 A R 12 F 0.8623 13 A R 13 M 0.9595 14 M R 15 A R 14 F 0.8344 3817 . g 3818 14 1 A L 1 F 0.8906 2 A L 2 F 0.8837 3 A L 3 M 1.0012 4 A R 4 M 0.9925 5 A R 5 F 0.9106 6 A R 6 F 0.8052 7 A R 7 M 0.9135 8 A R 8 M 1.0314 9 A R 9 M 0.9259 10 A R 10 F 0.8523 11 A R 11 F 0.8937 3819 14 1 A L 1 M 0.8560 2 A L 2 F 0.8368 3 A L 3 F 0.8758 4 A L 4 F 0.9340 5 A L 5 M 0.9200 6 A L 6 F 0.8570 7 A L 7 M 0.9478 8 A L 8 M 0.9177 9 E R 10 A R 9 F 0.9978 11 A R 10 F 0.9447 12 A R 11 M 1.0698 13 A R 12 F 0.9054Appendix II 25

Table A-4. Individual Embryo/Fetal Data (page 15 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

10000 3820 13 1 A L 1 F 0.7350 2 A L 2 M 0.7557 3 A L 3 F 0.8625 4 A L 4 M 0.9689 5 A L 5 F 0.8864 6 A L 6 F 0.9464 7 A L 7 M 0.9343 8 A R 8 F 0.9803 9 A R 9 F 0.8394 10 A R 10 M 1.0069 11 A R 11 M 0.9770 12 A R 12 F 0.9931 3821 . i 1 D L 0.7329 2 A L 1 M 0.9617 3 A L 2 M 0.8419 4 A L 3 M 0.8902 5 A L 4 F 0.9598 6 A L 5 M 0.9075 7 E R 8 A R 6 F 0.9211 9 A R 7 F 0.9019 10 A R 8 M 0.7350 11 A R 9 M 0.8225 12 L R 13 A R 10 M 0.7931 14 A R 11 M 0.8594 15 A R 12 M 0.8169 16 A R 13 F 0.9058 3822 13 1 A L 1 M 0.8384 2 E L 3 A L 2 M 0.8672 4 A L 3 F 0.8362 5 A L 4 F 0.9295 6 E L 7 A R 5 M 0.9220 8 A R 6 M 0.7331 9 A R 7 F 0.8611 10 A R 8 M 0.8935 11 A R 9 M 0.8065 12 A R 10 M 0.9002 13 A R 11 M 0.8964 14 A R 12 F 0.9067 15 A R 13 F 0.7866 External Variation Abnormal Rugae in Midline of Palate 3823 . g

20000 4801 15 1 A L 1 M 0.7756 External Variation Hematoma: Neck 2 A L 2 M 0.9303 3 A L 3 F 0.8679 4 A L 4 F 0.8982 5 A L 5 M 0.8411 6 A L 6 F 0.9145 7 A L 7 M 0.9662 8 A L 8 M 0.9387 9 A L 9 F 0.7659 10 A L 10 F 0.8141 11 A R 11 F 0.9080 12 A R 12 F 0.9201 13 A R 13 F 0.8216Appendix II 26

Table A-4. Individual Embryo/Fetal Data (page 16 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

20000 4802 13 1 A L 1 M 1.0719 2 A L 2 M 1.0172 3 E L 4 A L 3 F 0.9307 External Variation Hematoma: Neck 5 A L 4 F 0.7838 6 A L 5 F 1.0347 7 A R 6 F 0.9325 8 A R 7 F 0.9266 9 A R 8 F 1.0504 10 A R 9 F 0.9771 11 A R 10 F 1.0003 12 A R 11 M 0.9209 13 A R 12 F 1.0126 4803 12 1 A L 1 F 1.1729 2 A L 2 M 1.1987 3 A L 3 M 1.0925 4 A L 4 M 1.1984 5 A L 5 F 1.1336 6 A L 6 F 1.1188 7 A L 7 F 1.1314 8 A R 8 F 1.1490 9 A R 9 F 1.0659 10 A R 10 F 1.1253 11 A R 11 M 1.1589 12 A R 12 M 1.0840 13 A R 13 M 1.1359 4804 15 1 A L 1 M 1.1320 2 A L 2 M 1.0061 3 A L 3 M 1.0438 4 A L 4 F 1.0220 5 A L 5 M 1.0451 6 A R 6 F 0.9857 7 A R 7 M 0.9875 8 A R 8 M 0.9954 9 A R 9 F 0.9630 10 A R 10 F 0.9950 11 A R 11 F 0.7918 12 A R 12 M 1.0053 13 M R 14 A R 13 M 0.9716 4805 13 1 A L 1 M 0.8523 2 A L 2 F 0.8314 3 A L 3 F 0.9040 4 A L 4 F 0.8392 5 A L 5 M 0.7989 6 A L 6 M 0.8065 7 A L 7 M 0.8913 8 A R 8 M 0.9045 9 A R 9 M 0.9049 10 A R 10 F 0.9584 11 A R 11 M 0.9496 12 A R 12 F 0.8322 13 A R 13 M 0.9007Appendix II 27

