PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

AEPA

United States	Office of Chemical Safety and

Environmental Protection Agency	Pollution Prevention

Draft Risk Evaluation for
Carbon Tetrachloride

Supplemental File:

Benchmark Dose Modeling and Source Code for PBPK

Model for Derivation of the IRIS Reference
Concentration (POD for Chronic Inhalation Exposures)
and Inhalation Unit Risk

CASRN: 56-23-5

ci

C""C|

cr

ci

January 2020

1


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

BENCHMARK DOSE MODELING FOR DERIVING THE REFERENCE

CONCENTRATION

MALE RAT:

Incidence data for fatty changes of the liver

Male F344 rats exposed to carbon tetrachloride vapor for 104 weeks (6 hours/day,
5 days/week)

Exposure concentrations modeled: 0, 5, 25,125ppm
BMR = 10%

Model

Vmax = 0.4

vmax = 0.65

AIC

y?p-value3

BMC10

BMCL10

AIC

y2/;-value11

BMC10

BMCL10

MCA (jimol/L)

Gammab

144.336

0.0007

0.0793248

0.0551873

144.772

0.0005

0.0689847

0.051179

Logistic0

155.104

0.0000

0.170834

0.137191

156.51

0.0000

0.157857

0.126743

Log~Logisticc

137.403

0.4355

0.136715

0.0790319

137.463

0.4087

0.123076

0.0707077

Multistage
l-degreede

142.388

0.0074

0.0714015

0.0550523

142.778

0.0031

0.0665234

0.0511645

Probit0

169.521

0.0000

0.22329

0.17626

171.234

0.0000

0.21463

0.168317

Log-probitc

138.408

0.1761

0.124953

0.0755939

138.529

0.1581

0.112257

0.0803264

Quantal-linear

142.388

0.0074

0.0714017

0.0550523

142.778

0.0031

0.0665234

0.0511645

W eibullb

142.388

0.0074

0.0714016

0.0550523

142.778

0.0031

0.0665235

0.0511645

MRAMKL

fimol/hr-kg liver)

Gammab

137.468

0.4177

3.98707

2.6343

137.338

0.4760

5.31098

3.35649

Logistic0

136.747

0.3444

3.25675

2.58557

136.513

0.3671

4.60057

3.65284

Log-Logistic0

136.933

0.8012

4.56744

3.08461

136.996

0.7246

6.20422

4.00273

Multistage
2-degreeef

137.073

0.2702

3.55184

2.02617

138.991

0.0944

4.99656

2.5022

Probit0

138.891

0.0826

2.97807

2.41619

138.712

0.0728

4.23817

3.44383

Log-probitฐ

136.871

0.9538

4.27176

3.06539

136.872

0.9470

5.73628

3.97844

Quantal-linear

151.674

0.0008

1.01942

0.831472

148.898

0.0025

1.45532

1.18412

W eibullb

138.997

0.1316

3.34831

2.18252

138.601

0.1751

4.4781

2.81908

aValues <0.1 fail to meet conventional goodness-of-fit criteria;p value from the y} test.

''Power restricted to >1.
cSlope restricted to >1.

dUsed smallest degree polynomial available with an adequate fit; the 2-and 3-degree polynomials provided the
same fit as the 1-degree.
eBetas restrictedto >0.

fUsed smallest degree polynomial available with an adequate fit; the 3-degree polynomial providedthe same fit as
the 2-degree.

2


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

FEMALE RAT:

Incidence data for fatty changes of the liver

Female F344 rats exposed to carbon tetrachloride vapor for 104 weeks (6 hours/day,
5 days/week)

Exposure concentrations modeled: 0, 5, 25,125ppm
BMR = 10%

None of the models in BMDS provided an adequate fit of the female rat data.

Incidence data for fatty changes of the liver

Female F344 rats exposed to carbon tetrachloride vapor for 104 weeks (6 hours/day,
5 days/week)

Exposure concentrations modeled: 0, 5, 25ppm (high dose dropped)

BMR = 10%





v

* max

II

o

4^





V

'max

= 0.65



Model

AIC

y?p-value3

BMC10

BMCL10

AIC

y?p-value3

BMC10

BMCL10

MCA (jimol/L)

Gammab

92.9928

NA

0.187771

0.107455

92.9928

NA

0.170979

0.0971536

Logistic0

93.4185

0.1121

0.106984

0.0803379

93.3172

0.1201

0.0979754

0.0734707

Log-Logistic0

92.9928

a NA

0.182663

0.111838

92.9928

NA

0.166144

0.101213

Multistage*1'ฎ

2nd degree
92.4089

0.2442

0.123631

0.0851972

2nd degree
92.3049

0.2617

0.113721

0.0775873



3rd degree
94.9928

NA

0.213915

0.090506

3rd degree
92.9928

NA

0.195194

0.08177

Probit0

93.6833

0.0968

0.100288

0.0779817

93.5689

0.1043

0.0919928

0.0714911

Log-probit0

92.9928

NA

0.174053

0.112578

92.9928

NA

0.158234

0.101889

Quantal-linear

111.424

0.0000

0.0363563

0.0277405

111.025

0.0001

0.0332712

0.0253689

W eibullb

92.9928

NA

0.213201

0.102923

92.9928

NA

0.194228

0.0930656

MRAMKL

fimol/hr-kg liver)

Gammab

92.9928

NA

4.85516

3.42634

92.9928

NA

6.52318

4.43018

Logistic0

99.7262

0.0020

2.45785

1.90371

97.8675

0.0064

3.34536

2.58247

Log-Logistic0

92.9928

NA

4.84705

3.48106

92.9928

NA

6.48806

4.51798

Miltistaged'e

2nd degree
100.7

0.0039

2.43344

1.99357

2nd degree
98.1134

0.0124

3.42266

2.75565



3rd degree
92.2866

0.2650

3.76974

2.82488

3rd degree
91.5964

0.4421

5.42354

3.74923

Probit0

100.988

0.0013

2.16088

1.70134

98.8142

0.0044

2.98448

2.34695

Log-probit0

92.9928

NA

4.69168

3.49658

92.9928

NA

6.26103

4.54001

Quantal-linear

127.034

0.0000

0.817323

0.634088

123.548

0.0000

1.12472

0.870515

W eibullb

92.9928

NA

5.3798

3.29131

92.9928

NA

7.27174

4.24944

aValues <0.1 fail to meet conventional goodness-of-fit criteria;p value from the y} test.
''Power restricted to >1.
cSlope restricted to >1.

dUsed smallest degree polynomial available with an adequate fit.
eBetas restrictedto >0.

3


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Male Rat

Dose metric: MCA

Vmax = 0.4 mg/hour/kg BW0'07

Log-Logistic Model with 0.95 Confidence Level

0 0.5	1 1.5 2 2.5 3 3.5 4

dose

10:59 10/12 2007

Logistic Model. (Version: 2.9; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\MALE RAT\MCA-
VMAX= 0.4\RAT-FATTYLIVER-MCA-4.(d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\MALE RAT\MCA-
VMAX= 0.4\RAT-FATTYLIVER-MCA-4.pit

Fri Oct 12 10:59:34 2007

BMDS MODEL RUN

The form of the probability function is:

P[response] = background+(1-background)/[1+EXP(-intercept-slope*Log(dose))]

Dependent variable = FattyLiver
Independent variable = umol/L
Slope parameter is not restricted

Total number of observations = 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

User has chosen the log transformed model

Default Initial Parameter Values
background =	0.08

intercept =	1.42536

slope =	1.89476

4


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Asymptotic Correlation Matrix of Parameter Estimates

background intercept	slope

background	1	-0.077	0.34

intercept	-0.077	1	0.54

slope	0. 34	0.54	1

Parameter Estimates

95.0% Wald Confidence Interval
Variable	Estimate	Std. Err.	Lower Conf. Limit Upper Conf. Limit

background	0.073606	*	*	*

intercept	1.74202	*	*	*

slope	1. 97967	*	*	*

* - Indicates that this value is not calculated.

Analysis of Deviance Table

Model	Log(likelihood)	# Param's	Deviance Test d.f. P-value

Full model	-65.434	4

Fitted model	-65.7017	3	0.535433 1	0.4643

Reduced model	-138.619	1	146.371 ^	<.0001

AIC:	137.403

Goodness of Fit

Scaled

Dose	Est._Prob. Expected Observed	Size	Residual

0.0000	0.0736	3.680	4	50	0.173

0.1280	0.1559	7.796	7	50	-0.310

0.7080	0.7614	38.068	39	50	0.309

3.8920	0.9891	49.456	49	50	-0.621

Chi^2 = 0.61	d.f. = 1	P-value = 0.4355

Benchmark Dose Computation
Specified effect =	0.1

Risk Type	Extra risk

Confidence level =	0.95

BMD =	0.136715

BMDL =	0.07 9 0319

5


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Male Rat

Dose metric: MCA

Vmax = 0.65 mg/hour/kg BW0'07

Log-Logistic Model with 0.95 Confidence Level

dose

11:12 10/122007

Logistic Model. (Version: 2.9; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\MALE RAT\MCA-
VMAX= 0.65\RAT-FATTYLIVER-MCA-65. (d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\MALE RAT\MCA-
VMAX=0.65\RAT-FATTYLIVER-MCA-65.pit

Fri Oct 12 11:12:25 2007

BMDS MODEL RUN

The form of the probability function is:

P[response] = background+(1-background)/[1+EXP(-intercept-slope*Log(dose))]

Dependent variable = FattyLiver

Independent variable = umol/L

Slope parameter is restricted as slope >= 1

Total number of observations = 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

User has chosen the log transformed model

Default Initial Parameter Values
background =	0.08

intercept =	1.54201

slope =	1.8567 2

Asymptotic Correlation Matrix of Parameter Estimates
background intercept	slope


-------
background
intercept
slope

PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

1

-0.05
0. 33

-0. 05
1

0. 6

0. 33
0. 6
1

Variable
background
intercept
slope

Parameter Estimates

Estimate
0. 0733292
1.88323
1.94775

Std. Err.

Indicates that this value is not calculated.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

Model
Full model
Fitted model
Reduced model

Analysis of Deviance Table

Deviance Test d.f.

Log(likelihood)
-65.434
-65.7316
-138.619

Param's
4
3
1

P-value

0.595159
146.371

(51. 44^
C. 0001



AIC



137.463











Goodness of Fit















Scaled

Dose



Est. Prob.

Expected Observed

Size

Residual

0.0000



0.0733

3.666 4

50

0.181

0.1160



0.1568

7.841 7

50

-0.327

0.6530



0. 7603

38.017 39

50

0. 326

3.7750



0.9895

49.476 49

50

-0.661

Chi/N2 = 0.

68

d.f.

= 1 P-value = 0.4087





Benchmark Dose Computation
Specified effect =	0.1

Risk Type	=	Extra risk

Confidence level =	0.95

BMD =	0.12 3 07 6

BMDL =	0.0707077

7


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Male Rat

Dose metric: MRAMKL
Vmax = 0.4 mg/hour/kg BW0 07

Logistic Model with 0.95 Confidence Level

Logistic Model. (Version: 2.9; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\MALE RAT\MRAMKL-
VMAX=0.4\FATTY_LIVER_MRAMKL-4.(d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\MALE
RAT \ MRAMKL - VMAX= 0 . 4 \ FATTY_LIVER_MRAMKL-4 .pit

Fri Oct 12 11:17:49 2007

BMDS MODEL RUN

The form of the probability function is:

P[response] = 1/[1+EXP(-intercept-slope*dose)]

Dependent variable = FattyLiver
Independent variable = umol/hr-kgL
Slope parameter is not restricted

Total number of observations 3 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

—1—1—1—1—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—1—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—1—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—1—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—1—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—ฆ—r

Logistic 	

BMDL BVID.

