EPA/635/R -15/ 034b Public Comment Draft 

### www.epa.gov/iris



## Toxicological Review of **Hexahydro**

**- 1,3,5** 

**- trinitro** 

**- 1,3,5** 

**- triazine (RDX)** 
( CASRN 121

- 82 -4)
**In Support of Summary Information on the**  **Integrated Risk Information System (IRIS)**
**Supplemental Information**

March 2016

**NOTICE**

National Center for Environmental Assessment Office of Research and Development U.S. Environmental Protection Agency Washington, DC This document is a **Public Comment D**  **raft.** This information is distributed solely for the purpose of pre

- dissemination peer review under applicable information quality guidelines. It has not been formally disseminated by EPA. It does not represent and should not be construed to represent any Agency determination or policy. It is being circulated for review of its technical accuracy and science poli cy implications.

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**DISCLAIMER**
This document is a preliminary draft for review purposes only. This information is distributed solely for the purpose of pre

- dissemination peer review under applicable information quality guidelines. It has not been formally disseminated by EPA. It does not represent and should not be construed to represent any Agency determination or policy. Mention of trade names or commercial products does not constitute endorsement of recommendation for use.

This document is a draft for review purposes only and does not constitute Agency policy. 

### ii DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine



# CONTENTS

APPENDIX A. ASSESSMENTS BY OTHER NATIONAL AND INTERNATIONAL HEALTH AGENCIES

. A

1
APPENDIX B. ADDITIONAL DETAILS OF LITERATURE SEARCH STRATEGY | STUDY SELECTION AND EVALUATION

................................ ................................ ............................... B

- 1
B.1. DEFENSE TECHNICAL INFORMATION CENTER (DTIC) LITERATURE SEARCH AND SCREEN

................................ ................................ .............................. B

- 1
APPENDIX C. INFORMATION IN SUPPORT OF HAZARD IDENTIFICATION AND DOSE

- RESPONSE ANALYSIS

................................ ................................ ................................ ............. C

- 1
C.1. TOXICOKINETICS

................................ ................................ ........................ C

- 1
C.1.1. Absorption

................................ ................................ .................... C

- 1
C.1.2. Distribution

................................ ................................ .................. C

- 6
C.1.3. M etabolism

................................ ................................ .................. C

- 7
C.1.4. Excretion

................................ ................................ ...................... C

- 9
C.1.5. Physiologically Based Pharmacokinetic (PBPK) Models

............. C

- 11
C.2. HUMAN STUDIES

................................ ................................ ..................... C

- 31
C.3. OTHER PERTINENT TOXICITY INFORMATION

................................ .......... C

- 37
C.3.1. Mortality in Animals

................................ ................................ ... C

- 37
C.3.2. Other Noncancer Effects

................................ ............................ C

- 42
C.3.3. Genotoxicity

................................ ................................ ............... C

- 62
APPENDIX D. DOSE

- RESPONSE MODELING FOR THE DERIVATION OF REFERENCE VALUES FOR EFFECTS OTHER THAN CANCER AND THE DERIVATION OF CANCER RISK ESTIMATES

................................ ................................ ................................ ......... D

- 1
D.1. BENCHMARK DOSE MODELING SUMMARY FOR NONCANCER ENDPOINTS

................................ ................................ ............................... D

- 1
D.1.1. Evaluation of Model Fit and Model Selection

............................. D

- 3
D.1.2. Modeling Re sults

................................ ................................ ......... D

- 4
D.1.3. Mortality: Dose

- Response Analysis and BMD Modeling Documentation

................................ ................................ ......... D

- 36
D.2. BENCHMARK DOSE MODELING SUMMARY FOR CANCER ENDPOINTS

. D

- 46
D.2.1. Evaluation of Model Fit and Model Selection for Mouse Tumor Data

................................ ................................ ........................... D

- 46
D.2.2. Modeling Results for Mouse Tumor Data

................................ . D

- 47

This document is a draft for review purposes only and does not constitute Agency policy. 

### iii DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

D.2.3. Dose

- response Analysis and BMD Modeling Documentation for Other Tumor Data Sets

................................ ............................. D

- 61
REFERENCES FOR APPENDICES

................................ ................................ ................................ ..... R

- 1

This document is a draft for review purposes only and does not constitute Agency policy. 

### iv DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine


# TABLES

Table A

- 1. Assessments by other national and international health agencies

.......................... A

- 1 Table B

- 1. Summary of detailed search strategies for RDX (Pubmed, Toxline, Toxcenter, TSCATS)

................................ ................................ ................................ ................................ ........... B

- 3 Table B

- 2. Summary of detailed search strategies for RDX (DTIC)

................................ ............. B

- 10 Table B

- 3. Processes used to augment the search of core databases for RDX

........................ B

- 11 Table C

- 1. Distribution of RDX or radiola bel from administered RDX a



### ................................ ........ C

- 7 Table C

- 2. Principal urinary metabolites of RDX in miniature swine 24 hours after dosing with RDX

................................ ................................ ................................ ................................ ........... C

- 9 Table C

- 3. Elimination t 1/2 

### values for RDX or radiolabeled RDX

................................ ................... C

- 10 Table C

- 4. Parameter values used in the Sweeney et al. (2012a) and Sweeney et al. (2012b) PBPK models for RDX in rats, humans, and mice

................................ ......................... C

- 12 Table C

- 5. Parameters values used in the EPA application of the rat, human, and mouse models

................................ ................................ ................................ ................................ ............ C

- 15 Table C

- 6. Doses, dosing formulations, and absorption rate constants in animal and human studies

................................ ................................ ................................ ................................ ............ C

- 20 Table C

- 7. Sensitivity coefficients for rat and human RDX PBPK models

................................ . C

- 26 Table C

- 8. Summary of case reports of exposure to RDX

................................ ................................ . C

- 31 Table C

- 9. Occupational e pidemiologic studies of RDX: summary of methodologic features C

35 Table C

- 10. Evidence pertaining to mortality in animals a



### ................................ ............................... C

- 38 Table C

- 11. Evidence pertaining to other noncancer effects (hematological) in humans

. C

- 46 Table C

- 12. Evidence pertaining to other noncancer effects in animals a



### ................................ .. C

- 48 Table C

- 13. Summary of in vitro studies of the genotoxicity of RDX

................................ .......... C

- 63 Table C

- 14. S ummary of in vivo studies of the genotoxicity of RDX

................................ ........... C

- 67 Table C

- 15. Summary of in vitro and in vivo studies of the genotoxicity of RDX metabolites

................................ ................................ ................................ ................................ ... C

- 68 Table D

- 1. Noncancer endpoints selected for dose

- response modeling for RDX

.................... D

- 1 Table D

- 2. Convulsion or mortality endpoints from Crouse et al. (2006) selected for dose

response modeling for RDX

................................ ................................ ................................ ..... D

- 2 Table D

- 3. Model predictions for convulsions in female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006); BMR = 10% ER

................................ .......................... D

- 4 Table D

- 4. Model predictions for convulsions in female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006); BMR= 5% ER

................................ .............................. D

- 7 Table D

- 5. Model predictions for convulsions in female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006); BMR= 1% ER

................................ .............................. D

- 9 Table D

- 6. Model predictions for convulsions in male F344 rats exposed to RDX by gavage for 90 days (Crouse et al ., 2006); BMR = 10% ER

................................ ....................... D

- 11 Table D

- 7. Model predictions for convulsions in male F344 rats exposed to RDX by gavage for 90 days ( Crouse et al., 2006); BMR = 5% ER

................................ .......................... D

- 13 Table D

- 8. Model predictions for convulsions in male F344 rats exposed to RDX by gavage f or 90 days (Crouse et al., 2006); BMR = 1% ER

................................ .......................... D

- 15 Table D

- 9. Model predictions for convulsions in male and female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006); BMR = 10% ER

................................ .. D

- 17 Table D

- 10. Model predictions for convulsions in ma le and female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006); BMR = 5% ER

.......................... D

- 19 Table D

- 11. Model predictio ns for convulsions in male and female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006); BMR = 1% ER

.......................... D

- 21

This document is a draft for review purposes only and does not constitute Agency policy.