Table A-4. Individual Embryo/Fetal Data (page 17 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

20000 4806 13 1 A L 1 F 0.9073 2 A L 2 M 0.8205 3 A L 3 F 0.9315 4 A L 4 M 0.9512 5 A L 5 F 0.6950 6 A L 6 M 0.9079 7 A L 7 F 0.9127 8 A L 8 F 0.8739 9 A R 9 M 0.9375 10 A R 10 M 0.9311 11 A R 11 M 0.8928 12 A R 12 M 0.8862 13 A R 13 F 0.8459 4807 13 1 A L 1 M 0.8860 2 A L 2 F 0.9351 3 A L 3 F 0.9293 4 A L 4 M 0.9260 5 A L 5 F 0.9085 6 A R 6 F 0.8282 7 A R 7 M 0.8468 8 A R 8 F 0.7430 9 A R 9 F 0.7945 10 A R 10 F 0.8475 11 A R 11 M 0.9188 12 A R 12 M 0.8337 13 A R 13 F 0.8254 4808 14 1 A L 1 M 0.8765 2 A L 2 M 0.8406 3 A L 3 F 0.7655 4 A L 4 M 0.8192 5 A L 5 M 0.8560 6 A L 6 M 0.9250 7 A L 7 F 0.8788 8 A L 8 F 0.8942 9 A R 9 F 0.8507 10 A R 10 F 0.8718 11 A R 11 M 0.8882 12 A R 12 F 0.9554 13 A R 13 M 0.9473 14 A R 14 M 0.8788 4809 19 1 A L 1 M 0.8810 2 A L 2 M 0.8340 3 A L 3 F 0.7542 4 A L 4 M 0.8357 5 A L 5 M 0.8400 6 A L 6 F 0.8336 7 A L 7 M 0.8674 8 A L 8 M 0.8963 9 A R 9 F 0.7561 10 A R 10 F 0.7887 11 A R 11 M 0.7948 12 A R 12 F 0.8452 13 E R 14 A R 13 F 0.8102 15 A R 14 F 0.8396 16 A R 15 M 0.8383 17 A R 16 F 0.7813 18 A R 17 F 0.7973 19 A R 18 F 0.7978Appendix II 28

Table A-4. Individual Embryo/Fetal Data (page 18 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

20000 4810 13 1 A L 1 F 0.7567 2 A L 2 M 0.8425 3 A L 3 M 0.8637 4 A L 4 M 0.9020 5 A L 5 F 0.8044 6 A L 6 F 0.8863 7 A L 7 M 0.9203 8 A R 8 F 0.7931 9 A R 9 M 0.8795 10 A R 10 F 0.8196 11 A R 11 F 0.8692 12 A R 12 M 0.8416 13 A R 13 F 0.8331 4811 13 1 A L 1 F 1.1239 2 A L 2 F 1.1188 3 A L 3 F 1.1692 4 A L 4 M 1.2309 5 A L 5 F 1.1065 6 D L 0.8750 7 A L 6 F 1.1848 8 A R 7 F 1.2355 9 A R 8 F 1.2401 10 A R 9 F 1.1965 11 A R 10 F 1.1526 12 A R 11 M 1.2364 13 A R 12 M 1.2652 4812 14 1 A L 1 M 0.9838 2 A L 2 F 0.9007 3 A L 3 F 0.8484 4 A L 4 M 0.9993 5 A L 5 F 0.8709 6 A R 6 M 0.9528 7 A R 7 F 0.8973 8 A R 8 F 0.8975 9 A R 9 F 0.9014 10 A R 10 F 0.8079 11 A R 11 F 0.7973 12 A R 12 M 1.0232 4813 16 1 A L 1 M 1.1349 2 A L 2 M 1.0482 3 A L 3 M 1.1813 4 A L 4 F 1.1449 5 E L 6 A R 5 M 1.1067 7 A R 6 F 1.0247 8 A R 7 F 0.9778 9 A R 8 F 0.9348 10 A R 9 F 0.9969 11 A R 10 F 0.9783 12 A R 11 F 1.0145 13 A R 12 F 1.0523 14 A R 13 F 0.9525 15 A R 14 F 0.9687 16 A R 15 F 0.9585Appendix II 29