0	5

11:17 10/122007

Default Initial Parameter Values

background =	0 Specified

intercept =	-2.35241

slope =	0.249767

Asymptotic Correlation Matrix of Parameter Estimates

( *** The model parameter (s) -background

have been estimated at a boundary point, or have been specified by the user,
and do not appear in the correlation matrix )

8


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

intercept	slope

intercept	1	-0.82

slope	-0.82	1

Parameter Estimates

Variable
intercept
slope

Estimate
-2.68587
0.309634

95.0% Wald Confidence Interval
Std. Err.	Lower Conf. Limit Upper Conf. Limit

0.383165	-3.43685	-1.93488

0.0415113	0.228273	0.390994

Analysis of Deviance Table

Model
Full model
Fitted model
Reduced model

AIC:

Log(likelihood)
-65.434
-66.3737
-138.619

136. 747

Param's
4
2
1

Deviance Test d.f.

1. 87944
146.371

P-value

0.3907
C. 0001

Dose

Goodness of Fit

Est. Prob.

Expected

Observed

Scaled
Residual

0.0000
3.8130
12.0920
24.3200

Chi^2

2 .13

0.0638
0.1816
0.7424
0.9922

d.f.

3.191
9. 082
37 .118
4 9.609

4
7
39
49

2

P-value

0.3444

50
50
50
50

0. 468
-0.764
0|^9
-0. 979

Benchmark Dose Computation
Specified effect =	0.1

Risk Type	=	Extra risk

Confidence level =	0.95

BMD =	3.25 675

BMDL =	2.58 557

9


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Male Rat

Dose metric: MRAMKL
Vmax = 0.65 mg/hour/kg BW0'07

Logistic Model with 0.95 Confidence Level

1

0.8

"O
CD

0.6

a
o

o 0.4

CD

0.2

0

11:23

Logistic: Model. (Version: 2.9; Date: 02/20/2007)

Input Data File: G:\CARB0N TET\EMD\EMD MODELING 10-2007\RFC RAT LIVER\MALE RAT\MRAMKL-
VMAX=0. 65\MRAT_FATTY_LIVER_MRAMKL-65. (d)

Gnuplot Plotting File: G:\CARE0M TET\EMD\EMD MODELING 10-2007\RFC RAT LIVER\MALE
RAT\MRAMKL- VMAX= (J . 65\MRAT_FATTY_LIVER_MRAMKL-65 . pit

Fri Oct 12 11:23:29 2007

EMDS MODEL RUM

The form of the probability function is:

P[response] = 1/[1+EXP(-intercept-slope*dose)]

Dependent variable = FattyLiver
Independent variable = umol/hr-kgL
Slope parameter is not restricted

Total number of observations = 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

background =	0 Specified

intercept =	-2.28912

slope =	0.166325

Asymptotic Correlation Matrix of Parameter Estimates

(	The model parameter (s) -background

have been estimated at a boundary point, or have been specified by the user,
and do not appear in the correlation matrix )

dose

10/12 2007

10


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

intercept	slope

intercept	1	-0.8

slope	-0.8	1

Variable
intercept
slope

Parameter Estimates

Estimate
-2.59592
0.207777

Std. Err.

0. 370821
0.0278282

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit
-3.32272	-1.86913

0.153235	0.26232

Model
Full model
Fitted model
Reduced model

AIC:

Analysis of Deviance Table

Deviance Test d.f.

Log(likelihood)
-65.434
-66.2567
-138.619

136.513

Param's
4
2
1

1. 64 536 \
146.371	3

P-value

0.4393
C. 0001





Goodness of Fit













Scaled

Dose

Est. Prob.

Expected Observed

Size

Residual

0.0000

0.0694

3.470 4

50

0.|ฃ9 5

4.9910

0.1738

8.690 7

50

-0.631

17 . 6260

0.7439

37.195 39

50

0. 585

36 .2660

0.9929

49.645 49

50

-1.085

Chi ^2 = 2.

.00 d.f. =

2 P-value = 0.3671





Benchmark Dose Computation
Specified effect =	0.1

Risk Type	=	Extra risk

Confidence level =	0.95

BMD =	4.60 0 57

BMDL =	3.652 84

11


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Female Rat

Dose metric: MCA

Vmax = 0.4 mg/hour/kg BW0'07

1

0.8

"O
CD

% 0.6
| 0.4

Ll_

0.2

0

11:42

Multistage Model. (Version: 2.8; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\FEMALE RAT\MCA-
VMAX= 0.4\FRAT-FATTYLIVER-MCA-4.(d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\FEMALE
RAT\MCA-VMAX= 0.4\FRAT-FATTYLIVER-MCA-4.pit

Fri Oct 12 11:42:22 2007

BMDS MODEL RUN

The form of the probability function is:

P[response] = background + (1-background)*[1-EXP(
-betal*dose/sl-beta2*dose/s2 ) ]

The parameter betas are restricted to be positive

Dependent variable = FattyLiver
Independent variable = umol/L

Total number of observations =

Total number of records with missing values = 0
Total number of parameters in model = 3
Total number of specified parameters = 0
Degree of polynomial = 2

Multistage Model with 0.95 Confidence Level

dose

10/12 2007

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
Background = 0.0746099
Beta(1) =	0

Beta(2) =	7.64624

Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter (s) -Beta(l)

12


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

have been estimated at a boundary point, or have been specified by the user,
and do not appear in the correlation matrix )

Background	Beta(2)

Background	1	-0.21

Beta (2)	-0.21	1

Parameter Estimates

95.0% Wald Confidence Interval
Variable	Estimate	Std. Err.	Lower Conf. Limit Upper Conf. Limit

Background	0.0951491	*	*	*

Beta(1)	0	*	*	*

Beta(2)	6.89319	*	*	*

* - Indicates that this value is not calculated.

Analysis of Deviance Table

Model	Log(likelihood)	# Param's	Deviance Test d.f.	P-value

Full model	-43.4964	3

Fitted model	-44.2044	2	1.41613 1	0.234

Reduced model	-101.707	1	116.422 2	<.0001

AIC:	92 .4089

Goodness of Fit

Dose

Est. Prob.

Expected

Observed

Si ze

Scaled
Residual

0.0000

0.0951

4 .757

6

50

0.599

0.1280

0.1918

9. 589



50

-0.930

0.7080

0.9714

48.571

49

50

0. 364

Chi^2 = 1.36	d.f. = 1	P-value = 0.2442

Benchmark Dose Computation

Specified effect =	0.1

Risk Type =	Extra risk

Confidence level =	0.95

BMD =	0.12 3 631

BMDL =	0. 0851972

BMDU =	0.148857

Taken together, (0.0851972, 0.148857) is a 90	% two-sided confidence

interval for the BMD

13


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Female Rat

Dose metric: MCA

Vmax = 0.65 mg/hour/kg BW0'07

Multistage Model with 0.95 Confidence Level

1

0.8

"O
CD
O

& 0.6
<

| 0.4

Ll_

0.2

0

11:47

Multistage Model. (Version: 2.8; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\FEMALE RAT\MCA-
VMAX= 0.65\FRAT-FATTYLIVER-MCA-65. (d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\FEMALE
RAT\MCA-VMAX= 0.6 5\FRAT-FAT TYLIVE R-MCA-6 5.p11

Fri Oct 12 11:47:23 2007

BMDS MODEL RUN

The form of the probability function is:

P[response] = background + (1-background)*[1-EXP(
-betal*dose/sl-beta2*dose/N2 ) ]

The parameter betas are restricted to be positive

Dependent variable = FattyLiver
Independent variable = umol/L

Total number of observations ^ 3

Total number of records with missing values = 0

Total number of parameters in model = 3

Total number of specified parameters = 0

Degree of polynomial = 2

dose

10/12 2007

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
Background = 0.0765787
Beta(1) =	0

Beta(2) =	8.98383

Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter (s) -Beta(l)

14


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

have been estimated at a boundary point, or have been specified by the user,
and do not appear in the correlation matrix )

Background	Beta(2)

Background	1	-0.21

Beta(2)	-0.21	1

Parameter Estimates

95.0% Wald Confidence Interval
Variable	Estimate	Std. Err.	Lower Conf. Limit Upper Conf. Limit

Background	0.095736	*	*	*

Beta(1)	0	*	*	*

Beta(2)	8.14699	*	*	*

* - Indicates that this value is not calculated.

Analysis of Deviance Table

Model	Log(likelihood)	# Param's	Deviance Test d.f.	P-value

Full model	-43.4964	3

Fitted model	-44.1525	2	1.31215 1	0.252

Reduced model	-101.707	1	116.422 2	<.0001

AIC:	92.3049

Goodness of Fit

Dose

Est. Prob.

Expected

Observed

Si ze

Scaled
Residual

0.0000

0.0957

4 .787

6

50

0. 583

0.1160

0.1896

9. 481



50

-0.895

0.6530

0.9720

48.599

49

50

0. 344

Chi^2 = 1.26	d.f. = 1	P-value = 0.2617

Benchmark Dose Computation

Specified effect =	0.1

Risk Type =	Extra risk

Confidence level =	0.95

BMD =	0.113721

BMDL =	0. 07 7 58 73

BMDU =	0. 137047

Taken together, (0.0775873, 0.137047) is a 90	% two-sided confidence

interval for the BMD

15


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Female Rat

Dose metric: MRAMKL
Vmax = 0.4 mg/hour/kg BW0 07

Multistage Model with 0.95 Confidence Level

Multistage Model. (Version: 2.8; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\FEMALE RAT\MRAMKL-
VMAX=0.4\FRAT_FATTY_LIVER_MRAMKL-4.(d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\FEMALE
RAT \ MRAMKL - VMAX= 0 . 4 \ FRAT_FATTY_LIVER_MRAMKL- 4 .pit

Fri Oct 12 11:52:42 2007

BMDS MODEL RUN

Observation # < parameter # for Multistage model.

The form of the probability function is:

P[response] = background + (1-background)*[1-EXP(

-betal*dose/sl-beta2*doseL 2-beta3*dose/s3)

The parameter betas are restricted to be positive

Dependent variable = FattyLiver
Independent variable = umol/hr-kgL

Total number of observations = 3

Total number of records with missing values = 0

Total number of parameters in model = 4

Total number of specified parameters = 0

Degree of polynomial = 3

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
Background = 0.0769299

Beta(1)
Beta(2)
Beta(3)

0
0

0.00216647

16


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Asymptotic Correlation Matrix of Parameter Estimates

( *** The model parameter (s) -Beta(l) -Beta (2)

have been estimated at a boundary point, or have been specified by the user,
and do not appear in the correlation matrix )

Background
Beta(3)

Background
1

-0.21

Beta(3)
-0. 21
1

Parameter Estimates

Variable
Background
Beta(1)
Beta (2)
Beta(3)

Estimate
0.0958436
0
0

0. 00196673

Std. Err.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

Indicates that this value is not calculated.

Analysis of Deviance Table

Model
Full model

Log(likelihood)
-43.4964

Param's
3

Deviance Test d.f.

P-value

Fitted :
Reduced :

model
model



-44.

-101

1433 2 1.29386
.707 1 116.422

2

0.2553
r <.0001



AIC:



92 .

2866







Dose

Est

. Prob



Goodness of Fit
Expected Observed

Si ze



Scaled
Residual

0.0000
3.8130
12.0920

o o o

0958
1892
9721



4.792 6
9.462 7
48.603 49

L 50^

50
50



O CTi o
CD CD

lo co co
o o o

Chi ^2 = 1

.24

d.f.

= 1

P-value = 0.2650







Benchmark Dose Computation

Specified effect
Risk Type
Confidence level
BMD
BMDL
BMDU

0.1
Extra risk
0. 95
3.76974
2^82 4 88
4. 26949

Taken together, (2.82488, 4.26949) is a 90
interval for the BMD

two-sided confidence

17


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Female Rat

Dose metric: MRAMKL
Vmax = 0.65 mg/hour/kg BW0'07

1

0.8

"O
CD
O

0.6
| 0.4

Ll_

0.2

0

0 2 4 6 8 10 12 14 16 18

dose

11:57 10/12 2007

Multistage Model. (Version: *2 . 8; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\FEMALE RAT\MRAMKL-
VMAX= 0.65\FRAT_FATTY_LIVER_MRAMKL-65. (d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\RFC RAT LIVER\FEMALE
RAT \ MRAMKL - VMAX= 0 . 65 \ FRAT_FATTY_LIVER_MRAMKL-65 .pit

Fri Oct 12 11:57:06 2007

BMDS MODEL RUN

Observation # < parameter # for Multistage model.

