United States
kS^laMIjk Environmental Protection
^J^iniiil m11 Agency
EPA/690/R-11/012F
Final
3-30-2011
Provisional Peer-Reviewed Toxicity Values for
Bis(2-chloro-1 -methylethyl)ether
(CASRN 108-60-1)
Superfund Health Risk Technical Support Center
National Center for Environmental Assessment
Office of Research and Development
U.S. Environmental Protection Agency
Cincinnati, OH 45268
-------
AUTHORS, CONTRIBUTORS, AND REVIEWERS
CHEMICAL MANAGER
Chris Cubbison, PhD
National Center for Environmental Assessment, Cincinnati, OH
DRAFT DOCUMENT PREPARED BY
ICF International
9300 Lee Highway
Fairfax, VA 22031
PRIMARY INTERNAL REVIEWERS
Ambuja Bale, PhD, DABT
National Center for Environmental Assessment, Washington, DC
Paul G. Reinhart, PhD, DABT
National Center for Environmental Assessment, Research Triangle Park, NC
This document was externally peer reviewed under contract to
Eastern Research Group, Inc.
110 Hartwell Avenue
Lexington, MA 02421-3136
Questions regarding the contents of this document may be directed to the U.S. EPA Office of
Research and Development's National Center for Environmental Assessment, Superfund Health
Risk Technical Support Center (513-569-7300).
ii Bis(2-chloro-1 -methyl ethyl)ether
-------
TABLE OF CONTENTS
COMMONLY USED ABBREVIATIONS iv
BACKGROUND 5
HISTORY 5
DISCLAIMERS 5
QUESTIONS REGARDING PPRTVS 6
INTRODUCTION 6
REVIEW OF POTENTIALLY RELEVANT DATA (CANCER AND NONCANCER) 8
HUMAN STUDIES 12
Oral and Inhalation Exposure 12
ANIMAL STUDIES 12
Oral Exposure 12
Short-term Study 12
Sub chronic-duration Studies 12
Chronic-duration Studies 15
Inhalation Exposure 19
Other Studies 19
Developmental and Reproductive Toxicity Studies 19
Other Data (Short-Term Tests, Other Examination) 19
DERIVATION 01 PROVISIONAL VALUES 25
DERIVATION OF ORAL REFERENCE DOSE 25
Derivation of Subchronic p-RfD 25
Derivation of a Chronic RfD 28
DERIVATION OF INHALATION REFERENCE CONCENTRATIONS 29
CANCER WEIGHT-OF-EVIDENCE DESCRIPTOR 29
MODE-OF-ACTION DISCUSSION 31
DERIVATION OF ORAL SLOPE FACTOR 31
DERIVATION OF INHALATION UNIT RISK 31
APPENDIX A. PROVISIONAL SCREENING VALUES 32
APPENDIX B. DATA TABLES 34
APPENDIX C. BMD MODELING OUTPUTS FOR BCMEE 47
APPENDIX D. REFERENCES 114
in
Bis(2-chloro-1 -methyl ethyl)ether
-------
COMMONLY USED ABBREVIATIONS
BMC
benchmark concentration
BMCL
benchmark concentration lower bound 95% confidence interval
BMD
benchmark dose
BMDL
benchmark dose lower bound 95% confidence interval
HEC
human equivalent concentration
HED
human equivalent dose
IUR
inhalation unit risk
LOAEL
lowest-observed-adverse-effect level
LOAELadj
LOAEL adjusted to continuous exposure duration
LOAELhec
LOAEL adjusted for dosimetric differences across species to a human
NOAEL
no-ob served-adverse-effect level
NOAELadj
NOAEL adjusted to continuous exposure duration
NOAELhec
NOAEL adjusted for dosimetric differences across species to a human
NOEL
no-ob served-effect level
OSF
oral slope factor
p-IUR
provisional inhalation unit risk
POD
point of departure
p-OSF
provisional oral slope factor
p-RfC
provisional reference concentration (inhalation)
p-RfD
provisional reference dose (oral)
RfC
reference concentration (inhalation)
RfD
reference dose (oral)
UF
uncertainty factor
UFa
animal-to-human uncertainty factor
UFC
composite uncertainty factor
UFd
incomplete-to-complete database uncertainty factor
UFh
interhuman uncertainty factor
UFl
LOAEL-to-NOAEL uncertainty factor
UFS
subchronic-to-chronic uncertainty factor
WOE
weight of evidence
iv
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
PROVISIONAL PEER-REVIEWED TOXICITY VALUES FOR
BIS(2-CHLORO-l-METHYLETHYL)ETHER (CASRN 108-60-1)
BACKGROUND
HISTORY
On December 5, 2003, the U.S. Environmental Protection Agency's (EPA) Office of
Superfund Remediation and Technology Innovation (OSRTI) revised its hierarchy of human
health toxicity values for Superfund risk assessments, establishing the following three tiers as the
new hierarchy:
1) EPA's Integrated Risk Information System (IRIS)
2) Provisional Peer-Reviewed Toxicity Values (PPRTVs) used in EPA's Superfund
Program
3) Other (peer-reviewed) toxicity values, including
~ Minimal Risk Levels produced by the Agency for Toxic Substances and Disease
Registry (ATSDR);
~ California Environmental Protection Agency (CalEPA) values; and
~ EPA Health Effects Assessment Summary Table (HEAST) values.
A PPRTV is defined as a toxicity value derived for use in the Superfund Program when
such a value is not available in EPA's IRIS. PPRTVs are developed according to a Standard
Operating Procedure (SOP) and are derived after a review of the relevant scientific literature
using the same methods, sources of data, and Agency guidance for value derivation generally
used by the EPA IRIS Program. All provisional toxicity values receive internal review by a
panel of six EPA scientists and external peer review by three independently selected scientific
experts. PPRTVs differ from IRIS values in that PPRTVs do not receive the multiprogram
consensus review provided for IRIS values. This is because IRIS values are generally intended
to be used in all EPA programs, while PPRTVs are developed specifically for the Superfund
Program.
Because new information becomes available and scientific methods improve over time,
PPRTVs are reviewed on a 5-year basis and updated into the active database. Once an IRIS
value for a specific chemical becomes available for Agency review, the analogous PPRTV for
that same chemical is retired. It should also be noted that some PPRTV documents conclude that
a PPRTV cannot be derived based on inadequate data.
DISCLAIMERS
Users of this document should first check to see if any IRIS values exist for the chemical
of concern before proceeding to use a PPRTV. If no IRIS value is available, staff in the regional
Superfund and Resource Conservation and Recovery Act (RCRA) program offices are advised to
carefully review the information provided in this document to ensure that the PPRTVs used are
appropriate for the types of exposures and circumstances at the Superfund site or RCRA facility
in question. PPRTVs are periodically updated; therefore, users should ensure that the values
contained in the PPRTV are current at the time of use.
5
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
It is important to remember that a provisional value alone tells very little about the
adverse effects of a chemical or the quality of evidence on which the value is based. Therefore,
users are strongly encouraged to read the entire PPRTV document and understand the strengths
and limitations of the derived provisional values. PPRTVs are developed by the EPA Office of
Research and Development's National Center for Environmental Assessment, Superfund Health
Risk Technical Support Center for OSRTI. Other EPA programs or external parties who may
choose of their own initiative to use these PPRTVs are advised that Superfund resources will not
generally be used to respond to challenges of PPRTVs used in a context outside of the Superfund
Program.
QUESTIONS REGARDING PPRTVS
Questions regarding the contents of the PPRTVs and their appropriate use (e.g., on
chemicals not covered, or whether chemicals have pending IRIS toxicity values) may be directed
to the EPA Office of Research and Development's National Center for Environmental
Assessment, Superfund Health Risk Technical Support Center (513-569-7300), or OSRTI.
INTRODUCTION
Bis(2-chloro-lmethylethyl)ether (BCMEE) is used in paint and varnish removers as an
intermediate in dyes, resins, and pharmaceuticals, as a solvent for natural and synthetic resins, as
a soil fumigant, and as a nematocide in Japan (HSDB, 2010; NCI, 1979; IARC, 1986). It is also
used in spotting agents, cleaning solutions, and as a soap adjuvant in the textile industry
(OEHHA, 1999). It is reported to be formed as a by-product in some propylene oxide/propylene
glycol production processes (OEHHA, 1999; IARC, 1986). The empirical formula for BCMEE
is C6H12CI2O (see Figure 1). Table 1 provides the physical properties of BCMEE.
No Structure
Figure 1. BCMEE Structure
6
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table 1. Physical Properties Table (BCMEE)
Property (unit)
Value
Boiling point (°C)
187a
Melting point (°C)
-9.7 x I0la
Density (g/cm3 at 20°C)
1.103b
Vapor pressure (Pa at 20°C)
74.7a
pH (unitless)
Not available
Solubility in water (mg/L at 20°C)
1700a
Relative vapor density (air =1)
5.9b
Molecular weight (g/mol)
171. la
Octanol/water partition coefficient (unitless)
2.48a
aValues from ChemlDPlus Advanced;
http://chem.sis.nlm. nih.gov/chemidplus/jsp/common/ChemFull.jsp?calledFrom=null.
bValues from http://toxnet.nlm.nih.gov/cgi-bin/sis/htmlgen7HSDB.
The U.S. Environmental Protection Agency (EPA) Integrated Risk Information System
(IRIS) (U.S. EPA, 2010a) does not list a chronic reference concentration (RfC) or a cancer
assessment for 2(chloro-l-methyl)ether (CAS No. 108-60-1). A chronic oral reference dose
(RfD) of 4 x 10~2 mg/kg-day is included in the IRIS database (U.S. EPA, 2010a) based on the
critical endpoint of decreased hemoglobin and possible erythrocyte destruction in SPF-ICR mice
observed in a 104-week dietary BCMEE (purity 98.5%) study (Mitsumori et al, 1979). A
Federal Drinking Water Guideline of 300 |j,g/L is published by EPA Office of Water (OW)
(U.S. EPA, 2006). No subchronic or chronic RfD or RfC values are reported in the HEAST;
(U.S. EPA, 2010b). CalEPA (OEHHA, 1999) has not derived toxicity values for exposure to
BCMEE. The toxicity of BCMEE has not been reviewed by the Agency for Toxic Substances
and Disease Registry (ATSDR, 2008) or the World Health Organization (WHO, 2010). A
Health and Environmental Effects Profile (HEEP) (U.S. EPA, 1987) has not been developed for
BCMEE. No occupational exposure limits for BCMEE have been derived by the American
Conference of Governmental Industrial Hygienists (ACGIH, 2009), the National Institute of
Occupational Safety and Health (NIOSH, 2003), or the Occupational Safety and Health
Administration (OSHA, 1997).
The HEAST (U.S. EPA, 2010b) reports an EPA cancer weight-of-evidence (WOE)
classification of Group C (Possible Human Carcinogen), an oral slope factor (OSF) of
—2 _ 1 _ i
7x10 (mg/kg-day) and an inhalation slope factor (ISF) of 3.5 x 10 (mg/kg-day) for
BCMEE. Both values were based on increased incidences of liver and lung tumors in male and
female B6C3Fi mice in a 103-week gavage study (NTP, 1982). The ISF was derived via
route-to-route extrapolation from the oral dose in mice and assuming 50% inhalation
absoroption. The HEAST (U.S. EPA, 2010b) also reported oral unit risk (UR) and inhalation
UR values of 2 x 10~6 per |j,g/L and 1 x 10~5 per |~ig/m3, respectively, for BCMEE. The
inhalation values (ISF and IUR) were derived by a route-to-route extrapolation from the oral
mouse doses (NTP, 1982) and assuming 50% absorption via the lungs. The chemical BCMEE
has not been evaluated under the Guidelines for Carcinogen Risk Assessment (U.S. EPA, 2005).
7
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
The International Agency for Research on Cancer (IARC, 2000) has classified BCMEE as a
Group 3 agent (Not Classifiable as to Its Carcinogenicity to Humans). CalEPA (OEHHA, 1999)
has developed a qualitative document outlining evidence for the carcinogenicity of technical
grade BCMEE based on the development of liver and lung tumors in male mice and lung tumors
in female B6C3Fi mice treated with BCMEE by gavage.
Literature searches were conducted from 1900 through November 2010, for studies
relevant to the derivation of provisional toxicity values for BCMEE, CAS No. 108-60-1. The
EPA Health and Environmental Research Online (HERO) database of scientific literature that
searches the following databases was used: AGRICOLA; American Chemical Society; BioOne;
Cochrane Library; DOE: Energy Information Administration; DOE: Information Bridge; DOE:
Energy Citations Database; EBSCO: Academic Search Complete; GeoRef Preview; GPO:
Government Printing Office; Informaworld; IngentaConnect; J-STAGE: Japan Science &
Technology; JSTOR: Mathematics & Statistics; JSTOR: Life Sciences; NSCEP/NEPIS (EPA
publications available through the National Service Center for Environmental Publications
[NSCEP] and National Environmental Publications Internet Site [NEPIS] database); PubMed
(MEDLINE and CANCERLIT databases among others); SAGE; Science Direct; Scirus;
Scitopia; SpringerLink; TOXNET (Toxicology Data Network: ANEUPL; CCRIS; ChemlDplus;
CIS; CRISP; DART; EMIC; EPIDEM; ETICBACK; FEDRIP; GENE-TOX; HAPAB; HEEP;
HMTC; Hazardous Substances Data Bank (HSDB); IRIS; ITER; LactMed; Multi-Database
Search; NIOSH; NTIS; PESTAB; PPBIB; RISKLINE; TRI; and TSCATS); Virtual Health
Library; Web of Science (searches Current Content database among others); World Health
Organization; and Worldwide Science. The following databases outside of HERO were searched
for risk assessment values: ACGIH; AT SDR; CalEPA; EPA IRIS; EPA HEAST; EPA HEEP;
EPA OW; EPA TSCATS/TSCATS2; NIOSH; NTP; OSHA; and RTECS. A final search of the
published literature was conducted from January 2010 through November 2010 for recent
studies.
REVIEW OF POTENTIALLY RELEVANT DATA
(CANCER AND NONCANCER)
Table 2 provides information for all of the potentially relevant studies. Entries for the
principal studies are bolded and are labeled "PS".
8
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table 2. Summary of Potentially Relevant Data for BCMEE (CASRN 108-60-1)
Notes3
Category
Number of Male/Female
Species, Study Type, and
Duration
Dosimetryb
Critical Effects
NOAELb
BMDL/
BMCLb
LOAELbc
Reference
(Comments)
Human
1. Oral (mg/kg-day)b
None
2. Inhalation (mg/m3)b
None
Animal
1. Oral (mg/kg-day)b
PS
Subchronic
56M/56F SPF-ICR mice
in the diet, 7 d/wk, total
104 weeks; 7/7 sacrificed
at 13 weeks for analysis
0, 9.69,48.42,
242.18, 984.9
(males) and 0,
11.99,60.26,
305.80,1211.7
(females); intake
was author
determined
Decreased RBC count,
hematocrit, hemoglobin, and
particularly in total leukocyte
counts in male mice and blood
biochemical parameters
Not
identifiable
Run;
however,
results not
suitable for
POD
determination
9.69 (male
mice)
Mitsumori et al.,
1979
10M/10F F344 rats by
gavage, 7 d/wk, 13 weeks
0, 10, 25, 50, 100,
250 Purity 69.4%
Reduction in body weight in the
high dose group, particularly in
males
None
Not run
None
NCI, 1979
10M/10F B6C3FJ mice by
gavage, 7 d/wk, 13 weeks
0, 10, 25, 50, 100,
250 Purity approx.
69.4%
Respiratory lesions and focal
pneumonitis were seen in the
three highest doses
None
Not run
None
NTP, 1982
Chronic
50M/50F F344 rats by
gavage, 5 d/wk, 103 weeks
0,71.4, 142.9
Purity approx.
69.4%
Reduction in body weight and
survival was noted in the high
dose group
71.4
Not run
142.9
NCI, 1979
U.S.
EPA,
2010a
56M/56FSPF- ICR mice in
the diet, 7 d/wk, 104 weeks
0,8.41,40.1, 198,
927 (males) and 0,
7.58, 35.8, 194,
961 (females);
intake was author
determined
Hemosiderin deposition in
spleen, decrease in hemoglobin,
and erythrocyte (red blood cell
[RBC]) count
198 (males);
35.8
(females);
reported by
study authors
Not run
927 (males);
194
(females)
Mitsumori et al.,
1979
9
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table 2. Summary of Potentially Relevant Data for BCMEE (CASRN 108-60-1)
Notes3
Category
Number of Male/Female
Species, Study Type, and
Duration
Dosimetryb
Critical Effects
NOAELb
BMDL/
BMCLb
LOAELbc
Reference
(Comments)
50M/50F B6C3FJ mice
gavage, 5d/wk, 103 weeks
0,71.4, 142.9
Purity approx.
69.4%
No clinical observations; body-
weight changes were
comparable to the control group
None
Not run
None
NTP, 1982
Developmental
None
Reproductive
None
Carcinogenic
50M/50F F344 rats by
gavage, 5d/wk, 103 weeks
0,71.4, 142.9
Purity 69.4%
Significant dose-response trends
in tumor incidences were not
noted in either male or female
rats
None
Not run
None
NCI, 1979
In most instances,
the number of
tumors was higher
in the control group
compared to the
low and high dose
BCMEE treated
groups; the study
authors concluded
that these results
may partly be due
to lower survival
rates in the high-
dose group
10
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table 2. Summary of Potentially Relevant Data for BCMEE (CASRN 108-60-1)
Notes3
Category
Number of Male/Female
Species, Study Type, and
Duration
Dosimetryb
Critical Effects
NOAELb
BMDL/
BMCLb
LOAELbc
Reference
(Comments)
50M/50F B6C3FJ mice by
gavage, 5d/wk, 103 weeks
0,71.4, 142.9
Purity approx.
69.4%
A dose-related, statistically
significant increase in incidence
of alveolar/bronchiolar
adenomas were noted in male
and female mice (males: 5/50,
13/50, 11/50 in the control, low,
and high dose groups,
respectively; females: 1/50,
4/50, 8/50 in the control, low,
and high dose groups,
respectively); a statistically
significant increase in the
incidence of hepatocellular
carcinomas was noted in male
mice (5/50, 13/50, 17/50 in the
control, low-, and high-dose
groups, respectively)
None
Not run
None
NTP, 1982
Carcinogenic
56M/56F SPF-ICR mice in
the diet, 7 d/wk, 104 weeks
0,8.41,40.1, 198,
927 (males) and 0,
7.58,35.8, 194,
961 (females);
intake was author
determined
No significant (p < 0.05)
difference between controls and
treated mice for any tumor type
927 (males);
961 (females)
Not run
None
Mitsumori et al.,
1979
This was the only
chronic-duration
study that used
relatively pure
BCMEE (98.5%)
2. Inhalation (mg/m3)b
None
''IRIS = utilized by IRIS, date of last update; PS = principal study in bold text; POD dose also in bold font.
''Dosimetry, NOAEL, BMDL/BMCL, and LOAEL values are converted to human equivalent dose (HED in mg/kg-day) or human equivalent concentration (HEC in mg/m3).
Noncancer oral data are only adjusted for continuous exposure.
'Not reported by the study author but determined from data.
11
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
HUMAN STUDIES
Oral and Inhalation Exposure
No published studies investigating the effects of subchronic- or chronic-duration oral or
inhalation exposure to BCMEE in humans have been identified. HSDB (2010) reports that the
toxicity of BCMEE is less than that of dichloroethyl ether (isomer not specified). Damage is
reported to occur in the liver and kidneys rather than in the lungs. The report also stated that the
central nervous system (CNS) depressant action of chlorinated ethers leads to loss of
consciousness following high exposures (concentration estimates not provided).
ANIMAL STUDIES
Oral Exposure
The effects of oral exposures to BCMEE have been evaluated in subchronic- and
chronic-duration animal studies (NCI, 1979; NTP, 1982; Mitsumori et al., 1979). Published
studies pertaining to developmental and reproductive effects of BCMEE have not been
identified.
Short-term Study
NTP (1982) conducted a short-term 14-day study as part of its chronic-duration
carcinogenesis assay using an isomeric mixture of 69.4% BCMEE and 30%
2-chloro-l-methylethyl(2-chloropropyl)ether. Groups of five male and five female B6C3Fi mice
were administered this mixture of BCMEE isomers at 17.8-, 31.6-, 56.2-, 100-, 178-, 316-, or
562-mg/kg-body weight (BW) per day (mg/kg-day) via corn oil gavage for 14 consecutive days.
A control group was not used in this study. Mice were observed daily for mortality and were
weighed on Days 0, 7, and 14. Necropsies were performed on all animals at study termination.