Table A-4. Individual Embryo/Fetal Data (page 19 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

20000 4814 9 1 M L 2 A L 1 F 1.2799 3 A L 2 F 1.3151 4 A L 3 M 1.3496 5 E L 6 A L 4 F 1.1992 7 A R 5 M 1.3070 8 A R 6 M 1.2653 9 E R 4815 . g 4816 13 1 A L 1 M 0.8639 2 A L 2 M 0.9429 3 A L 3 F 0.8220 4 A L 4 M 0.8379 5 A L 5 M 0.9266 6 A L 6 F 0.9481 7 A L 7 F 0.9420 8 A L 8 F 0.9747 9 A R 9 F 1.0099 10 A R 10 F 0.8801 11 A R 11 F 0.9603 12 A R 12 F 0.9483 13 A R 13 M 0.9909 4817 14 1 A L 1 M 1.0301 2 E L 3 A L 2 M 1.0935 4 A L 3 M 1.0339 5 A L 4 M 1.0177 6 A L 5 M 0.9907 7 A L 6 M 0.9161 8 A L 7 F 1.0390 9 A R 8 M 1.0381 10 A R 9 F 1.0077 11 A R 10 M 1.0401 12 A R 11 F 0.9537 13 A R 12 F 0.9633 4818 14 1 I L 2 I L 3 I L 4 I L 5 I L 6 I L 7 I R 8 I R 9 I R 10 I R 11 I R 12 I R 13 I R 14 I R 15 I R 16 I R 17 I RAppendix II 30

Table A-4. Individual Embryo/Fetal Data (page 20 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

20000 4819 5 1 I L 2 I L 3 I L 4 I R 5 I R 6 I R 7 I R 8 I R 9 I R 10 I R 4820 13 1 A L 1 F 0.9712 2 A L 2 F 0.9335 3 A L 3 F 0.9432 4 A L 4 F 0.9313 5 A L 5 F 0.7709 6 A L 6 M 1.0237 7 A L 7 F 0.9602 8 A L 8 M 0.9823 9 A R 9 M 0.9386 10 E R 11 A R 10 F 0.9344 12 A R 11 F 0.9133 4821 15 1 A L 1 M 0.8292 2 A L 2 F 0.8331 3 A L 3 F 0.7993 4 A R 4 M 0.8301 5 A R 5 M 0.7856 6 A R 6 M 0.6891 7 A R 7 F 0.7089 External Malformation Cleft Palate 8 A R 8 M 0.5875 9 A R 9 M 0.7899 10 D R 0.6560 11 A R 10 M 0.8427 12 A R 11 M 0.8427 13 A R 12 M 0.8158 14 A R 13 F 0.7604 15 A R 14 M 0.7674 4822 12 1 A L 1 F 0.9253 2 A L 2 M 0.9153 3 A L 3 F 0.9479 External Variation Abnormal Rugae in Midline of Palate 4 A L 4 M 0.9499 5 A R 5 F 0.9415 6 A R 6 F 0.8508 7 A R 7 M 0.9713 8 A R 8 M 0.9437 9 A R 9 F 0.8754 10 A R 10 M 0.8642 11 A R 11 F 0.8712 4823 12 1 A L 1 F 0.7820 2 A L 2 F 0.8031 3 A L 3 M 0.8638 4 E L 5 A L 4 M 0.9099 6 A L 5 M 0.6567 7 A R 6 F 0.9373 8 A R 7 F 0.8679 9 A R 8 F 0.9426 10 A R 9 F 0.9749 11 A R 10 F 0.8428 12 A R 11 M 0.9194Appendix II 31

Table A-4. Individual Embryo/Fetal Data (page 21 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