### v DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Tabl e D

- 12. Model predictions for convulsions in female F344 rats exposed to RDX by gavage on gestational days (GDs) 6−19 (Cholakis et al., 1980); BMR = 10% ER

. D

23 Table D

- 13. Model predictions for convulsions in female F344 rats exposed to RDX by gavage on GDs 6−19 (Cholakis et al., 1980); BMR = 5% ER

................................ .... D

- 25 Table D

- 14. Model predictions for convulsions in female F344 rats exposed to RDX by gavage on GDs 6−19 (Cholakis et al., 1980); BMR = 1% ER

................................ .... D

- 27 Table D

- 15. Model predictions for combined incidence of convulsion and mortality in male and female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006); BMR = 1% ER

................................ ................................ ................................ ............... D

- 29 Table D

- 16. Model predictions for testicular degeneration in male B6C3F 1 

### mice exposed to RDX by die t for 24 months (Lish et al., 1984); BMR = 10% ER

.............................. D

- 31 Table D

- 17. Model predictions for prostate suppurative inflammation in ma le F344 rats exposed to RDX by diet for 24 months (Levine et al., 1983b); BMR = 10% ER

.... D

34 Table D

- 18. Mortality data selected for dose

- response modeling for RDX

............................. D

- 36 Table D

- 19. Model predictions for combined mortality in male and female F344 rats exposed to RDX by diet for 13 weeks (Levine et al., 1981); BMR = 1% ER

...... D

- 39 Table D

- 20. Summary of BMD modeling results for model predicti ons for mortality (number found dead) in rats exposed to RDX in the diet for 13 weeks (von Oettingen et al., 1949)

................................ ................................ ................................ ................................ .............. D

- 41 Table D

- 21 . Model predictions for combined mortality (number found dead) in male and female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006); BMR= 1% ER

................................ ................................ ................................ ............................... D

- 42 Table D

- 22. Model predictions for mortality in female Sprague

- Dawley rats exposed to RDX by gavage on gestation days 6

– 15 (Angerhofer, 1986); BMR = 1% ER

.............. D

- 44 Table D

- 23. Cancer endpoints selected for dose

- response modeling for RDX

....................... D

- 46 Table D

- 24. Model predictions for combined alveolar/bronchiolar adenoma and carcinoma in female B6C3F 1 

### mice exposed to RDX by diet for 24 months (Lish et al., 1984); BMR = 10% ER

................................ ................................ ................................ ........................... D

- 47 Table D

- 25. Model predictions for combined alveolar/bronchiolar adenoma and carcinoma in female B6C3F 1 

### mice exposed to RDX by diet for 24 months (Lish et al., 1984); BMR = 5% ER

................................ ................................ ................................ .............................. D

- 50 Table D

- 26. Model predictions for combined hepatocellular adenoma and carcinoma in female B6C3F 1 

### mice exposed to RDX by diet for 24 months (Parker et al., 2006); BMR = 10% ER

................................ ................................ ................................ ........................... D

- 52 Table D

- 27. Model predictions for B6C3F 1 

### female mouse combined hepatocellular adenoma and carcinoma in mice exposed to RDX by diet for 24 months (Parker et a l., 2006); BMR = 5% ER

................................ ................................ ................................ ............... D

- 54 Table D

- 28. Model predictions for combined hepatocellular adenoma and carcinoma in female B6C3F 1 

### mice exposed to RDX by diet for 24 months, using incidence frequencies from Parker et al. (2006) and sample sizes from Lish et al. (1984); BMR = 10% ER

................................ ................................ ................................ ........................... D

- 58 Table D

- 29. Liver carcinoma data from Levine et al. (1983b)

................................ ...................... D

- 61 Table D

- 30. Model predictions and oral slope factor for alveolar/bronchiolar carcinomas in male B6C3F 1 

### mice exposed to RDX by diet for 2 years (Lish et al., 1984)

........ D

- 62 Table D

- 31. Summary of BMD modeling results for model predictions for alveolar/bronchiolar carcinoma in male B6C3F1 mice exposed to RDX by diet for 2 years (Lish et al., 1984); BMR = 10% ER

................................ .............................. D

- 62

This document is a draft for review purposes only and does not constitute Agency policy. 

### vi DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Table D

- 32. Model predictions and oral slope factor for hepatocellular carcinomas in male F344 rats administered RDX in the diet for 2 years (Levine et al., 1983b)

...... D

- 65 Table D

- 33. Model predictions for combined hepatocellular adenoma and carcinoma in F344 rats exposed to RDX by diet for 24 months (Levine et al., 1983b); BMR = 5% ER

................................ ................................ ................................ ................................ ............ D

- 65

This document is a draft for review purposes only and does not constitute Agency policy. 

### vii DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine


# FIGURES

Figure C

- 1. PBPK model structure for RDX in rats and humans.

................................ .................. C

- 11 Figure C

- 2. EPA rat PBPK model predictions fitted to observed RDX blood concentrations in male and female Sprague

- D awley rats following i.v. exposure.

............................. C

- 17 Figure C

- 3. EPA rat PBPK model predictions fitted to observed RDX blood concentrations following oral exposure to RDX dissolved in water.

................................ ................... C

- 18 Figure C

- 4. EPA rat model predictions fitted to observed RDX blood concen trations following oral exposure to RDX in dry capsules.

................................ .......................... C

- 18 Figure C

- 5. Effect of varying oral absorption parameters on EPA rat model predictions fitted to observed RDX blood concentrations following oral exposure to coarse

- grain RDX.

................................ ................................ ................................ ................................ ................. C

- 19 Figure C

- 6. EPA rat model predictions fitted to observed RDX brain tissue concentrations following oral exposure to RDX.

................................ ................................ .......................... C

- 19 Figure C

- 7. EPA r at model predictions fitted to observed RDX blood concentrations following oral exposure to fine

- grain RDX in a saline slurry.

................................ .. C

- 21 Figure C

- 8. Comparison of EPA rat model predictions with data from Schneider et al. (1978) for the subchronic gavage study.

................................ ................................ ......................... C

- 22 Figure C

- 9. Comparison of EPA rat model predictions with data from Schneider et al. (1978) for the subchronic drinking water study.