Compound-related deaths were noted at the two highest doses (316 and 562 mg/kg-day). One
male mouse dosed with 56.2-mg/kg-day BCMEE was found dead on Day 7, and five male mice
died following the first 562-mg/kg-day dose of BCMEE. One female mouse each in the 100-
and 316-mg/kg-day dose groups were found dead on Days 8 and 6, respectively, and all 5 female
mice were dead on Day 1 following treatment with 562-mg/kg-day BCMEE. Animals (number
not specified) receiving 562 mg/kg-day exhibited a hunched appearance. No other signs of overt
toxicity were observed at the other dose levels. No compound-related gross lesions were noted
at any of the administered doses at necropsy (data not provided). Body-weight changes as result
of BCMEE exposure were not reported. Because an isomeric mixture of BCMEE was used in
this assay and detailed results from short-term BCMEE exposures were not reported in the
technical report, this study is of limited use for deriving toxicity values.
Subchronic-duration Studies
The study by Mitsumori et al. (1979) is selected as the principal study for deriving
the subchronic p-RfD. Mitsumori et al. (1979) conducted a 104-week chronic-duration toxicity
study in which groups of 56 male and 56 female specific-pathogen-free (SPF)-ICR mice were
fed a diet containing 0-, 80-, 400-, 2000-, or 10,000-ppm BCMEE (purity 98.5%) for 104 weeks.
Adjusted for continuous exposure, these levels correspond to doses of 0, 9.69, 48.42, 242.18, and
984.9 mg/kg-day in human males and 0, 11.99, 60.26, 305.80, and 1211.7 mg/kg-day in human
females. Animal body weights were determined weekly from Weeks 0 to 26, once every
2 weeks from Weeks 27 to 52, and once every 4 weeks from Weeks 52 to 104. During Week 13,
7 mice/sex/group were sacrificed after removing blood for testing. Blood for hematological and
biochemical testing was obtained from the posterior vena cava while the animals were under
anesthesia. Hematological examinations included determination of erythrocyte count,
12 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
hemoglobin concentration, leukocyte count, and hematocrit measurement. Tail vein blood was
collected for the determination of differential leukocyte count (%). Blood biochemical
examinations included determination of plasma glutamic-oxaloacetic transaminase (GOT),
glutamic-pyruvic transaminase (GPT), alkaline phosphatase (ALP), glucose, total protein (TP),
urea nitrogen (UN), cholesterol, and bilirubin. Urinalysis included determination of pH, protein,
glucose, ketone bodies, and occult blood. Mice that were moribund were euthanized and
examined in a manner similar to that which was used for mice sacrificed by design. All animals
that were euthanized by design or in extremis received a necropsy examination. Following
necropsy, the following organs were weighed: brain, pituitary, thyroid, heart, thymus, liver,
kidneys, spleen, adrenals, gonads (testes and ovaries), and muscle (triceps surae muscle of hind
leg). In addition to these organs, the salivary glands, lungs, lymph nodes, pancreas, stomach,
duodenum, jejunum, ileum, cecum, colon, seminal vesicles, prostrate, uterus, bladder, bone
marrow (femur), and other regions considered to present abnormalities following necropsy were
fixed for further examination.
Treatment-related mortalities occurred in both sexes in the 10,000-ppm dose group at
8 weeks after BCMEE administration. At Week 13, in the 10,000-ppm dose group, besides the
7 males and females that were euthanized by design, 8 male and 12 female mice were either dead
or euthanized in extremis. In contrast, besides the 7 males and females that were euthanized by
design, none of the animals in the other dose groups were dead or euthanized in extremis.
Although the duration of the observation period was not specified, the authors reported that the
general condition of animals in the 10,000-ppm dose group revealed smaller body size and
emaciation, which they primarily attributed to undernutrition due to food aversion rather than an
effect of BCMEE toxicity. Hematological examinations performed at 13 weeks indicated
dose-related reductions in the erythrocyte (red blood cell [RBC]) count, percent hematocrit (Ht),
and hemoglobin (Hb) levels in male mice, but this trend was not observed in female mice. As
outlined in Table B.l, the authors reported statistically significant drops in RBC counts
(p < 0.05), percent Ht (p < 0.05), and Hb (p < 0.01) levels in males beginning at the lowest
administered dose when compared to the control group. In contrast, statistically significant
drops in percent Ht (p< 0.05) and Hb (p < 0.01) levels were reported only at the highest dose in
female mice when compared to the concurrent controls.
Leukocyte counts exhibited a decreasing and statistically significant (p < 0,05, p < 0.01,
orp< 0.001) dose-response in males (see Table B.l); however, this trend was not noted in
females. The study authors reported a statistically significant (p < 0.05) increase in leukocyte
counts in the 2,000-ppm females compared to controls and a statistically significant (p < 0.01)
decrease in neutrophils in the 10,000-ppm females (see Table B. 1). Because a clear
dose-response trend was not observed in females, the toxicological significance of changes in
these leukocyte measurements in the female mice is unclear. In contrast to the hematological
results, blood biochemical examinations indicated a statistically significant (p < 0.05 orp < 0.01)
difference in GOT, GPT, ALP(males only), glucose, TP, and UN(females only) mainly in the
10,000-ppm males and females when compared to the control group (see Table B.2).
Absolute and relative organ weights and histopathological examination results of animals
euthanized by design or in extremis at 13 weeks were not reported by the study authors. The
LOAEL for the 13-week oral exposure is identified as an average daily dose of 80 ppm
(9.69 mg/kg-day) in male SPF-ICR mice for significant changes in hematological endpoints,
13 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
including significant reduction in total number of leukocytes. A NOAEL is not established for
this study.
NCI (1979) conducted a 13-week rat study as part of its chronic carcinogenesis assay.
Groups of 10 Fisher 344 (F344) rats/sex were administered 0, 10, 25, 50, 100, or 250 mg/kg-day
of 69.4% BCMEE, 2.1% bis(2-chloro-//-propyl)ether, and 28.5% of the mixed iso- and normal
ether (referred to as BCMEE mixture in the study summary below) in corn oil 7 days/week via
gavage for 13 weeks. A control group composed of 10 rats/sex received only corn oil. All
animals were checked daily for mortality; body-weight data were also collected. At the end of
13 weeks, all surviving animals were sacrificed, and necropsies were performed on all animals.
No treatment-related mortalities occurred. An adverse effect on body weight was
observed only at the 250-mg/kg-day dose, with males exhibiting a 20% drop and females
exhibiting an 8% drop in mean body weight compared with the corresponding controls. Detailed
results on histopathological evaluations, if conducted, were not presented in the NCI (1979)
technical report. A statistical analysis of these results could not be performed because
body-weight data for control animals were not provided in the study report. Because detailed
results from the 13-week exposure to the BCMEE mixture are not presented in the technical
report, and, also, because an isomeric mixture containing only 69.4% BCMEE was used in this
assay, a LOAEL and a NOAEL for the pure compound cannot be identified from this study.
Studies using chemicals of high purity are preferred because of the possibility that the observed
effects of exposure are caused by an impurity or by an interaction between BCMEE and the
impurity.
In a 13-week study conducted by the National Toxicology Program (NTP, 1982) as part
of its chronic carcinogenesis assay, groups of 10 B6C3Fi mice/sex were administered 0, 10, 25,
50, 100, or 250 mg/kg-day of 69.4% BCMEE and 30% 2-chloro-l-methyl ethyl (2-
chloropropyl)ether (referred to as BCMEE mixture in the study summary below) in corn oil
7 days/week via gavage for 13 weeks. All animals were checked daily for mortality and
morbidity and were observed weekly for overt signs of toxicity. Body-weight data were
collected on a weekly basis. At the end of 13 weeks, all surviving animals were sacrificed, and
necropsies were performed on all mice. Gross lesions, tissue masses, abnormal lymph nodes,
skin, mandibular lymph nodes, mammary gland, salivary gland, bone marrow, sternebrae, femur,
thymus, larynx, trachea, lungs and bronchi, and other organs from the control and high-dose
groups (high-dose groups not specified) were preserved for analysis.
The study authors stated that no compound-related changes in mean body weights were
observed in any of the animals. None of the treated animals died as a result of exposure to the
BCMEE mixture. While detailed results of histopathological evaluations were not presented in
the NTP (1982) technical report, histopathological changes were noted in the respiratory system.
Focal pneumonitis was observed in 3/10 males and 1/10 females, 2/10 males and 3/10 females,
and 8/10 males and 4/10 females in the 50-, 100-, and 250-mg/kg-day exposure groups,
respectively. Statistical analysis of these data indicates that the incidence of focal pneumonitis
was statistically significant (p = 0.0349) only in the high-dose male mice compared to the
low-dose male mice. Statistical analysis between the dosed and control groups could not be
performed because control data were not provided. Because an isomeric mixture containing only
69.4% BCMEE was used in this assay and detailed results from the 13-week exposure to the
14 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
BCMEE mixture are not presented in the technical report, the LOAEL and NOAEL for the
mixture (see Table 2) cannot represent pure BCMEE in a quantitative toxicity assessment.
Chronic-duration Studies
NCI (1979) conducted a 103-week chronic-duration toxicity and carcinogenicity study in
F344 rats. Groups of 50 F344 rats/sex were administered 0, 100, or 200 mg/kg-day of 69.4%
BCMEE, 2.1% bis(2-chloro-n-propyl)ether, and 28.5% of the mixed iso- and normal ether
(referred to as BCMEE mixture in the study summary below) in corn oil 5 days/week via gavage
for 103 weeks. The corresponding daily average doses for continuous exposure to the BCMEE
mixture were 0, 71.4, or 142.9 mg/kg-day, respectively. Two control groups composed of
50 animals/sex served as corn oil and untreated controls. All animals were observed twice daily
for overt signs of toxicity, and the presence of palpable masses was recorded on a weekly basis.
Mean body weights of the animals were recorded once every 2 weeks for the first 12 weeks of
the study, then monthly until Week 72, and then every 2 weeks until study termination. Animals
that were moribund and those that survived until study termination were sacrificed, and gross
and microscopic examinations were performed on major tissues. Presence of gross lesions was
not reported for animals that were sacrificed or animals that died during the study. Microscopic
examinations were performed on many tissues including sections from the lungs, bronchi,
trachea, kidneys, and liver.
Mean body weights of the male and female rats exhibited a dose-related trend, and dosed
animals had lower mean body weights than those of the control groups throughout the exposure
duration. Additionally, animals treated with the BCMEE mixture exhibited a hunched
appearance. The study authors stated that a departure from linear trend was noted in each sex
due to a relatively steep decrease in survival in the 200-mg/kg-day dose group. In male rats,
56% of animals in the high-dose group were alive at Week 78 of the study compared to 92% in
the low-dose group and 88% in the corresponding control groups. In females, 50% of animals in
the high-dose group were alive at Week 78 of the study compared to 88% in the low-dose group
and 96%) in the corresponding control groups. The study authors also reported that, except in the
high-dose group males and females, there were sufficient numbers of rats of each sex that were
at risk for the development of late-appearing tumors. As outlined in Table B.3, a notable
increase in the incidence of esophageal hyperkeratosis (82% and 65% in male and female rats,
respectively) was noted in male and female rats dosed with 200 mg/kg-day compared to the
corresponding control groups. Additionally, 10% of females treated with 200 mg/kg-day had an
increased incidence of esophageal acanthosis. In contrast, the incidence of gastric hyperkeratosis
was higher in vehicle controls compared to animals dosed with the BCMEE mixture. In addition
to these effects, a dose-related increase in the incidence of aspiration pneumonia was noted in
low- and high-dose male and female rats. Males exhibited a 14% and 24% increase in aspiration
pneumonia at the low- and high-doses, respectively, compared to 2% in vehicle control animals.
Females exhibited a 33% and 46% increase at the low- and high-doses, respectively, compared to
2% in the vehicle controls. Because an isomeric mixture of BCMEE was used in this assay and
the effects described above cannot be attributed exclusively to BCMEE exposure, a LOAEL and
a NOAEL cannot be identified for the pure compound from this study.
Evidence of carcinogenic activity of BCMEE was not observed in male or female
F344 rats. Tumor incidences in dosed groups were not significantly higher than those noted in
the vehicle controls. Significant results, using the one-tailed Fisher's exact test in the negative
direction, were reported in the incidences of hematopoietic tumors, tumors of the adrenal,
15 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
preputial gland, and testes in the males, and in the tumors of the pituitary gland, uterus, and
pancreatic islets in females. The NCI (1979) report stated that the apparent negative
dose-response relationships may be attributed to the relatively low survival of rats in the
200-mg/kg-day group. The study authors stated that, although tumors outlined above were noted
in animals dosed with the BCMEE mixture, the incidences of these tumors were lower in the
dosed groups than in the corresponding control groups.
Two male rats, one in the high-dose group and one in the low-dose group, died during
Week 15 of the study with malignant lymphoma affecting multiple organs. The NCI (1979)
report stated that these early deaths with tumors were not considered to be treatment related
because F344 rats are known to be prone to juvenile lymphoid tumors. The NCI (1979)
concluded that, under the conditions of the bioassay, BCMEE was not carcinogenic to F344 rats
of either sex. However, the NCI report also stated that BCMEE cannot be considered adequately
tested until additional bioassays have been conducted in other animal species.
NTP (1982) conducted a 103-week chronic-duration toxicity and carcinogenicity study in
B6C3Fi mice. Groups of 50 mice/sex were administered 0, 100, or 200 mg/kg-day of 69.4%
BCMEE and 30% 2-chloro-l-methylethyl(2-chloropropyl)ether (referred to as BCMEE mixture
in the study summary below) in corn oil 5 days/week via gavage for 103 weeks. Groups of
50 mice/sex received corn oil alone and served as vehicle controls. The corresponding daily
average doses adjusted for continuous exposure were 0, 71.4, or 142.9 mg/kg-day, respectively,
for the BCMEE mixture. All animals were observed twice daily for mortality and morbidity.
Clinical signs were recorded on a monthly basis. Body weights were recorded on a weekly basis
for the first 13 weeks and once a month thereafter until study termination. Moribund animals
and animals surviving until the end of study were sacrificed and necropsied. All major tissues
and organs were examined for grossly visible lesions. Microscopic examinations were also
performed on the mammary gland, salivary gland, bone marrow, thymus, larynx, trachea, lungs
and bronchi, heart, thyroid, parathyroid, esophagus, stomach, duodenum, liver, gallbladder,
pancreas, spleen, kidneys, adrenals, urinary bladder, seminal vesicles/prostate/testes (males) or
ovaries/uterus (females), brain, and pituitary. Additionally, sections of the nasal turbinates were
examined in male mice treated with 200 mg/kg-day of the BCMEE mixture.
Treatment with the BCMEE mixture had no effects on clinical observations or body
weights throughout the duration of the study. Although there were no significant differences in
animal survival between the dosed groups and control group, in males, 82%, 88%, and 74% of
animals survived until study termination in the control, low-dose, and high-dose groups,
respectively. In females, 62%, 68%, and 56% of animals survived until study termination in the
control, low-dose, and high-dose groups, respectively. Nonneoplastic lesions observed included
a 60% increase in chronic inflammation of the nasal cavity in male mice treated with
200 mg/kg-day of the BCMEE mixture, as well as a 12% and 28% increase in fatty
metamorphosis of the liver in the 100- and 200-mg/kg-day dose groups, respectively, compared
to a 2% increase in the control group (per data reported by NTP). Additionally, a 56% increase
in chronic inflammation was noted in the naso-lacrimal duct of male mice treated with
200 mg/kg-day of the BCMEE mixture. A similar analysis of nonneoplastic lesions in females
indicated a 71% increase in the incidence of cystic hyperplasia in the uterus/endometrium of the
100- and 200-mg/kg-day treated female mice, but the effect was comparable to a 61% increase in
the control group (per data reported in NTP [1982]). Because an isomeric mixture of BCMEE
16 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
was used in this assay and the effects described above cannot be attributed to BCMEE exposure,
a LOAEL and a NOAEL cannot be identified for the pure substance from this study.
Evidence of carcinogenic activity was well supported in both male and female mice (see
Table B.4). Statistical significance was evaluated by the authors using both the Incidental Tumor
Test and the Fischer Exact Test (see Table B.5). In the lung, statistically significant (p < 0.05)
increases in alveolar/bronchiolar adenomas occurred in male and female mice with a positive
trend. In male mice, 13/50 and 11/50 exhibited these tumors in the 100- and 200-mg/kg-day
groups, respectively, compared to 5/50 in the control group. In female mice, 4/50 and
8/50 exhibited these tumors in the 100- and 200-mg/kg-day groups, respectively, compared to
1/50 in the control group. The tumor incidence was significantly (p < 0.029) higher in the
high-dose females than in the control group. The combined incidence of alveolar/ bronchiolar
adenomas and carcinomas indicated a significant (males,/? < 0.062; femalesp < 0.004; high dose
group) positive trend with the males and females treated with the BCMEE mixture exhibiting
significantly (males,/? < 0.035; females,/? < 0.008; high dose group) higher incidences of the
combined tumors compared to the control group (see Table B.5). In the liver, hepatocellular
carcinomas exhibited a statistically significant (p < 0.004; high dose group) positive trend in
male mice with the incidence of tumors significantly (p < 0.007) higher in mice treated with
200-mg/kg-day BCMEE mixture compared to the corresponding control group (see Table B.5).
The combined incidence of hepatocelluar adenomas and carcinomas was significant (p < 0.003)
in trend tests in male mice, with male mice treated with the 200-mg/kg-day BCMEE mixture
exhibiting a significantly (p < 0.005 in all tests) higher incidence of these tumors compared to
the control group (see Table B.5). Metastases to the lung was reported in 1/50, 4/50, and
3/50 mice in the control, low dose, and high dose groups, respectively. In contrast, incidences of
livers tumors in female mice were not statistically significant. In the hematopoietic system,
incidence of histiocytic lymphoma was noted in 3/50 (6%) male micei treated with
200 mg/kg-day with a positive trend (p < 0.086). However, incidences of other types of
lymphoma were not observed in the male mice, and female mice did not exhibit any type of
malignant lymphoma at a statistically significant level. Additionally, squamous-cell papillomas
were seen in 2/49 female mice treated with 200 mg/kg-day of the BCMEE mixture and in
1/50 low-dose and 1/50 high-dose male mice. Squamous-cell carcinoma was observed in one
high-dose female mouse that did not have squamous-cell papillomas.
Mistumori et al. (1979) investigated the chronic toxicity of BCMEE using SPF-ICR mice.
Groups of 56 mice/sex were fed a diet containing 0-, 80-, 400-, 2000-, or 10,000-ppm BCMEE
(purity 98.5%) seven days/week for 104 weeks. The total average daily intake of BCMEE as
calculated by the study authors was 0, 8.41, 40.1, 198, and 927 mg/kg-day in male mice and 0,
7.58, 35.8, 194, and 961 mg/kg-day in female mice. Body weights were determined weekly
from Weeks 0 to 26, once every 2 weeks from Weeks 27 to 52, and once every 4 weeks from
Weeks 53 to 104. After study initiation, following the removal of blood samples for analysis,
7 mice/sex/group were sacrificed by design at Weeks 13, 26, and 52, and 6 mice/sex/group were
sacrificed at Week 78. All remaining surviving animals were followed through Week 104 prior
to sacrifice. Blood samples obtained prior to sacrifice were used for hematological and
biochemical examinations. Necropsies were performed on all animals that were sacrificed by
design and in extremis. Animals that died during the course of the study were also necropsied.
Organs weights for brain, pituitary, thyroid, heart, thymus, liver, kidneys, spleen, adrenals,
gonads (testes, ovaries), and muscle (triceps surae muscle of hind leg) were recorded. In
addition to these organs, the salivary gland, lungs, lymph nodes, pancreas, stomach, duodenum,
17 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
jejunum, ileum, cecum, colon, seminal vesicles, prostrate, uterus, bladder, bone marrow, and
other regions were examined for chemical-specific abnormalities.
The study authors reported smaller body size and emaciation in both male and female
mice in the 10,000-ppm dose group, which they attributed to undernutrition. At Week 52, the
cumulative death in the 10,000-ppm BCMEE dose group was 14 males and 22 females,
compared to 3 males and 4 females in the control group. There was a remarkable inhibition in
weight gain beginning Week 1 in the 10,000-ppm treatment group that continued until study
termination at 104 weeks. A similar tendency of lower body-weight gain was also noted in
female mice treated with 2000-ppm BCMEE. Additionally, the study authors also reported a
significant (significance level not reported) difference in body weight during certain weeks
(weeks not specified) in other treated groups. Food consumption was markedly lower in male
and female mice treated with 10,000-ppm BCMEE throughout the study period. In contrast, in
other treatment groups, generally stable values in consumption were observed throughout the
study period, though some fluctuations were noted. Based on the study authors' evaluation, food
aversion is the likely cause of reduced body weight in the 10,000-ppm dose group and not due to
the toxicity of BCMEE.