30000 5801 . g 5802 13 1 A L 1 M 0.9804 2 A L 2 M 1.1120 3 A L 3 M 1.0901 4 A R 4 M 1.0564 5 A R 5 M 1.0590 6 A R 6 F 0.9079 7 A R 7 F 0.9483 8 A R 8 F 0.9521 9 A R 9 F 0.9818 10 A R 10 F 0.9639 11 A R 11 M 1.0117 12 L R 13 A R 12 M 0.9321 5803 13 1 A L 1 F 1.1267 2 A L 2 M 1.1515 3 A L 3 M 1.0179 4 A L 4 M 1.1013 5 A L 5 M 1.0478 6 A L 6 M 1.1700 7 A R 7 F 1.1515 8 A R 8 M 1.2303 9 A R 9 F 1.1074 10 A R 10 F 1.2296 11 M R 12 A R 11 M 1.0830 13 A R 12 M 1.1170 5804 15 1 A L 1 M 1.1464 2 A L 2 M 1.0855 3 A L 3 M 0.7745 External Malformation Cleft Palate 4 A L 4 M 1.0201 5 A L 5 F 1.1951 6 A R 6 F 0.9255 7 A R 7 M 1.2088 8 M R 9 A R 8 M 1.1263 10 A R 9 F 1.0717 11 A R 10 M 1.1267 12 A R 11 M 1.0228 13 A R 12 F 1.1004 5805 13 1 A L 1 M 1.0774 2 A L 2 F 0.9531 3 A L 3 F 0.9565 4 A L 4 M 0.9550 5 A L 5 F 0.8124 6 A L 6 M 1.0683 7 A L 7 F 1.0170 8 A L 8 M 1.0931 9 A R 9 F 1.0089 10 A R 10 M 1.0940 11 A R 11 F 1.0375 12 A R 12 M 1.0142 13 A R 13 M 1.0256Appendix II 32

Table A-4. Individual Embryo/Fetal Data (page 22 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

30000 5806 18 1 A L 1 M 0.9533 2 A L 2 F 0.9073 3 A L 3 M 0.9066 4 A L 4 F 0.9136 5 A L 5 F 1.0194 6 L L 7 A L 6 F 0.9655 8 A L 7 M 1.0357 9 A L 8 F 0.9715 10 A L 9 M 0.9788 11 A L 10 M 1.0303 12 A R 11 F 1.0017 13 A R 12 F 0.9086 14 A R 13 M 1.0746 15 A R 14 M 1.1079 16 E R 17 A R 15 F 0.9868 18 A R 16 F 0.9189 5807 13 1 A L 1 F 0.9649 2 A L 2 F 0.9225 3 A L 3 F 1.0054 4 A L 4 F 1.0138 5 A L 5 M 1.0280 6 A L 6 F 1.0273 7 A R 7 F 0.9359 8 A R 8 F 0.9338 9 A R 9 F 0.9303 10 A R 10 M 1.0376 11 A R 11 F 0.9072 12 A R 12 F 0.8520 13 A R 13 M 0.9406 5808 13 1 A L 1 M 0.9483 2 A L 2 F 1.0104 3 A L 3 M 1.0077 4 A R 4 M 0.9191 5 A R 5 M 0.8189 6 A R 6 M 0.9226 7 A R 7 M 0.7506 8 A R 8 M 0.8968 9 A R 9 M 0.9360 10 A R 10 F 0.9469 11 A R 11 F 0.9655 12 A R 12 F 0.9790 13 A R 13 M 0.9588 5809 16 1 A L 1 M 0.8884 2 A L 2 M 0.8127 3 A L 3 M 0.8462 4 A L 4 M 0.9215 5 A L 5 F 0.9343 6 E L 7 A R 6 F 0.8106 8 A R 7 F 0.8068 9 A R 8 M 0.8612 10 L R 11 A R 9 F 0.9078 12 A R 10 F 0.8592 13 A R 11 M 0.9218 14 L R 15 A R 12 F 0.8428 16 A R 13 M 0.8899Appendix II 33

Table A-4. Individual Embryo/Fetal Data (page 23 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