The form of the probability function is:

P[response] = background + (1-background)*[1-EXP(

-betal*dose/sl-beta2*dose/N2-beta3*dose/N3) ]

The parameter betas are restricted to be positive

Dependent variable = FattyLiver
Independent variable = umol/hr-kgL

Total number of observations = 3

Total number of records with missing values = 0

Total number of parameters in model = 4

Total number of specified parameters = 0

Degree of polynomial = 3

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
Background =	0

Beta(1) =	0

Beta(2) =	0

Beta(3) = 0.000714264

Asymptotic Correlation Matrix of Parameter Estimates

Multistage Model with 0.95 Confidence Level

Multistage







T









""^1





I	

[ SMI)[



BMD	



18


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Background
Beta(3)

*** The model parameter(s) -Beta(l) -Beta(2)

have been estimated at a boundary point, or have been specified by the user,
and do not appear in the correlation matrix )

Background	Beta(3)

1	-0.19

-0.19	1

Parameter Estimates

Variable
Background
Beta(1)
Beta (2)
Beta(3)

Estimate
0.101433
0
0

0.000660435

Std. Err.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

- Indicates that this value is not calculated.

Model
Full model
Fitted model

Reduced model
AIC:

Analysis of Deviance Table

Param's Deviance Test d.f.

Log(likelihood
-43.4964
-43.7982

-101.707
91.5964

0.603632
116.422

P-value

0.4372
C. 0001

Goodness of Fit

Dose Est. Prob.

Expected Observed Size

Scaled
Residual

0.0000 0.1014
4.9910 0.1723
17.6260 0.9759

5.072 6 50
8.613 7 50
48.793 49 50

0. 435
-0.604
0.191

Chi/N2 = 0.59 d.f.

= 1 P-value = 0.4421



Benchmark Dose Computation



Specified effect =

^0.1



Risk Type =

Extra risk



Confidence level =

0. 95



BMD =

5. 42354



BMDL =

3.74923



BMDU =

6. 17189



Taken together, (3.74923
interval for the BMD

, 6.17189) is a 90 % two-sided

confidence

19


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

CANCER ASSESSMENT: BMD MODELING OUTPUTS FOR LOW-DOSE LINEAR

EXTRAPOLATION APPROACH

Benchmark Dose Analysis

Liver tumors (adenoma or

carcinoma)













Female F344 rats exposed to carbon tetrachloride vapor for 104 weeks (6 hours/day,



5 days/week)



















Exposure concentrations modeled: 0, 5, 25,125ppm











Multistage; MCA: 2-stage model

MRAMKL: 4-stage model









Vmax = 0.4

Vmax= 0.65

BMR









BMR/









BMR/

(extrarisk)

AIC

X2/?-value1'

BMC

BMCL

BMCL

AIC

y?p-value3

BMC

BMCL

BMCL











MCA



















(jimol/L)











0.05

61.6602

0.9842

0.609955

0.387377

0.129

61.5904

0.9916

0.588686

0.354766

0.141









MRAMKL



















(jimol/hr-kg liver)









0.05

63.3399

0.6503

9.8151

8.40334

0.00595

62.8343

0.7440

14.582

12.2867

0.00407

aValues <0.1 fail to meet conventional goodness-of-fit criteria; /J-value from the x2 test.

Liver tumors (adenoma or

carcinoma)













Female F344 rats exposed to carbon tetrachloride vapor for 104 weeks (6 hours/day,



5 days/week)



















Exposure concentrations modeled: 0, 5, 25ppm











Multistage; 2-stage model



















Vmax = 0.4

Vmax= 0.65

BMR









BMR/









BMR/

(extrarisk)

AIC

X2/?-value1'

BMC

BMCL

BMCL

AIC

y?p-value3

BMC

BMCL

BMCL











MCA



















(jimol/L)











0.05

24.8957

0.9507

0.655398

0.345984

0.144

24.8889

0.9523

0.604144

0.317726

0.157









MRAMKL



















(jimol/hr-kg liver)









0.05

25.2825

0.8571

11.5604

6.92352

0.00722

25.1734

0.8831

16.6986

9.76339

0.00512

aValues <0.1 fail to meet conventional goodness-of-fit criteria;/>-value from the y} test.
Note: 3-stage model did not proudc a sufficiently improved model fit.

20


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Liver tumors (adenoma or carcinoma)













Female BDF1 mouse exposed to carbon tetrachloride vapor for 104 weeks (6 hours/day,



5 days/week)



















Exposure concentrations modeled: 0, 5, 25ppm











Multistage; MCA: 2-stage model

MRAMKL: 2-stage model







BMR

Fisher

Thrall

(extra









BMR/









BMR/

risk)

AIC

y?p-value3

BMC

BMCL

BMCL

AIC

X2/?-value3

BMC

BMCL

BMCL









MCA



















(jimol/L)











0.1

117.307

NA

0.10186

0.0467576

2.14

117.307

NA

0.194624

0.0885305

1.13









MRAMKL



















(jimol/hr-kg liver)









0.1

115.912

0.4437

9.70893

6.3204

0.0158

117.341

0.1654

10.4557

7.59255

0.0132

aValues <0.1 fail to meet conventional goodness-of-fit criteria; /J-value from the %2 test.
Note: 3-stage model did not provide a sufficiently improved model fit.

Liver tumors (adenoma or

carcinoma)













Female BDF1 mouse exposed to carbon tetrachloride vapor for 104 weeks (6 hours/day,

5 days/week)



















Exposure concentrations modeled: 0, 5ppm











Multistage; 2-stage model



















Fisher

Thrall

BMR









BMR/









BMR/

(extrarisk)

AIC

y?p-value3

BMC

BMCL

BMCL

AIC

y?p-value3

BMC

BMCL

BMCL











MCA





















(jimol/L)









0.1

80.6149

NA

0.101967

0.044224

2.26

80.6149

NA

0.195666

0.0848621

1.18











MRAMKL

















(jimol/hr-kg liver)









0.1

80.6149

NA

11.6352

5.04631

0.0198

80.6149

NA

14.1982

6.15788

0.0162

aValues <0.1 fail to meet conventional goodness-of-fit criteria; /J-value from the x2 test.

21


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Liver tumors (adenoma or

carcinoma)













Male BDF1 mouse exposed to carbon tetrachloride vapor for 104 weeks (6 hours/day,



5 days/week)



















Exposure concentrations modeled: 0, 5, 25ppm











Note: models could not fit data with all 4 dose groups; highest dose group dropped



BMR = 0.1



















Multistage; 3-stage model



















Fisher

Thrall

BMR









BMR/









BMR/

(extrarisk)

AIC

y?p-value3

BMC

BMCL

BMCL

AIC

X2 p-value3

BMC

BMCL

BMCL











MCA





















(jimol/L)









0.1

151.192

0.3562

0.191106

0.063650

1.57

151.158

0.3660

0.388392

0.122027

0.819











MRAMKL

















(junol/hr-kg liver)









0.1

152.089

0.1864

13.3804

7.30705

0.0137

152.924

0.1086

14.185

8.82145

0.0113

aValues <0.1 fail to meet conventional goodness-of-fit criteria;/>-value from the x2 test.

22


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Pheochromocytomas

Female BDF1 mouse exposed to carbon tetrachloride vapor for 104 weeks (6 hours/day,
5 days/week)

Exposure concentrations modeled: 0, 5, 25,125ppm
Multistage; 2-stage model
BMR = 10%

BMR

(extrarisk)

Fisher

Thrall

AIC

X2 p-value3

BMC

BMCL

BMR/
BMCL

AIC

y?p-value3

BMC

BMCL

BMR/
BMCL

MCA
(jimol/L)

0.1

71.4077

0.7947

1.42662

1.13753

0.0879

71.3358

0.8039

2.94801

2.34113

0.0427

aValues <0.1 fail to meet conventional goodness-of-fit criteria; /J-value from the x2 test.
Note: 3-stage model did not provide a sufficiently improved model fit.

Pheochromocytomas

MaleBDFl mouse exposed to carbon tetrachloride vapor for 104 weeks (6 hours/day,
5 days/week)

Exposure concentrations modeled: 0, 5, 25,125ppm
Cancer Multistage
BMR = 10%

Cancer Multistage (restricted mode) model did not provide an adequate fit of the male
pheochromocytoma data (1, 2, and 3 stage models provided the same outputs); therefore other
models inBMDS were used (see table below).

BMR

(extrarisk)

Fisher

Thrall

AIC

X2 p-value3

BMC

BMCL

BMR/
BMCL

AIC

X2 p-value3

BMC

BMCL

BMR/
BMCL

MCA
(jimol/L)

1st. 2nd & 3rd
0.1

139.129

0.0513

0.292123

0.230102

0.435

139.077

0.0488

0.600117

0.472644

0.212

aValues <0.1 fail to meet conventional goodness-of-fit criteria; /J-value from the x2 test.

23


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Pheochromocytomas

MaleBDFl mouse exposed to carbon tetrachloride vapor for 104 weeks (6 hours/day,
5 days/week)

Exposure concentrations modeled: 0, 5, 25,125ppm
Models other than Multistage
BMR = 0.1

Model

Fisher

Thrall

AIC

X2 p-value3

BMC

BMCL

BMR/
BMCL

AIC

X2 p-value3

BMC

BMCL

BMR/
BMCL

MCA ((amol/L)

Gammab

139.129

0.0513

0.292124

0.230102

0.435

139.077

0.0488

0.600118

0.472644

0.212

Gamma ~
unrestricted

140.755

0.0401

0.238028

0.10463

0.956

140.587

0.0428

0.473653

0.204957

0.488

Logistic0

161.228

0.0000

0.929566

0.75614

0.132

161.353

0.0000

1.9184

1.56019

0.064

Logistic —
unrestricted

161.228

0.0000

0.929566

0.75614

0.132

161.353

0.0000

1.9184

1.56019

0.064

Log-logistic0

138.661

0.0978

0.24731

0.147398

0.678

138.467

0.1050

0.492945

0.297393

0.336

Log-logistic
unrestricted

138.661

0.0978

0.247311

0.130943

0.764

138.467

0.1050

0.492945

0.257935

0.388

Probit0

159.808

0.0000

0.851235

0.702221

0.142

159.949

0.0000

1.75643

1.44878

0.069

Pro bit -
unrestricted

159.808

0.0000

0.851235

0.702221

0.142

159.949

0.0000

1.75643

1.44878

0.069

Log-probit0

141.637

0.0044

0.423924

0.340228

0.294

141.988

0.0035

0.867906

0.696011

0.144

Log-probit —
unrestricted

137.136

0.1533

0.264859

0.150882

0.663

136.945

0.1648

0.527758

0.297349

0.336

Quantal-linear

139.129

0.0513

0.292124

0.230102

0.435

139.077

0.0488

0.60012

0.472644

0.212

W eibullb

139.129

0.0513

0.292124

0.230102

0.435

139.077

0.0488

0.60012

0.472644

0.212

Weibull --
unrestricted

140.513

0.0497

0.226525

0.10562

0.947

140.316

0.0535

0.45102

0.207636

0.482

aValues <0.1 fail to meet conventional goodness-of-fit criteria;/>-value from the x2 test.
''Power restricted to >1.
cSlope restricted to >1.

24


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Female F344 rat — hepatocellular adenomas or carcinomas (0, 5, 25,125 ppm dose groups)

Dose metric: MRAMKL
Vmax = 0.4 mg/hour/kg BW0 07

Multistage Cancer Model with 0.95 Confidence Level

1

0.8

1 0.6

% 0.4
5

LL

0.2

0

0	5	10	15	20	25

dose

10:00 10/162007

Multistage Cancer Model. (Version: 1.5; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS FEMALE RAT LIVER\MRAMKL-
VMAX=0.4\FRAT_LIVER_ADCAR_MRAMKL-4.(d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS FEMALE RAT
LIVER\ MRAMKL-VMAX=0 . 4 \ FRAT_LIVER_ADCAR_MRAMKL- 4 .pit

Tue Oct X16 10:00:27 2007

BMDS MODEL RUN

Observation # < parameter # for Multistage Cancer model.