Evaluation of hematological parameters indicated a mild, but statistically significant
(p < 0.05) decrease in erythrocyte count in males beginning at the lowest dose level of 80 ppm at
Weeks 13 and 26, and in females during Weeks 26 and 52 (significance level p < 0.05 at
Week 52 beginning at 2000 ppm; see Table B.l for 13-week results). Mild depression of
hematocrit levels, compared to the control group, was noted in both sexes of the same dose
groups at Weeks 13 (see Table B.l) and 52 (see Table B.6), and in females at Week 26 (data not
provided in article). Additionally, hemoglobin concentration in the 10,000-ppm group was
reduced in male and female mice at Weeks 13 (see Table B. 1) and 26, and in females at Week 52
(see Table B.6). Leukocyte counts exhibited a decreasing trend in 10,000-ppm males at each
assigned period of sacrifice by design. Additionally, 400- and 2000-ppm group male mice and
10,000-ppm group female mice showed a small decrease in leukocyte counts at Weeks 13 (see
Table B. 1). Differential leukocyte counts exhibited reduction in leukocytes and increases in
polymorphonuclear neutrophils in 10,000-ppm males at Weeks 13 (see Table B.l) and 26 and in
females euthanized in extremis during the first 13 weeks of treatment (see Tables B.l).
Blood biochemical examination showed a significant (p < 0.01) increase in plasma GOT
and GPT levels in both male and female mice treated with 10,000-ppm BCMEE during
Week 13. UN levels were significantly (p < 0.05 orp< 0.01) increased in female mice treated
with 10,000-ppm BCMEE during Weeks 13 and 52. In male mice, statistically significant
(p < 0.0.1 orp< 0.05) increases in UN levels were seen only during Week 52 at the 400-, 2000-,
and 10,000-ppm dose levels. In addition to GOT and GPT levels, ALP levels were increased in
male mice during Week 13 and in both sexes during Weeks 26 and 52 (data not provided in
article). Minor decreases in total protein were also seen in both sexes, primarily in animals
treated with 10,000-ppm BCMEE during Weeks 13 (males only), 26, and 52. The study authors
also reported reductions in blood glucose levels in both sexes during Weeks 13 (see Table B.2),
26, and 52 (see Table B.8), and in females during Week 78 (data not provided in article).
Though some significant changes in organ weights in male mice treated with 10,000-ppm
BCMEE were noted, in general, the absolute and relative organ weights of animals treated with
BCMEE did not show marked adversity and weight changes corresponding to decreased body
18 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
weights. Table B.9 presents absolute and relative organ weights for select organs in SPF-ICR
mice.
Examination of nonneoplastic endpoints indicated an increased incidence of hemosiderin
deposition in the spleen. Seventeen males and 17 females in the 10,000-ppm dose group
exhibited increased hemosiderin deposition in the spleen compared to only 1 male and 2 females
in the control group. Additionally, splenic hemosiderin deposition was also noted in females
during Week 13 and in males during Weeks 13, 26, and 52. The study authors also reported
mild-to-moderate increases in extramedullar hematopoieses in the spleen in males during
Week 13 (numerical data not reported). Though not statistically significant, this effect was
associated with a high number of mice exhibiting erythroblastic hyperplasia compared to the
control group at 104 weeks (see Table B. 10). The study authors identified a NOAEL for
BCMEE of 2,000 ppm (198 mg/kg-day) in male mice and 400 ppm (35.8 mg/kg-day) in females
for hematological changes. Based on these results, a chronic LOAEL of 10,000 ppm
(927 mg/kg-day) for males and 2,000 ppm (194 mg/kg-day) for females is identified in this
study.
No evidence of BCMEE-related carcinogenicity was observed in either male or female
mice. Adenomas of the lung, lymphatic leukemia, reticulum cell sarcomas, and other types of
tumors were observed at a relatively high incidence in each of the groups. However, there was
no difference in the incidence of these tumors, age of onset, and histological findings between
the treatment groups and the control group. Sporadic tumors, such as benign papilloma of the
forestomach in one male mouse, and a granulose cell tumor, two adenomas of the ovary, three
pituitary adenomas, and a uterine leiomyoma in females were observed, but the incidences of
these tumors in male and female mice were low and independent of BCMEE dose. Similarly, the
occurrences of malignant tumors such as carcinoma of the lung, subcutaneous leiomyosarcoma,
subcutaneous osteogenic sarcoma, and subcutaneous undifferentiated tumor in males, and
undifferentiated tumors in the uterus or peritoneum in females were sporadic, and the incidences
were low. The study authors concluded that because the occurrence of benign and malignant
tumors were sporadic with low incidence, the evidence of carcinogenic activity in male and
female SPF-ICR mice was negative.
Inhalation Exposure
No studies investigating the effects of subchronic- or chronic-duration inhalation
exposure to BCMEE in animals were identified.
Other Studies
Developmental and Reproductive Toxicity Studies
No studies pertaining to the developmental and reproductive toxicity of BCMEE were
identified.
Other Data (Short-Term Tests, Other Examination)
Little information is available on the toxicokinetics of BCMEE. Results of available
studies (i.e., U.S. EPA, 1987; Smith et al., 1978) show evidence of saturation of absorption
mechanisms at high doses, with concentrations of radioactivity peaking in blood 2-4 hours after
treatment at the lower doses. A ti/2 for the elimination of BCMEE and its metabolites from blood
in monkeys was reported to be about 5 hours in the a-phase, while, in rats, the tm was
approximately 48 days. Beyond 24 hours, elimination curves for blood in monkeys and rats
19 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
were stated to be identical (Smith et al., 1978). Smith et al. (1978) also studied tissue
distribution of BCMEE by administering a single parenteral dose of 30 mg/kg to rats and
monkeys. In rats, a significant (exact percentage not reported in study summary) amount of
BCMEE was excreted via the bile with reabsorption into the intestine. In contrast, in monkeys,
there seemed to be a statistically significant (28.8 mg/kg) accumulation of BCMEE in the liver
with very little excretion of BCMEE or its metabolites, via the bile into the intestines.
Additionally, much higher concentrations of radioactivity were noted in the brain and muscle
mass (3.3 mg/kg in both tissues) of the monkeys compared to the rats. Elimination of BCMEE is
primarily in urine and is rapid, and is composed of BCMEE and its metabolites (1-chloro-
2-propanol [CIP], and propylene) with the rats excreting approximately twice (63.36% of the
administered dose) as much BCMEE compared to the monkeys (28.61% of the administered
dose). Excretion in the feces ranged from 1% in the monkeys to 6% in the rats.
The genotoxicity of BCMEE has been tested in a select number of studies (e.g. Zeiger,
1987; Moriya et al., 1983; Mirsalis et al., 1989; Jorgenson et al., 1977; and McGregor et al.,
1988) using in vitro and in vivo test systems. Test results were equivocal, with some results
indicating genotoxicity, while others were negative.
Table 3 summarizes the toxicokinetics and genotoxicity studies for BCMEE.
20
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table 3. Other Studies
Tests
Materials and Methods
Results
Conclusions
References
Toxicokinetic
Three female CD rats were
administered (route not specified) a
single dose of 0.2 |ig/kg to 300 mg/kg
of BCMEE (purity >95%); tissue
distribution was studied by treating
three rats with 30 mg/kg of BCMEE
via intraperitoneal injection and
treating one monkey with 30 mg/kg of
BCMEE via intravenous injection.
Tail vein blood from rats was collected
at specified intervals between
15 minutes and 48 hours after BCMEE
administration, and animals were
sacrificed after 48 hours; monkey
blood was also collected (intervals not
specified); tissues from various organs
were collected from rats and monkeys
that were sacrificed 7 days after
BCMEE administration; urine and
feces were collected up to 168 hours
after BCMEE administration along
with expired air (collection times not
specified).
There was evidence of saturation
of absorption mechanisms at high
doses. In the rat, a large
proportion of material seemed to
be excreted via the bile, with
reabsorption by the intestine.
Tissue distribution indicated that
monkeys had an accumulation of
28.8 mg/kg of BCMEE in the
liver. Monkeys also had
substantially higher concentrations
in the brain and muscle mass
(3.3 mg/kg in both tissues)
compared to rats. Elimination of
BCMEE was primarily in urine
and was rapid, and was composed
of BCMEE and its metabolites,
with the rats excreting
approximately twice as much
(63.36% of the administered dose)
BCMEE compared to the monkeys
(28.61%) of the administered
dose). Excretion in the feces
ranged from 1% in the monkeys to
6% in the rats. Two metabolites
of BCMEE were identified in the
urine: l-chloro-2-propanol (CIP)
and propylene oxide (PO).
BCMEE is well absorbed from the
gastrointestinal tract in the rats—
but not monkeys. Tissue
distribution indicated higher levels
of BCMEE in the fat, urine, and
feces of the rat, whereas higher
quantities were found in the
muscle and liver of the monkey.
Elimination is rapid,
predominantly as BCMEE and its
metabolites in urine. Excretion in
feces was minimal.
Smith et al., 1978
21
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table 3. Other Studies
Tests
Materials and Methods
Results
Conclusions
References
Toxicokinetic
Seven adult male Sprague-Dawley rats
were administered 14C BCMEE in corn
oil at 90 mg/kg via gavage.
Radioactivity was measured in expired
air, urine, feces, carcass, and cage wash
48 hours after BCMEE administration.
Total 14C recovery was
73.3 ± 7.7% of the administered
BCMEE dose. Fecal excretion
accounted for 3.8% of the
administered BCMEE dose.
Excretion rates in expired air and
urine were not provided.
The authors state that the results
suggest that gastrointestinal
absorption of BCMEE was nearly
complete.
U.S. EPA, 1987
Genotoxicity
Salmonella mutagenicity study results
for BCMEE were obtained from two
programs. In one program, Salmonella
strains TA98, TA100, TA1535, and
TA1537 were used in a standard plate
assay without metabolic activation and
with activation by liver S9 preparations
from uninduced and Aroclor
1254-induced male Fischer 344 rats,
B6C3Fi mice, and Syrian hamsters. In
the second program, strains TA98,
TA100, TAI535, and either TA97 or
TAI537 were used in a preincubation
assay without activation and with liver
S9 preparations from Aroclor
1254-induced male Sprague-Dawley
rats and Syrian hamsters.
The author concluded that
BCMEE is mutagenic. BCMEE
was reported to be mutagenic in
TA98 and TA100—but not
mutagenic in TA97, TA1535, and
TA1537 strains. The study does
not state whether mutagenicity
was observed both with and
without S9 activation.
This is a review article outlining
the carcinogenicity and
mutagenicity of 224 chemicals.
BCMEE was characterized as a
bacterial mutagen by the review
author.
Zeiger, 1987
22
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table 3. Other Studies
Tests
Materials and Methods
Results
Conclusions
References
Genotoxicity
Ames mutagenicity assay was used to
test for mutagenic potential.
Salmonella strains, TA98, TA100,
TA1535, TA1537, and TA1538 were
used along with a WP2 her strain of
Escherichia coli with and without S9
activation. Though specific tested
doses are not reported, the authors state
that BCMEE, along with several other
pesticides, was tested up to a dose of
5000 |ig/plate.
The authors concluded that
BCMEE had a negative response
in the mutagenicity assay
(strain-specific information not
provided).
The article reports mutagenicity
results for 228 pesticides
including BCMEE. The study
authors concluded that BCMEE is
not mutagenic in the Ames assay.
Moriya et al.,
1983
Genotoxicity
Male F344 and male and female
B6C3Fi mice were treated with
BCMEE in corn oil as a single bolus
dose via gavage. Doses of 20, 100, and
400 mg/kg were used in male and
female mice to study unscheduled
DNA synthesis (UDS) induction.
Induction of S-phase synthesis (SPS) in
male and female mice was examined
using doses of 75-, 300-, and
400-mg/kg BCMEE.
Induction of UDS was not
observed in mice treated with
BCMEE. The study authors
reported that the SPS induction
was positive, particularly at higher
doses in male mice, but equivocal
in female mice.
In this study, 19 chemicals were
evaluated for their potential to
cause UDS and SPS. UDS was
not induced in either male or
female mice as a result of BCMEE
exposure. SPS induction was
positive in male mice but
equivocal in female mice.
Mirsalis et al.,
1989
23
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table 3. Other Studies
Tests
Materials and Methods
Results
Conclusions
References
Genotoxicity
Male mice (strain not specified) were
treated daily via gavage for 8 weeks
with three doses of BCMEE (doses not
specified) to examine the mutagenic
potential of BCMEE via the heritable
translocation test. After treatment,
each male was mated with two virgin
females to produce an F1 generation.
Upon maturity, 100 F1 males per
treatment group then were selected and
bred to three virgin females. Pregnant
females were evaluated against a set of
predetermined selection criteria (not
specified) to identify compromised
males. These males were rebred with
three additional virgin females.
Presumptive F1 males were examined
cytogenetically after two breedings.
Preliminary evaluations indicated
that heritable translocations did
not occur in animals exposed to
BCMEE.
Detailed results were unavailable
because the study results were
retrieved from an abstract. A
publication outlining detailed
study results could not be located.
Jorgenson et al.,
1977
Genotoxicity
Mouse lymphoma L5178 tk+/tk" cells
were used to test the mutagenic
potential of BCMEE. Cultures cells
(6 x 106) were treated with BCMEE (0,
DMSO, 62.5, 125, 250, 500, 1000,
2000 (J,g/ml) without S9 activation.
Mutant-forming colonies were
significantly elevated (p < 0.05) at
250, 500, and 1000 (J,g/ml in a
significant (p < 0.05)
dose-response trend.
BCMEE is mutagenic under the
conditions of the test.
McGregor et al.,
1988
24
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
DERIVATION OF PROVISIONAL VALUES
Table 4 presents a summary of noncancer reference values. Table 5 presents a summary
of cancer values. The toxicity values were converted to HEC/HED units, and the conversion
process is presented in the text below. IRIS data are indicated in the table if available.
DERIVATION OF ORAL REFERENCE DOSE
Derivation of Subchronic p-RfD
Three publications (NTP, 1982; NCI, 1979; and Mitsumori et al., 1979) were considered
as principle studies. Both NCI (1979) and NTP (1982) used a mixture of approximately
69.4% BCMEE with about 30% of related compounds, thus making them unsuitable for
evaluating the effects of high purity BCMEE. Because they used relatively high purity (98.5%)
BCMEE, the study by Mistumori et al. (1979) is selected as the principal study for the derivation
of the subchronic p-RfD. The critical endpoints are statistically significant (p < 0.05) changes in
RBC count, hematocrit, hemoglobin, and total leukocytes in male SPF-ICR mice at the lowest
dose tested (9.96 mg/kg-day). This study, published in a peer-reviewed journal, was conducted
with multiple doses (0, 80, 400, 2000, 10,000 ppm in the diet) with a variety of toxicologic
endpoints that demonstrated a statistically significant dose response in male and female rats, with
interim sacrifice of seven animals per sex per dose at 13, 26, 52, and 104 weeks. Among the
available subchronic-duration studies, the Mitsumori et al. (1979) study is the only one that
provides information for the determination of a credible point of departure (POD) for deriving a
subchronic p-RfD using relatively pure BCMEE. Thus, the Mitsumori et al. (1979) study
provides a LOAEL (9.69 mg/kg-day) as the only POD.
25 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Table 4. Summary of Noncancer Reference Values for BCMEE (CASRN 108-60-1)
Toxicity Type (Units)
Species/
Sex
Critical Effect
p-Reference
Value
POD Method
POD
UFc
Principal Study
Subchronic p-RfD
(mg/kg-day)—
Screening Value
Mouse/M
Decreased RBC count,
hematocrit,
hemoglobin, and
particularly in total
leukocyte counts in
male mice and blood
biochemical
parameters
1 x 1(T3
LOAEL
9.69
10,000
Mitsumori et al. (1979)
Chronic RfDa
(mg/kg-day) (IRIS)
Mouse/F
Decreased hemoglobin
concentration and
possible erythrocyte
destruction
4 x 10~2
NOAEL
35.8
1000
Mitsumori et al. (1979)
Subchronic p-RfC
(mg/m3)
None
None
None
None
None
None
None
Chronic p-RfC
(mg/m3)
None
None
None
None
None
None
None
"Value from IRIS (EPA, 2010a).
Table 5. Summary of Cancer Values for BCMEE (CASRN 108-60-1)
Toxicity Type
Species/Sex
Tumor Type
Cancer Value
Principal Study
p-OSF
None
None
None
None
p-IUR
None
None
None
None
26
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table 6. Summary of Potentially Relevant Oral Systemic Subchronic Toxicity Studies for
BCMEE
#/Sex (M/F)
Critical Endpoint
Exposure
(ppm)
Frequency/
Duration
NOAEW
(mg/kg-day)
LOAEW
(mg/kg-day)
References
56/56, mice;
7/7
evaluated at
13 weeks
Decreased RBC
count, hematocrit,
hemoglobin, and
particularly in total
leukocyte counts in
male mice and
blood biochemical
parameters
0, 80, 400,
2000, 10,000
Daily for a total
of 104 weeks
None
9.69 (males)
Mitsumori et al.,
1979
10/10, F344
rats
Adverse effect on
body weight at the
highest dose; an
isomeric mixture of
BCMEE was used
which precludes the
identification of a
NOAEL and
LOAEL
0, 10, 25, 50,
100, 250
7 d/wk for
13 weeks
None
None
NCI, 1979
10/10,
B6C3Fi
mice
Focal pneumonitis at
the three highest
doses administered;
an isomeric mixture
of BCMEE was
used which
precludes the
identification of a
NOAEL and
LOAEL
0, 10, 25, 50,
100, 250
7 d/wk for 13
weeks
None
None
NTP, 1982
aNOAELADj or LOAELadj = Dose (NOAEL or LOAEL) x Food Consumption Value ^ day x (1 -f- BW Value) x
Days Dosed ^ Total Days in Study.
27
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
A benchmark dose (BMD) analysis of the various hematological and biochemical
parameters was conducted for the Mitsumori et al. (1979) study to determine if a credible
benchmark dose lower bound (BMDL) could be established for the derivation of a p-RfD.
Modeling was performed with—and without—the highest dose (984.9 mg/kg-day) because the
reduced body weight at that dose may not represent an effect of BCMEE toxicity. BMD models
were run, but an adequate fit could not be attained because, in most instances, consistent,
monotonic dose-response relationships were not observed at the doses that were modeled. All
model runs failed a visual inspection and one or more of the four BMD model test. Additionally,
not all BMD modeling criteria were met with many of the resulting BMDLs being extremely
small (e.g., BMDL = 7.4 x io~6 mg/kg-day; Table C.l shows these values as a zero). This can
occur when the dose range in the study does not adequately cover the selected benchmark
response level (BMRisd) This was true in the Mitsumori et al. (1979) study. Consequently, a
traditional NOAEL/LOAEL approach has been used for the derivation of a subchronic p-RfD. A
POD of 80 ppm or 9.69 mg/kg-day in male SPF-ICR mice has been identified using the
conventional NOAEL/LOAEL approach from the Mitsumori et al. (1979) study.
Since a NOAEL could not be determined from the Mitsumori et al. (1979) study, the
BMD analysis did not provide an acceptable POD, and because no acceptable multigeneration
reproduction or developmental studies were identified, the composite uncertainty factor (UFC)
exceeds 3000 (see Table A.l). A very high level of uncertainty (UFc > 3000) precludes
derivation of a subchronic p-RfD. Hence, a screening value is presented in Appendix A.
Derivation of a Chronic RfD
A chronic RfD of 4 x 10~2 is included in the IRIS database (U.S. EPA, 2010a) based on
the critical endpoint of decreased hemoglobin and possible erythrocyte destruction observed in a
104-week study, in which SPF-ICR mice were exposed to BCMEE in the diet. A NOAEL of
35.8 mg/kg-day and a LOAEL of 198 mg/kg-day were identified by the study authors
(Mitsumori et al., 1979) with the NOAEL serving as a POD for chronic RfD derivation.
"3
It should be noted that the screening subchronic p-RfD value (1 x 10" mg/kg-day; see
Appendix A) is lower than the chronic RfD (4 x 10" mg/kg-day) because hematopoietic effects
were more significant during the 13-week observation period compared to the observations at
104 weeks (see Table B. 1) that were used to derive the chronic RfD (U.S. EPA, 2010a). After
13 weeks of exposure, significant changes in hematological endpoints in the male mice appeared
at the lowest administered BCMEE dose, precluding the identification of a NOAEL. This led to
the application of an additional uncertainty factor (UFL) of 10 in the derivation of a subchronic
p-RfD. The chronic RfD is based on the chronic-duration study of Mitsumori et al. (1979) in
SPF-ICR mice. The chronic-duration study does not show direct evidence of hematological
effects after 52 weeks of the study, but does provide a description of splenic effects—namely
hemosiderin deposition. Although hemosiderin deposition may be regarded as a sequelae of
hematological effects, the dose-response relationship between them is not characterized. Also,
other chronic-duration studies of BCMEE exposure in mice, most notably that of the NTP (1982)
study that used B6C3Fi mice, showed no evidence of chemical-related effects in the spleen.