30000 5810 15 1 A L 1 F 0.8985 2 A L 2 F 0.9065 3 A L 3 F 0.9001 4 A L 4 F 0.9440 5 E L 6 A L 5 F 0.9382 External Malformation Cleft Palate 7 A L 6 F 0.6057 External Malformation Cleft Palate External Malformation Gastroschisis 8 A L 7 M 0.8893 9 A R 8 F 0.8080 10 A R 9 F 0.8684 11 A R 10 F 0.8034 12 A R 11 M 0.8844 13 A R 12 M 0.8406 External Malformation Cleft Palate 14 A R 13 F 0.8456 15 A R 14 F 0.9768 5811 16 1 A L 1 M 0.9681 2 A L 2 F 0.9535 3 L L 4 A L 3 F 1.0411 5 A L 4 F 0.9981 6 A L 5 F 1.0156 7 E L 8 A L 6 M 1.1714 9 A L 7 M 1.1660 10 A R 8 F 1.1421 11 A R 9 M 1.1165 12 A R 10 M 1.0938 13 A R 11 M 1.1929 14 A R 12 F 1.1948 15 A R 13 M 1.2758 16 A R 14 F 1.2000 5812 11 1 A L 1 M 1.0452 2 A L 2 M 0.9058 3 A L 3 F 0.9278 4 A L 4 F 1.0021 5 A L 5 M 0.9996 6 A L 6 M 0.9291 7 A L 7 M 1.0208 8 A R 8 M 1.0233 9 A R 9 M 1.0146 10 A R 10 F 0.9597 11 A R 11 M 1.0220 5813 4 1 I L 2 I L 3 I L 4 I L 5 I L 6 I R 7 I R 8 I R 9 I R 10 I R 11 I R 12 I RAppendix II 34

Table A-4. Individual Embryo/Fetal Data (page 24 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

30000 5814 12 1 A L 1 M 0.9903 2 A L 2 F 0.9792 3 A L 3 M 1.0668 4 A L 4 F 1.0714 5 A L 5 F 1.0852 6 A R 6 F 0.8905 7 A R 7 M 0.8522 8 A R 8 F 0.8659 9 A R 9 M 1.0235 10 A R 10 F 0.9251 11 A R 11 F 1.0418 12 A R 12 M 1.0593 5815 12 1 A L 1 M 1.1130 2 A L 2 F 1.0309 3 A L 3 M 1.0280 4 A L 4 F 1.0562 5 A L 5 F 1.0166 6 A L 6 F 1.0871 7 A R 7 F 1.0025 8 A R 8 M 0.7835 9 A R 9 F 1.0818 10 A R 10 F 0.9947 11 A R 11 F 1.0768 12 A R 12 F 0.9457 5816 14 1 A L 1 M 1.3655 2 A L 2 F 1.2790 3 D L 0.8132 4 A L 3 F 1.2630 5 A L 4 F 1.1545 6 A L 5 F 1.2992 7 A L 6 M 1.2512 8 A R 7 M 1.3040 9 A R 8 M 1.2248 10 A R 9 F 1.2875 11 A R 10 M 1.3026 12 E R 13 A R 11 F 1.2087 14 A R 12 F 1.3550 5817 . g 5818 16 1 A L 1 F 0.8573 2 A L 2 F 0.8314 3 A L 3 M 0.8302 4 A L 4 F 0.8590 5 A L 5 F 0.7868 6 A L 6 F 0.9320 7 A L 7 F 0.9167 8 A L 8 F 0.8636 9 A R 9 F 0.8699 10 A R 10 F 0.8666 11 A R 11 M 0.7944 12 A R 12 M 0.8529 13 A R 13 F 0.8398 14 A R 14 M 0.8448 15 A R 15 M 0.8584Appendix II 35

Table A-4. Individual Embryo/Fetal Data (page 25 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