................................ ................................ ........ C

- 22 Figure C

- 10. EPA human model predictions fitted to observed RDX blood concentrations resulting from an accidental ingestion of RDX by a 14.5

- kg boy (Woody et al., 1986).

................................ ................................ ................................ ................................ .............. C

- 24 Figure C

- 11. EPA human model predictions fitted to observed RDX blood concentrations resulting from accidental exposure to adults assumed to be 70 kg (Özhan et al., 2003 ).

................................ ................................ ................................ ................................ .............. C

- 24 Figure C

- 12. Comparison of EPA mouse PBPK model predictions with data from oral exposure to RDX dissolved in water.

................................ ................................ ................. C

- 27 Figure D

- 1. Plot of incidence rate by dose, with fitted curve for selected model, for convulsions in female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006).

................................ ................................ ................................ ................................ .... D

- 5 Figure D

- 2. Plot of incidence rate by dose, with fitted curve for selected model, for convulsions in female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006).

................................ ................................ ................................ ................................ .... D

- 7 Figure D

- 3. Plot of i ncidence rate by dose, with fitted curve for selected model, for convulsions in female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006).

................................ ................................ ................................ ................................ .... D

- 9 Figure D

- 4. Plot of incidence rate by dose, with fitted curve for selected model, for convulsions in male F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006).

................................ ................................ ................................ ................................ ...... D

- 11 Figure D

- 5. Plot of incidence rate by dose, with fitted curve for selected model, for convulsions in male F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 20 06).

................................ ................................ ................................ ................................ ...... D

- 13 Figure D

- 6. Plot of incidence rate by dose, with fitted curve for selected model, for convulsions in male F344 rats exposed to RDX by gavage for 90 days (Cro use et al., 2006).

................................ ................................ ................................ ................................ ...... D

- 15 Figure D

- 7. Plot of incidence rate by dose, with fitted curve for selected model, for convulsions in male and female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006).

................................ ................................ ................................ ................ D

- 17

This document is a draft for review purposes only and does not constitute Agency policy. 

### viii DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Figure D

- 8. Plot of incidence rate by dos e, with the fitted curve of the selected model, for convulsions in male and female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006).

................................ ................................ ................................ ................ D

- 19 Figure D

- 9. Plot of incidence rate by dose, with the fitted curve of the multistage 3° model, for convulsions in male and female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006). BMR = 1% ER; dose shown in mg/kg

- day.

.............. D

- 21 Figure D

- 10. Plot of incidence rate by dose, with the fitted curve of the selected model, for convulsions in female F344 rats exposed to RDX by gavage on GDs 6−19 (Cholakis et al., 1980).

................................ ................................ ................................ ............. D

- 23 Figure D

- 11. Plot of incidence rate by dose, with the fitted curve of the selected model, for convulsions in female F344 rats exposed to RDX by gavage on GDs 6−19 (Cholakis et al., 1980).

................................ ................................ ................................ ............. D

- 25 Figure D

- 12. Plot of incidence rate by dose, with the fitted curve of the selected model, for convulsions in female F344 rats exposed to RDX by gavage on GDs 6−19 (Cholakis et al., 1980).

................................ ................................ ................................ ............. D

- 27 Figure D

- 13. Plot of incidence rate by dose with fitted curve for Multistage 3° model for Model predictions for combined incidence of convulsion and mortality in male and fe male F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006).

................................ ................................ ................................ ................................ ............. D

- 29 Figure D

- 14. Plot of incidence rate by dose, with fitted curve for selected model, for testicular degeneration in male B6C3F 1 

### mice exposed to RDX by diet for 24 months (Lish et al., 1984).

................................ ................................ ................................ ..... D

- 32 Figure D

- 15. Plot of incidence rate by dose, with fitted curve for selected model, for prostate suppurative inflammation in male F344 rats exposed to RDX by diet for 24 months (Levine et al., 1983b).

................................ ................................ ....................... D

- 35 Figure D

- 16. Plot of incidence rate by dose, with the fitted curve of the multista ge 2° model, for combined mortality in male and female F344 rats exposed to RDX by diet for 13 weeks (Levine et al., 1981); BMR = 1% ER.

................................ ............................. D

- 39 Figure D

- 17. Plot of incidence rate by dose with fitted curve for Dichotomous

- Hill model for Model predictions for mortality (number found dead) in rats exposed to RDX in the diet for 13 weeks (von Oettingen et al., 1949); dose shown in mg/kg

- d ay.

... D

41 Figure D

- 18. Plot of incidence rate by dose, with the fitted curve of the multistage 2° model, for mortality in male and female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006). BMR = 1% ER; dose shown in mg/kg

- day.

.............. D

- 43 Figure D

- 19. Plot of incidence rate by dose, with the fitted curve of the multistage 3° model, for mortality in female Sprague

- Dawley rats exposed to RDX by gavage on gestation days 6

– 15 (Angerhofer, 1986); BMR = 1% ER.

................................ ........ D

- 45 Figure D

- 20. Plot of incidence rate by dose, with the fitted curve for the selected model, for combined alveolar/bronchiolar adenoma and carcinoma in female B6C3F 1 

### mic e exposed to RDX by diet for 24 months (Lish et al., 1984).

................................ ....... D

- 48 Figure D

- 21. Plot of incidence rate by dose, with fitted curve for selected model, for combined alveolar/bronchiolar adenoma and carcinoma in female B6C3F 1 

### mice exposed to RDX by diet for 24 months (Lish et al., 1984).

................................ ....... D

- 50 Figure D

- 22. Plot of incidence rate by dose, with fitted curve for selected model, for combined hepatocellular adenoma and carcinoma in female B6C3F 1 

### mice exposed to RDX by diet for 24 months (Parker et al., 2006).

................................ . D

- 52 Figure D

- 23. Plot of incidence rate by dose, with fitted curve for selected model, for B6C3F 1 

### female mouse combined hepatocellular adenoma and carcinoma in mice exposed to RDX by diet for 24 months (Parker et al., 2006).

................................ . D

- 54

This document is a draft for review purposes only and does not constitute Agency policy. 

### ix DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Figure D

- 24. Plot of incidence rate by dose with fitted curve for Multistage

- Cancer 1° model for model predictions for combined hepatocellular adenoma and carcinoma in female B6C3F1 mice exposed to RDX by diet for 24 months, using incidence frequencie s from Parker et al. (2006) and sample sizes from (Parker et al., 2006).

................................ ................................ ................................ ................................ ............. D

- 58 Figure D

- 25. Plot of incidence rate by dose with fi tted curve for Multistage

- Cancer 1° model for Model predictions for alveolar/bronchiolar carcinoma in male B6C3F1 mice exposed to RDX by diet for 24 months (Lish et al., 1984).

................................ ....... D

- 62 Figure D

- 26. Plot of incidence rate by dose, with fitted curve for selected model, for combined hepatocellular adenoma and carcinoma in F344 rats exposed to RDX by diet for 24 months (Levine et al., 1983 b).

................................ ................................ D

- 66

This document is a draft for review purposes only and does not constitute Agency policy.