The form of the probability function is:

P[response] = background + (1-background)*[1-EXP(

-betal*dose/sl-beta2*dose/N2-beta3*dose/s3-beta4*doseA4 ) ]

The parameter betas are restricted to be positive

Dependent variable = IncLiverTumor
Independent variable = umol/hr-kgL

Total number of observations = 4

Total number of records with missing values = 0
Total number of parameters in model = 5
Total number of specified parameters = 0
Degree of polynomial = 4

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

Background =	0

Beta(1) =	0

Beta(2) =	0

Beta(3) =	0

Beta(4) = 6.11699e-006
Asymptotic Correlation Matrix of Parameter Estimates

25


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

( *** The model parameter (s) -Background -Beta(l) -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 )

Beta(4)

Beta(4)	1

Parameter Estimates

Variable
Background
Beta(1)
Beta (2)
Beta(3)
Beta(4)

Estimate
0
0
0
0

5.52 68 9e-006

Std. Err.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

Indicates that this value is not calculated.

Model
Full model
Fitted model
Reduced model

AIC:

Analysis of Deviance Table

Deviance Test d.f.

Log(likelihood)
-2 9.6 94 6
-30.67
-109.05

63 .3399

Param's
4
1
1

1.95065
158.71

P-value

0.5827^
C. 0001

Dose

Est. Prob.

Goodness of Fit
Expected Observed	Size

Scaled
Residual

0. 0000
3. 8130
12.0920
24.3200

Chi^2

0.0000
0.0012
0.1114
0.8554

1. 64

d.f.

0. 000
0. 058
5. 572
42.768

0
0
3
44

P-value

0.6503

50
50
50
50

0. 000
-0.242
-1.156
0. 495

Benchmark Dose Computation

Specified effect :
Risk Type
Confidence level :
BMD =
BMDL =
BMDU =

0. 05
Extra risk
0. 95
9.8151
8. 40334
10 .5331

Taken together, (8.40334, 10.5331) is a 90
interval for the BMD

two-sided confidence

Multistage Cancer Slope Factor

0.00595002

26


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Dose metric: MRAMKL
Vmax = 0.65 mg/hour/kg BW0'07

Multistage Cancer Model with 0.95 Confidence Level

Multistage Cancer Model. (Version: 1.5; Date: 02/20/2007)

Input Data File: G:\CARBON TET\EMD\EMD MODELING 1 0-2 0 07 \ TUMORS FEMALE RAT LIVER\MRAMKL-
VMAX=0.65\FRAT_LIVER_ADCAR_MRAMKL-65.(d)

Gnuplot Plotting File: G:\CARBON TET\EMD\EMD MODELING 10-2 0 07 \ TUMORS FEMALE RAT
LIVER\MRAMKL-VMAX=0 . 65\ FRAT_LIVER_ADCAR_MRAMKL-65 . pit

Tue Oct 16 10:05:09 2007

BMDS MODEL RUN

Multistage Cancer
Linear extrapolation

Dependent variable = I
Independent variable = umo

Total number of observations = 4

Total number of records with missing values = 0
Total number of parameters in model = 5
Total number of specified parameters = 0
Degree of polynomial = 4

0	5

13:09 12/142007

Observation
The form o

nd)*[1-EXP(
-beta3*doseA3-beta4*doseA 4)

The parameter betas

positive

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
Background =	0

Beta(1) =	0

Beta(2) =	0

Beta(3) =	0

Beta(4) = 1.23526e-006

27


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Asymptotic Correlation Matrix of Parameter Estimates

( *** The model parameter (s) -Background -Beta(l) -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 )

Beta(4)

Beta(4)	1

Parameter Estimates

Variable
Background
Beta(1)
Beta (2)
Beta(3)
Beta (4)

Estimate
0
0
0
0

1.13446e-006

Std. Err.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

Indicates that this value is not calculated.

Model
Full model
Fitted model
Reduced model

AIC:

Analysis of Deviance Table

Deviance

Log(likelihood)
-2 9. 6 94 6
-30.4171
-109.05

62 .8343

Param's
4
1
1

Test d.f.

1. 44504	3X

158.71	3

P-value

0. 695
:. 0001





Goodness of Fit















Scaled

Dose

Est. Prob.

Expected

Observed

Size

Residual

0.0000

0.0000

^oAa

0

50

0. 000

4.9910

0.0007

0. 035

0

50

-0.188

17 . 6260

0.1037

5.186

3

50

-1.014

36 .2660

0.8595

42.97 4

44

50

0. 418

Chi ^2

1.24

d.f.

P-value

0. 7440

Benchmark Dose Computation

Specified effect	=

Risk Type	=

Confidence level	=

BMD	=

BMDL	=

BMDU	=

0. 05
Extra risk
0. 95
14 .582
12 .28 67
15.6526

Taken together, (12.2867, 15.6526) is a 90
interval for the BMD

two-sided confidence

Multistage Cancer Slope Factor

0.00406945

28


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Female F344 rat — hepatocellular adenomas or carcinomas (0, 5, 25 ppm dose groups)

Dose metric: MRAMKL
Vmax = 0.4 mg/hour/kg BW0 07

Multistage Cancer Model with 0.95 Confidence Level

"O
CD
O
.(D

<

ฃ=
O

~o

CT3

0	2	4	6	8	10	12

dose

08:23 10/12 2007

Multistage Cancer Model. (Version: 1.5; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS FEMALE RAT LIVER\MRAMKL-
VMAX=0.4\FRAT_LIVER_ADCAR_MRAMKL-4.(d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS FEMALE RAT
LIVER\ MRAMKL-VMAX=0 . 4 \ FRAT_LIVER_ADCAR_MRAMKL- 4 .pit

Fri Oct 12 08:23:17 2007

BMDS MODEL RUN

The form of the probability function isx:

P[response] = background + (1-background)*[1-EXP(
-betal\*dose/sl-beta2*dose/N2 ) ]

The parameter betas are restricted to be positive

Dependent variable = IncLiverTumor
Independent variable = umol/hr-kgL

Total number of observations = 3

Total number of records with missing values = 0

Total number of parameters in model = 3

Total number of specified parameters = 0

Degree of polynomial = 2

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

Background =	0

Beta(1) =	0

Beta(2) =	0.00044169

Asymptotic Correlation Matrix of Parameter Estimates

( *** The model parameter (s) -Background -Beta(l)

have been estimated at a boundary point, or have been specified by the user,

29


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

and do not appear in the correlation matrix
Beta(2)

Beta(2)	1

Parameter Estimates

Variable
Background
Beta(1)
Beta (2)

Estimate
0
0

0.000383811

95.0% Wald Confidence Interval
Std. Err.	Lower Conf. Limit Upper Conf. Limit

Indicates that this value is not calculated.

Model
Full model
Fitted model
Reduced model

AIC:

Analysis of Deviance Table

Deviance Test d.f.

Log(likelihood
-11.3484
-11.6412
-14.7059

25.2825

Param's
3
1
1

0. 585705
6. 71498

P-value

0. 7461
0.03482

Goodness of Fit

Dose Est. Prob.

Expected Observed

Si ze

Scaled
Residual

0.0000 0.0000

0.000 0

50

0^00

3.8130 0.0056

0.278 0

50

^0. 529

12.0920 0.0546

2.729 3

50

0.169

Chi/N2 = 0.31 d.f.

= 2 P-value = 0.8571





Benchmark Dose Computation





Specified effect =

o
Cn





Risk Type =

Extra risk





Confidence level =

0. 95





BMD =

11.5604





BMDI^^

6. 92 3 52





BMDU =

30 .5183





Taken together, (6.92352

, 30.5183) is a 90 % two-

-sided

confidence

interval for the BMD







Multistage Cancer Slope

Factor = 0.00722176





30


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Dose metric: MRAMKL
Vmax = 0.65 mg/hour/kg BW0'07

Multistage Cancer Model with 0.95 Confidence Level

Multistage Cancer
Linear extrapolation

T3
o

ฃ
<

o
CO

0.15

0.1

0.05

08:35 10/12 2007

Multistage Cancer Model. (Version: 1.5; Date: 02/20/2007)

Input Data File: G:\CARB0N TET\EMD\EMD MODELING 1 0-2 0 07 \ TUMORS FEMALE RAT LIVER\MRAMKL-
VMAX=0. 65\FRAT_LIVER_ADCAR_MRAMKL-65. (d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS FEMALE RAT
LIVER\ MRAMKL-VMAX=0 . 65 \ FRAT_LIVER_ADCAR_MRAMKL - 65 .pit

Fri Oct 12 08:35:44 2007

BMDS MODEL RUN

The form of the probability function is:

background + (1-background)*[1-EXP(
-betal*dose/sl-beta2*dose/s2 ) ]

P[response]

The parameter betas are restricted to be positive

Dependent variable = IncLiverTumor
Independent variable = umojL/hr-kgL

Total number of observations =

Total number of records with missing values = 0
Total number of parameters in model = 3
Total number of specified parameters = 0
Degree of polynomial = 2

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

Background =	0

Beta(1) =	0

Beta(2) =	0.000206402

Asymptotic Correlation Matrix of Parameter Estimates

( *** The model parameter (s) -Background -Beta(l)

have been estimated at a boundary point, or have been specified by the user,
and do not appear in the correlation matrix )

Beta(2)

31


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Beta(2)

Variable
Background
Beta(1)
Beta (2)

Parameter Estimates

Estimate
0
0

0. 000183949

Std. Err.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

Indicates that this value is not calculated.

Model
Full model
Fitted model
Reduced model

AIC:

Analysis of Deviance Table

Deviance Test d.f.

Log(likelihood)
-11.3484
-11.5867
-14.7059

25.1734

Param's
3
1
1

0.476667
6. 71498

P-value

0. 7879
0.03482

Goodness of Fit

Dose

Est. Prob.

Expected

Observed

Si ze

Scaled
Residual

0.0000

0.0000

0. 000

0



0. 000

4.9910

0.0046

0. 229

0

50

-0.479

17.6260

0.0555

2 .777

3

50

0.137

Chi ^2

0. 25

d.f.

P-value

0.8831

Benchmark Dose Computation

Specified effect
Risk Type
Confidence level
BMD
BMDL
BMDU

0. 05
Extra risk
0. 95
16 . 6986
9. 76339
43 .9237

Taken together, (9.76339, 43.9237) is a 90
interval for the BMD

two-sided confidence

Multistage Cancer Slope Factor

0.00512117

32


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Multistage Cancer Model. (Version: 1.5; Date: 02/20/2007)

Input Data File: G:\CARBON TET\EMD\EMD MODELING 10-2 0 07 \ TUMORS FEMALE MOUSE LIVER\MRAMKL-
FISHER\FMOUSE_LIVER_ADCAR_MRAMKL-FISHER.(d)

Gnuplot Plotting File: G:\CARBON TET\EMD\EMD MODELING 10-2 0 07 \ TUMORS FEMALE MOUSE
LIVER\MRAMKL-FISHER\FMOUSE_LIVER_ADCAR_MRAMKL-FISHER.pit

Fri Oct 12 08:54:44 2007

BMDS MODEL RUN

Female BDF1 mouse — hepatocellular adenomas or carcinomas (0, 5, 25 ppm dose groups)

Dose metric: MRAMKL
Fisher model

Multistage Cancer Model with 0.95 Confidence Level

The form of th^\probability function is:

P[response] = background + (1-background)*[1-EXP(
-betal*dose/sl-beta2*dose/N2 ) ]

The parameter betas are restricted to be positive

Dependent variable = IncLiverTumor
Independent variable = umol/hr-kgL

Total number of observations = 3

Total number of records with missing values = 0

Total number of parameters in model = 3

Total number of specified parameters = 0

Degree of polynomial = 2

							

M u Itistage Cancer —
Linear extrapolation —

0	5

12:04 10/15 2007

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 Background =
0.0482072

Beta(1) =	0

Beta(2) = 0.00119035

Asymptotic Correlation Matrix of Parameter Estimates

( *** The model parameter (s) -Beta(l)

have been estimated at a boundary point, or have been specified by the user,

33


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Background
Beta (2)

and do not	appear in the correlation matrix

Background	Beta(2)

1	-0.38

-0.38	1

Variable
Background
Beta(1)
Beta (2)

Parameter Estimates

Estimate
0. 0693295
0

0.00111772

Std. Err.