This observation raises the possibility of a species-specific effect whose relevance to other
strains of mice, and to humans, may also be uncertain.
28 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
DERIVATION OF INHALATION REFERENCE CONCENTRATIONS
No published studies investigating the effects of subchronic- or chronic-duration
inhalation exposure to BCMEE in humans or animals were identified. This precludes the
derivation of subchronic and chronic inhalation toxicity values.
CANCER WEIGHT-OF-EVIDENCE DESCRIPTOR
Table 7 identifies the cancer WOE descriptor for BCMEE.
29
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table 7. Cancer WOE Descriptor for BCMEE
Possible WOE
Descriptor
Designation
Route of Entry (Oral,
Inhalation, or Both)
Comments
"Carcinogenic to
Humans "
N/A
N/A
No human studies are available.
"Likely to be
Carcinogenic to
Humans "
N/A
N/A
No strong animal cancer data are available.
"Suggestive Evidence of
Carcinogenic Potential"
X
Oral administration by
gavage only
Under the 2005 Guidelines for Carcinogenic Risk Assessment (U.S. EPA, 2005), the
available evidence from oral exposure to BCMEE is suggestive of carcinogenic
potential based on evidence of carcinogenicity in male and female mice in the NTP
(1982) gavage bioassay. Results of NTP (1982) show statistically significant
increases in incidences of alveolar/bronchiolar adenomas and carcinomas in treated
male and female mice compared to study and historical controls. Additionally,
incidences of hepatocellular adenomas and carcinomas were significantly increased
in treated males compared to the control group, with metastases occurring in the
lung. Rare forms of squamous cell papillomas were seen in the stomach or
forestomach of females and males. The NTP study authors report that because
these stomach tumors are rare in B6C3Fi mice, the presence of these tumors,
particularly in high-dose female mice, were probably related to administration of
BCMEE. Because this study utilized an isomeric mixture of 69.4% BCMEE and
30% 2-chloro-l-methylethyl (2-chloropropyl)ether, tumor occurrence in B6C3Fi
mice cannot be firmly associated with exposure to BCMEE. Exposure-related
tumors have not been observed in male and female rats exposed via gavage to
BCMEE for 103 weeks (NCI, 1979). There was no evidence of carcinogenicity in
male and female SPF-ICR mice fed diets containing high purity (98.5%) BCMEE
(Mitsumori et al., 1979). Studies evaluating the carcinogenic potential of inhaled
BCMEE in humans or animals were not located.
"Inadequate Information
to Assess Carcinogenic
Potential"
N/A
N/A
Available information adequate to assess carcinogenic potential.
"Not Likely to be
Carcinogenic to
Humans "
N/A
N/A
No strong evidence of noncarcinogenicity in humans is available.
30
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
MODE-OF-ACTION DISCUSSION
The Guidelines for Carcinogenic Risk Assessment (U.S. EPA, 2005) define
mode-of-action as "a sequence of key events and processes starting with the interaction of an
agent with a cell, proceeding through operational and anatomical changes, and resulting in
cancer formation. Because the mechanism of potential carcinogenicity of BCMEE has not yet
been investigated, a discussion of the mode-of-action is not applicable.
DERIVATION OF ORAL SLOPE FACTOR
No published studies demonstrating carcinogenic effects of chronic-duration oral
exposure to relatively pure BCMEE in humans or animals were identified. An obsolete oral
slope factor (OSF), of 7 x 10~2 reported in the HEAST (U.S. EPA, 2010b), was derived from an
NTP (1982) gavage study in mice. Existing studies showing a positive dose-response
relationship between BCMEE exposure and tumor formation in mice (NTP, 1982)—but not in
rats (NCI, 1979)—used a mixture of 69.4% BCMEE and 30% other isomers and could not be
used to derive an OSF for pure BCMEE. Consequently no p-OSF is developed.
DERIVATION OF INHALATION UNIT RISK
No published studies demonstrating carcinogenic effects of chronic-duration inhalation
exposure to relatively pure BCMEE in humans or animals were identified. This precludes the
derivation of inhalation unit risk (IUR) values. An obsolete IUR of 3.5 x 10 2, reported in the
HEAST (U.S. EPA, 2010b), is derived by route-to-route extrapolation from an NTP (1982)
gavage study in mice. The EPA methodology (U.S. EPA, 2005) allows for such extrapolation—
but sufficient information from metabolic studies and pharmacokinetic/pharmacodynamic studies
is not available for reliable route-to-route extrapolation. Existing studies showing a positive
dose-response relationship between BCMEE exposure and tumor formation in mice (NTP,
1982)—but not in rats (NCI, 1979)—used a mixture of 69.4% BCMEE and 30% other isomers
and could not be extrapolated to an IUR for pure BCMEE. Consequently no p-IUR is developed.
31
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
APPENDIX A. PROVISIONAL SCREENING VALUES
DERIVATION OF SCREENING PROVISIONAL ORAL REFRENCE DOSES
Derivation of Screening Subchronic Provisional RfD (subchronic p-RfD)
For reasons noted in the main document, it is inappropriate to derive a provisional
subchronic p-RfD for BCMEE. However, information is available which, although insufficient
to support derivation of a provisional toxicity value under current guidelines, may be of limited
to use to risk assessors. In such cases, the Superfund Health Risk Technical Support Center
summarizes available information in a supplement and develops a screening value. Appendices
receive the same level of internal and external scientific peer review as the main document to
ensure their appropriateness within the limitation detailed in the document. Users of the
screening toxicity values in a supplement to a PPRTV assessment should understand that there is
considerably more uncertainty associated with the derivation of a supplement screening toxicity
value than for a value presented in the body of the assessment. Questions or concerns about the
appropriate use of screening values should be directed to the Superfund Health Risk Technical
Support Center.
The study by Mitusmori et al. (1979) is selected as the principal study for the derivation
of a screening subchronic p-RfD. The critical endpoint is a statistically significant reduction in
several hematological parameters, including significant reduction in the total number of
leukocytes in male SPF-ICR mice. This study is a peer-reviewed journal publication and, though
not stated in the article, seems to be performed in general accordance with good laboratory
practice (GLP) principles. It was conducted with multiple doses (0, 80, 400, 2000, 10,000 ppm
in the diet) with a variety of toxicologic endpoints that demonstrated a statistically significant
dose response. Details on the study are provided in the Review of Potentially Relevant Data
section. Among the available, acceptable studies (see Table 6), this study represents the lowest
POD for developing a subchronic p-RfD.
Adjusted doses for daily exposure:
The following dosimetric adjustments were made for each dose in the principal study for
food intake. Dosimetric adjustment for 80 ppm (mg/kg) is presented below.
(DOSEadj) = DOSEcitation x Food Consumption Value ^ day x (l -h BW Value) x
Days Dosed ^ Total Days in Study
= 80 ppm (mg/kg) x 4.6 g/day (males) x (1 -h 38.0228 g) x 91 days ^
91 days
(DOSEadj) = 368 mg/kg-day x 0.0263
(DOSEadj) = 9.69 mg/kg-day
Food consumption values and animal body weights were obtained by digitizing the food
intake rates and body weights presented in Figure 2 in the Mitsumori et al. (1979) article using
the GetData graph digitizer tool (http://www.getdata.com.ru). Average food intake values and
body-weight values from these digitized results were used to determine the daily average dose of
BCMEE. Because food intake rates at 80 and 400 ppm were not provided in Figure 2
(Mitsumori et al., 1979), average food intake rates from the 0-ppm dose level were used to
calculated daily average doses for the 80- and 400-ppm dose groups.
32 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
The screening subchronic p-RfD for BCMEE based on the LOAEL of 9.69 mg/kg-day in
the male ICR mouse (Mitsumori et al., 1979) is derived as follows:
Screening Subchronic p-RfD = LOAEL UFc
= 9.69 -h 10,000
= 0.000969 mg/kg-day or 1 x 10 3 mg/kg-day
Table A. 1 summarizes the UFs for the subchronic p-RfD for BCMEE.
Table A.l. Uncertainty Factors for Subchronic p-RfD for BCMEE
UF
Value
Justification
UFa
10
A UFa of 10 is applied for interspecies extrapolation to account for
potential toxicokinetic and toxicodynamic differences between mice
and humans. There are no data to determine whether humans are
more or less sensitive than mice to hematological effects of BCMEE.
UFd
10
A UFd of 10 is selected because there are no acceptable
two-generation reproduction studies or developmental studies, and
there is no indication of any other studies that may be relevant for the
database UF.
UFh
10
A UFh of 10 is applied for intraspecies differences to account for
potentially susceptible individuals in the absence of information on
the variability of response to humans.
UFl
10
A UFl of 10 is applied for using a POD based on a LOAEL because
a NOAEL cannot be determined from the available database.
UFs
1
A UFS of 1 is applied because results from a subchronic duration
(Mitsumori et al., 1979) were utilized as the principal study.
UFC
10,000
33
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
APPENDIX B. DATA TABLES
Table B.l. Results of Hematological Examination in SPF-ICR Mice Fed BCMEE in the Diet for 13 Weeksa'b'c
Sex and Dose
Group
(mg/kg-BW)d
No. of
Mice
Mean RBC
(x 106/mm3)
Mean Ht (%)
Mean Hb (g/dl)
Leukocyte
Total
(x 106/mm3)
Differential %
Mean L
Mean N
Mean M
Mean E
Mean Others
Stab.
Seg.
Males—13 Weeks
0
7
7.2 ±0.6
41.8 ±3.1
14.1 ±0.9
6.5 ± 1.7
78.1
2.1
17.0
1.9
0.9
0
9.69
7
6.3 ±0.7* (5)
37.7 ±2.8*
11.8 ± 1.2** (5)
4.3 ± 1.2*
73.1
1.3
22.7
1.4
1.4
0
48.42
7
6.2 ±0.6*
38.3 ±2.9*
12.5 ±0.7**
3.4 ± 1.2**
69.9
2.3
22.9
2.4
2.6
0
242.18
7
6.8 ±0.5
40.4 ±2.0
13.0 ±0.6*
3.4 ± 1.1**
68.1
2.0
24.9
2.6
2.3
0.1
984.9
7
5.1 ± 1.3**
34.9 ± 3.2**
11.5 ±0.7***
1.5 ±0.5***
63.3
4.0
29.1
2.1
1.4
0
Females—13 Weeks
0
7
6.9 ±0.7
36.9 ±2.7
13.4 ± 1.0
6.8 ±3.0
75.7
3.0
18.9
1.1
1.3
0
11.99
7
7.4 ±0.5
37.6 ± 1.4
13.9 ±0.5
8.5 ±5.4
77.4
1.7
18.0
1.4
1.4
0
60.26
7
7.1 ± 0.5
37.8 ±2.8
13.7 ± 1.0
6.9 ±2.4
80.3
2.7
13.6
1.3
2.1
0
305.80
7
7.0 ±0.5
39.0 ± 1.9
13.2 ±0.3
3.9 ± 1.2
85.1*
1.6
11.1*
0.7
1.4
0
1211.7
7
6.2 ±0.5
33.4 ±2.8*
11.0 ± 1.1**
2.6 ± 1.1* (6)
80.7
0 9**
14.9
1.0
1.1
0
Males—52 Weeks
0
7
7.9 ±0.5
38.6 ±3.4
13.4 ± 1.1
6.5 ±2.1
53.4
1.1
43.6
0.9
0.6
0.4
9.69
7
7.7 ± 1.3
37.7 ±3.0
12.7 ±2.3
6.4 ± 1.5 (6)
69.3 (6)
2.2 (6)
26.3* (6)
1.0 (6)
1.2 (6)
0(6)
48.42
7
7.9 ±0.6
37.3 ± 1.7
13.3 ±0.9
5.5 ± 1.2
50.1
0.4
47.3
1.3
0.9
0
242.18
7
7.8 ±0.7
35.3 ±3.5
12.5 ± 1.2
6.4 ±3.8
72.6**
0.7
24.6**
0.3
1.7
0.1
984.9
7
7.3 ±0.7
33.9 ±3.0*
11.6 ± 1.1
5.7 ±2.7
62.3
1.3
35.0
0.4
1.0
0
34
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table B.l. Results of Hematological Examination in SPF-ICR Mice Fed BCMEE in the Diet for 13 Weeksa'b'c
Sex and Dose
Group
(mg/kg-BW)d
No. of
Mice
Mean RBC
(x 106/mm3)
Mean Ht (%)
Mean Hb (g/dl)
Leukocyte
Total
(x 106/mm3)
Differential %
Mean L
Mean N
Mean M
Mean E
Mean Others
Stab.
Seg.
Females—52 Weeks
0
7
8.2 ±0.5
38.9 ±2.0
13.3 ±0.9
4.3 ± 1.9
59.1
4.6
32.0
1.7
1.9
0.7
11.99
7
8.2 ±0.5
38.5 ± 1.9
13.5 ±0.9
3.9 ±0.8
63.9
4.1
30.7
0.3*
0.9
0.1
60.26
7
7.7 ±0.7 (6)
38.9 ±2.5 (6)
13.4 ± 1.0 (6)
3.6 ± 1.3 (5)
61.1
4.6
32.7
o**
1.4
0.1
305.80
7
6.7 ± 1.3*
33.8 ±5.6
11.2 ± 1.7*
3.9 ± 1.8(6)
62.5 (6)
4.2 (6)
31.3 (6)
0.3* (6)
1.0 (6)
0.7 (6)
1211.7
7
6.7 ±0.4*** (4)
32.9 ± 1.9*** (4)
11.0 ± 1.0** (4)
5.1 ±1.6 (4)
67.4
5.0
24.4
0.1*
2.4
0.6
Males—104Weeks
0
8
7.1 ± 0.9
36.3 ±5.7
11.2 ± 1.5
5.8 ±2.7
55.8
4.0
37.1
0.5
2.4
1.3
9.69
5
6.6 ±1.1
35.5 ±4.6
10.7 ± 1.3
7.6 ±2.6
49.0
3.6
44.2
1.0
2.2
0
48.42
8
7.0 ±0.7
35.7 ±4.1
10.6 ± 1.4
5.2 ± 1.6
52.6
1.9
40.9
0.9
3.8
0
242.18
5
6.8 ±0.5
35.1 ±2.2
10.4 ±0.7
4.0 ± 1.2
58.4
6.2
32.8
0.6
2.0
0
984.9
6
6.9 ±0.6
34.4 ±4.4
10.2 ±0.9
3.4 ± 1.3
52.2
3.3
41.8
1.2
1.5
0
Females—104 Weeks
0
5
7.3 ±0.5
36.9 ±3.8
10.8 ± 1.1
4.5 ±2.4
63.2
4.0
30.0
2.2
0.6
0
11.99
9
7.2 ±0.8
34.7 ±2.3
10.5 ± 1.2
3.7 ± 1.4
49.1
5.3
40.3
2.8
1.3
1.1
60.26
9
6.4 ±1.1 (8)
33.6 ±5.1
10.0 ± 1.6 (8)
2.8 ±0.8 (8)
47.7
7.6
42.7
1.1
0.7
0.3
305.80
7
6.6 ±0.9
34.7 ±5.0
10.0 ± 1.4
3.1 ±2.3
47.7*
3.0
46.0
1.6
1.6
0.1
1211.7
1
6.8
34.0
9.8
3.2
86
1
13
0
0
0
aMitsumori et al. (1979).
bValues are mean± SD or means (differential leukocytes).
°Parentheses values: number of mice examined for that specific dose.
dAdjusted doses determined using digitized results for body weight and food intake from the graph provided by the authors; these doses were used in the BMD analysis
for total leukocytes.
RBC: erythrocyte count; Ht: hematocrit; Hb: hemoglobin; L: leukocytes; N: neutrophils; M: monocytes; E: eosinophils.
*p < 0.05, **p < 0.01, ***p < 0.001 based on Student's /-test.
35
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table B.2. Results of Blood Biochemical Examination in Mice Fed BCMEE in the Diet for 13 Weeksa'b
Dose Group
(mg/kg-day)
No. of
Animals
Mean GOT
(K-unit)
Mean GPT
(K-unit)
Mean ALP
(K-A unit)
Mean Glucose
(mg/dl)
Mean TP (g/dl)
Mean UN
(mg/dl)
Mean CHO
(mg/dl)
Mean Bil
(mg/dl)
Male
0
7
45 ±6
26 ±7
4.0 ±0.9
232 ± 13
5.0 ±0.2
27 ±3
110 ± 10
0.3 ±0.05
9.69
7
44 ± 8(5)
24 ± 7(5)
3.7 ±0.9(5)
218 ± 13(5)
4.8 ±0.3(4)
24 ± 3(4)
121 ±31(3)
0.2 ±0.06(3)
48.42
7
45 ±6
23 ±6
3.6 ± 1.1
244 ± 45
4.9 ±0.3
29 ±3
123 ± 15
0.2 ±0.04
242.18
7
53 ± 11
37 ± 18
4.4 ±0.8
252 ± 30
4.9 ±0.3
26 ±4
102 ± 26
0.2 ±0
984.9
7
87 ±25**
64 ±26**
11.8 ± 7.1*
184 ±32*
4.5 ±0.3** (6)
35 ± 12(6)
98 ± 12(4)
0.3 ± 0.07(2)
Female
0
7
45 ± 3(6)
21 ±3(6)
4.7 ± 1.3(6)
194 ± 29(6)
4.8 ±0.4(6)
20 ± 3(6)
68 ± 24(4)
0.2 ±0.05(4)
11.99
7
46 ±6
24 ± 10
5.0 ±0.8
216 ±36
4.8 ±0.1
20 ±4
61 ± 14
0.2 ±0.05
60.26
7
51 ± 15
26 ± 18
4.9 ±0.5
217 ±40(6)
4.8 ±0.3
20 ±3
63 ± 15(5)
0.2 ± 0(5)
305.8
7
48 ±5
20 ±3
4.7 ± 1.4
205 ± 19
4.8 ±0.3
22 ±5
79 ± 12(5)
0.2 ±0.05(5)
1211.7
7
70±14**
35 ± 14*
5.2 ± 1.8
147 ± 32*(6)
4.5 ± 0.3*(6)
28 ±7*
106 ± 42(3)
0.2 ±0.06(3)
aValues obtained from Mitsumori et al. (1979).
bParentheses values: number of mice examined for that specific dose.
GOT: glutamic-oxaloacetic transaminase; GPT: glutamic-pyruvic transaminase; ALP: alkaline phosphatase; TP: total protein; UN: urea nitrogen; CHO:
cholesterol; Bil: bilirubin.
*p < 0.05, ** p < 0.01, *** p < 0.001.(Student's / test)
36
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table B.3. Results of Microscopic Examination of Selected Tissues in F344 Rats Treated
with BCMEE Mixture via Gavage for 103 Weeks"
Title
Male
Female
Untreated
Control
Vehicle
Control
100
mg/kg-day
200
mg/kg-day
Untreated
Control
Vehicle Control
100
mg/kg-day
200
mg/kg-day
No. of tissues
examined
microscopically
47
50
50
49
49
50
49
48
Esophageal
hyperkeratosis
0 (0%)
9 (18%)
10 (20%)
40 (82%)
0 (0%)
13 (26%)
10 (20%)
31 (65%)
Esophageal
acanthosis
0 (0%)
0 (0%)
1 (2%)
1 (2%)
0 (0%)
1 (2%)
0 (0%)
5 (10%)
Gastric
hyperkeratosis
0 (0%)
13 (26%)
5 (10%)
10 (20%)
0 (0%)
21 (42%)
14 (29%)
11 (23%)
Gastric acanthosis
1 (2%)
6 (12%)
4 (8%)
9 (18%)
0 (0%)
8 (16%)
5 (10%)
9 (19%)
aValues obtained from NCI (1979).
37
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table B.4. Primary Tumors in B6C3Fi Mice Administered
BCMEE Mixture via Gavage for 103 Weeks"
Tumor Type
Vehicle Controlb
Low Doseb—100
(71.4 mg/kg-day)
High Doseb—200
(142.9m g/kg- day)
Male
Lung: Alveolar/Bronchiolar Adenomas
5/50
13/50
11/50
Lung: Alveolar/Bronchiolar Adenoma or
Carcinoma
6/50
15/50
13/50
Hematopoietic System: All Malignant
Lymphoma
6/50
3/50
7/50
Liver Adenoma
8/50
10/50
13/50
Liver Carcinoma
5/50
13/50
17/50
Liver Adenoma or Carcinoma
13/50
23/50
27/50
Forestomach: Squamous Cell Papilloma
0/49
1/50
0/50
Female
Lung: Alveolar/Bronchiolar Adenomas
1/50
4/50
8/50
Lung: Alveolar/Bronchiolar Adenoma or
Carcinoma
1/50
4/50
10/50
Stomach/Forestomach: Squamous Cell
Papilloma/ Carcinoma
0/50
0/49
3/49
aValues obtained from NTP (1982).
bNumber of tumor-bearing animals/number of animals examined at the site.