### x DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine


# ABBREVIATIONS

AAP Army ammunition plant ACGIH American Conference of Governmental Industrial Hygienists AChE acetylcholinesterase ADAF age
- dependent adjustment factor AIC Akaike’s information criterion ALP alkaline phosphatase ALT alanine aminotransferase AST aspartate aminotransferase atm atmosphere ATSDR Agency for Toxic Substances and Disease Registry AUC area under the curve BDNF brain

- derived neurotrophic facto r BHC beta

- hexachlorocyclohexane BMC benchmark concentration BMCL benchmark concentration lower confidence limit BMD benchmark dose BMDL benchmark dose lower confidence limit BMDS Benchmark Dose Software BMDU benchmark dose upper bound BMR benchmar k response BUN blood urea nitrogen BW body weight CASRN Chemical Abstracts Service Registry Number CCL Contaminant Candidate List CI confidence interval CICAD Concise International Chemical Assessment Document CNS central nervous system CSF cerebro spinal fluid CYP450 cytochrome P450 DAF dosimetric adjustment factor DDT dichlorodiphenyltrichloroethane d.f. degrees of freedom DMSO dimethylsulfoxide DNA deoxyribonucleic acid DNX 1

- nitro

- 3,5

- dinitroso

1,3,5

- triazacyclohexane DTIC Defense Technical Information Center EEG electroencephalogram EHC Environmental Health Criteria EPA Environmental Protection Agency ER extra risk FDA Food and Drug Administration FOB functional observational battery FUDS Formerly Used Defense Sites GAB A gamma
- amino butyric acid GD gestational day GI gastrointestinal GLP good laboratory practices HED human equivalent dose HERO Health and Environmental Research Online HGPRT hypoxanthine

- guanine phosphoribosyltransferase HMX octahydro

- 1,3,5,7

- tetranitro

1,3,5,7

- tetrazocine IARC International Agency for Research on Cancer i.p. intraperitoneal IPCS International Programme on Chemical Safety IRIS Integrated Risk Information System IUR inhalation unit risk i.v. intravenous LDH lactate dehydrogenase LOAEL lowest

- ob served

- adverse

- effect level LOD limit of detection miRNA microRNA MNX hexahydro

- 1

- nitroso

- 3,5

- dinitro

1,3,5

- triazine MOA mode of action MRL Minimal Risk Level NAPDH nicotinamide adenine dinucleotide phosphate NAS National Academy of Science NCE normochromatic erythrocyte NCEA National Center for Environmental Assessment NCI National Cancer Institute NCTR National Center for Toxicological Research NHANES National Health and Nutrition Examination Survey NICNAS National Industrial Chemicals Notifi cation and Assessment Scheme NIEHS National Institute of Environmental Health Sciences NIOSH National Institute for Occupational Safety and Health NOAEL no
- observed

- adverse

- effect level NOEL no
- observed

- effect level NPL National Priorities List NRC Nucle ar Regulatory Commission NSCEP Natio n al Service Center for Environmental Publications NTP National Toxicology Program NZW New Zealand White OR odds ratio

This document is a draft for review purposes only and does not constitute Agency policy. 

### xi DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

ORD Office of Research and Development OSF oral slope factor OSHA Occupational Safety and Health A dministration PBPK physiologically based pharmacokinetic PCB polychlorinated biphenyl PCE polychromatic erythrocyte PEL Permissible Exposure Limit PND postnatal day POD point of departure PWG Pathology Working Group RBC red blood cell RDX Royal Demolition eXplosive (hexahydro

- 1,3,5

- trinitro

1,3,5- triazine) REL Recommended Exposure Limit RfC inhalation reference concentration RfD oral reference dose SDMS spontaneous death or moribund sacrifice SDWA Safe Drinking Water Act SGOT glutamic oxaloacetic transaminase, also known as AST SGPT glutamic pyruvic transaminase, also known as ALT SLE systemic lupus erythematosus SS scheduled sacrifice TLV Threshold Limit Value TNT trinitrotoluene TNX hexahydro -1,3,5- trinitroso

1,3,5- triazine TSCATS Toxic Substances Control Act Test Submissions TWA time

- weighted average U.S. United States of America UCM Unregulated Contaminant Monitoring UF uncertainty factor UF A animal

- to

- human uncertainty factor UF D database deficiencies uncertainty factor UF H h uman variation uncertainty factor UF L LOAEL

- to

- NOAEL uncertain factor UF S subchronic

- to

- chronic uncertainty factor WBC white blood cell WHO World Health Organization

Th is document is a draft for review purposes only and does not constitute Agency policy. 

### xii DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine



# APPENDIX A. ASSESSMENTS BY OTHER NATIONAL AND INTERNATIONAL HEALTH AGENCIES

**Table A**

**- 1.**  **Assessments by o**  **ther n**  **ational and i**  **nternational h**  **ealth a**  **gencies** 1

Organization Toxicity value Agency for Toxic Substances and Disease Registry ( ATSDR, 2012 ) Acute oral M inimal R isk L evel (MRL)

— 0.2 mg/kg

- d Basis: tremors and convulsions in rats ( Crouse et al., 2006 ) ; application of a composite uncertainty factor (UF) of 30 (3 for extrapolation from animals to humans with dosimetric adjustments [ physiologically based pharmacokinetic or PBPK modeling] and 10 for human variability) Intermediate oral MRL

— 0.1 mg/kg -d B as is: convulsions in rats ( Crouse et al., 2006 ) ; application of a composite UF of 30 (3 for extrapolation from animals to humans with dosimetric adjustments [PBPK modeling] and 10 for human variability) Chronic oral MRL

— 0.1 mg/kg -d Ba sis: tremors and convulsions in rats ( Levine et al., 1983b ) ; application of a composite UF of 30 (3 for extrapolation from animals to humans with dosimetric adjustments [PBPK modeling] and 10 for human variabili ty)
National Institute for Occupational Safety and Health ( NIOSH, 2012 ) Recommended Exposure Limit (REL)

— 1.5 mg/m 3 TWA for up to a 10

- hr workday during a 40

- hr workweek ; short

- term (15

- min) limit

— 3 mg/m 3 Basis: agreed with Occupational Safety and Health Administration ( OSHA )- proposed P ermissible E xposure L imit (PEL) in 1988 PEL Hearings Skin designation indicates potential for dermal absorption Basis: agreed with OSHA proposal for skin notation in 1988 PEL Hearings

O ccupational S afety and H ealth A dministration ( OSHA, 2012a , b ) PEL

— 1.5 mg/m 3 time

- weighted average ( TWA ) for an 8

- hr workday in a 40

- hr workweek Basis: adopted from the American Conference of Governmental Industrial Hygienists ( ACGIH ) Threshold Limit Value ( TLV ) established in 1969 Skin designation indicates that cutaneous exposure may contribute to overall exposure and measures should be taken to prevent skin absorption Basis: adopted from ACGIH
Hazardous Substances Information System ( Safe Work Australia, 2014 ) Exposure standard

— 1.5 mg/m 3 TWA for an 8

- hr workday Basis: adopted from the ACGIH TLV established in 1991 Skin absorpt ion notice indicates that absorption through the skin may be a significant source of exposure Basis: adopted from ACGIH

This document is a draft for review purposes only and does not constitute Agency policy. 