Indicates that this value is not calculated.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

Model
Full model
Fitted model
Reduced model

AIC:

Analysis of Deviance Table

Deviance Test d.f.

Log(likelihood)
-55.6537
-55.9559
-99.1295

115.912

Param's
3
2
1

P-value

0. 604318
86 . 9516

0.4369
C. 0001

Goodness of Fit

Dose

Est. Prob.

Expected

Observed

Si ze

Scaled
Residual

0.0000

0.0693

3. 466

4

50

0. 297

12.6660

0.2221

10.883

9

49

-0.647

41.6750

0.8664

43.321

44

50

0. 282

Chi ^2

0.59

d.f.

P-value

0.4437

Benchmark Dose Computation

Specified effect :
Risk Type
Confidence level :
BMD =
BMDL =
BMDU =

0.1

Extra risk
0. 95
9. 70893
6.3204
11^2 94 2

Taken together, (6.3204 , 11.2942) is a 90
interval for the BMD

two-sided confidence

Multistage Cancer Slope Factor

0.0158218

34


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Dose metric: MRAMKL
Thrall model

ง

0.8

0.6

tj 0.4

0.2

0

Multistage Model with 0.95 Confidence Level

1 Multistage

BMDL . BMP

0 5
12:10 10/152007

10 15 20 25 30 35 40
dose

45

Multistage Model. (Version: 2.8; Date: 02/20/2007)

Input Data File: G:\CARBON TET\EMD\EMD MODELING 10-2 0 07 \ TUMORS FEMALE
MOUSE LIVER\MRAMKL-THRALL\ FMOUSE_LIVER_ADCAR_MRAMKL-THRALL .(d)

Gnuplot Plotting File: G:\CARBON TET\EMD\EMD MODELING 10-2 0 07 \ TUMORS
FEMALE MOUSE LIVER\MRAMKL-THRALL\ FMOUSE_LIVER_ADCAR_MRAMKL-THRALL . pit

Fri Oct 12 09:01:03 2007

BMDS MODEL RUN

The form of the probability function is:

background + (1-background)*[1-EXP(
-betal*dose/sl-beta2*dose/s2 ) ]

P[response]

The parameter betas are restricted to be positive

Dependent variable = IncLiverTumor
Independent variable = umol/hr-kgL

Total number of observations =

Total number of records with missing values = 0
Total number of parameters in model = 3
Total number of specified parameters = 0
Degree of polynomial = 2

Maximum number of iterations = 250

Relative Function Convergence has been set to: le-00ฃ
Parameter Convergence has been set to: le-008

Default Initial Parameter Values
Background = 0.0162478
Beta(1) =	0

Beta(2) = 0.00110173

35


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Asymptotic Correlation Matrix of Parameter Estimates

( *** The model parameter (s) -Beta(l)

have been estimated at a boundary point, or have been specified by the

36


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

and do not	appear in the correlation matrix

Background	Beta(2)

Background 1	-0.4

Beta (2) -0.4	1

Limit

Variable

Background
Beta(1)
Beta (2)

Parameter Estimates

Estimate

0.0643165
0

0.000963757

Std. Err.

- Indicates that this value is not calculated.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf.

Model
Full model
Fitted model

Reduced model
AIC:

Analysis of Deviance Table

Deviance Test d.f.

Log(likelihood
-55.6537
-56.6705

-99.1295
117.341

Param's
3
2

1

2.03362
86 .9516

P-value

0.1539
C.0001

Goodness of Fit

Dose Est. Prob.

Expected Observed Size

Scaled
Residual

0.0000 0.0643
15.4560 0.2567
43.5990 0.8502

3.216 4 50
12.580 9 49
42.510 44 50

0. 452
-1.171
0. 590

Chi/N2 = 1. 92 d.f. :

= 1 P-value = 0.1654



Benchmark Dose Computation



Specified effect =

0.1



Risk Type =

Extra risk



Confidence level^=

0. 95



BMD =

10 .4557



BMDL =

7.59255



BMDU =

12.107



Taken together, (7.59255
interval for the BMD

, 12^107 ) is a 90 % two-sided

confidence

37


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Female BDF1 mouse — hepatocellular adenomas or carcinomas (0, 5 ppm dose groups)

Dose metric: MRAMKL
Fisher model

0.35

Multistage Cancer Model with 0.95 Confidence Level

Multistage Cancer
Linear extrapolation

12:49 10/15 2007

Multistage Cancer Model. (Version: 1.5; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS FEMALE MOUSE LIVER\MRAMKL-
FISHER\FMOUSE_LIVER_ADCAR_MRAMKL-KISHER.(d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS FEMALE MOUSE
LIVER\MRAMKL-FISHER\FMOUSE_LIVER_ADCAR_MRAMKL-FISHER.pit

Fri Oct 12 09:15:17 2007

BMDS MODEL RUN

Observation # < parameter # for Multistage Cancer model.
The form of the probability function is:

P[response]

background + (1-background)*[1-EXP(
-betal*dose/sl-beta2*dose/s2 ) ]

The parameter betas are restricted to be positive

Dependent variable = IncLiverTumor
Independent variable = umol/hr-kgL

Total number of observations = 2

Total number of records with missing values = 0

Total number of parameters in model = 3

Total number of specified parameters = 0

Degree of polynomial = 2

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
Background =	0.24898

Beta(1) = 0.0160225
Beta(2) =	0.001265

38


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Asymptotic Correlation Matrix	of Parameter Estimates

Background Beta(l)	Beta (2)

Background 1 -2.2e-008	8.3e-009

Beta (1) -6e-0 09 1	-1

Beta (2) -3.2e-009 -1	1

Parameter Estimates

95.0% Wald Confidence Interval
Variable	Estimate	Std. Err.	Lower Conf. Limit Upper Conf. Limit

Background	0.08	*	*	*

Beta (1)	0. 00471969	*	*	*

Beta (2)	0.000372627	*	*

* - Indicates that this value is not calculated.

Error in computing chi-square; returning 2

Analysis of Deviance Table

Model	Log(likelihood)	# Param's Deviance	Test d.f.	P-value

Full model	-37.3075	2

Fitted model	-37.3075	3 2.84217e-014	-1	NA

Reduced model	-38.4987	1 2.38238	1	0.1227

AIC:	80.6149

Goodness of Fit

Scaled

Dose	Est._Prob. Expected Observed	Size	Residual

0.0000	0.0800	4.000	4	50	-0.000

12.6660	0.1837	9.000	9	49	0.000

Chi^2 = 0.00	d.f. = -1	P-value =	NA

Benchmark Dose Computation
Specified effect =	0.1

Risk Type	=	Extra risk

Confidence level =	0.95

BMD =	11^3 52

BMDL =	5.04 6 31

BMDU did not converge for BMR = 0.100000
BMDU calculation failed

BMDU = 3.56605e+007

Multistage Cancer Slope Factor =	0.0198165

39


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Dose metric: MRAMKL
Thrall model

Multistage Model with 0.95 Confidence Level

Multistage Model. (Version: 2. 8; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS FEMALE MOUSE LIVER\MRAMKL-
THRALL\FMOUSE_LIVER_ADCAR_MRAMKL-THRALL.(d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS FEMALE MOUSE
LIVER\MRAMKL-THRALL\FMOUSE_LIVER_ADCAR_MRAMKL-THRALL.pit

Fri Oct 12 09:17:46 2007

BMDS MODEL RUN

Dependent variable = I
Independent variable = umo

Total number of observations = 2^B
Total number of records with missing values = 0
Total number of parameters in model = 3
Total number of specified parameters = 0
Degree of polynomial = 2

0.35

0.25

0.05

0	2

12:50 10/152007

Observation
The form o

P[response] =

The parameter betas

BMDL

positive

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 Background =

0.24898

Beta(1) = 0.0131302
Beta(2) = 0.000849523

Asymptotic Correlation Matrix of Parameter Estimates

Background	Beta(l)

Beta(2)

40


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Background
Beta(1)
Beta(2)

NA
NA

NA
NA

NA
NA

NA - This parameter's variance has been estimated as zero or less.
THE MODEL HAS PROBABLY NOT CONVERGED!!!

Parameter Estimates

Variable
Background
Beta(1)
Beta (2)

Estimate
0. 08
0.00386773
0.000250241

Std. Err.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

* - Indicates that this value is not calculated.

At least some variance estimates are negative.

THIS USUALLY MEANS THE MODEL HAS NOT CONVERGED!
Try again from another starting point.

Error in computing chi-sguare; returning 2

Analysis of Deviance Table

Model
Full model
Fitted model
Reduced model

AIC:

Log(likelihood)
-37.3075
-37.3075
-38.4987

80.6149

Param's Deviance	Test d.f.

2

3	2.84217e-014	-1
1 2.38238	1

P-value

Est. Prob.

Goodness of Fit
Expected Observed	Size

0.0000
15.4560

Chi ^2

0. 00

0.0800
0.1837

d.f.

4 . 000
9. 000

50
49

NA
0.1227

Scaled
Residual

-0.000
0. 000

-1

P-value

NA

Benchmark Dose Computation

Specified effect	=

Risk Type	=

Confidence level	=

BMD	=

BMDL	=

BMDU	=

0. 1
Extra risk

14 .1982
6.15788
2. 64 632e+ 014

Taken together, (6.15788, 2.64632e+014) is a 90
interval for the BMD

two-sided confidence

41


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Male BDF1 mouse — hepatocellular adenomas or carcinomas (0, 5, 25 ppm)

Dose metric: MRAMKL
Fisher model

Multistage Cancer Model with 0.95 Confidence Level

BMDS MODEL RUN

Multistage Cancer Model. (Version: 1.5; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS MALE MOUSE
FISHER\MMOUSE_LIVER_ADCAR_MRAMKL-FISHER.(d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS MALE
LIVER\MRAMKL-FISHER\MMOUSE_LIVER_ADCAR_MRAMKL-FISHER.pit

Tue Dec 04 12:03:25 2007

LIVER\MRAMKL-
MOUSE

Observation # < parameter # for Multistage Cancer model.
The form of the probability function is:

P[response] = background + (1-background)*[1-EXP(

-betal*dose/sl-beta2*dose/N2-beta3*dose/s3) ]

The parameter betas are restricted to be positive

Dependent variable = IncLiverTumor
Independent variable = umol/hr-kgL

Total number of observations = 3

Total number of records with missing values = 0

Total number of parameters in model = 4

Total number of specified parameters = 0

Degree of polynomial = 3

Multistage Cancer
Linear extrapolation

BiyPL

02
12:03

0	5

12/04 2007

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
Background =	0.352068

Beta(1) =	0

Beta(2) =	0

Beta(3) = 4.77425e-005

Asymptotic Correlation Matrix of Parameter Estimates

42


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Background
Beta(3)

*** The model parameter(s) -Beta(l) -Beta(2)

have been estimated at a boundary point, or have been specified by the user,
and do not appear in the correlation matrix )

Background	Beta(3)

1	-0.22

-0.22	1

Parameter Estimates

Variable
Background
Beta(1)
Beta (2)
Beta(3)

Estimate
0.41973
0
0

4 . 398 18e-005

Std. Err.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

Indicates that this value is not calculated.

Model
Full model
Fitted model

Reduced model
AIC:

Analysis of Deviance Table

Log(likelihood)
-73.1699
-74.0443

-9 9.6 09 6
152.089

Param's Deviance Test d.f.

1. 74874
52 .87 95

P-value

0.186
:. 0001

Goodness of Fit

Dose

Est. Prob.

Expected

Observed

Si ze

Scaled
Residual

0.0000

0.4197

20.987

24

50

0. 864

12.6660

0.4693

23.467

20

50

-0.982

41.6750

0.9760

48.798

49

50

0.187

Chi ^2

1.75

d.f.