30000 5819 14 1 A L 1 F 1.3938 2 A L 2 M 1.4852 3 A R 3 F 1.2505 4 A R 4 M 1.3148 5 A R 5 M 1.1664 6 A R 6 F 0.8750 7 A R 7 M 1.3726 8 E R 9 A R 8 M 1.1908 10 A R 9 M 1.4493 11 A R 10 M 1.3387 12 A R 11 M 1.3200 5820 13 1 L L 2 A L 1 F 0.9656 3 A L 2 M 1.0512 4 A L 3 F 0.9488 5 A L 4 F 0.9156 6 A L 5 F 1.0114 7 A R 6 F 0.9614 8 E R 9 A R 7 F 0.9043 10 A R 8 F 0.8433 11 A R 9 F 0.8728 12 A R 10 F 0.9826 13 M R 14 A R 11 F 0.9827 5821 17 1 A L 1 F 1.0300 2 A L 2 F 0.9014 3 A L 3 M 0.9976 4 A L 4 M 1.0325 5 E L 6 A L 5 F 0.8470 7 A L 6 M 1.0474 8 E L 9 A R 7 M 1.0702 10 A R 8 F 1.0475 11 A R 9 F 0.9996 12 A R 10 M 1.0595 13 A R 11 F 0.9625 14 A R 12 F 1.0545 5822 11 1 A L 1 F 1.0140 2 A L 2 F 1.0250 3 A R 3 F 0.9620 4 A R 4 M 0.9893 5 A R 5 F 0.8717 6 A R 6 M 0.9655 7 A R 7 M 0.9312 8 A R 8 F 1.0055 9 A R 9 M 1.0297 10 A R 10 F 0.9577 11 A R 11 F 0.9520Appendix II 36

Table A-4. Individual Embryo/Fetal Data (page 26 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

30000 5823 13 1 A L 1 M 1.5475 2 A L 2 F 1.3667 3 A L 3 F 1.2577 4 A L 4 M 1.4539 5 A L 5 M 1.3758 6 A L 6 F 1.4071 7 A L 7 F 1.3181 8 A L 8 F 1.3916 9 A R 9 F 1.4486 10 A R 10 M 1.3247 11 A R 11 F 1.2217 12 A R 12 M 1.4355 13 A R 13 M 1.4881 5824 5 1 I R 2 I R 3 I R 4 I R 5825 11 1 A L 1 M 1.0203 2 A L 2 F 0.9395 3 A L 3 F 0.9975 4 A L 4 F 0.9981 5 A L 5 F 1.0025 6 A L 6 M 0.8743 7 A L 7 F 0.9600 8 A R 8 F 1.1533 9 E R 10 A R 9 M 1.0698 11 A R 10 M 1.0630 5826 16 1 A L 1 M 0.8428 2 A L 2 M 0.9350 3 A L 3 F 0.9731 4 A L 4 M 1.0226 5 A L 5 M 0.9213 6 A L 6 M 0.8604 7 A R 7 F 0.8411 8 A R 8 F 0.9784 9 A R 9 M 0.8939 10 A R 10 F 0.8954 11 A R 11 F 0.8163 12 A R 12 M 0.9026 13 A R 13 M 0.8072 14 A R 14 F 0.8687 5827 14 1 A L 1 M 0.9385 2 A L 2 F 0.9648 3 A L 3 M 0.9358 4 A L 4 F 0.7844 5 A R 5 F 0.9408 6 A R 6 F 0.9805 7 A R 7 F 1.0502 8 A R 8 F 0.8167 9 A R 9 M 1.0225 10 A R 10 F 0.9819 11 A R 11 F 0.9894Appendix II 37

Table A-4. Individual Embryo/Fetal Data (page 27 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

30000 5828 11 1 A L 1 F 0.9467 2 A L 2 F 0.9957 3 A L 3 F 0.8725 4 A L 4 F 0.8279 5 A L 5 F 0.9775 6 A L 6 F 1.0164 External Malformation Cleft Palate 7 A L 7 M 0.8595 8 A L 8 F 0.9852 9 A R 9 F 0.8664 10 A R 10 F 0.9977 11 E R 12 A R 11 F 0.9024 5829 14 1 A L 1 F 1.1204 2 E L 3 A L 2 F 1.1703 4 A L 3 F 1.1694 5 A R 4 M 1.1893 6 A R 5 M 1.1602 7 A R 6 M 1.1989 8 A R 7 M 1.1962 9 A R 8 F 1.1228 10 E R 11 A R 9 M 1.1192 5830 15 1 A L 1 M 1.0950 2 A L 2 F 1.1618 3 A L 3 M 1.0445 4 A L 4 F 1.0395 5 A R 5 F 1.2304 6 A R 6 F 1.0734 7 A R 7 M 0.9946 External Malformation Cleft Palate 8 A R 8 M 1.1270 9 A R 9 M 0.9994 10 A R 10 F 1.1301 11 A R 11 F 1.0525 External Malformation Cleft Palate 12 A R 12 M 0.9878 13 A R 13 F 1.0706 5831 14 1 A L 1 F 0.9588 2 A L 2 M 1.0149 3 A L 3 M 0.9975 4 A L 4 F 0.8350 5 A L 5 F 0.9145 6 A L 6 M 1.0151 7 A R 7 M 0.9522 8 A R 8 F 0.9081 9 A R 9 F 0.8420 10 A R 10 M 0.9452 11 A R 11 F 0.8850 12 A R 12 M 0.9097 13 A R 13 M 0.9792Appendix II 38