### A-1 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine



# APPENDIX B. ADDITIONAL DETAILS OF LITERATURE SEARCH STRATEGY | STUDY SELECTION AND EVALUATION

The literature search for hexahydro- 1,3,5

- trinitro

- 1,3,5

- triazine ( RDX ) was conducted in 1
2
3
4
5 6
7 8
9

10 11
12 13
14
15 16
17 18
19 20
21
22 23
24
25 five online scientific databases through January, 2015 . The detailed search strategy used to sea rch four of these databases

— PubM ed, Toxline, Toxcenter, and Toxic Substances Control Act Test Submissions ( TSCATS )— is provided in Table B

- 1. The search strategy used to search the Defense Technical Information Center (DTIC) database is described in Table B

- 2. The computerized database searches were augmented by review of online regulatory sources
, as well as “forward” and “backward” Web o f Science searches of two recent reviews (Table B- 3). Forward searching was used to identi fy articles that cite d the selected studies (i.e., the two reviews identified in Table B-3 ), and backward searching was used to identify articles that the selected s tud ies cite d.



## B.1. **DEFENSE TECHNICAL INFORMATION CENTER (DTIC) LITERATURE SEARCH AND SCREEN**  Among the RDX

- related citations that were identified in the DTIC database, 826 (722 after duplicate removal within DTIC) were classified with the distribution "Approv ed for Public Release", 239 (217 after duplicate removal) were classified as “distribution limited to U.S. Government agencies and their Contractors,” and 199 (181 after duplicate removal) were classified as “distribution limited to U.S. Government agencie s only.” A preliminary screen of the 1,120 unique citations was performed; 85 citations with unlimited distribution and 10 citations with limited distribution were selected for further review as potential sources of health effects data or supporting infor mation. The remaining 1
, 025 unlimited and limited

- distribution DTIC references not selected for further consideration were not studies of RDX or did not contain information pertinent to the assessment of the health effects of RDX (e.g., documents were rel ated to environmental properties such as leaching, explosive properties, fuel and propellant properties, weapons systems, treatment of wastewater containing explosives, and disposal technologies). The 85 unique citations with unlimited distribution were up loaded to the Health and Environmental Research Online (HERO) website 1 

### ( http://hero.epa.gov ). The 10 citations with

1 HERO is a database of scientific studies and other references used to develop EPA’s assessments aimed at understanding the health and environmental effects of pollutants and chemicals. It is d eveloped and managed in EPA’s Office of Research and Development (ORD) by the National Center for Environmental Assessment (NCEA). The database includes more than 1.6 million scientific references, including articles from the peer

- reviewed literature. Ne w studies are added continuously to HERO.
This document is a draft for review purposes only and does not constitute Agency policy. 

### A-1 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

l imited distribution were subject to a more in

- depth screen to determine whether the references 1
2 3
4
5

6 7 8

9 10 11 12 13 14 15

16 17

18
19
20 provided additional primary health effects data and whether the U.S. Environmental Protection Agency (EPA) should seek authorization for public distribution and upload to HERO. A review of the abstract or full

- text of the documents associated with the lim ited

- distribution citations resulted in the following determinations:

• o ne c itation was excluded because it did not provide additional primary health effects data. The citation reported data from a study that was subsequently published ( Hathaway and B uck, 1977 ) and had already be en identified by the literature search strategy

.

• one citation (dated 1944 ) provided human and animal inhalation data and was considered pertinent, but was not brought forward for further review because flaws in the design of both the human and animal stud i es were such that results would not be considered credible. Experimental animal study design issues included lack of a control group, small numbers of animals, incomplete information on dosage or exposure levels, and inadequate reporting

. The human study described a case series and lacked a referent group and measures of RDX exposure.

• eight citations were regulatory documents, reviews, or risk assessments that did not specifically identify RDX and did not appear to contain primary health effects data.
Based on these determinations, none of the 10 limited distribution citations that were subject to further review were selected for further consideration or added to HERO.

This document is a draft for review purposes only and does not constitute Agency policy. 

### A-2 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**T able B** 

**- 1. Summary of detailed search strategies for RDX (Pubmed**  **, Toxline,** 1 2 **Toxcenter, TSCATS)**

Database Terms Hits

PubMed Date: 4/ 2012 ((((121

- 82

- 4) OR (Cyclonite[tw] OR Cyclotrimethylenetrinitramine[tw] OR "cyclotrimethylene trinitramine"[tw] OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazine"[tw] OR "Hexahydro

- 1,3,5

- trinitro -s- triazine"[tw] OR Hexogen[tw] OR "1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR "1,3,5

- Triaza

1,3,5

- trinitrocyclohexane"[tw] OR "1,3,5

- Trinitro

- 1,3,5

- triazacyclohexane"[tw] OR "1,3,5

- Trinitrohexahydro

- 1,3,5

- triazine"[tw] OR "1,3,5

- Trinit rohexahydro

s- triazine"[tw] OR "1,3,5

- Trinitroperhydro

- 1,3,5

- triazine"[tw] OR "Esaidro

1,3,5

- trinitro

- 1,3,5

- triazina"[tw] OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazin"[tw] OR "Perhydro

- 1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR Cyclotrimethylenenitramine[tw] OR T rimethylenetrinitramine[tw] OR "Trimethylene trinitramine"[tw] OR Trimethyleentrinitramine[tw] OR "Trinitrocyclotrimethylene triamine"[tw] OR Trinitrotrimethylenetriamine[tw] OR "CX 84A"[tw] OR Cyklonit[tw] OR Geksogen[tw] OR Heksogen[tw] OR Hexogeen[tw] O R Hexolite[tw] OR "KHP 281"[tw] OR "PBX (af) 108"[tw] OR "PBXW 108(E)"[tw] OR "Pbx(AF) 108"[tw]) OR (rdx[tw])) NOT medline[sb]) OR (((121

- 82

- 4) OR (Cyclonite[tw] OR Cyclotrimethylenetrinitramine[tw] OR "cyclotrimethylene trinitramine"[tw] OR "Hexahydro

- 1,3 ,5

- trinitro

1,3,5

- triazine"[tw] OR "Hexahydro

- 1,3,5

- trinitro -s- triazine"[tw] OR Hexogen[tw] OR "1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR "1,3,5

- Triaza

1,3,5

- trinitrocyclohexane"[tw] OR "1,3,5

- Trinitro

- 1,3,5

- triazacyclohexane"[tw] OR "1,3,5

- Trinitrohexahydro -1 ,3,5

- triazine"[tw] OR "1,3,5

- Trinitrohexahydro

s- triazine"[tw] OR "1,3,5

- Trinitroperhydro

- 1,3,5

- triazine"[tw] OR "Esaidro

1,3,5

- trinitro

- 1,3,5

- triazina"[tw] OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazin"[tw] OR "Perhydro