P-value

0.18 64

Benchmark Dose Computation

Specified effect
Risk Type
Confidence level
BMD
BMDL
BMDU

0.1
Extra risk
0. 95
13.3804
7.30705
15.6428

Taken together, (7.30705, 15.6428) is a 90
interval for the BMD

two-sided confidence

Multistage Cancer Slope Factor

0.0136854

43


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Dose metric: MRAMKL
Thrall model

Multistage Cancer Model with 0.95 Confidence Level

0 5 10 15 20 25 30 35 40 45

dose

13:12 12/142007

Multistage Cancer Model. (Version: 1.5; Date: 02/20/2007)

Input Data File: G:\CARBON TET\EMD\EMD MODELING 1 0-2 0 07 \ TUMORS MALE MOUSE LIVER\MRAMKL-
THRALL\MMOUSE_LIVER_ADCAR_MRAMKL-THRALL.(d)

Gnuplot Plotting File: G:\CARBON TET\EMD\EMD MODELING 10-2 0 07 \ TUMORS MALE MOUSE
LIVER\MRAMKL-THRALL\MMOUSE_LIVER_ADCAR_MRAMKL-THRALL . P'lt

Tue Dec 04 12:19:57 2007

BMDS MODEL RUN

Observation # < parameter # for Multistage Cancer model.
The form of the probability function is:

P[response] = background + (1-background)*[1-EXP(

-betal*dose/sl-beta2*dose/N2-beta3*dose/N3) ]

The parameter betas are restricted to be positive

Dependent variable = IncLiverTumor
Independent variable = umol/hr-kgL

Total number of observations^a3

Total number of records with missing values = 0

Total number of parameters in model = 4

Total number of specified parameters = 0

Degree of polynomial = 3

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 Background =

0.317881

Beta(1) =	0

Beta(2) =	0

Beta(3) = 4.21166e-005

Asymptotic Correlation Matrix of Parameter Estimates

( *** The model parameter (s) -Beta(l) -Beta (2)

have been estimated at a boundary point, or have been specified by the user,

44


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

and do	not appear in the correlation matrix

Background	Beta(3)

Background 1	-0.26

Beta(3) -0.26	1

Variable
Background
Beta(1)
Beta (2)
Beta(3)

Parameter Estimates

Estimate
0.410703
0
0

3. 69143e-0 05

Std. Err.

Indicates that this value is not calculated.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

Model
Full model
Fitted model
Reduced model

AIC:

Analysis of Deviance Table

Deviance Test d.f.

Log(likelihood)
-73.1699
-74.462
-9 9.6 09 6

152.924

Param's
3
2
1

2.58426
52 .87 95

P-value

0.1079
C. 0001

Goodness of Fit

Dose

Est. Prob.

Expected

Observed

Si ze

Scaled
Residual

0.0000

0.4107

20.535

24

50

0.996

15.4560

0.4858

24 .289

20

50

-1.214

43.5990

0.9724

48.618

49

50

0. 330

Chi ^2

2 . 57

d.f.

P-value

0.1086

Benchmark Dose Computation

Specified effect
Risk Type
Confidence level
BMD
BMDL
BMDU

0.1
Extra risk
0. 95
14 .185
8. 82145
16 .5171

Taken together, (8.82145, 16.5171) is a 90
interval for the BMD

two-sided confidence

Multistage Cancer Slope Factor

0.011336

45


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

BDF1 mouse (female) — pheochromocytomas
Dose metric: MCA
Fisher model

Multistage Cancer Model with 0.95 Confidence Level

BMDL

Multistage Cancer Model. (Version: 1.5; Date: 02/20/2007)

Input Data File: G:\CARBON TET\EMD\EMD MODELING 10-2 0 07 \ TUMORS FEMALE
PHEOCHROMOCYTOMAS\FISHER\FMOUSE_PHEOCHROMOCYTOMA_MCA-FISHER.(d)

Gnuplot Plotting File: G:\CARBON TET\EMD\EMD MODELING 10-2 0 07 \ TUMORS FEMALE
PHEOCHROMOCYTOMAS\FISHER\FMOUSE_PHEOCHROMOCYTOMA_MCA-FISHER.pit

Fri Oct 12 09:49:11 2007

BMDS MODEL RUN

Multistage Cancer
Linear extrapolation

The form of the probability function is:

P[response] = background + (1-background)*[1-EXP(
-betal*dose/sl-beta2*dose'\2 ) ]

The parameter betas are restricted to be positive

Dependent variable = Pheochrom
Independent variable = umol/L

0	0.5

09:49 10/12 2007

Total number of observations = 4

Total number of records with missing values = 0
Total number of parameters in model = 3
Total number of specified parameters = 0
Degree of polynomial = 2

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
Background =	0

Beta(1) =	0

Beta(2) = 0.0548062

46


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Asymptotic Correlation Matrix of Parameter Estimates

( *** The model parameter (s) -Background -Beta(l)

have been estimated at a boundary point, or have been specified by the user,
and do not appear in the correlation matrix )

Beta(2)

Beta(2)	1

Parameter Estimates

Variable
Background
Beta(1)
Beta (2)

Estimate
0
0

0.0517683

Std. Err.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

Indicates that this value is not calculated.

Model
Full model
Fitted model
Reduced model

Analysis of Deviance Table

Log(likelihood)
-33.7087
-34.7039
-69.0688

Param's
4
1
1

Deviance Test d.f.

1.99041
70 .7202

P-value

0.5744
C. 0001

AIC:

71.4077

Dose

Est. Prob.

Goodness of Fit
Expected Observed

Scaled
Residual

0. 0000
0.1110
0.6030
3.3150

Chi^2

1. 03

0.0000
0.0006
0.0186
0.4338

d.f.

0. 000
0. 031
93^
21.259

0
0
0
22

50

49

50
49

0. 000
-0.177
-0.975
0. 214

P-value

0. 7947

Benchmark Dose Computation

Specified effect	=

Risk Type	=

Confidence level	=

BMD	=

BMDL	=

BMDU	=

0.1
Extra risk
M. 95
1. 42662
1. 13753
1. 72224

Taken together, (1.13753, 1.72224) is a 90	% two-sided confidence

interval for the BMD

Multistage Cancer Slope Factor =	0.08791

47


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Dose metric: MCA
Thrall model

Multistage Cancer Model with 0.95 Confidence Level

Multistage Cancer Model. (Version: 1.5; Date: 02/20/2007)

Input Data File: G:\CARBON TET\EMD\EMD MODELING 10-2 0 07 \ TUMORS FEMALE
PHEOCHROMOCYTOMAS\THRALL\FMOUSE_PHEOCHROMOCYTOMA-MCA-THRALL.(d)

Gnuplot Plotting File: G:\CARBON TET\EMD\EMD MODELING 10-2 0 07 \ TUMORS FEMALE
PHEOCHROMOCYTOMAS\THRALL\FMOUSE_PHEOCHROMOCYTOMA-MCA-THRALL.pit

Fri Oct 12 09:53:23 2007

BMDS MODEL RUN

T3

-S3

O

ฃ
<

0.4

The form of the probability function is:

P[response] = background + (1-background)*[1-EXP(
-betal\*dose/sl-beta2*dose/N2 ) ]

The parameter betas are restricted to be positive

Dependent variable = Pheochrom
Independent variable = umol/L

Total number of observations = 4

Total number of records with missing values = 0
Total number of parameters in model = 3
Total number of specified parameters = 0
Degree of polynomial = 2

Multistage Cancer 	

Linear extrapolation 	

BVCL

HVD

0	1

09:53 10/12 2007

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
Background =	0

Beta(1) =	0

Beta(2) = 0.0128084

Asymptotic Correlation Matrix of Parameter Estimates

48


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

( *** The model parameter (s) -Background -Beta(l)

have been estimated at a boundary point, or have been specified by the user,
and do not appear in the correlation matrix )

Beta(2)

Beta(2)	1

Parameter Estimates

Variable
Background
Beta(1)
Beta (2)

Estimate
0
0

0. 0121232

Std. Err.

Indicates that this value is not calculated.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

Model
Full model
Fitted model
Reduced model

AIC:

Analysis of Deviance Table

Log(likelihood)
-33.7087
-34. 6679
-69.0688

71.3358

Param's
4
1
1

Deviance Test d.f.

1.91847
70 .7202

P-value

0.5895
C. 0001

Dose

Goodness of Fit

Est. Prob.

Expected

Observed

Scaled
Residual

0.0000
0.2130
1.2260
6.8560

Chi^2

0. 99

0.0000
0.0005
0.0181
0.4344

d.f.

0. 000
0. 027
0. 903
21 hgQ5

0
0
0

22

P-value

0.8039

50

49

50
49

0. 000
-0.164
-0.959
0. 206

Benchmark Dose Computation

Specified effect	=

Risk Type	=

Confidence level	=

BMD	=

BMDL	=

BMDU	=

0.1
Extra risk
0. 95
2.94801
2.34113
3.55893

Taken together, (2.34113, 3.55893) is a 90
interval for the BMD

two-sided confidence

Multistage Cancer Slope Factor

0.0427144

49


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

BDF1 mouse (male) — pheochromocytomas

Dose metric: MCA
Fisher model

Probit Model with 0.95 Confidence Level

Probit Model. (Version: 2.8; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS MALE
PHEOCHROMOCYTOMAS\FISHER\MMOUSE_PHEOCHROMOCYTOMA_MCA-FISHER.(d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS MALE
PHEOCHROMOCYTOMAS\FISHER\MMOUSE_PHEOCHROMOCYTOMA_MCA-FISHER.pit

Fri Nov 30 12:55:04 2007

BMDS MODEL RUN

The form of the probability function is:

P[response] = Background

+ (1-Background) * CumNorm(Intercept+Slope*Log(Dose) ) ,

where CumNorm(.) is the cumulative normal distribution function

Dependent variable = Pheochrom
Independent variable = umol/L
Slope parameter is not restricted

Total number of observations = 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

Probit

0	0.5

12:55 11/30 2007

User has chosen the log transformed model

Default Initial	(and Specified) Parameter Values

background =	0

intercept =	-0.416734

slope =	0.792244

Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter (s) -background

50


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

intercept
slope

have been estimated at a boundary point, or have been specified by the user,
and do not appear in the correlation matrix )

intercept
1

-0 .092

slope
-0.092
1

Variable
background
intercept
slope

Estimate
0

-0.358995
0.694404

Parameter Estimates

Std. Err.

NA

0.125298
0.110458

NA - Indicates that this parameter has hit a bound
implied by some inequality constraint and thus
has no standard error.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

-0.604574
0.47791

-0.113416
0. 910899

Model
Full model
Fitted model
Reduced model

Analysis of Deviance Table

Log(likelihood)
-64.0144
-66.5682
-110.216

Param's
4

2 t

1

Deviance Test d.f.

5.10756	2

92.4032	3

137.136

Dose

Est. Prob.

Goodness of Fit
Expected Observed	Size

P-value

0.07779
<.0001

Scaled
Residual

0.0000
0.1110
0.6030
3.3150

Chi ^2

3.75

0.0000
0.0297
0.2388
0.6820

d.f.

0. 000
1.484
11.939
34.099

0
0
16
32

2

P-value

0.1533

50
50
50
50

0.	000
-1.237

1.	347
-0.637

Benchmark Dose Computation
Specified effect =	0.1

Risk Type	=	Extra risk

Confidence level =
BMD =
BMDL =

0. 95
0.264859
0.150882

51


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

Dose metric: MCA
Thrall model

Probit Model with 0.95 Confidence Level

Probit Model. (Version: 2.8; Date: 02/20/2007)

Input Data File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS MALE
PHEOCHROMOCYTOMAS\THRALL\MMOUSE_PHEOCHROMOCYTOMA_MCA-THRALL.(d)

Gnuplot Plotting File: G:\CARBON TET\BMD\BMD MODELING 10-2007\TUMORS MALE
PHEOCHROMOCYTOMAS\THRALL\MMOUSE_PHEOCHROMOCYTOMA_MCA-THRALL.pit

Fri Nov 30 13:15:12 2007

The form of the probability function is:

P[response] = Background

+ (1-Background) * CumNorm(Intercept+Slope*Log(Dose)),

where CumNorm(.) is the cumulative normal distribution function

Dependent variable = Pheochrom
Independent variable = umol/L
Slope parameter is not restricted

Total number of observations = 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

Probit

BMDL

BMD

0	1

13:15 11/30 2007

E'.MDS MODEL RUM

User has chosen the log transformed model

Default Initial	(and Specified) Parameter Values

background =	0

intercept =	-0.965049

slope =	0.776315

Asymptotic Correlation Matrix of Parameter Estimates

( *** The model parameter (s) -background

have been estimated at a boundary point, or have been specified by the user,
and do not appear in the correlation matrix )

52


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

intercept	slope

intercept	1	-0.58

slope	-0.58	1

Parameter Estimates

Variable
background
intercept
slope

Estimate
0

-0.844448
0. 683918

Std. Err.