38
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table B.5. Incidence of Neoplastic Tumors in a 103-Week Gavage Study of BCMEE
Mixture in B6C3Fi Mice"
Exposure Group (Daily Average Dose, mg/kg-day)
Lesion Type
0
100 (71.4)
200 (142.9)
Male
Alveolar/Bronchiolar adenomas
Tumor rates
Overallb
5/50 (10%)
13/50 (26%)
11/50 (22%)
Adjusted0
12.2%
28.8%
28.9%
Terminald
5/41 (12%)
12/44 (27%)
10/37 (27%)
Statistics: Incidental tumor test
p = 0.045
p = 0.035
p = 0.067
Statistics: Fisher's exact test
p = 0.083
p = 0.033
p = 0.086
Alveolar/Bronchiolar adenomas or carcinomas
Tumor rates
Overallb
6/50 (12%)
15/50 (30%)
13/50 (26%)
Adjusted0
14.1%
33.2%
34.2%
Terminal
5/41 (12%)
14/44 (32%)
12/37 (32%)
Statistics: Incidental tumor test
p = 0.024
p = 0.019
p = 0.035
Statistics: Fisher's exact test
p = 0.061
p = 0.024
p = 0.062
Female
Alveolar/Bronchiolar adenomas
Tumor rates
Overallb
1/50 (2%)
4/50 (8%)
8/50 (16%)
Adjusted0
2.8%
11.8%
24.2%
Terminal
0/31 (0%)
4/34 (12%)
5/28 (18%)
Statistics: Incidental tumor test
p = 0.016
p = 0.148
p = 0.029
Statistics: Fisher's exact test
p = 0.011
p = 0.181
p = 0.015
Alveolar/Bronchiolar adenomas or carcinomas
Tumor rates
Overallb
1/50 (2%)
4/50 (8%)
10/50 (20%)
Adjusted0
2.8%
11.8%
30.8%
Terminal
0/31 (0%)
4/34 (12%)
7/28 (25%)
Statistics: Incidental tumor test
p = 0.004
p = 0.148
p = 0.008
Statistics: Fisher's exact test
p = 0.003
p = 0.181
p = 0.004
Male
Liver carcinomas
Tumor rates
Overallb
5/50 (10%)
13/50 (26%)
17/50 (34%)
Adjusted0
11.5%
27.6%
40.1%
Terminal
3/41 (7%)
10/44 (23%)
12/37 (32%)
Statistics: Incidental tumor test
p = 0.004
p = 0.023
p = 0.007
Statistics: Fisher's exact test
p = 0.004
p = 0.033
p = 0.004
3 9 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Table B.5. Incidence of Neoplastic Tumors in a 103-Week Gavage Study of BCMEE
Mixture in B6C3Fi Mice"
Exposure Group (Daily Average Dose, mg/kg-day)
Lesion Type
0
100 (71.4)
200 (142.9)
Female
Liver adenoma or carcinoma
Tumor rates
Overallb
13/50
23/50
27/50
Adjusted0
29.5%
48.9%
64.0%
Terminal
10/41 (24%)
20/44 (45%)
22/37 (59%)
Statistics: Incidental tumor test
p =0.003
p = 0.030
p = 0.005
Statistics: Fisher's exact test
p = 0.003
p = 0.030
p = 0.004
aValues obtained from NTP (1982).
bNumber of tumor-bearing animals/number of animals examined at the site.
°Kaplan-Meier estimated lifetime tumor incidence after adjusting for intercurrent mortality.
dObserved tumor incidence at terminal kill.
40
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table B.6. Results of Hematological Examination in SPF-ICR Mice Fed BCMEE
in the Diet for 104 Weeks"
Sex and Dose Group
Mean RBC
Mean Ht
Mean Hb
(mg/kg-day)
No. of Mice
(x 106/mm3)
(%)
(g/dl)
Male-52 Weeksb
0
7
7.9 ±0.5
38.6 ±3.4
13.4 ± 1.1
8.41
7
7.7 ± 1.3
37.7 ±3.0
12.7 ±2.3
40.1
7
7.9 ±0.6
37.3 ± 1.7
13.3 ±0.9
198
7
7.8 ±0.7
35.3 ±3.5
12.5 ± 1.2
927
7
7.3 ±0.7
33.9 ±3.0*
11.6 ± 1.1
Male-104 Weeksb
0
8
7.1 ±0.9
36.3 ±5.7
11.2 ± 1.5
8.41
5
6.6 ±1.1
35.5 ±4.6
10.7 ± 1.3
40.1
8
7.0 ±0.7
35.7 ±4.1
10.6 ± 1.4
198
5
6.8 ±0.5
35.1 ±2.2
10.4 ±0.7
927
6
6.9 ±0.6
34.4 ±4.4
10.2 ±0.9
Female-52 Weeksb
0
7
8.2 ±0.5
38.9 ±2.0
13.3 ±0.9
7.58
7
8.2 ±0.5
38.5 ± 1.9
13.5 ±0.9
35.8
7
7.7 ± 0.7(6)
38.9 ±2.5(6)
13.4 ± 1.0(6)
194
7
6.7 ± 1.3*
33.8 ±5.6
11.2 ± 1.7*
961
7
6.7 ± 0.4***(4)
32.9 ± 1.9***(4)
11.0 ± 1.0**(4)
Female-104 Weeksb
0
5
7.3 ±0.5
36.9 ±3.8
10.8 ± 1.1
7.58
9
7.2 ±0.8
34.7 ±2.3
10.5 ± 1.2
35.8
9
6.4 ± 1.1(8)
33.6 ±5.1
10.0 ± 1.6(8)
194
7
6.6 ±0.9
34.7 ±5.0
10.0 ± 1.4
961
1
6.8
34.0
9.8
aValues obtained from Mitsumori et al. (1979). Results for males and females at 13 weeks are provided in
Table B.l.
bAdjusted doses reported by the study authors.
Parentheses values = number of mice examined for that specific dose.
RBC: erythrocyte count; Ht: hematocrit; Hb: hemoglobin.
*p < 0.05, ** p < 0.01, *** p < 0.001.
41 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table B.7. Results of Hematological Examination in SPF-ICR Mice Fed BCMEE in the
Diet at 52 Weeks and 104 Weeksa b c
Sex and Dose
Group
(mg/kg-BW)
No. of
Mice
Leukocyte
Total
(xl06/mm3)
Differential %
Mean L
Mean N
Mean M
Mean E
Mean Others
Stab.
Seg.
Male-52 Weeksd
0
7
6.5 ±2.1
53.4
1.1
43.6
0.9
0.6
0.4
8.41
7
6.4 ± 1.5(6)
69.3(6)
2.2(6)
26.3*(6)
1.0(6)
1.2(6)
0(6)
40.1
7
5.5 ± 1.2
50.1
0.4
47.3
1.3
0.9
0
198
7
6.4 ±3.8
72.6**
0.7
24.6**
0.3
1.7
0.1
927
7
5.7 ±2.7
62.3
1.3
35.0
0.4
1.0
0
Male-104 Weeks'1
0
8
5.8 ±2.7
55.8
4.0
37.1
0.5
2.4
0.3
8.41
5
7.6 ±2.6
49.0
3.6
44.2
1.0
2.2
0
40.1
8
5.2 ± 1.6
52.6
1.9
40.9
0.9
3.8
0
198
5
4.0 ± 1.2
58.4
6.2
32.8
0.6
2.0
0
927
6
3.4 ± 1.3
52.2
3.3
41.8
1.2
1.5
0
Female-52 Weeks'1
0
7
4.3 ± 1.9
59.1
4.6
32.0
1.7
1.9
0.7
7.58
7
3.9 ±0.8
63.9
4.1
30.7
0.3*
0.9
0.1
35.8
7
3.6 ± 1.3(5)
61.1
4.6
32.7
o**
1.4
0.1
194
7
3.9 ± 1.8(6)
62.5(6)
4.2(6)
31.3(6)
0.3*(6)
1.0(6)
0.7(6)
961
7
5.1 ± 1.6(4)
67.4
5.0
24.4
0.1*
2.4
0.6
Female-104 Weeks'1
0
5
4.5 ±2.4
63.2
4.0
30.0
2.2
0.6
0
7.58
9
3.7 ± 1.4
49.1
5.3
40.3
2.8
1.3
1.1
35.8
9
2.8 ±0.8(8)
47.7
7.6
42.7
1.1
0.7
0.3
194
7
3.1 ± 2.3
47.7*
3.0
46.0
1.6
1.6
0.1
961
1
3.2
86
1
13
0
0
0
aMitsumori et al. (1979).
bValues are mean± SD or means (differential leukocytes).
°Parentheses values: number of mice examined for that specific dose.
dAdjusted doses determined using digitized results for body weight and food intake from the graph provided by
the authors; these doses were used in the BMD analysis for total leukocytes.
RBC: erythrocyte count; Ht: hematocrit; Hb: hemoglobin; L: Leukocytes; N: Neutrophils; M: Monocytes;
E: Eosinophils.
*p < 0.05, ** p < 0.01, *** p < 0.001 based on Student's /-test.
42 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table B.8. Results of Blood Biochemical Examination in SPF-ICR Mice Fed BCMEE in the Diet for 104 Weeks"'b
Dose Group
(mg/kg-BW)
No. of
Animals
Mean GOT
(K-unit)
Mean GPT
(K-unit)
Mean ALP
(K-A unit)
Mean Glucose
(mg/dl)
Mean TP (g/dl)
Mean UN
(mg/dl)
Mean CHO
(mg/dl)
Mean Bil
(mg/dl)
Male-52 Weeks
0
7
62 ±25
43 ±49
2.3 ±0.7
218 ±34
5.3 ±0.4
21 ± 4
125 ±29
0.2 ± 0.22(5)
8.41
7
65 ±21
57 ±72
3.8 ± 1.5*
206 ± 50
5.2 ±0.4
26 ±6
112 ± 19
0.2 ±0.19(6)
40.1
7
52 ± 14
27 ± 17
2.5 ± 1.1
196 ±33
4.9 ±0.4
29 ±5**
119 ±20
0.2 ± 0.05(6)
198
7
49 ± 14
22 ±9
2.9 ± 1.0
184 ± 27
5.0 ± 0.2(6)
26 ±4*
116 ±9(6)
0.1 ±0.08(6)
927
7
49 ± 13
18 ±8
5.6 ±2.3**
130 ±35***
4.4 ±0.3***
28 ±8*
101 ±28
0.1 ±0(3)
Male-104 Weeks
0
8
50 ± 10
37 ±28
12.7 ±25.6
143 ± 26
4.7 ±0.7
24 ±6
185 ± 97(7)
0.2 ±0.11(7)
8.41
5
50 ±8
31 ± 19
4.7 ±0.9
166 ±21
4.7 ±0.2
27 ±4
184 ±68
0.2 ±0.07
40.1
8
51 ± 5
29 ± 11
7.7 ±7.1
164 ± 28
5.0 ±0.3
28 ±8
150 ±30
0.2 ±0.06
198
5
54 ±8
24 ±8
3.7 ± 1.0
169 ± 12
5.5 ±0.2*
33 ± 11
139 ±36
0.2 ±0.07
927
6
59 ±28
59 ±87
6.5 ±4.8
166 ±35
4.9 ±0.3
29 ±5
129 ±32
0.2 ±0.04
Female-52 Weeks
0
7
56 ±6
18 ±3(6)
4.0 ± 1.2(6)
172 ± 18(6)
5.4 ±0.3(5)
22 ± 3(6)
86 ± 9(3)
-
7.58
7
67 ±24
22 ± 11
3.2 ±0.8
164 ± 15
5.4 ±0.3
21 ± 7
78 ± 6(6)
-
35.8
7
122 ± 109
86 ± 117
4.0 ±0.7
205 ± 42
5.0 ±0.4*
25 ±8
130 ± 74(5)
-
194
7
80 ±47
23 ± 14
10.9 ± 16.3(6)
178 ± 18(6)
5.5 ±0.3(6)
27 ± 9(6)
87 ± 13(4)
-
961
7
97 ± 54(5)
50 ± 6(5)
6.7 ± 1.5**(5)
124 ± 22**(5)
4.6 ± 0.3**(4)
32 ± 4**(5)
84 ± 18(4)
-
43
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table B.8. Results of Blood Biochemical Examination in SPF-ICR Mice Fed BCMEE in the Diet for 104 Weeks"'b
Dose Group
(mg/kg-BW)
No. of
Animals
Mean GOT
(K-unit)
Mean GPT
(K-unit)
Mean ALP
(K-A unit)
Mean Glucose
(mg/dl)
Mean TP (g/dl)
Mean UN
(mg/dl)
Mean CHO
(mg/dl)
Mean Bil
(mg/dl)
Female-104 Weeks
0
5
56 ±21
41 ±44
4.7 ± 1.5
152 ±20
5.1 ±0.3
26 ±7
133 ±51
0.2 ±0.09
7.58
9
53 ± 12
33 ± 12
5.1 ±2.3
142 ±30
5.2 ±0.4
25 ± 11
101 ±27
0.2 ±0.04
35.8
9
58 ±22
37 ±23(8)
5.0 ± 1.9
148 ± 17
5.1 ± 0.3
25 ±8
120 ±33(8)
0.2 ± 0.08(8)
194
7
60 ±20
32 ± 14
4.6 ± 1.9
131 ± 29
5.1 ±0.4
22 ±6
102 ± 12
0.1 ±0.05
961
1
42
17
3.6
129
4.6
28
93
0.1
aValues obtained from Mitsumori et al. (1979). Results for males and females at 13 weeks are provided in Table B.2.
bParentheses values: number of mice examined for that specific dose.
GOT: glutamic-oxaloacetic transaminase; GPT: glutamic-pyruvic transaminase; ALP: alkaline phosphatase; TP: total protein; UN: urea nitrogen; CHO: cholesterol;
Bil: bilirubin.
*p < 0.05, **p < 0.01, ***p < 0.001.
44
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table B.9. Absolute and Relative Organ Weights in SPF-ICR Mice Fed BCMEE in the Diet for
104 Weeks a'b'c
Dose Group
(mg/kg-day)
No. of
Animals
Body Weight
(g)
Absolute and Relative Organ Weights (mg)d
Brain
Thyroid
Kidney
Heart
Male
0
8
48.0 ±6.4
485 ± 18
[1.02 ±0.13]
3.1 ±0.15(6)
[0.006 ± 0.003]
796 ± 92
[1.67 ±0.20]
212 ±34
[0.44 ± 0.06]
8.41
5
43.7 ±4.5
508 ±9*
[1.17 ± 0.11]
4.4 ±0.2
[0.010 ±0.001]
748 ± 78(4)
[1.72 ±0.08]
211 ± 22
[0.48 ± 0.02]
40.1
8
45.0 ±3.2
495 ± 14
[1.11 ±0.09]
3.1 ± 1.1(7)
[0.007 ± 0.003]
742 ± 74
[1.65 ±0.17]
203 ± 24
[0.45 ± 0.05]
198
5
44.2 ±2.7
526 ±27**
[1.19 ±0.06]
4.9 ±0.7*
[0.011 ±0.002]
845 ± 89
[1.92 ±0.25]
224 ± 29
[0.51 ±0.10]
927
6
37.2 ±2.6**
485 ±38
[1.30 ±0.07]
3.2 ± 1.1
[0.009 ± 0.003]
636 ±21**
[1.72 ±0.14]
180 ±21
[0.48 ± 0.05]
Female
0
5
42.1 ±6.5
518 ±27
[1.25 ±0.19]
3.1 ± 1(3)
[0.008 ± 0.002]
506 ± 48(4)
[1.22 ±0.33]
179 ±23
[0.44 ±0.10]
7.58
9
38.1 ±6.4
510 ±45
[1.37 ±0.27]
3.8 ±0.7
[0.010 ±0.003]
468 ± 58
[1.27 ±0.32]
151± 23*
[0.40 ± 0.08]
35.8
9
42.2 ±2.6
517 ±33
[1.23 ±0.07]
3.4 ± 1.0
[0.008 ± 0.002]
474 ± 59
[1.12 ± 0.13]
157 ±29
[0.37 ± 0.06]
194
7
36.5 ±2.9
510 ± 35
[1.41 ±0.15]
3.6 ± 1.0
[0.010 ±0.003]
477 ± 67
[1.31 ±0.17]
157 ± 18
[0.43 ± 0.06]
961
1
28.6
440
[1.54]
4.7
[0.016]
384
[1.34]
128
[0.45]
aValues obtained from Mitsumori et al. (1979).
bValues are means ± SD.
°Parentheses values: number of mice examined for that specific dose.
dRelative organ weight (100 x organ weight/body weight); presented in brackets [].
*p < 0.05, **p < 0.01.
45
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table B.10. Incidence of Nonneoplastic Histological Findings in Male and Female
SPF-ICR mice Fed BCMEE for 104 Weeksa b
Nonneoplastic
Lesion
Dose Group
Male (Dose Groups, ppm)
Female (Dose Groups, ppm)
0
80
400
2000
10,000
0
80
400
2000
10,000
Spleen:
Hemosiderin
Deposition
1/56
0/56
0/56
3/56
17/56
2/56
3/56
1/56
8/56
17/56
Spleen: Increased
Extramedullary
Hematopoiesis
9/56
10/56
7/56
6/56
10/56
9/56
7/56
6/56
6/56
7/56
aValues obtained from Mitsumori et al. (1979).
bGroups of 56 mice/sex/group were examined. This includes animals sacrificed by design and animals euthanized in
extremis or found dead.
46
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
APPENDIX C. BMD MODELING OUTPUTS FOR BCMEE
Table C.l. BMD Modeling Output Summary for BCMEEa
Endpoint
Species
Sex
Figure
Model
Homogeneity
Variance
/7-Value
Goodness-of-Fit
p-V alueb
AIC for
Fitted Model
bmd1sd
(mg/kg-day)
BMDL1sd
(mg/kg-day)
Conclusions
Hemoglobin (Hb)
Mouse
F
C.l
Power
(nonconstant
variance)
0.009
0.974
12.21
308.39
243.29
Lowest AIC
Poor variance model
Hemoglobin (Hb)
Mouse
M
C.2
Hill
(nonconstant
variance)
0.381
0.041
27.03
0.00
0.00
Lowest BMDL
p-score 4 < 0.1
Wrong variance model
hit bound (n= 1)
Hematocrit (Ht)
Mouse
F
C.3.1
Linear
(constant
variance)
0.260
0.829
76.09
367.54
188.75
Lowest AIC
Lowest BMDL
C.3.2
Polynomial
(constant
variance)
0.260
0.829
76.09
367.54
188.75
Lowest AIC
Lowest BMDL
Maximum order beta = 0
(32 = 0
(33 =0
C.3.3
Power
(constant
variance)
0.260
0.829
76.09
367.54
188.75
Lowest AIC
Lowest BMDL
hit bound (power =1)
Hematocrit (Ht)
Mouse
M
C.4
Hill
(constant
variance)
0.700
0.043
92.05
0.00
0.00
Lowest AIC
Lowest BMDL
p-score 4 < 0.1
Erythrocytes
(RBC)
Mouse
F
C.5
Power
(nonconstant
variance)
0.726
0.352
-0.53
1032.70
307.91
Lowest AIC
BMD/BMDL ratio > 3
Wrong variance model
hit bound (power =1)
Erythrocytes
(RBC)
Mouse
M
C.6
Hill
(constant
variance)
0.906
0.038
6.95
0.00
0.00
Lowest AIC
Lowest BMDL
p-score 4 < 0.1
47
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table C.l. BMD Modeling Output Summary for BCMEEa
Endpoint
Species
Sex
Figure
Model
Homogeneity
Variance
/7-Value
Goodness-of-Fit
/7-Valueb
AIC for
Fitted Model
bmd1sd
(mg/kg-day)
BMDLisd
(mg/kg-day)
Conclusions
Mouse
F
Data were not
modeled
because the
values for all
doses were the
same
Total Leukocytes
(WBC)
Mouse
F
C.7.1
Linear
(nonconstant
variance)
0.003
0.113
90.52
311.05
204.12
Lowest AIC
Lowest BMDL
C.7.2
Polynomial
(nonconstant
variance)
0.003
0.113
90.52
311.05
204.12
Lowest AIC
Lowest BMDL
Maximum order beta = 0
(32 = 0
(33 =0
C.7.3
Power
(nonconstant
variance)
0.003
0.113
90.52
311.05
204.12
Lowest AIC
Lowest BMDL
hit bound (power =1)
Total Leukocytes
(WBC)
Mouse
M
C.8
Hill
(constant
variance)
0.632
NA
49.27
7.56
0.00
Lowest AIC
Lowest BMDL
p-score 4 < 0.1
BMD/BMDL ratio > 3
Total Leukocytes
(WBC)—
includes high
dose
Mouse
F
C.9.1
Linear
(nonconstant
variance)
0.000
0.002
109.52
1020.34
713.21
Lowest AIC
p-score 4 < 0.1
C.9.2
Polynomial
(nonconstant
variance)
0.000
0.002
109.52
1020.34
713.21
Lowest AIC
p-score 4 < 0.1
(33 =0
C.9.3
Power
(nonconstant
variance)
0.000
0.002
109.52
1020.34
713.21
Lowest AIC
p-score 4 < 0.1
hit bound (power =1)
48
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table C.l. BMD Modeling Output Summary for BCMEEa
Endpoint
Species
Sex
Figure
Model
Homogeneity
Variance
/7-Value
Goodness-of-Fit
/7-Valueb
AIC for
Fitted Model
bmd1sd
(mg/kg-day)
BMDLisd
(mg/kg-day)
Conclusions
Total Leukocytes
(WBC)—
includes high
dose
Mouse
M
C.10
Hill
(constant
variance)
0.068
0.014
58.99
5.11
1.71
Lowest BMDL
p-score 4 < 0.1
Poor variance model
Wrong variance model
hit bound (n= 1)
aAll endpoints modeled from data in Mitsumori et al. (1979).
bValues <0.10 fail to meet conventional goodness-of-fit criteria.