- 1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR Cyclotrimethylenenitramine[tw] OR Trimethylenetrinitramine[tw] OR "Trimethylene trinitramine"[tw] OR Trimethyleentrinitramine[tw] OR "Trinitrocyclotrimethylene triamine"[tw] OR Trinitrotrimethylenetriamine[tw] OR "CX 84A"[tw] OR Cyklonit[tw] OR Geksogen[tw ] OR Heksogen[tw] OR Hexogeen[tw] OR Hexolite[tw] OR "KHP 281"[tw] OR "PBX (af) 108"[tw] OR "PBXW 108(E)"[tw] OR "Pbx(AF) 108"[tw]) OR (rdx[tw])) AND (to[sh] OR po[sh] OR ae[sh] OR pk[sh] OR (me[sh] AND (humans[mh] OR animals[mh])) OR ci[sh] OR bl[sh] OR cf[sh] OR ur[sh] OR ((pharmacokinetics[mh] OR metabolism[mh]) AND (humans[mh] OR mammals[mh])) OR "dose

- response relationship, drug"[mh] OR risk[mh] OR "toxicity tests"[mh] OR noxae[mh] OR cancer[sb] OR "endocrine system"[mh] OR "endocrine disruptors"[mh] O R "Hormones, Hormone Substitutes, and Hormone Antagonists"[mh] OR triazines/ai OR ("Inhalation Exposure"[Mesh] OR "Maternal Exposure"[Mesh] OR "Maximum Allowable Concentration"[Mesh] OR "Occupational Exposure"[Mesh] OR "Paternal Exposure"[Mesh] OR "Environ mental Exposure"[Mesh:noexp])))) NOT (((((121

- 82

- 4) OR (Cyclonite[tw] OR Cyclotrimethylenetrinitramine[tw] OR "cyclotrimethylene trinitramine"[tw] OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazine"[tw] OR "Hexahydro

- 1,3,5

- trinitro -s- triazine"[tw] OR Hexogen[tw] OR "1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR "1,3,5

- Triaza

1,3,5

- trinitrocyclohexane"[tw] OR "1,3,5

- Trinitro

- 1,3,5

- triazacyclohexane"[tw] OR "1,3,5

- Trinitrohexahydro

- 1,3,5

- triazine"[tw] OR "1,3,5

- Trinitrohexahydro

s- triazine"[tw] OR "1,3,5

- Trinitroperhydro

- 1,3,5

- triazine"[tw] OR "Esaidro

1,3,5

- trinitro

- 1,3,5

- triazina"[tw] OR "Hexahydro

- 1,3,5

- trinitro

337

This document is a draft for review purposes only and does not constitute Agency policy. 

### B-3 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Database Terms Hits
1,3,5

- triazin"[tw] OR "Perhydro

- 1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR Cyclotrimethylenenitramine[tw] OR Trimethylenetrinitramine[tw] OR "Trimethylene trinitramine"[tw] OR Trimethyleentrinitramine[tw] OR "Trinitrocyclotrimethylene triamine"[tw] OR Trinitrotrimethylenetriamine[tw] OR "CX 84A"[tw] OR Cyklonit[tw] OR Geksogen[tw] OR Heksogen[tw] OR Hexogeen[tw] OR Hexolite[tw] OR "KHP 281"[tw] OR "PBX (af) 108"[tw] OR "PBXW 108(E)"[tw] OR "Pbx(AF) 108"[tw]) OR (rdx[tw])) NOT medline[sb]) OR (((121

- 82

- 4) OR (Cyclonite[tw] OR Cyclotrimethylenetrinitramine[tw] OR "cyclotrimethylene trinitramine"[tw] OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazine"[tw] OR "Hexahydro

- 1,3,5

- trini tro -s- triazine"[tw] OR Hexogen[tw] OR "1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR "1,3,5

- Triaza

1,3,5

- trinitrocyclohexane"[tw] OR "1,3,5

- Trinitro

- 1,3,5

- triazacyclohexane"[tw] OR "1,3,5

- Trinitrohexahydro

- 1,3,5

- triazine"[tw] OR "1,3,5

- Trinitrohexahydro

s- triazine "[tw] OR "1,3,5

- Trinitroperhydro

- 1,3,5

- triazine"[tw] OR "Esaidro

1,3,5

- trinitro

- 1,3,5

- triazina"[tw] OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazin"[tw] OR "Perhydro

- 1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR Cyclotrimethylenenitramine[tw] OR Trimethylenetrinitramin e[tw] OR "Trimethylene trinitramine"[tw] OR Trimethyleentrinitramine[tw] OR "Trinitrocyclotrimethylene triamine"[tw] OR Trinitrotrimethylenetriamine[tw] OR "CX 84A"[tw] OR Cyklonit[tw] OR Geksogen[tw] OR Heksogen[tw] OR Hexogeen[tw] OR Hexolite[tw] OR "KHP 281"[tw] OR "PBX (af) 108"[tw] OR "PBXW 108(E)"[tw] OR "Pbx(AF) 108"[tw]) OR (rdx[tw])) AND (to[sh] OR po[sh] OR ae[sh] OR pk[sh] OR (me[sh] AND (humans[mh] OR animals[mh])) OR ci[sh] OR bl[sh] OR cf[sh] OR ur[sh] OR ((pharmacokinetics[mh] OR metabolism[m h]) AND (humans[mh] OR mammals[mh])) OR "dose

- response relationship, drug"[mh] OR risk[mh] OR "toxicity tests"[mh] OR noxae[mh] OR cancer[sb] OR "endocrine system"[mh] OR "endocrine disruptors"[mh] OR "Hormones, Hormone Substitutes, and Hormone Antagonists "[mh] OR triazines/ai OR ("Inhalation Exposure"[Mesh] OR "Maternal Exposure"[Mesh] OR "Maximum Allowable Concentration"[Mesh] OR "Occupational Exposure"[Mesh] OR "Paternal Exposure"[Mesh] OR "Environmental Exposure"[Mesh:noexp])))) AND (invertebrates OR aquatic organisms OR fish OR fishes OR amphibians OR earthworm*))
PubMed Date limit: 1/2012

− 2/2013 (Cyclonite[tw] OR RDX[tw] OR Cyclotrimethylenetrinitramine[tw] OR "cyclotrimethylene trinitramine"[tw] OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazine"[tw] OR "Hexahydro

- 1,3,5

- trinitro -s- triazine"[tw] OR Hexogen[tw] OR "1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR "1,3,5

- Triaza

1,3,5

- trinitrocyclohexane"[tw] OR "1,3,5

- Trinitro

- 1,3,5

- triazacyclohexane"[tw] OR "1,3,5

- Trinitrohexahydro

- 1,3,5

- triazine"[tw] OR "1,3,5

- Tri nitrohexahydro

s- triazine"[tw] OR "1,3,5

- Trinitroperhydro

- 1,3,5

- triazine"[tw] OR "Esaidro

1,3,5

- trinitro

- 1,3,5

- triazina"[tw] OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazin"[tw] OR "Perhydro

- 1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR Cyclotrimethylenenitramine[tw] OR Trimethylenetrinitramine[tw] OR "Trimethylene trinitramine"[tw] OR Trimethyleentrinitramine[tw] OR "Trinitrocyclotrimethylene triamine"[tw] OR Trinitrotrimethylenetriamine[tw] OR "CX 84A"[tw] OR Cyklonit[tw] OR Geksogen[tw] OR Heksogen[tw] OR Hexogeen[tw ] OR Hexolite[tw] OR "KHP 281"[tw] OR "PBX (af) 108"[tw] OR "PBXW 108(E)"[tw] OR "Pbx(AF) 108"[tw]) AND ((" 2012/01/01"[Date - MeSH] : "3000"[Date - MeSH]) OR (" 2012/01/01"[Date - Entrez] : "3000"[Date - Entrez]) OR (" 2012 /01/01"[Date

- Create] : "3000"[Dat e

- Create])) 112

This document is a draft for review purposes only and does not constitute Agency policy. 