NA

0.153761
0.109119

NA - Indicates that this parameter has hit a bound
implied by some inequality constraint and thus
has no standard error.

95.0% Wald Confidence Interval
Lower Conf. Limit Upper Conf. Limit

-1.14581
0. 470048

-0.543082
0.897787

Model
Full model
Fitted model
Reduced model

AIC:

Analysis of Deviance Table

Deviance Test d.f.

Log(likelihood)
-64.0144
-66.4723
-110.216

136.945

Param's
4
2
1

4.91585
92 .4 032

P-value

0.08561
k<. 0001





Goodness of Fit















Scaled

Dose

Est. Prob.

Expected

Observed

Size

Residual

0.0000

0.0000

0. 000

0

50

0. 000

0.2130

0.0286

1. 429

0

L 50

-1.213

1.2260

0.2404

12.019

16

50

1. 318

6.8560

0.6816

34 .ฃ18 0

32

50

-0.631

Chi ^2

3. 61

d.f.

P-value

0.1648

Benchmark Dose Computation

Specified effect =
Risk Type	=

Confidence level =
BMD =
BMDL =

0.1
Extra risk
0. 95
0.527758

0.29734 9

53


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

SOURCE CODE FOR PHYSIOLOGICALLY BASED
PHARMACOKINETIC MODELS

%	

%

% File: HUMINH.M

0/

/O

% Programmed by Gary Diamond
% Syracuse Research Corporation, 02/2005

0/

/O

%Tliis run time file implements CCL4.CSL for inhalation exposure
%(human parameters)

0/

/O	

%Prepare time history variables
prepare @clear @all

%Set communication interval
CINT=1.;

%Set simulation stop (lir)

TSTOP=17250.;

%hitegration error check
!!SET WESITG=.F.

%Air Exposure Parameters
AIRC=2.27; %ppm
AIRON=0.; %lir

AIROFF=l 000000.; %17520.; %hr
CIOFF=0.; %ppm
APERl=24.;%lir
AWID1=24.; %hr
APER2= 168.; %lir
AWID2= 168.; %lir

%Oral Exposure Parameters

RGIL=0.0;

FGIL=1.0;

%Human Parameters

BW=70.;

VLC=0.04;

WC=0.30; %Revisedfrom 0.1 (03/2007)

VSC=0.62;

VRC=0.05;

54


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

QCC=15.;

QPC=15.;

QSF=0.74;

QLC=0.25;

QFC=0.06;

QSC=0.18;

QRC=0.51;

PBLD=2.64;

PL=3.14;

PF=79.42;

PS=1.0;

PR=3.14;

%Chemical Parameters

MW=153.8;

VMAXC=1.49;

KMX =0.25;

VMAXSF=0.7;

Al=0.065;

A2=0.095;

A3=0.84;

Kl=0.25;

K2=0.03;

K3=0.025;

K4=0.;

K5=0.0004;

!! START /NC

%Output

HUMl=|_time day air ca ramkb mcl _cf_af, _cl];

HUM2=rot90(fliplr(HUMl));
out=fopen('HUMAN.out','w');

fprintf(out '%f,%f,%f,%e ,%e ,%e,%e ,%e,%e\ii",HUM2);
fclose(out);

55


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

0/

/o	

%

% File: HUMOR.M

0/

/O

% Programmed by Gary Diamond
% Syracuse Research Corporation, 10/2007

%

%Tliis run time file implements CCL4.CSL for "oral" exposure (RGIL)
%(human parameters)

0/

/o	

%Prepare time history variables
prepare @clear @all

%Set communication interval
CINT=24.;

%Set simulation stop (lir)

TSIDP=5000.;

%hitegration error check
!!SET WESITG=.F.

%Air Exposure Parameters
AIRC=0.; %ppm
AIRON=0.; %lir

AIROFF= 1000000. ;%17520.; %hr
CIOFF=0.; %ppm
APERl=24.;%lir
AWID1=24.; %hr
APER2=168.;%lir
AWID2=168.;%lir

%Oral Exposure Parameters

RGILC=10.0;

FGIL=1.0;

%Human Parameters

BW=70.;

VLC=0.04;

WC=0.30; %Revisedfrom0.1

VSC=0.62;

VRC=0.05;

QCC=15.;

QPC=15.;

QSF=0.74;

56


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

QLC=0.25;

QFC=0.06;

QSC=0.18;

QRC=0.51;

PBLD=2.64;

PL=3.14;

PF=79.42;

PS=1.0;

PR=3.14;

%Chemical Parameters

MW=153.8;

VMAXC=0.40;

KMX =0.25;

VMAXSF=0.7;

Al=0.065;

A2=0.095;

A3=0.84;

Kl=0.25;

K2=0.03;

K3=0.025;

K4=0.;

K5=0.0004;

!! START /NC

%Output

HUM 1=[_time _rgil mca mramkl];

HUM2=rot90(fliplr(HUMl));
out=fopen('HUMAN.out7w');
fprintf(out '%e ,%e ,%e ,%e\n",HUM2);
fclose(out);

57


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

0/

/o	

%

% File: MOUINH_JF.M

0/

/O

% Programmed by Gary Diamond
% Syracuse Research Corporation, 10/2007

0/

/O

%Tliis run time file implements CCL4.CSL for inhlation exposure
%(mouse parameters. Fisher et al. 2004)

0/

/o	

%Prepare time history variables
prepare @clear @all

%Set communication interval
CINT=24.;

%Set simulation stop (lir)

TSTOP=17520.;

%Integration error check
!!SET WESnXKF.

%Air Exposure Parameters:

AIRC=125.; %ppm
AIRON=0.; %lir
AIROFF=17520;%lir
CIOFF=0.; %ppm
APERl=24.;%lir
AWID1=6.; %lir
APER2= 168.; %lir
AWID2= 120.; %lir

%Mouse Parameters:

BW=0.036;

VLC=0.04;

VFC=0.04;

VSC=0.69;

VRC=0.14;

QCC=30.;

QPC=30.;

QSF=0.75;

QLC=0.24;

QFC=0.05;

QSC=0.17;

QRC=0.54;

58


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

PBLD=3.8;

PL=4.8;

PF=91.4;

PS=2.5;

PR=4.8;

%Chemical Parameters:

MW=153.8;

VMAXC=1.;

KMX = 0.3;

VMAXSF=0.75;

%From Hirall et al 2000

Al=0.065;

A2=0.095;

A3=0.84;

Kl=0.25;

K2=0.03;

K3=0.025;

K4=0.;

K5=0.00042;

!! START /NC
%Output

%MOU=[AIR MCA MRAMKB MCL MRAMKL]

MOU1=|_time day air mramkl mca mramkb mcl mramkl]; %output matrix

%formatingof output for printed comma-dilimited file:

MOU2=rot90(fliplr(MOU 1));
out=fopen('MOU.out7w');

%fprintf(out,'%f,%f,%f,%f,%f,%f,%f\n",MOU2);

%fprintf(out. "%f,%f,%f,%e ,%e ,%e,%e ,%e\n",MOU2);
fprintf(out '%e\n",MOU);
fclose(out);

59


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

0/

/o	

%

% File: MOUINHJCT.M

0/

/O

% Programmed by Gary Diamond
% Syracuse Research Corporation, 3/2007

0/

/O

%Tliis run time file implements CCL4.CSL for inhlation exposure
%(mouse parameters. Thrall et al. 2000)

0/

/o	

%Prepare time history variables
prepare @clear @all

%Set communication interval
CINT=24.;

%Set simulation stop (lir)

TSTOP=17520.;

%Integration error check
!!SET WESITG=.F.

%Air Exposure Parameters:

AIRC=2.5; %ppm
AIRON=0.; %lir
AIROFF=17520;%lir
CIOFF=0.; %ppm
APERl=24.;%lir
AWID1=6.; %lir
APER2= 168.; %lir
AWID2= 120.; %hr

%Mouse Parameters:

BW=0.036;

VLC=0.04;

VFC=0.04;

VSC=0.78;

VRC=0.05;

QCC=28.;

QPC=28.;

QSF=0.74;

QLC=0.24;

QFC=0.05;

QSC=0.19;

QRC=0.52;

60


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

PBLD=7.83;

PL=2.08;

PF=23.0;

PS=0.61;

PR=2.08;

%Chemical Parameters:

MW=153.8;

VMAXC=0.79;

KMX =0.46;

VMAXSF=0.7;

Al=0.065;

A2=0.095;

A3=0.84;

Kl=0.25;

K2=0.03;

K3=0.025;

K4=0.;

K5=0.00042;

!! START /NC

%Output

%MOU=[AIR MCA MRAMKB MCL MRAMKL]
MOUl=[_time dayairmramklmcamcl]; %output matrix

%formatingof output for printed comma-dilimited file:

MOU2=rot90(fliplr(MOU 1));

out=fopen('MOU.out7w');

fprintf(out "%f,%f,%e ,%e ,%e ,%e\n",MOU2);

fprintf(out "%e\n",MOU);

fclose(out);

61


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

0/

/o	

%

% File: RATTNH.M

0/

/O

% Programmed by Gary Diamond
% Syracuse Research Corporation, 02/2005

0/

/O

%Tliis run time file implements CCL4.CSL for inhalation exposure
%(rat parameters)

0/

/o	

%Prepare time history variables
prepare @clear @all

%Set communication interval
CINT=1.;

%Set simulation stop (lir)

TSTOP=17250.;

%hitegration error check
!!SET WESITG=.F.

%Air Exposure Parameters:

AIRC=4.; %ppm
AIRON=0.; %lir
AIROFF=17520;%lir
CIOFF=0.; %ppm
APERl=24.;%lir
AWID1=6.; %6.; %hr
APER2=168.;%lir
AWID2= 120.; %120.; %hr

%Rat Parameters:

BW=0.452;

VLC=0.04;

WC=0.08;

VSC=0.74;

VRC=0.05;

QCC=15.;

QPC=15.;

QSF=0.74;

QLC=0.25;

QFC=0.04;

QSC=0.20;

QRC=0.51;

62


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

PBLD=4.52;

PL=3.14;

PF=79.42;

PS=1.0;

PR=3.14;

%Chemical Parameters:

MW=153.8;

VMAXC=0.4;

KMX =0.25;

VMAXSF=0.7;

Al=0.065;

A2=0.095;

A3=0.84;

Kl=0.25;

K2=0.03;

K3=0.025;

K4=0.;

K5=0.00042;

!! START /NC
%Output

RATI=|_timc day air mca mramkl mcl]; %outputmatrix

%formating of output for printed comma-dilimited file:
%RAT2=rot90(fliplr(RATl));

%out=fopen('RAT.out7wl);

%fprintf(out, '%f,%f,%f,%e ,%e ,%e\n",RAT2);

%fclose(out);

63


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

PROGRAM: CCL4R

IThis program simulates the pharmacokinetics of carbon tetrachloride

!The program is based onlTRICCTAACSL, developed by KD THRALL 9/98; ACSL code provided to
GDiamond, 04/2004

!The above code was translated with minor modifications, by GDiamond, 05/2004

INITIAL

VARIABLE TIME = 0.0 ! Set independent variable to be TIME
ALGORITHM IALG = 2 [Numerical integration algorithm - Gear for stiff systems
CINTERVAL CINT=100. [Communication interval
NSTP = 1000 ! Set initital integration cycle length at CINT/1000
MERROR AL=0.0001 !Set error tolerance for Gear

I*****BODyAND TISSUE MASSES*****

CONSTANT BW = 0.2 IBody weiglit (kg)

CONSTANT VLC =0.04 ! Li\cr fraction of body weight
CONSTANT VFC = 0.08 ! Adipose fraction of body weiglit
CONSTANT VSC =0.74 ! Slowly-perfused fraction of body weiglit
CONSTANT VRC = 0.05 IRapdily-perfused fraction of body weight

VL=VLC*BW ILivcr (kg
W=WC*BW !Adipose (kg)