AIC = Akaike's Information Criteria; BMD = benchmark dose; BMDL lower confidence limit (95%) on the benchmark dose; 1SD = 1 standard deviation; M = Male;
F = Female.
49
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Models Considered for the Derivation of a Subchronic p-RfD for
Bis(2-chloro-l-methylethyl)ether
Power Model with 0.95 Confidence Level
dose
14:18 02/11 2010
Figure C.l. Power Nonconstant Variance BMD Model for Female Hemoglobin Data
(Mitsumori et al., 1979)
Text Output for Power Nonconstant Variance BMD Model for Female Hemoglobin Data
(Mitsumori et al., 1979)
Power Model. (Version: 2.15; Date: 04/07/2008)
Input Data File: C:\BCMEE\Mitsumori 197 9 13wk Hb female Power 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori 197 9 13wk Hb female Power l.plt
Thu Feb 11 14:18:24 2010
Table3 13wks Hb females
The form of the response function is:
50 Bis(2-chloro-l-methyl ethyl)ether
-------
FINAL
3-30-2011
Y[dose] = control + slope * doseApower
Dependent variable = Mean
Independent variable = Dose
The power is restricted to be greater than or equal to 1
The variance is to be modeled as Var(i) = exp(lalpha + log(mean(i)) * rho)
Total number of dose groups = 4
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
Default Initial Parameter Values
lalpha = -0.536143
rho = 0
control = 13.2
slope = 1.17467
power = -0.208394
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -rho -power
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
lalpha control slope
lalpha 1 -0.52 0.22
control -0.52 1 -0.66
slope 0.22 -0.66 1
Parameter Estimates
Confidence Interval
Variable
Upper Conf. Limit
lalpha
-47.1047
rho
control
14.0672
slope
4.11304e-046
power
Estimate
-47.7331
18
13.7454
-1.18724e-045
18
Std. Err.
0.320638
NA
0. 164149
3. 95895e-046
NA
95.0% Wald
Lower Conf. Limit
-48 . 3616
13.4237
-1. 96318e-045
NA - Indicates that this parameter has hit a bound
51 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
implied by some inequality constraint and thus
has no standard error.
Table of Data and Estimated Values of Interest
Dose
Res .
Obs Mean
Est Mean Obs Std Dev Est Std Dev
Scaled
0
11. 99
60.26
305. 8
13.4
13. 9
13.7
13.2
13.7
13.7
13.7
13. 1
1
0.5
1
0.3
0.756
0.756
0.756
0.489
-1.21
0.541
-0.159
0.562
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = exp(lalpha + rho*ln(Mu(i)))
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
-4.335882
1.437949
-2.994674
-3.106005
-6.226495
# Param's
5
8
6
3
2
AIC
18 . 671765
13. 124101
17.989348
12.212010
16.452991
Test 1:
Test
Test
Test
Explanation of Tests
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
52 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Test
2*log(Likelihood Ratio) Test df
p-value
Test 1
Test 2
Test 3
Test 4
15.3289
11.5477
8.86525
0.222662
6
3
2
3
0.01785
0.009105
0.01188
0.9739
The p-value for Test 1 is less than .05. There appears to be a
difference between response and/or variances among the dose levels
It seems appropriate to model the data
The p-value for Test 2 is less than .1. A non-homogeneous variance
model appears to be appropriate
The p-value for Test 3 is less than .1. You may want to consider a
different variance model
The p-value for Test 4 is greater than .1. The model chosen seems
to adequately describe the data
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0.95
BMD
308 . 389
BMDL = 243.2 85
53
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Hill Model with 0.95 Confidence Level
O)
w
c
0
Q.
W
0)
01
CO
0)
Hill
15
14
13
12
11
10BMDLBMD
50
100
150
200
250
dose
15:14 02/11 2010
Figure C.2. Hill Nonconstant Variance BMD Model for Male Hemoglobin Data
(Mitsumori et al., 1979)
Text Output for Hill Nonconstant Variance BMD Model for Male Hemoglobin Data
(Mitsumori et al., 1979)
Hill Model. (Version: 2.14; Date: 06/26/2008)
Input Data File: C:\BCMEE\Mitsumori 1979 13wk Hb male Hill 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori_197 9_13wk_Hb_male_Hill_l.pit
~Thu Feb 11 15:14:59 2010
Table3 13wks Hb males
The form of the response function is:
Y[dose] = intercept + v*doseAn/(kAn + doseAn)
Dependent variable = Mean
54 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Independent variable = Dose
Power parameter restricted to be greater than 1
The variance is to be modeled as Var(i) = exp(lalpha + rho * In(mean(i)))
Total number of dose groups = 4
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
Default Initial Parameter Values
lalpha = -0.336109
rho = 0
intercept = 14.1
v = -2.3
n = 0.179559
k = 4 . 845
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -n -k
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
lalpha rho intercept v
lalpha 1 -1 -0.039 0.049
rho -1 1 0.039 -0.049
intercept -0.039 0.039 1 -0.84
v 0.049 -0.049 -0.84 1
Parameter Estimates
Confidence Interval
Variable Estimate
Upper Conf. Limit
lalpha 2.54557
28.6113
rho -1.09987
9.0871
intercept 14.1
14.7173
v -1.6
-0.864292
n 1
k
Std. Err.
13.2991
5.19753
0.314934
0.375368
NA
NA
2 . 4218e-013
NA - Indicates that this parameter has hit a bound
95.0% Wald
Lower Conf. Limit
-23.5202
-11.2868
13.4827
-2 . 33571
5 5 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
implied by some inequality constraint and thus
has no standard error.
Table of Data and Estimated Values of Interest
Dose
Res .
Obs Mean
Est Mean Obs Std Dev Est Std Dev Scaled
0
9. 69
48 . 42
242.2
14 . 1
11. 8
12 . 5
13
14 . 1
12 . 5
12 . 5
12 . 5
0.9
1.2
0.7
0.6
0. 833
0.89
0.89
0.89
1. 87e-008
-1.76
-2 . 78e-008
1.49
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = exp(lalpha + rho*ln(Mu(i)))
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
-6.458884
-4 . 925139
-6.321516
-9.515180
-16.105402
# Param's
5
8
6
4
2
AIC
22 . 917768
25. 850278
24.643032
27.030360
36.210805
Test 1:
Test
Test
Test
Explanation of Tests
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
56 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Test
2*log(Likelihood Ratio) Test df
p-value
Test 1
Test 2
Test 3
Test 4
22 . 3605
3. 06749
2 .79275
6. 38733
6
3
2
2
0. 001042
0.3813
0.2475
0.04102
The p-value for Test 1 is less than .05. There appears to be a
difference between response and/or variances among the dose levels
It seems appropriate to model the data
The p-value for Test 2 is greater than .1. Consider running a
homogeneous model
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
The p-value for Test 4 is less than .1. You may want to try a different
model
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0. 95
BMD
2 . 63176e-013
BMDL
2.6317 6e-013
57
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Linear Model with 0.95 Confidence Level
41
40
39
38
37
36
35
34
14:20 02/11 2010
BMD
Figure C.3.1. Linear Constant Variance BMD Model for Female Hematocrit Data
(Mitsumori et al., 1979)
Text Output for Linear Constant Variance BMD Model for Female Hematocrit Data
(Mitsumori et al., 1979)
Polynomial Model. (Version: 2.13; Date: 04/08/2008)
Input Data File:
C:\BCMEE\Mitsumori 197 9 13wk Ht female LinearCV 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori 197 9 13wk Ht female LinearCV l.plt
Thu Feb 11 14:20:08 2010
Table3 13wks Ht females
The form of the response function is:
Y[dose] = beta 0 + beta l*dose + beta 2*doseA2 + ...
5 8 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Dependent variable = Mean
Independent variable = Dose
rho is set to 0
Signs of the polynomial coefficients are not restricted
A constant variance model is fit
Total number of dose groups = 4
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
Default Initial Parameter Values
alpha = 5.175
rho = 0 Specified
beta_0 = 37.2798
beta 1 = 0.0057687
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -rho
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
alpha beta 0 beta 1
alpha 1 8.le-010 4.1e-013
beta_0 8.le-010 1 -0.61
beta 1 4.le-013 -0.61 1
Parameter Estimates
Confidence Interval
Variable Estimate
Upper Conf. Limit
alpha 4.49546
6.85028
beta_0 37.2798
38.2671
beta_l 0.0057687
0.0120993
Std. Err.
1.20146
0.503728
0.00322994
95.0% Wald
Lower Conf. Limit
2.14064
36.2925
-0. 000561875
Table of Data and Estimated Values of Interest
Dose N Obs Mean Est Mean Obs Std Dev Est Std Dev Scaled
Res .
59 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
0 7 36.9 37.3 2.7 2.12 -0.474
11.99 7 37.6 37.3 1.4 2.12 0.313
60.26 7 37.8 37.6 2.8 2.12 0.215
305.8 7 39 39 1.9 2.12 -0.0547
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
-34.855641
-32.850272
-34.855641
-35.042945
-36.553365
# Param's
5
8
5
3
2
AIC
79.711282
81.700543
79.711282
76.085889
77 .106729
Test 1:
Test
Test
Test
Explanation of Tests
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
Test
-2*log(Likelihood Ratio) Test df
p-value
Test 1
Test 2
Test 3
Test 4
7 .40619
4 . 01074
4 . 01074
0.374607
6
3
3
2
0.2849
0.2603
0.2603
0.8292
The p-value for Test 1 is greater than .05. There may not be a
diffence between responses and/or variances among the dose levels
60 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Modelling the data with a dose/response curve may not be appropriate
The p-value for Test 2 is greater than .1. A homogeneous variance
model appears to be appropriate here
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
The p-value for Test 4 is greater than .1. The model chosen seems
to adequately describe the data
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0.95
BMD
367.544
BMDL
188.749
61
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Polynomial Model with 0.95 Confidence Level
41
40
39
O)
w
0 38
w
O)
01
c
TO 07
CD
36
35
34
0 50 100 150 200 250 300 350
dose
14:20 02/11 2010
Figure C.3.2. Polynomial Constant Variance BMD Model for Female Hematocrit Data
(Mitsumori et al., 1979)
Text Output for Polynomial Constant Variance BMD Model for Female Hematocrit Data
(Mitsumori et al., 1979)
Polynomial Model. (Version: 2.13; Date: 04/08/2008)
Input Data File: C:\BCMEE\Mitsumori 197 9 13wk Ht female PolyCV 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori 197 9 13wk Ht female PolyCV l.plt
Thu_Feb 11 14:20:08 2010
Table3 13wks Ht females
The form of the response function is:
Y[dose] = beta 0 + beta l*dose + beta 2*doseA2 + ...
62 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Dependent variable = Mean
Independent variable = Dose
rho is set to 0
The polynomial coefficients are restricted to be positive
A constant variance model is fit
Total number of dose groups = 4
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
Default Initial Parameter Values
alpha = 5.175
rho = 0 Specified
beta 0 = 36.9
beta~l = 0.0713065
beta 2 = 0
beta 3 = 2.95163e-006
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -rho -beta 2 -beta 3
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
alpha beta 0 beta 1
alpha 1 -4.le-008 2.le-008
beta_0 -4.le-008 1 -0.61
beta 1 2.le-008 -0.61 1
Parameter Estimates
Confidence Interval
Variable Estimate
Upper Conf. Limit
alpha 4.49546
6.85028
beta_0 37.2798
38.2671
beta_l 0.0057687
0.0120993
beta 2 0
beta 3 0
Std. Err.
1.20146
0.503728
0. 00322994
NA
NA
95.0% Wald
Lower Conf. Limit
2 . 14064
36.2925
-0.000561875
NA - Indicates that this parameter has hit a bound
63 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
implied by some inequality constraint and thus
has no standard error.
Table of Data and Estimated Values of Interest
Dose
Res .
Obs Mean
Est Mean Obs Std Dev Est Std Dev
Scaled
0
11. 99
60.26
305. 8
36.9
37 . 6
37 . 8
39
37 . 3
37 . 3
37 . 6
39
2 . 7
1.4
2 . 8
1.9
2 . 12
2 . 12
2 . 12
2 . 12
-0.474
0.313
0.215
-0. 0547
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
-34.855641
-32.850272
-34.855641
-35.042945
-36.553365
# Param's
5
8
5
3
2
AIC
79.711282
81.700543
79.711282
76.085889
77 .106729
Test 1:
Test
Test
Test
Explanation of Tests
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
64 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Test
2*log(Likelihood Ratio) Test df
p-value
Test 1
Test 2
Test 3
Test 4
7 .40619
4.01074
4.01074
0.374607
6
3
3
2
0.2849
0.2603
0.2603
0.8292
The p-value for Test 1 is greater than .05. There may not be a
diffence between responses and/or variances among the dose levels
Modelling the data with a dose/response curve may not be appropriate
The p-value for Test 2 is greater than .1. A homogeneous variance
model appears to be appropriate here
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
The p-value for Test 4 is greater than .1. The model chosen seems
to adequately describe the data
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0.95
BMD
367.544
BMDL
188.749
65
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Power Model with 0.95 Confidence Level
dose
14:20 02/11 2010
Figure C.3.3. Power Constant Variance Model for Female Hematocrit Data
(Mitsumori et al., 1979)
Text Output for Power Constant Variance Model for Female Hematocrit Data
(Mitsumori et al., 1979)
Power Model. (Version: 2.15; Date: 04/07/2008)
Input Data File: C:\BCMEE\Mitsumori 197 9 13wk Ht female PowerCV 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori 197 9 13wk Ht female PowerCV l.plt
Thu Feb 11 14:20:08 2010
Table3 13wks Ht females
The form of the response function is:
Y[dose] = control + slope * doseApower
Dependent variable = Mean
66 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Independent variable = Dose
rho is set to 0
The power is restricted to be greater than or equal to 1
A constant variance model is fit
Total number of dose groups = 4
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
Default Initial Parameter Values
alpha = 5.175
rho = 0 Specified
control = 36.9
slope = 0.301101
power = 0.339379
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -rho -power
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
alpha control slope
alpha 1 8.4e-010 -3.3e-009
control 8.4e-010 1 -0.61
slope -3.3e-009 -0.61 1
Parameter Estimates
Confidence Interval
Variable Estimate
Upper Conf. Limit
alpha 4.49546
6.85028
control 37.2798
38.2671
slope 0.0057 687
0.0120993
power 1
Std. Err.
1.20146
0.503728
0.00322994
NA
NA - Indicates that this parameter has hit a bound
implied by some inequality constraint and thus
has no standard error.
95.0% Wald
Lower Conf. Limit
2 . 14064
36.2925
-0. 000561875
67 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table of Data and Estimated Values of Interest
Dose
Res .
Obs Mean
Est Mean Obs Std Dev Est Std Dev
Scaled
0
11. 99
60.26
305. 8
36.9
37 . 6
37 . 8
39
37 . 3
37 . 3
37 . 6
39
2 . 7
1.4
2 . 8
1.9
2 . 12
2 . 12
2 . 12
2 . 12
-0.474
0.313
0.215
-0. 0547
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
-34.855641
-32.850272
-34.855641
-35.042945
-36.553365
# Param's
5
8
5
3
2
AIC
79.711282
81.700543
79.711282
76.085889
77 .106729
Explanation of Tests
Test 1:
Test 2
Test 3
Test 4
(Note:
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
When rho=0 the results of Test 3 and Test 2 will be the same.)
Test
Tests of Interest
-2*log(Likelihood Ratio) Test df
p-value
Test 1
Test 2
Test 3
7 .40619
4.01074
4.01074
0.2849
0.2603
0.2603
68
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Test 4
0.374607
2
0.8292
The p-value for Test 1 is greater than .05. There may not be a
diffence between responses and/or variances among the dose levels
Modelling the data with a dose/response curve may not be appropriate
The p-value for Test 2 is greater than .1. A homogeneous variance
model appears to be appropriate here
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
The p-value for Test 4 is greater than .1. The model chosen seems
to adequately describe the data
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0.95
BMD = 367.544
BMDL
188 . 749
69
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Hill Model with 0.95 Confidence Level
44
42
O)
w
I 40
01
c
CO
0)
38
36
Bf
0 50 100 150 200 250
dose
15:16 02/11 2010
Figure C.4. Hill Constant Variance BMD Model for Male Hematocrit Data
(Mitsumori et al., 1979)
Text Output for Hill Constant Variance BMD Model for Male Hematocrit Data
(Mitsumori et al., 1979)
Hill Model. (Version: 2.14; Date: 06/26/2008)
Input Data File: C:\BCMEE\Mitsumori 1979 13wk Ht male HillCV 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori 197 9 13wk Ht male HillCV l.plt
Thu Feb 11 15:16:07 2010
Table3 13wks Ht males
The form of the response function is:
Y[dose] = intercept + v*doseAn/(kAn + doseAn)
Hill
/IDL
BMD
70 Bis(2-chloro-l-methyl ethyl)ether
-------
FINAL
3-30-2011
Dependent variable = Mean
Independent variable = Dose
rho is set to 0
Power parameter restricted to be greater than 1
A constant variance model is fit
Total number of dose groups = 4
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
Default Initial Parameter Values
alpha = 7.465
rho = 0 Specified
intercept = 41.8
v = 0
n = 1
k = 0
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -rho -k
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
alpha intercept v n
alpha 1 -1.2e-008 2.7e-009 -6.2e-007
intercept -1.2e-008 1 -0.87 8e-007
v 2.7e-009 -0.87 1 -2.3e-007
n -6.2e-007 8e-007 -2.3e-007 1
Parameter Estimates
Confidence Interval
Variable
Upper Conf. Limit
alpha
11.2817
intercept
43.8157
v
-0.672503
n
40553
k
Estimate
7 .40357
41.8
-3
1.31004
2.4218e-013
Std. Err.
1.97869
1. 02842
1.18752
20690
NA
95.0% Wald
Lower Conf. Limit
3.52541
39.7843
-5.3275
-40550.4
71 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
NA - Indicates that this parameter has hit a bound
implied by some inequality constraint and thus
has no standard error.
Table of Data and Estimated Values of Interest
Dose
Res .
Obs Mean
Est Mean Obs Std Dev Est Std Dev Scaled
0
9. 69
48 . 42
242.2
41.8
37 . 7
38 . 3
40.4
41.
38 .
38 .
38 .