### B-4 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Database Terms Hits

PubMed Date limit: 11/ 2012

− 1/ 2014 (Cyclonite[tw] OR RDX[tw] OR Cyclotrimethylenetrinitramine[tw] OR "cyclotrimethylene trinitramine"[tw] OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazine"[tw] OR "Hexahydro

- 1,3,5

- trinitro

- s

- triazine"[tw] OR He xogen[tw] OR "1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR "1,3,5

- Triaza

1,3,5

- trinitrocyclohexane"[tw] OR "1,3,5

- Trinitro

- 1,3,5

- triazacyclohexane"[tw] OR "1,3,5

- Trinitrohexahydro

- 1,3,5

- triazine"[tw] OR "1,3,5

- Trinitrohexahydro

s

- triazine"[tw] OR "1,3,5

- Trinitroperhydro

- 1,3,5

- triazine"[tw] OR "Esaidro

1,3,5

- trinitro

- 1,3,5

- triazina"[tw] OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazin"[tw] OR "Perhydro

- 1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR Cyclotrimethylenenitramine[tw] OR Trimethylenetrinitramine[tw] OR "Trimethylene trinitramine"[tw] OR Trimethyleentrinitramine[tw] OR "Trinitrocyclotrimethylene triamine"[tw] OR Trinitrotrimethylenetriamine[tw] OR "CX 84A"[tw] OR Cyklonit[tw] OR Geksogen[tw] OR Heksogen[tw] OR Hexogeen[tw] OR Hexolite[tw] OR "KHP 281"[tw] OR "PBX (af) 108"[tw] OR "PBXW 108(E)"[tw] OR "Pbx(AF) 108"[tw]) AND ((" 2012 /11/01"[Date

- MeSH] : "3000"[Date

- MeSH]) OR (" 2012 /11/01"[Date

- Entrez] : "3000"[Date

- Entrez]) OR (" 2012 /11/01"[Date

- Create] : "3000"[Date

- Create])) 138

PubMed Date limit: 11/ 2013

− 1/ 2015 ("cyclonite"[nm] OR Cyclonite[tw] OR RDX[tw] OR Cyclotrimethylenetrinitramine[tw] OR "cyclotrimethylene trinitramine"[tw] OR "Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR "Hexahydro

- 1,3,5

- trinitro

s

- triazine"[tw] OR Hexogen[tw] OR "1,3,5

- trinitro

- 1,3,5

- triazine"[tw] OR "1,3,5

- Triaza

- 1,3,5

- trinitrocyclohexane"[tw] OR "1,3,5

- Trinitro

1,3,5

- triazacyclohexane"[tw] OR "1,3,5

- Trinitrohexahydro

- 1,3,5

- triazine"[tw] OR "1,3,5

- Trinitrohexahydro

- s

- triazine"[tw] OR "1,3,5

- Trinitroperhydro

1,3, 5

- triazine"[tw] OR "Esaidro

- 1,3,5

- trinitro

- 1,3,5

- triazina"[tw] OR "Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazin"[tw] OR "Perhydro

- 1,3,5

- trinitro

1,3,5

- triazine"[tw] OR Cyclotrimethylenenitramine[tw] OR Trimethylenetrinitramine[tw] OR "Trimethylene trinitramine"[ tw] OR Trimethyleentrinitramine[tw] OR "Trinitrocyclotrimethylene triamine"[tw] OR Trinitrotrimethylenetriamine[tw] OR "CX 84A"[tw] OR Cyklonit[tw] OR Geksogen[tw] OR Heksogen[tw] OR Hexogeen[tw] OR Hexolite[tw] OR "KHP 281"[tw] OR "PBX (af) 108"[tw] OR "P BXW 108(E)"[tw] OR "Pbx(AF) 108"[tw]) AND ( 2013 /11/01 : 3000[crdat] OR 2013 /11/01 : 3000[edat]) 76

Toxline Date: 4/ 2012 Notes: Searched CASRN or synonyms; removed invertebrates, aquatic organisms, amphibians, earthworms

. 507

Toxline Date limit: 2011

− 2/ 2013 @OR+("Cyclonite"+"RDX"+"Cyclotrimethylenetrinitramine"+"cyclotrimethylen e trinitramine"+"Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine"+"Hexahydro

1,3,5

- trinitro

- s

- triazine"+"Hexogen"+"1,3,5

- trinitro

1,3,5triazine"+"1,3,5

- Triaza

- 1,3,5

- trinitrocyclohexane"+" 1,3,5

- Trinitro

1,3,5

- triazacyclohexane"+"1,3,5

- Trinitrohexahydro

- 1,3,5

- triazine"+ "1,3,5

- Trinitrohexahydro

- s

- triazine"+@term+@rn+121

- 82

- 4)+ @AND+@range+yr+ 2011 + 2013 +@NOT+@org+pubmed+pubdart+crisp+ tscats 5

@OR+("1,3,5

- Trinitroperhydro

- 1,3,5

- triazine"+"Esaidro

- 1,3,5

- trinitro

1,3,5

- triazina"+"Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazin"+"Perhydro

1,3,5

- trinitro

- 1,3,5

- triazine"+ "Cyclotrimethylenenitramine"+ "Trimethylenetrinitramine"+ "Trimethylene+trinitramine"+

" Trimethyleentrinitramine"+ "Trinitrocyclotrimethylene+ triamine"+ "Trinitrotrimethylenetriamine"+ "CX+84A"+ "Cyklonit"+ "Geksogen"+ 0

This document is a draft for review purposes only and does not constitute Agency policy. 

### B

- 5 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Database Terms Hits "Heksogen"+"Hexogeen"+"Hexolite"+"KHP+281")+@AND+@range+yr+ 2011 + 2013 + @NOT+ @org+ pubmed+ pubdart+ crisp+tscats
Toxline Date limit: 2012

− 1/ 2014 @OR+("Cyclonite"+"RDX"+"Cyclotrimethylenetrinitramine"+"cyclotrimethylen e trinitramine"+"Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine"+"Hexahydro

1,3,5

- trinitro

- s

- triazine"+"Hexogen"+"1,3,5

- trinitro

- 1,3,5

- triazine"+ "1,3,5

- Triaza

- 1,3,5

- trinitrocyclohexane"+"1,3,5

- Trinitro

1,3,5

- triazacyclohexane"+ "1,3,5

- Trinitrohexahydro

- 1,3,5

- triazine"+ "1,3,5

- Trinitrohexahydro

- s

- triazine"+@term+@rn+121

- 82

- 4)+ @AND+ @range+yr+ 2012 + 2014 +@NOT+@org+pubmed+pubdart+crisp+tscats 10

@OR+("1,3,5

- Trinitroperhydro

- 1,3,5

- triazine"+"Esaidro

- 1,3,5

- trinitro

1,3,5

- triazina"+"Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazin"+"Perhydro