Table A-4. Individual Embryo/Fetal Data (page 28 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

30000 5832 14 1 A L 1 M 1.1667 2 A L 2 F 1.0426 3 A L 3 F 1.0582 4 A L 4 F 0.9516 5 A L 5 F 0.9912 6 A L 6 F 0.9265 7 A L 7 M 1.0659 8 A L 8 M 1.1146 9 A L 9 F 1.0422 10 A R 10 F 1.0611 11 A R 11 M 1.1725 12 A R 12 M 1.1148 13 A R 13 F 1.1144 14 A R 14 M 1.0441 5833 16 1 A L 1 M 0.9488 2 A L 2 F 0.8791 3 E L 4 A L 3 F 0.8688 5 A L 4 F 0.7998 6 E L 7 A L 5 M 0.8976 8 A L 6 F 0.9030 9 A R 7 F 0.8906 10 A R 8 F 0.8321 11 A R 9 F 0.8715 12 A R 10 F 0.8348 13 A R 11 M 0.8665 14 A R 12 F 0.9051 5834 10 1 A L 1 M 0.9527 2 A L 2 F 0.9792 3 A L 3 M 0.9705 4 A L 4 M 0.8064 5 A L 5 M 1.0083 6 A L 6 M 0.9922 7 A L 7 F 0.9961 8 A R 8 M 1.0352 9 A R 9 M 0.9722 10 A R 10 F 1.0271 5835 12 1 A L 1 F 1.5416 2 E L 3 A L 2 M 1.5103 4 A L 3 M 1.4931 5 A L 4 M 1.1562 6 A R 5 M 1.2581 7 A R 6 F 1.4263 8 A R 7 F 1.5269 9 A R 8 F 1.4091 10 A R 9 M 1.5567 11 A R 10 F 1.3408 12 A R 11 F 1.3742 13 A R 12 M 1.5313Appendix II 39

Table A-4. Individual Embryo/Fetal Data (page 29 of 29)

Implant Dam Posi- Fetus Defect f

Dose a ID# NCL b # Type c tion d # Sex Wt. e Exam Type Description

30000 5836 13 1 A L 1 F 0.9538 2 A L 2 M 0.9240 3 A L 3 F 0.8944 4 A L 4 M 0.9606 5 A L 5 F 0.9417 6 A L 6 M 0.8582 7 A L 7 M 0.9532 8 A R 8 F 0.8749 9 A R 9 F 1.0230 10 A R 10 F 0.9640 11 A R 11 M 0.8196 12 A R 12 M 0.9105 13 A R 13 F 0.8231 14 A R 14 M 1.0398 5837 15 1 I L 2 I L 3 I L 4 I L 5 I L 6 I R 7 I R 8 I R 9 I R 10 I R 11 I R 12 I R 13 I R 5838 13 1 A L 1 M 1.0087 2 A L 2 F 0.9374 3 A L 3 M 0.8882 4 A L 4 M 0.9084 5 A L 5 F 0.8730 6 L L 7 A L 6 M 0.9249 8 A L 7 M 0.9987 9 A L 8 F 0.9615 10 A L 9 F 0.9365 11 A R 10 M 1.0784 12 A R 11 F 1.0335 13 A R 12 F 1.0152 14 E R

a Mg/m 3 of gasoline MTBE vapor condensate. b Number of corpora lutea. c Implant type codes are as follows: A - Live Fetus; D - Dead Fetus; F - Full Resorption; L - Late Resorption; M -

Middle Resorption; E - Early Resorption and I - Implantation Site. d Position refers to uterine horn (R - right, L - left). e Weight is in grams. f Absence of entries under "Exam", "Type" and "Description " for "Defect" indicates no external malformation or

variation observed for that fetus. g Female was not pregnant. h Female was removed due to a preexisting condition. At necropsy she was found to have an undescended testis on

the right and seminal vesicles and prostate to the right of the vagina and cervix. i Right ovary was inadvertently lost pr ior to the corpora lutea being counted.

# Appendix III:

Protocol and 2 Amendments