VS=VSC*BW !Slowly-perfused (kg)

VR=VRC*BW IRapidly-perfused (kg)

l*****BLOOD FLOWS*****

CONSTANT QCC=14 ICardiac output (L/lir-BWASF)

CONSTANT QPC=14 ! Alveolar ventilation (L/lir-BWASF)

CONSTANT QLC = 0.25 ILiver fraction of cardiac output
CONSTANT QFC = 0.09 ! Adipose fraction of cardiac output
CONSTANT QSC = 0.15 ! Slowly-perfused fraction of cardiac output
CONSTANT QRC = 0.51 IRapidly-perfused fraction of cardiac output

CONSTANT QSF = 0.74 !QC and QP scaling factor (SF)

QC=QCC*BW**QSF!Cardiac output (L/lir)

QP=QPC*BW**QSF! Alveolar ventilation (L/lir)

QL = QLC*QC ILiver (L/lir)

QF = QFC*QC lAdipose (L/lir)

QS = QSC*QC ! Slowly-perfused (L/lir)

QR= QRC*QC IRapidly-perfused (L/lir)

!***** PARTITION COEFFICIENTS* * * * *

CONSTANT PBLD = 4.52 !Blood:air partition coefficient

64


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

CONSTANT PL = 3.14 !Liver:bloodpartitioncoefficient
CONSTANT PF = 79.42 !Adipose:bloodpartitioncoefficient

CONSTANT PS = 1.0 !Slowly-perfused:blood partition coefficient
CONSTANT PR = 3.14 !Rapidly-perfused:bloodpartitioncoefficient

!***** METABOLISM and EXCRETION* * * * *

CONSTANT MW=153.8 IMolecular weight of CC14

CONSTANT VMAXC = 0.40 !VMAX for metabolism in liver (mg/lir-BWASF)

CONSTANT KMX= 0.25 !KM for metabolism in liver (mg/L)

CONSTANT VMAXSF=0.7 ! Scaling factor for VMAXC (SF)

CONSTANT Al=0.085 IFraction of metabolism rate to Ml pool
CONSTANT A2=0.095 IFraction of metabolism rate to M2 pool
CONSTANT A3=0.84 IFraction of metabolism rate to M3 pool

CONSTANT Kl=0.123 IRate constantforconversionofMl to exhaled metabolite (C02) (lir-1)
CONSTANT K2=0.03 IRate constant for conversion ofM2 to urinary metabolite (lir-1)
CONSTANT K3=0.0252 IRate constant for conversion of M3 to fecal metabolite (hr-1)
CONSTANT K4=0. IRate constantforconversionofM2toMl (lir-1)

CONSTANT K5=0.00042 IRate constantfor conversion of M3 to Ml (hr-1)

VMAX = 1000* VMAXC* BW**VMAXSF/MWI Maximum rate of metabolism in liver (umol/hr)
KM = 1000*KMX/MW IMichaelis constant for metabolism in liver (umol/L)

|*****EXPOSURE-AIR*****

CONSTANT AIRC = 1.1 Air exposure concentration (ppm)

AIR = AIRC; I Air exposure concentration (ppm)

AIRCM = AIR/24.45 I Air exposure (umol/L)

mgAIR = AIR*MW/24.45 I Air exposure (mg/m3)
ugAIR=AIR*MW/24.45 I Air exposure concentration (ug/L)

CONSTANT TSTOP = 700. ILengthofsimulaion(lir)

CONSTANT AIRON=0. ITime air exposure starts (lir)

CONSTANT AIROFF=700. ITime air exposure stops (lir)

CONSTANT CIOFF=0. IConcentrationininlialedairwhen exposure is off (ppm)

CONSTANT APER1=24. IPulse period 1 for air exposure (e.g., hours in a day)

CONSTANT AWID1=24. IPulse width 1 for air exposure (e.g., 6 hours each day)

CONSTANT APER2=168. IPulse period 2 for air exposure (e.g., hours in a day)

CONSTANT AWID2=168. IPulse width 2 for air exposure (e.g., 6 hours each day)
!*****EXPOSURE-ORAL ******GD08/2007

CONSTANT RGILC=0. IRate of uptake from GI to liver (umol/hr)

RGIL=RGILC IRate of uptake from GI to liver (umol/hr)

lUse for simuulating constant rate of uptake from Gl-tract
MGRGIL=RGIL*MW/1000 IRate of uptake from GI to liver (mg/lir)
MGRGILKGD=RGIL*24*MW/(1000*BW)IRate ofuptake to liver (mg/kg-day)

CONSTANT GILF=1. I Absorption fraction
ICONSTANT POINTS = 96.

ICINT = TSTOP/POINTS ISets communication for 96 times in the simulation

65


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

END !of INITIAL section of program

DYNAMIC

DERIVATIVE

DAY=TTME/24
YEAR=DAY/365

I * * * * * CONCENTRATION IN INHALED AIR (umol/L)*****

CION = AIRCM*PULSE( AIRON, APER1, AWID1) * PULSE( AIRON, APER2, AWID2)

CI = RSW(TIME.LE.AIROFF,CION,CIOFF)

RAI = QP*(CA/PBLD-CI)!Rate inhaled (uinol/lir)

CP =CI*24.45 IConcentration in chamber (ppm)

l*****AMOUNTTAKEN IN BY ONE ANIMAL (umol)*****

RIN = QP*CI

AIN = INTEG(RIN,0.0)

I***** LIVER* * * * *

!Use for simulation of constant rate of uptake from Gl-tract (GD 08/2007)
RAL = QL*(CA-CVL)+RGIL-RAM !Rate of change in amount (umol/hr)

!Use for simulation of inhalation exposure

!RAL = QL*(CA-CVL)-RAM !Rate of change in amount (umol/lir)

AL = WIEG(RAL, 0.0)!Amount (umol)

CVL = CL/PL IConcentration in venous blood (umol/L)

CL = AL/VL IConcentration (umol/L)

AUCCL = INIEG(CL,0.) IAUC concentration (umol/L xlir)

I Average concentration in li\er (umol/L) -MCL(GD 05/2004)

IF (TIME .GT. 0.) THEN

MCL = AUCCL/TTME

ELSE

MCL= 0.

END IF

ugCL = CL*MW/1000 IConcentration (ug/g)

MugCL = MCL*MW/1000 I Average concentration (ug/g)

AUCugCL = AUCCL*MW/1000 IAUC concentration (ug/g xlir)

I ***** METABOLIZED* *** *

RAM = (VMAX*CVL)/(KM+CVL) IRate, total (umol/lir)

RAMKB= RAM/BW IRate, total (umol/hr xkq body)

RAMKL=RAM/VL IRate. total (umol/hr x kg liver)

AUCRAM=INIEG(RAM,0.0) IAUC rate (umol/hr xlir)

66


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

! Ave rage rate of metabolism (umol/hr) - MRAM (GD 05/2004)

IF (TIME .GT. 0.) THEN
MRAM = AUCRAM/UME
ELSE

MRAM= 0.

END IF

MRAMKB=MRAM/BW ! Average rate, total (umol/hr x kg body) (GD 05/2004)
MRAMKL=MRAM/VL ! Ave rage rate, total (umol/hr x kg liver) (GD 05/2004

RAXA = RAM*A1-K1*AXA+K4*AXU+K5*AXF !Rate to air pool (umol/hr)

RAXF = RAM*A3-K3* AXF-K5* AXF !Rate to feces pool (umol/hr)

RAXU = RAM*A2-K2*AXU-K4* AXU !Rate to urine pool(umol/hr)

AM = INTEG(RAM,0.) lAmount total (umol)

AMK = AM/BW ! Amount total (umol/kgbw)

AMKL=AM/VL !Amount total (umol/kg liver)

AXA = WIEG(RAXA0.) ! Amount in air pool (umol)

AXF = INTEG(RAXF,0.) ! Amount in feces pool (umol)

AXU = WIEG(RAXU,0.) ! Amount in urine pool (umol)

RA = AXA*kl !Rate to air (umol/hr)

RU = AXU*k2 !Rate to urine (umol/hr)

RF = AXF*k3 !Rate to feces (umol/hr)
ugRA = RA*MW !Rate to air (ug/lir)
ugRU = RU*MW !Rate to urine (ug/lir)
ugRF = RF*MW !Rate to feces (ug/lir)

CAX = RA/((3/2)*QP) IConcentration of metabolite inexlialedair (umol/L)
CAXM = CAX*24.45 IConcentration of metabolite in exhaled air (ppm)

(***** pAT* * * * *

RAF = QF*(CA-CVF) IRate of change in amount (umol/hr)

AF = WIEG(RAF, 0.0) lAmount (umol)

CVF = CF/PF IConcentration in venous blood (umol/L)

CF=AF/VF IConcentration (umol/L)

AUCCF =WIEG(CF, 0.) IAUC concentration (umol/L xlir)

lAverage concentration in fat (umol/L) - MCF (GD 03/2007)

IF (TIME .GT. 0.) THEN

MCF = AUCCF/UME

ELSE

MCF=0.

END IF

ugCF = CF*MW/1000 !Concentration(ug/g)

MugCF =MCF*MW/1000 I Average concentration (ug/g)

AUCugCF = AUCCF*MW/1000 IAUC concentration (ug/g x lir)

67


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

!***** SLOWLY PERFUSED TISSUES* * * * *

RAS = QS*(CA-CVS) !Rate of change in amount (uinol/lir)

AS = INTEG(RAS, 0.0)! Amount (uinol)

CVS = CS/PS IConcentration in venous blood (uinol/L)

CS = AS/VS IConcentration in uinol/L
ugCS = CS*MW/1000 IConcentration inug/g

l*****RICHLYPERFUSED TISSUES*****

RAR = QR*(CA-CVR) IRate of change in amount (umol/hr)

AR = WIEG(RAR0.0) !Amount (umol)

CVR = CR/PR IConcentration in venous blood (umol/L)

CR= AR/VR IConcentration in umol/L

ugCR = CR*MW/1000 !Concentrationinug/g

|*****MIXED VENOUS BLOOD*****

CV = (QF*CVF+QL*CVL+QS*CVS+QR*CVR)/QC IConcentration (umol/L)
AUCCV=INTEG(CVO.) IAUC concentration (umol/L xlir)

lAverage concentration in venous blood (umol/L) - MCV (GD 05/2004)

IF (TIME .GT. 0.) THEN

MCV = AUCCV/TTME

ELSE

MCV=0.

END IF

ugCV = CV*MW/1000 IConcentration (ug/g)

MugCV =MCV*MW/1000 lAverage concentration (ug/g)
AUCugCV = AUCCV*MW/1000 IAUC concentration (ug/g xlir)

!*****ARTERIAL BLOOD*****

CA= (QC*CV+QP*CI)/(QC+(QP/PBLD)) IConcentration(umol/L)
AUCCA=INTEG(CA0.) IAUC concentration (umol/L xlir)

I Average concentration in atrerial blood (umol/L) - MCA (GD 05/2004)

IF (TIME .GT. 0.) THEN

MCA = AUCCA/TTME

ELSE

MCA=0.

END IF

ugCA=CA*MW/1000 IConcentration (ug/g)

MugCA=MCA*MW/1000 lAverage concentration (ug/g)
AUCugCA=AUCCA*MW/1000 IAUC concentration (ug/g xlir)

68


-------
PEER REVIEW DRAFT-DO NOT CITE OR QUOTE

!***** AMOUNT EXHALED*****

CX=CA/PBLD IConcentration in alveolar air (umol/L)

CXPPM= (0.7*CX+0.3*CI)*24.45 IConcentration in exhaled air (ppm)

ITotal ventilation is 0.7 of alveolar ventilation
RAX= QP*CX !Rae of change in amount (umol/hr)

AX=INTEG(RAX,0.) lAmount (uinol)

!**** *NET AMOUNT ABOSORBED* * * * *

DOSEX = AIN-AX !Net amount absorbed(umol)

BODY=AL+AF+AS+AR ! Amount in body (umol)

MASSB = BODY+AM+AX !Mass balance (umol)

END !of DERIVATIVE section of program

TERMT(HME .GE. TSTOP) [Termination condition

END !of DYNAMIC section of program

TERMINAL

END !of TEMINAL section of program
END !of program

69


-------