3.1
2 . 8
2 . 9
2
2 . 72
2 . 72
2 . 72
2 . 72
4 . 39e-008
-1. 07
-0.486
1.56
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
-39.985047
-39.274047
-39.985047
-42.027475
-44.902099
# Param's
5
8
5
4
2
AIC
89.970093
94 . 548093
89. 970093
92 . 054950
93. 804198
Test 1:
Test
Test
Test
Explanation of Tests
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
72 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Test
2*log(Likelihood Ratio) Test df
p-value
Test 1
Test 2
Test 3
Test 4
4 . 08486
11.2561
1.422
1.422
6
3
3
1
0.08078
0.7004
0.7004
0.04327
The p-value for Test 1 is greater than .05. There may not be a
diffence between responses and/or variances among the dose levels
Modelling the data with a dose/response curve may not be appropriate
The p-value for Test 2 is greater than .1. A homogeneous variance
model appears to be appropriate here
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
The p-value for Test 4 is less than .1. You may want to try a different
model
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0. 95
BMD
1. 37755e-012
BMDL
1. 37755e-012
73
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
0 200 400 600 800 1000
dose
14:21 02/11 2010
Figure C.5. Power Nonconstant Variance BMD Model for Female Erythrocyte Data
(Mitsumori et al., 1979)
Text Output for Power Nonconstant Variance BMD Model for Female Erythrocyte Data
(Mitsumori et al., 1979)
Power Model with 0.95 Confidence Level
Power Model. (Version: 2.15; Date: 04/07/2008)
Input Data File: C:\BCMEE\Mitsumori 197 9 13wk RBC female Power 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori 197 9 13wk RBC female Power l.plt
Thu_Feb 11 14:21:49 2010
Table3 13wks RBC females
The form of the response function is:
Y[dose] = control + slope * doseApower
74 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Dependent variable = Mean
Independent variable = Dose
The power is restricted to be greater than or equal to 1
The variance is to be modeled as Var(i) = exp(lalpha + log(mean(i)) * rho)
Total number of dose groups = 4
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
Default Initial Parameter Values
lalpha = -1.17118
rho = 0
control = 6.9
slope = 1. 65359
power = -0.496845
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -rho -power
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
lalpha control slope
lalpha 1 -0.63 0.11
control -0.63 1 -0.56
slope 0.11 -0.56 1
Parameter Estimates
Confidence Interval
Variable
Upper Conf. Limit
lalpha
-35.7907
rho
control
7.39513
slope
0.000528273
power
Estimate
-36.5206
18
7 . 15536
-0. 000558929
1
Std. Err.
0.372436
NA
0. 122337
0. 000554705
NA
NA - Indicates that this parameter has hit a bound
implied by some inequality constraint and thus
has no standard error.
95.0% Wald
Lower Conf. Limit
-37.2506
6.91558
-0. 00164613
7 5 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Table of Data and Estimated Values of Interest
Dose
Res .
Obs Mean
Est Mean Obs Std Dev Est Std Dev Scaled
0
11. 99
60.26
305. 8
6.9
7 . 4
7 . 1
7
7 . 16
7 . 15
7 . 12
6.98
0.7
0.5
0.5
0.5
0.577
0.572
0.553
0.464
-1. 17
1. 16
-0.104
0.0887
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = exp(lalpha + rho*ln(Mu(i)))
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
4 . 554671
5.210925
4 . 903201
3.266896
2 . 822326
# Param's
5
8
6
3
2
AIC
0. 890657
5.578150
2.193599
-0.533793
-1.644651
Explanation of Tests
Test 1: Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Test 2: Are Variances Homogeneous? (A1 vs A2)
Test 3: Are variances adequately modeled? (A2 vs. A3)
Test 4: Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
Test -2*log(Likelihood Ratio) Test df p-value
Test 1 4.7772 6 0.5727
Test 2 1.31251 3 0.7262
76
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Test 3
Test 4
0.615449
3.27261
2
3
0.7351
0.3515
The p-value for Test 1 is greater than .05. There may not be a
diffence between responses and/or variances among the dose levels
Modelling the data with a dose/response curve may not be appropriate
The p-value for Test 2 is greater than .1. Consider running a
homogeneous model
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
The p-value for Test 4 is greater than .1. The model chosen seems
to adequately describe the data
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0.95
BMD = 1032.7
BMDL = 307.908
77
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Hill Model with 0.95 Confidence Level
7.5
7
O)
w
c
o
Q_
(/)
a)
* 6.5
TO
0)
6
5.5
Br
0 50 100 150 200 250
dose
15:17 02/11 2010
Figure C.6. Hill Constant Variance BMD Model for Male Erythrocyte Data
(Mitsumori et al., 1979)
Text Output for Hill Constant Variance BMD Model for Male Erythrocyte Data
(Mitsumori et al., 1979)
Hill Model. (Version: 2.14; Date: 06/26/2008)
Input Data File: C:\BCMEE\Mitsumori 197 9 13wk RBC male HillCV 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori_197 9_13wk_RBC_male_HillCV_l.pit
Thu Feb 11 15:17:29 2010
Table3 13wks RBC males
The form of the response function is:
Y[dose] = intercept + v*doseAn/(kAn + doseAn)
Hill
/IDL
BMD
78 Bis(2-chloro-l-methyl ethyl)ether
-------
FINAL
3-30-2011
Dependent variable = Mean
Independent variable = Dose
rho is set to 0
Power parameter restricted to be greater than 1
A constant variance model is fit
Total number of dose groups = 4
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
Default Initial Parameter Values
alpha = 0.353636
rho = 0 Specified
intercept = 7.2
v = 0
n = 1
k = 0
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -rho -k
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
alpha intercept v n
alpha 1 -2.2e-009 1.5e-009 -4.2e-009
intercept -2.2e-009 1 -0.85 4.5e-009
v 1.5e-009 -0.85 1 1.7e-009
n -4.2e-009 4.5e-009 1.7e-009 1
Parameter Estimates
Confidence Interval
Variable
Upper Conf. Limit
alpha
0.545446
intercept
7.64036
v
-0.2375
n
6.22437e+006
k
Estimate
0.35336
7.2
-0.752632
1.76817
2 . 4218e-013
Std. Err.
0.0980045
0.224678
0.262827
3.17576e+006
NA
95.0% Wald
Lower Conf. Limit
0.161275
6.75964
-1.26776
-6.22437e+006
79 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
NA - Indicates that this parameter has hit a bound
implied by some inequality constraint and thus
has no standard error.
Table of Data and Estimated Values of Interest
Dose
Res .
Obs Mean
Est Mean Obs Std Dev Est Std Dev Scaled
0
9. 69
48 . 42
242.2
7.2
6.3
6.2
6.8
7.2
6.45
6.45
6.45
0.6
0.7
0.6
0.5
0.594
0. 594
0.594
0.594
7 . 91e-009
-0.554
-1.1
1.57
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
2 . 685023
2 . 963405
2 . 685023
0.523471
-3. 040290
# Param's
5
8
5
4
2
AIC
4 . 629955
10.073191
4.629955
6.953058
10.080580
Test 1:
Test
Test
Test
Explanation of Tests
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
80 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Test
2*log(Likelihood Ratio) Test df
p-value
Test 1
Test 2
Test 3
Test 4
12 . 0074
0.556764
0.556764
4 . 3231
6
3
3
1
0.0618
0.9063
0.9063
0.0376
The p-value for Test 1 is greater than .05. There may not be a
diffence between responses and/or variances among the dose levels
Modelling the data with a dose/response curve may not be appropriate
The p-value for Test 2 is greater than .1. A homogeneous variance
model appears to be appropriate here
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
The p-value for Test 4 is less than .1. You may want to try a different
model
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0. 95
BMD
5. 12028e-013
BMDL
5. 12028e-013
81
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Linear Model with 0.95 Confidence Level
0 50 100 150 200 250 300
dose
15:58 02/12 2010
Figure C.7.1. Linear Nonconstant Variance BMD Model for Female Leukocyte Data
(Mitsumori et al., 1979)
Text Output for Linear Nonconstant Variance BMD Model for Female Leukocyte Data
(Mitsumori et al., 1979)
Polynomial Model. (Version: 2.13; Date: 04/08/2008)
Input Data File: C:\BCMEE\Mitsumori 197 9 13wk WBC female Linear 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori 197 9 13wk WBC female Linear l.plt
Fri Feb 12 15:58:18 2010
Table3 13wks WBC females
The form of the response function is:
Y[dose] = beta 0 + beta l*dose + beta 2*doseA2 + ...
Dependent variable = Mean
82 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Independent variable = Dose
Signs of the polynomial coefficients are not restricted
The variance is to be modeled as Var(i) = exp(lalpha + log(mean(i)) * rho)
Total number of dose groups = 4
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
Default Initial Parameter Values
lalpha = 2.42834
rho = 0
beta 0 = 7.68 64
beta~l = -0.0122883
Asymptotic Correlation Matrix of Parameter Estimates
lalpha rho beta 0 beta 1
lalpha 1 -0.99 -0.037 0.052
rho -0.99 1 0.036 -0.05
beta_0 -0.037 0.036 1 -0.91
beta 1 0.052 -0.05 -0.91 1
Parameter Estimates
Confidence Interval
Variable
Upper Conf. Limit
lalpha
-1.53236
rho
5.57691
beta 0
9.36423
beta 1
-0.00614039
Estimate
-4 . 94521
3.74478
7.69041
-0.0123644
Std. Err.
1.74128
0.934776
0.854003
0.00317559
95.0% Wald
Lower Conf. Limit
-8.35806
1.91266
6.0166
-0. 0185885
Table of Data and Estimated Values of Interest
Dose N Obs Mean Est Mean Obs Std Dev Est Std Dev Scaled
Res .
6.8 7.69 3 3.85 -0.613
83
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
11. 99
7
8 . 5
7 . 54
5.4
3.71
0. 683
60.26
7
6.9
6. 95
2 . 4
3. 18
-0. 0377
305. 8
7
3.9
3. 91
1.2
1. 08
-0.0229
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = exp(lalpha + rho*ln(Mu(i)))
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model Log(likelihood) # Param's AIC
A1 -45.838599 5 101.677197
A2 -38.741501 8 93.483002
A3 -39.074696 6 90.149391
fitted -41.257910 4 90.515820
R -49.328666 2 102.657331
Explanation of Tests
Test 1:
Test
Test
Test
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
Test -2*log(Likelihood Ratio) Test df p-value
Test 1 21.1743 6 0.001707
Test 2 14.1942 3 0.002652
Test 3 0.666389 2 0.7166
Test 4 4.36643 2 0.1127
The p-value for Test 1 is less than .05. There appears to be a
difference between response and/or variances among the dose levels
It seems appropriate to model the data
The p-value for Test 2 is less than .1. A non-homogeneous variance
model appears to be appropriate
84 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
The p-value for Test 4 is greater than .1. The model chosen seems
to adequately describe the data
Benchmark Dose Computation
Specified effect = 1
Risk Type = Estimated standard deviations from the control mean
Confidence level = 0.95
BMD = 311.05
BMDL = 204.122
85
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Polynomial Model with 0.95 Confidence Level
0 50 100 150 200 250 300
dose
15:58 02/12 2010
Figure C.7.2. Polynomial Nonconstant Variance BMD Model for Female Leukocyte Data
(Mitsumori et al., 1979)
Text Output for Polynomial Nonconstant Variance BMD Model for Female Leukocyte
Data (Mitsumori et al., 1979)
Polynomial Model. (Version: 2.13; Date: 04/08/2008)
Input Data File: C:\BCMEE\Mitsumori 197 9 13wk WBC female Poly 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori 197 9 13wk WBC female Poly l.plt
Fri Feb 12 15:58:19 2010
Table3 13wks WBC females
The form of the response function is:
Y[dose] = beta 0 + beta l*dose + beta 2*doseA2 + ...
Dependent variable = Mean
86 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Independent variable = Dose
The polynomial coefficients are restricted to be negative
The variance is to be modeled as Var(i) = exp(lalpha + log(mean(i)) * rho)
Total number of dose groups = 4
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
Default Initial Parameter Values
lalpha = 2.42834
rho = 0
beta 0 = 6.8
beta 1 = 0
beta 2 = -0.00360565
beta 3 = 0
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -beta 2 -beta 3
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
lalpha rho beta 0 beta 1
lalpha 1 -0.99 -0.037 0.052
rho -0.99 1 0.036 -0.05
beta_0 -0.037 0.036 1 -0.91
beta 1 0.052 -0.05 -0.91 1
Parameter Estimates
Confidence Interval
Variable
Upper Conf. Limit
lalpha
-1.53236
rho
5.57691
beta 0
9.36423
beta 1
-0.00614039
beta 2
beta 3
Estimate
-4 . 94521
3.74478
7.69041
-0.0123644
0
0
Std. Err.
1.74128
0.934776
0.854003
0.00317559
NA
NA
95.0% Wald
Lower Conf. Limit
-8 . 35806
1. 91266
6. 0166
-0.0185885
NA - Indicates that this parameter has hit a bound
87 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
implied by some inequality constraint and thus
has no standard error.
Table of Data and Estimated Values of Interest
Dose
Res .
Obs Mean
Est Mean Obs Std Dev Est Std Dev
Scaled
0
11. 99
60.26
305. 8
6.8
8 . 5
6.9
3.9
7 . 69
7 . 54
6. 95
3. 91
3
5.4
2 . 4
1.2
3. 85
3.71
3. 18
1. 08
-0. 613
0. 683
-0. 0377
-0.0229
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = exp(lalpha + rho*ln(Mu(i)))
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
-45.838599
-38.741501
-39.074696
-41.257910
-49.328666
# Param'
5
8
6
4
2
AIC
101.677197
93.483002
90.149391
90.515820
102.657331
Test 1:
Test
Test
Test
Explanation of Tests
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
88 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Test
2*log(Likelihood Ratio) Test df
p-value
Test 1
Test 2
Test 3
Test 4
21.1743
14.1942
0.666389
4.36643
6
3
2
2
0.001707
0.002652
0.7166
0.1127
The p-value for Test 1 is less than .05. There appears to be a
difference between response and/or variances among the dose levels
It seems appropriate to model the data
The p-value for Test 2 is less than .1. A non-homogeneous variance
model appears to be appropriate
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
The p-value for Test 4 is greater than .1. The model chosen seems
to adequately describe the data
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0. 95
BMD
311.05
BMDL
204.122
89
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Power Model with 0.95 Confidence Level
dose
15:58 02/12 2010
Figure C.7.3. Power Nonconstant Variance BMD Model for Female Leukocyte Data
(Mitsumori et al., 1979)
Text Output for Power Nonconstant Variance BMD Model for Female Leukocyte Data
(Mitsumori et al., 1979)
Power Model. (Version: 2.15; Date: 04/07/2008)
Input Data File: C:\BCMEE\Mitsumori 197 9 13wk WBC female Power 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori_197 9_13wk_WBC_female_Power_l.pit
Fri_Feb 12 15:58:19 2010
Table3 13wks WBC females
The form of the response function is:
Y[dose] = control + slope * doseApower
Dependent variable = Mean
90 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Independent variable = Dose
The power is restricted to be greater than or equal to 1
The variance is to be modeled as Var(i) = exp(lalpha + log(mean(i)) * rho)
Total number of dose groups = 4
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
Default Initial Parameter Values
lalpha = 2.42834
rho = 0
control = 3.9
slope = 8.87895
power = -0.264738
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -power
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
lalpha rho control slope
lalpha 1 -0.98 0.39 -0.57
rho -0.98 1 -0.51 0.65
control 0.39 -0.51 1 -0.9
slope -0.57 0.65 -0.9 1
Parameter Estimates
Confidence Interval
Variable
Upper Conf. Limit
lalpha
-0.474956
rho
6.24025
control
9.35501
slope
-0.00622005
power
Estimate
-4 . 94521
3.74478
7.69041
-0.0123644
1
Std. Err.
2.28079
1.27322
0.8493
0.00313495
NA
NA - Indicates that this parameter has hit a bound
implied by some inequality constraint and thus
has no standard error.
95.0% Wald
Lower Conf. Limit
-9.41547
1.24931
6. 02582
-0. 0185088
91 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Table of Data and Estimated Values of Interest
Dose
Res .
Obs Mean
Est Mean Obs Std Dev Est Std Dev Scaled
0
11. 99
60.26
305. 8
6.8
8 . 5
6.9
3.9
7 . 69
7 . 54
6. 95
3. 91
3
5.4
2 . 4
1.2
3. 85
3.71
3. 18
1. 08
-0. 613
0. 683
-0. 0377
-0.0229
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = exp(lalpha + rho*ln(Mu(i)))
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
-45.838599
-38.741501
-39.074696
-41.257910
-49.328666
# Param'
5
8
6
4
2
AIC
101.677197
93.483002
90.149391
90.515820
102.657331
Explanation of Tests
Test 1: Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Test 2: Are Variances Homogeneous? (A1 vs A2)
Test 3: Are variances adequately modeled? (A2 vs. A3)
Test 4: Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
Test -2*log(Likelihood Ratio) Test df p-value
92
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Test 1
Test 2
Test 3
Test 4
21.1743
14.1942
0.666389
4.36643
6
3
2
2
0.001707
0.002652
0.7166
0.1127
The p-value for Test 1 is less than .05. There appears to be a
difference between response and/or variances among the dose levels
It seems appropriate to model the data
The p-value for Test 2 is less than .1. A non-homogeneous variance
model appears to be appropriate
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
The p-value for Test 4 is greater than .1. The model chosen seems
to adequately describe the data
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0.95
BMD = 311.05
BMDL = 204.122
93
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Hill Model with 0.95 Confidence Level
Hill
-Br
7IDL
BMD
15:19 02/11 2010
50
100 150
dose
200
250
Figure C.8. Hill Constant Variance BMD Model for Male Leukocyte Data
(Mitsumori et al., 1979)
Text Output for Hill Constant Variance BMD Model for Male Leukocyte Data
(Mitsumori et al., 1979)
Hill Model. (Version: 2.14; Date: 06/26/2008)
Input Data File: C:\BCMEE\Mitsumori 197 9 13wk WBC male HillCV 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori_197 9_13wk_WBC_male_HillCV_l.pit
Thu Feb 11 15:19:43 2010
Table3 13wks WBC males
The form of the response function is:
Y[dose] = intercept + v*doseAn/(kAn + doseAn)
Dependent variable = Mean
94 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Independent variable = Dose
rho is set to 0
Power parameter restricted to be greater than 1
A constant variance model is fit
Total number of dose groups = 4
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
Default Initial Parameter Values
alpha = 1.745
rho = 0 Specified
intercept = 6.5
v = -3.1
n = 2.55229
k = 6.82705
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -rho
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
alpha intercept v n k
alpha 1 3.5e-008 -2.7e-005 -6.9e-005 -6.9e-005
intercept 3.5e-008 1 -0.75 -4.7e-005 -0.0011
v -2.7e-005 -0.75 1 0.38 0.38
n -6.9e-005 -4.7e-005 0.38 1 1
k -6.9e-005 -0.0011 0.38 1 1
Parameter Estimates
Confidence Interval
Variable Estimate
Upper Conf. Limit
alpha 1.49571
2.2792
intercept 6.5
7.40599
v -3.10012
-1.8986
n 5.3383
2429.53
Std. Err.
0.399746
0.462248
0.613029
1236.86
95.0% Wald
Lower Conf. Limit
0.712226
5.59401
-4.30163
-2418 .86
95 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
8 . 19631
317.522
-614.136
630.528
Table of Data and Estimated Values of Interest
Dose
Res .
Obs Mean
Est Mean Obs Std Dev Est Std Dev Scaled
0
9. 69
48 . 42
242.2
6.5
4 . 3
3.4
3.4
6.5
4 . 3
3.4
3.4
1.7
1.2
1.2
1.1
1.22
1.22
1.22
1.22
-1.26e-007
3 . 73e-007
-0. 000256
0. 000255
Degrees of freedom for Test A3 vs fitted <= 0
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
-19.636454
-18 . 775961
-19.636454
-19.636454
-29. 842855
# Param's
5
8
5
5
2
AIC
49.272909
53.551923
49.272909
49.272909
63.685710
Explanation of Tests
Test 1:
Test
Test
Test
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
96 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Tests of Interest
Test
2*log(Likelihood Ratio) Test df
p-value
Test 1
Test 2
Test 3
Test 4
1.3047 6e-007
22.1338
1.72099
1.72099
6
3
3
0
0.001145
0.6323
0.6323
NA
The p-value for Test 1 is less than .05. There appears to be a
difference between response and/or variances among the dose levels
It seems appropriate to model the data
The p-value for Test 2 is greater than .1. A homogeneous variance
model appears to be appropriate here
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
NA - Degrees of freedom for Test 4 are less than or equal to 0. The Chi-
Square
test for fit is not valid
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0. 95
BMD
7.5642
BMDL
7 . 43034e-006
97
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Linear Model with 0.95 Confidence Level
14
Linear
12
10
8
4
6
2
BMDL
BMD
0
200
400
600
dose
800
1000
1200
14:04 04/29 2010
Figure C.9.1. Linear Nonconstant Variance Model for Female Mouse Total Leukocyte
Data (includes high dose) (Mitsumori et al., 1979)
Text Output for Linear Nonconstant Variance Model for Female Mouse Total Leukocyte
Data (includes high dose) (Mitsumori et al., 1979)
Polynomial Model. (Version: 2.13; Date: 04/08/2008)
Input Data File:
C:\BCMEE\Mitsumori 1979 13wk WBC f hidose Linear 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori 1979 13wk WBC f hidose Linear l.plt
~Thu Apr 29 14:04:39 2010
Table3 13wks WBC females including high dose
The form of the response function is:
Y[dose] = beta 0 + beta l*dose + beta 2*doseA2 + ...
Dependent variable = Mean
Independent variable = Dose
Signs of the polynomial coefficients are not restricted
The variance is to be modeled as Var(i) = exp(lalpha + log(mean(i)) * rho)
Total number of dose groups = 5
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
98 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Default Initial Parameter Values
lalpha = 2.23152
rho = 0
beta_0 = 7.01907
beta 1 = -0.00402286
Asymptotic Correlation Matrix of Parameter Estimates
lalpha
rho
beta 0
beta 1
lalpha
1
-0.91
0. 047
-0. 041
rho
-0.91
1
-0.051
0. 047
beta 0
0. 047
-0. 051
1
-0.88
beta 1
-0.041
0. 047
-0.88
1
Parameter Estimates
Confidence Interval
Variable
Upper Conf. Limit
lalpha
-0.472588
rho
3.85284
beta 0
8.10022
beta 1
-0.00217083
Estimate
-2 . 54051
2.65055
6.76048
-0.0034637
Std. Err.
1.05508
0.613422
0.683554
0.000659636
95.0% Wald
Lower Conf. Limit
-4.60843
1.44827
5.42074
-0. 00475656
Table of Data and Estimated Values of Interest
Dose
Res .
Obs Mean
Est Mean Obs Std Dev Est Std Dev
Scaled
0
11. 99
60.26
305. 8
1212
6.8
8 . 5
6.9
3.9
2 . 6
6.76
6. 72
6. 55
5.7
2 . 56
3
5.4
2 . 4
1.2
1.1
3.53
3.51
3.39
2 . 82
0. 978
0. 0296
1.34
0.272
-1. 69
0.0987
Model Descriptions for likelihoods calculated
99 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = exp(lalpha + rho*ln(Mu(i)))
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model Log(likelihood) # Param's AIC
A1 -53.853939 6 119.707879
A2 -42.369145 10 104.738290
A3 -43.457904 7 100.915808
fitted -50.761704 4 109.523407
R -61.908764 2 127.817528
Explanation of Tests
Test 1:
Test
Test
Test
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
Test -2*log(Likelihood Ratio) Test df p-value
Test 1 39.0792 8 <.0001
Test 2 22.9696 4 0.0001284
Test 3 2.17752 3 0.5364
Test 4 14.6076 3 0.002185
The p-value for Test 1 is less than .05. There appears to be a
difference between response and/or variances among the dose levels
It seems appropriate to model the data
The p-value for Test 2 is less than .1. A non-homogeneous variance
model appears to be appropriate
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
The p-value for Test 4 is less than .1. You may want to try a different
model
100 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Benchmark Dose Computation
Specified effect = 1
Risk Type = Estimated standard deviations from the control mean
Confidence level = 0.95
BMD = 1020.34
BMDL = 713.21
101
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Polynomial Model with 0.95 Confidence Level
dose
14:04 04/29 2010
Figure C.9.2. Polynomial Nonconstant Variance Model for Female Mouse Total Leukocyte
Data (includes high dose) (Mitsumori et al., 1979)
Text Output for Polynomial Nonconstant Variance Model for Female Mouse Total
Leukocyte Data (includes high dose) (Mitsumori et al., 1979)
Polynomial Model. (Version: 2.13; Date: 04/08/2008)
Input Data File: C:\BCMEE\Mitsumori 1979 13wk WBC f hidose Poly 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori 1979 13wk WBC f hidose Poly l.plt
~Thu Apr 29 14:04:39 2010
Table3 13wks WBC females including high dose
The form of the response function is:
Y[dose] = beta 0 + beta l*dose + beta 2*doseA2 + ...
Dependent variable = Mean
Independent variable = Dose
The polynomial coefficients are restricted to be negative
The variance is to be modeled as Var(i) = exp(lalpha + log(mean(i)) * rho)
Total number of dose groups = 5
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
102 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Default Initial Parameter Values
lalpha = 2.23152
rho = 0
beta 0 = 6.8
beta 1 = 0
beta~2 = -0.00378976
beta 3 = 0
beta"4 = -8.06922e-009
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -beta 2 -beta 3 -beta 4
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
lalpha rho beta 0 beta 1
lalpha 1 -0.97 0.047 -0.041
rho -0.97 1 -0.051 0.047
beta_0 0.047 -0.051 1 -0.88
beta 1 -0.041 0.047 -0.88 1
Parameter Estimates
Confidence Interval
Variable
Upper Conf. Limit
lalpha
-0.472586
rho
3.85284
beta 0
8.10022
beta 1
-0.00217084
beta 2
beta 3
beta 4
Estimate
-2 . 54051
2 . 65055
6. 76048
-0.0034637
-7.67 647e-138
0
-5.22843e-144
Std. Err.
1.05508
0.613422
0.683554
0.000659636
NA
NA
NA
95.0% Wald
Lower Conf. Limit
-4 . 60843
1.44827
5.42074
-0. 00475656
NA - Indicates that this parameter has hit a bound
implied by some inequality constraint and thus
has no standard error.
Table of Data and Estimated Values of Interest
103 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Dose N Obs Mean Est Mean Obs Std Dev Est Std Dev Scaled
Res .
0
11. 99
60.26
305. 8
1212
6.8
8 . 5
6.9
3.9
2 . 6
6.76
6. 72
6. 55
5.7
2 . 56
3
5.4
2 . 4
1.2
1.1
3.53
3.51
3.39
2 . 82
0. 978
0. 0296
1.34
0.272
-1. 69
0.0987
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = exp(lalpha + rho*ln(Mu(i)))
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
-53.853939
-42.369145
-43.457904
-50.761704
-61.908764
# Param's
6
10
7
4
2
AIC
119.707879
104.738290
100.915808
109.523407
127 .817528
Test 1:
Test
Test
Test
Explanation of Tests
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
Test
-2*log(Likelihood Ratio) Test df
p-value
Test 1
Test 2
Test 3
Test 4
39.0792
22.9696
2.17752
14.6076
<.0001
0. 0001284
0.5364
0. 002185
104
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
The p-value for Test 1 is less than .05. There appears to be a
difference between response and/or variances among the dose levels
It seems appropriate to model the data
The p-value for Test 2 is less than .1. A non-homogeneous variance
model appears to be appropriate
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
The p-value for Test 4 is less than .1. You may want to try a different
model
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0.95
BMD
1020.34
BMDL
713.21
105
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Power Model with 0.95 Confidence Level
dose
14:04 04/29 2010
Figure C.9.3. Power Nonconstant Variance Model for Female Mouse Total Leukocyte
Data (includes high dose) (Mitsumori et al., 1979)
Text Output for Power Nonconstant Variance Model for Female Mouse Total Leukocyte
Data (includes high dose) (Mitsumori et al., 1979)
Power Model. (Version: 2.15; Date: 04/07/2008)
Input Data File:
C:\BCMEE\Mitsumori 1979 13wk WBC f hidose Power 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori 197 9 13wk WBC f hidose Power l.plt
~Thu Apr 29 14:04:40 2010
Table3 13wks WBC females including high dose
The form of the response function is:
Y[dose] = control + slope * doseApower
Dependent variable = Mean
Independent variable = Dose
The power is restricted to be greater than or equal to 1
The variance is to be modeled as Var(i) = exp(lalpha + log(mean(i)) * rho)
Total number of dose groups = 5
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: le-008
Parameter Convergence has been set to: le-008
106 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
Default Initial Parameter Values
lalpha = 2.23152
rho =
control =
slope =
power =
0
2 . 6
21.803
-0.467282
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -power
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
lalpha
rho
control
slope
lalpha
1
-0.91
0.3
-0.53
rho
-0.91
1
-0.44
0. 63
control
0.3
-0.44
1
-0.89
slope
-0.53
0. 63
-0.89
1
Confidence Interval
Variable
Upper Conf. Limit
lalpha
0.18512
rho
4.28712
control
8.12322
slope
-0. 00215302
power
Estimate
-2 . 54051
2 . 65055
6. 76048
-0.0034637
1
Parameter Estimates
Std. Err.
1.39065
0.834998
0.695291
0.000668727
NA
NA - Indicates that this parameter has hit a bound
implied by some inequality constraint and thus
has no standard error.
95.0% Wald
Lower Conf. Limit
-5.26614
1.01399
5.39773
-0. 00477438
Dose
Res .
Table of Data and Estimated Values of Interest
N Obs Mean Est Mean Obs Std Dev Est Std Dev Scaled
107 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
0
7
6.8
6.76
3
3.53
11. 99
7
8 . 5
6. 72
5.4
3.51
60.26
7
6.9
6. 55
2 . 4
3.39
305. 8
7
3.9
5.7
1.2
2 . 82
1212
7
2 . 6
2 . 56
1.1
0. 978
0. 0296
1.34
0.272
-1. 69
0.0987
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = exp(lalpha + rho*ln(Mu(i)))
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
-53.853939
-42.369145
-43.457904
-50.761704
-61.908764
# Param's
6
10
7
4
2
AIC
119.707879
104.738290
100.915808
109.523407
127 .817528
Test 1:
Test
Test
Test
Explanation of Tests
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
Test
-2*log(Likelihood Ratio) Test df
p-value
Test 1
Test 2
Test 3
Test 4
39.0792
22.9696
2.17752
14.6076
<.0001
0. 0001284
0.5364
0. 002185
The p-value for Test 1 is less than .05. There appears to be a
difference between response and/or variances among the dose levels
It seems appropriate to model the data
108 Bis(2-chloro-1 -m ethyl ethyl)ether
-------
FINAL
3-30-2011
The p-value for Test 2 is less than .1. A non-homogeneous variance
model appears to be appropriate
The p-value for Test 3 is greater than .1. The modeled variance appears
to be appropriate here
The p-value for Test 4 is less than .1. You may want to try a different
model
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0.95
BMD = 1020.34
BMDL = 713.21
109
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Hill Model with 0.95 Confidence Level
Bli/IDL
200
400
600
800
1000
dose
14:07 04/29 2010
Figure C.10. Hill Constant Variance Model for Male Mouse Total Leukocyte Data
(includes high dose) (Mitsumori et al., 1979)
Text Output for Hill Constant Variance Model for Male Mouse Total Leukocyte Data
(includes high dose) (Mitsumori et al., 1979)
Hill Model. (Version: 2.14; Date: 06/26/2008)
Input Data File:
C:\BCMEE\Mitsumori 1979 13wk WBC m hidose HillCV 1.(d)
Gnuplot Plotting File:
C:\BCMEE\Mitsumori 1979 13wk WBC m hidose HillCV l.plt
~Thu Apr 29 14:07:28 2010
Table3 13wks WBC males with high dose
The form of the response function is:
Y[dose] = intercept + v*doseAn/(kAn + doseAn)
Dependent variable = Mean
Independent variable = Dose
rho is set to 0
Power parameter restricted to be greater than 1
A constant variance model is fit
Total number of dose groups = 5
Total number of records with missing values = 0
Maximum number of iterations = 250
110 Bis(2-chloro-l-methyl ethyl)ether
-------
Relative Function Convergence has been set to: le-OOf
Parameter Convergence has been set to: le-008
FINAL
3-30-2011
Default Initial Parameter Values
alpha =
rho =
intercept =
v =
n =
k =
1.446
0
6.5
-5
0.377889
22 . 6
Specified
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -rho -n
have been estimated at a boundary point, or have been
specified by the user,
and do not appear in the correlation matrix )
alpha
intercept
alpha
1
1.8e-007
4 . 6e-007
-6e-007
intercept
1. 8e-007
1
-0.76
-0.5
v
4.6e-007
-0.76
1
0. 016
k
-6e-007
-0.5
0. 016
1
Confidence Interval
Variable Estimate
Upper Conf. Limit
alpha 1.57915
2.31902
intercept 6.43276
7.39388
v -4.06868
-2.94916
n 1
k 11.4417
25.9509
Parameter Estimates
Std. Err.
0.37749
0.490376
0.571192
NA
7 .40277
NA - Indicates that this parameter has hit a bound
implied by some inequality constraint and thus
has no standard error.
95.0% Wald
Lower Conf. Limit
0. 839287
5.47164
-5.18819
-3. 06747
Table of Data and Estimated Values of Interest
111 Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
Dose N Obs Mean Est Mean Obs Std Dev Est Std Dev Scaled
Res .
0 7 6.5 6.43 1.7 1.26 0.142
9.69 7 4.3 4.57 1.2 1.26 -0.562
48.42 7 3.4 3.14 1.2 1.26 0.544
242.2 7 3.4 2.55 1.1 1.26 1.79
984.9 7 1.5 2.41 0.5 1.26 -1.92
Model Descriptions for likelihoods calculated
Model A1: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A2: Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)A2
Model A3: Yij = Mu(i) + e(ij)
Var{e(ij)} = SigmaA2
Model A3 uses any fixed variance parameters that
were specified by the user
Model R: Yi = Mu + e(i)
Var{e(i)} = SigmaA2
Likelihoods of Interest
Model
A1
A2
A3
fitted
R
Log(likelihood)
-21.256383
-16.884404
-21.256383
-25.495563
-41. 177561
# Param's
6
10
6
4
2
AIC
54.512766
53.768807
54 . 512766
58 . 991126
86.355121
Test 1:
Test
Test
Test
Explanation of Tests
Do responses and/or variances differ among Dose levels?
(A2 vs. R)
Are Variances Homogeneous? (A1 vs A2)
Are variances adequately modeled? (A2 vs. A3)
Does the Model for the Mean Fit? (A3 vs. fitted)
(Note: When rho=0 the results of Test 3 and Test 2 will be the same.)
Tests of Interest
Test
-2*log(Likelihood Ratio) Test df
p-value
Test 1
Test 2
Test 3
Test 4
48.5863
8.74396
8 . 74396
8.47836
<.0001
0. 06783
0. 06783
0.01442
112
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
The p-value for Test 1 is less than .05. There appears to be a
difference between response and/or variances among the dose levels
It seems appropriate to model the data
The p-value for Test 2 is less than .1. Consider running a
non-homogeneous variance model
The p-value for Test 3 is less than .1. You may want to consider a
different variance model
The p-value for Test 4 is less than .1. You may want to try a different
model
Benchmark Dose Computation
Specified effect
1
Risk Type
Estimated standard deviations from the control mean
Confidence level
0. 95
BMD
5. 11308
BMDL
1.71103
113
Bis(2-chloro-1 -methyl ethyl)ether
-------
FINAL
3-30-2011
APPENDIX D. REFERENCES
ACGIH (American Conference of Governmental Industrial Hygienists). (2009) 2009 TLVs and
BEIs: Threshold limit values for chemical substances and physical agents and biological
exposure indices. Cincinnati, OH: ACGIH. 594528
ATSDR (Agency for Toxic Substances and Disease Registry). (2008) Toxicological profile
information sheet. Toxicological profile information sheet. Atlanta, GA: U.S. Department of
Health and Human Services, Public Health Service. Available online at
http://www.atsdr.cdc.gov/toxprofiles/index.asp Accessed 2/25/2010. 595415
HSDB (Hazardous Substance Data Bank). (2010) Bis(2-chloro-l-methylethyl) ether. Bethesda,
MD: National Library of Medicine. Available online at http://toxnet.nlm.nih.gov/cgi-bin/sis/
search/f?./temp/~FejoKs:2. Last updated on 2-14-2003.
IARC (International Agency for Research on Cancer). (1986) Bis(2-Chloro-l-methylethyl)ether.
In: Some halogenated hydrocarbons and pesticide exposures. IARC monographs on the
evaluation of carcinogenic risks to humans, volume 41. Geneva, Switzerland: World Health
Organization, pp 149-160. 201656
IARC (International Agency for Research on Cancer). (2000) Some industrial chemicals.
IARC monographs on the evaluation of carcinogenic risks to humans, volume 77. Geneva,
Switzerland: World Health Organization. Available online at http://monographs.iarc.fr/ENG/
Monographs/vol77/index.php. 201837
Jorgenson, TA; Rushbrook, CJ; Newell, GW; et al. (1977) Study of the mutagenic potential of
bis (2-chloroethyl) and bis (2-chloroisopropyl) ethers in mice by the heritable translocation test.
ToxicolApplPharmacol 41:196-197. 200858
McGregor, DB; Brown, A; Cattanach, P; et al. (1988) Responses of the L5178Y tk+!tk~ mouse
lymphoma cell forward mutation assay:III. 72 coded chemicals. EnvironMolMutat 12:85-154.
Mirsalis, JC; Tyson, CK; Steinmetz, KL; et al. (1989) Measurement of unscheduled DNA
synthesis and S-phase synthesis in rodent hepatocytes following in vivo treatment: Testing of 24
compounds. Environ Mol Mutagen 14(3): 155-164. 200781
Mitsumori, K; Usui, T; Takahashi, K;et al. (1979) Twenty-four month chronic toxicity studies
of dichlorodiisopropyl ether in mice. J Pesticide Sci 4:323-335. Available online at
http://rms 1 .agsearch.agropedia.affrc,go.jp/contents/JASI/pdf/society/19-3606.pdf. 200783
Moriya, M; Ohta, T; Watanabe, K; et al. (1983) Further mutagenicity studies on pesticides in
bacterial reversion assay systems. Mutat Res 1 16(3-4): 185-216. 200489
NCI (National Cancer Institute). (1979) Bioassay of technical grade bis(2-chloro-l-
methylethyl) ether for possible carcinogenicity (CAS No. 108-60-1). U.S. Department of Health,
Education, and Welfare. Bethesda, MD. NIH-79-1747. Available online at
http://ntp.niehs.nih.gov/ntp/htdocs/LT_rpts/tr 191 .pdf 201646
114 Bis(2-chloro-l-methyl ethyl)ether
-------
FINAL
3-30-2011
NIOSH (National Institute for Occupational Safety and Health). (2003). NIOSH pocket guide to
chemical hazards. Atlanta, GA: Department of Health and Human Services, Centers for Disease
Prevention and Control. 081445
NTP (National Toxicology Program). (1982) Carcinogenesis bioassay of Bis(2-chloro-l-
methylethyl)ether (-70%) (CAS No. 108-60-1) containing 2-Chloro-l-methyl ethyl-(2-
chloropropyl) ether (-30%) (CAS No. 83270-31-9) in B6C3F1 mice (gavage study). TR 239.
NTP. Research Triangle Park, NC. NTP-81-55. Available online at http://ntp.niehs.nih.gov/ntp/
htdocs/LT_rpts/tr23 9.pdf
OEHHA (Office of Environmental Health Hazard Assessment). (1999) Evidence on the
carcinogenicity of technical grade bis(2-chloro-l-methylethyl)ether. California Environmental
Protection Agency. http://oehha.ca.gov/prop65/pdf/BCMEEf.pdf.
OSHA (Occupational Safety and Health Administration). (1997) Air contaminants:
occupational safety and health standards for shipyard employment, subpart Z, toxic and
hazardous substances. U.S. Department of Labor, Washington, DC. OSHA Standard
1915.1000. Available online at http://www.osha.gov/pls/oshaweb/owadisp.show_document?
p table=STANDARDS&p_id= 10286. Accessed on 6/10/2004. 194391
Smith, CC; Lingg, RD; Tardiff, RG. (1978) Comparative metabolism of haloethers. Ann NY
AcadSci 298:111-123. 200778
U.S. EPA (Environmental Protection Agency). (1987) Health and environmental effects
document for haloethers. Office of Research and Development, Washington, DC. ECAO-CIN-
G014. 200805
U.S. EPA (Environmental Protection Agency). (2005) Guidelines for carcinogen risk
assessment, Final Report. Risk Assessment Forum, U.S. Environmental Protection Agency.
Washington, DC. EPA/630/P-03/001F. Available online at http://cfpub.epa.gov/ncea/cfm/
recordisplay.cfm?deid= 1 16283. 086237
U.S. EPA (Environmental Protection Agency). (2006) 2006 Edition of the drinking water
standards and health advisories. Office of Water, Washington, DC. EPA 822-R-06-013.
Available online at http://vvvvvv.epa.gov/vvaterscience/criteria/drinking/dvvstandards.pdf. 091193
U.S. EPA (Environmental Protection Agency). (2010a) Integrated risk information system
(IRIS). Available online at http://cfpub.epa.gov/ncea/iris/index.cfm. Accessed on 6/24/2009.
192196
U.S. EPA (Environmental Protection Agency). (2010b) Health effects assessment summary
tables (HEAST). Available online at http://epa-heast.oml.gov/. Accessed 2/25/2010. 595422
WHO (World Health Organization). (2010) Online catalogs for the Environmental Health
Criteria Series. Available online at http://www.who.int/ipcs/publications/ehc/en/. Accessed on
2/25/2010. 595424
Zeiger, E. (1987) Carcinogenicity of mutagens: predictive capability of the Salmonella
mutagenesis assay for rodent carcinogenicity. Cancer Res 47:1287-1296. 200779
115 Bis(2-chloro-1 -methyl ethyl)ether
------- |