1,3,5

- trinitro

- 1,3,5

- triazine"+ "Cyclotrimethylenenitramine"+ "Trimethylenetrinitramine"+"Trimethylene+trinitramine"+ "T rimethyleentrinitramine"+"Trinitrocyclotrimethylene+triamine"+ "Trinitrotrimethylenetriamine"+"CX+84A"+"Cyklonit"+"Geksogen"+ "Heksogen"+"Hexogeen"+"Hexolite"+"KHP+281")+@AND+@range+yr+ 2012 + 2014 +@NOT+@org+pubmed+pubdart+crisp+tscats 0

Toxline Date limit: 2013

- 1/ 2015 @SYN0+@OR+(RDX+"cyclotrimethylene+trinitramine"+"1,3,5

- trinitro

1,3,5

- triazine"+"1,3,5

- Triaza

- 1,3,5

- trinitrocyclohexane"+"1,3,5

- Trinitro

1,3,5

- triazacyclohexane" +"1,3,5

- Trinitrohexahydro

- 1,3,5

- triazine"+ "1,3,5

- Trinitrohexahydro

- s

- t riazine"+"1,3,5

- T rinitroperhydro

- 1,3,5

- triazine"+ "CX+84A")+@AND+@range+yr+ 2012 + 2015 +@NOT+@org+pubmed+pubdart+" nih+reporter"+tscats+crisp 19

@SYN0+@OR+("Cyclonite"+"Cyclotrimethylenenitramine"+"Cyclotrimethylene trinitramine"+"Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine"+"Hexahydro

1,3,5

- trinitro

- s

- triazine"+ "Hexogen"+ "Hexolite"+ "KHP+281"+ "PBX+(af)+108"+ "PBXW+108(E)"+"Pbx(AF)+108"+"Perhydro

- 1,3 ,5

- trinitro

- 1,3,5

- triazine")+ @AND+@range+yr+ 2012 + 2015 +@NOT+@org+pubmed+pubdart+ "nih+reporter"+tscats+crisp 9

@SYN0+@OR+("Research+Development+Explosive"+"Royal+Demolition+eXpl osive+"Trimethylenetrinitramine"+"Trinitrocyclotrimethylene+triamine"+"Trini trotrimethylenetriamine"+"sym

- Trimethylene+trinitramine"+ @term+ @rn+121

- 82

- 4+ @term+@rn+ 2046 55

- 61

- 8+@term+@rn+505 79

- 23

- 2+@term+@rn+53800

- 53

- 6+@term+@rn+57608

- 45

- 4+@term+ @rn+8 2030

- 42

- 0)+ @AND+ @range+yr+ 2012 + 2015 + @NOT+ @org+ pubmed+ pubdart+ "nih+reporter"+tscats+crisp 0

TSCATS Date: 2/ 2013 @term+@rn+121

- 82

- 4+@AND+@org+tscats 4

TSCATS 2 Date: 1/ 2015 121

- 82

- 4 from EPA receipt date 01/01/ 2000 0

TSCATS 8e/FYI Date: 1/ 2015 ("121

- 82

- 4" OR "1,3,5

- Triazine, hexahydro

- 1,3,5

- trinitro

- ") tsca (8e OR FYI) 0

This document is a draft for review purposes only and does not constitute Agency policy. 

### B

- 6 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Database Terms Hits

Toxcenter Date: 4/ 2012 ((121

- 82

- 4 OR Cyclonite OR RDX OR Cyclotrimethylenetrinitramine OR "cyclotrimethylene trinitramine" OR "Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine" OR "Hexahydro

- 1,3,5

- trinitro

- s

- triazine" OR Hexogen OR "1,3,5

- trinitro

1,3,5

- triazine" OR "1,3,5

- Triaza

- 1,3,5

- trinitrocyclohexane" OR "1,3,5

- Trinitro

1,3,5

- triazacyclohexane" OR "1,3,5

- Trinitrohexa hydro

- 1,3,5

- triazine" OR "1,3,5

- Trinitrohexahydro

- s

- triazine" OR "1,3,5

- Trinitroperhydro

- 1,3,5

- triazine" OR "Esaidro

- 1,3,5

- trinitro

- 1,3,5

- triazina" OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazin" OR "Perhydro

- 1,3,5

- trinitro

- 1,3,5

- triazine" OR Cyclotrimethylene nitramine OR Trimethylenetrinitramine OR "Trimethylene trinitramine" OR Trimethyleentrinitramine OR "Trinitrocyclotrimethylene triamine" OR Trinitrotrimethylenetriamine OR "CX 84A" OR Cyklonit OR Geksogen OR Heksogen OR Hexogeen OR Hexolite OR "KHP 281" OR "PBX (af) 108" OR "PBXW 108(E)" OR "Pbx(AF) 108") NOT (patent/dt OR tscats/fs))AND ((chronic OR immunotox? OR neurotox? OR toxicokin? OR biomarker? OR neurolog? OR pharmacokin? OR subchronic OR pbpk OR epidemiology/st,ct,it) OR acute OR subacute OR ld50# OR lc50# OR (toxicity OR adverse OR poisoning)/st,ct,it OR inhal? OR pulmon? OR nasal? OR lung? OR respir? OR occupation? OR workplace? OR worker? OR oral OR orally OR ingest? OR gavage? OR diet OR diets OR dietary OR drinking(w)water OR (maximum and

conce ntration? and (allowable OR permissible)) OR (abort? OR abnormalit? OR embryo? OR cleft? OR fetus? OR foetus? OR fetal? OR foetal? OR fertil? OR malform? OR ovum OR ova OR ovary OR placenta? OR pregnan? OR prenatal OR perinatal? OR postnatal? OR reproduc? OR steril? OR teratogen? OR sperm OR spermac? OR spermag? OR spermati? OR spermas? OR spermatob? OR spermatoc? OR spermatog? OR spermatoi? OR spermatol? OR spermator? OR spermatox? OR spermatoz? OR spermatu? OR spermi? OR spermo? OR neonat? OR newborn OR d evelopment OR developmental? OR zygote? OR child OR children OR adolescen? OR infant OR wean? OR offspring OR age(w)factor? OR dermal? OR dermis OR skin OR epiderm? OR cutaneous? OR carcinog? OR cocarcinog? OR cancer? OR precancer? OR neoplas? OR tumor? OR tumour? OR oncogen? OR lymphoma? OR carcinom? OR genetox? OR genotox? OR mutagen? OR genetic(w)toxic? OR nephrotox? OR hepatotox? OR endocrin? OR estrogen? OR androgen? OR hormon? OR rat OR rats OR mouse OR mice OR muridae OR dog OR dogs OR rabbit? OR ham ster? OR pig OR pigs OR swine OR porcine OR goat OR goats OR sheep OR monkey? OR macaque? OR marmoset? OR primate? OR mammal? OR ferret? OR gerbil? OR rodent? OR lagomorpha OR baboon? OR bovine OR canine OR cat OR cats OR feline OR pigeon? OR occupation? O R worker? OR epidem?) AND ((biosis/fs AND py> 1999 ) OR caplus/fs))

Notes: Duplicates were removed; Biosis subfile results were date limited to avoid extensive overlap with Toxline

. 337 titles screened (20 selected for full records and added to HERO)

Toxce nter

Date limit: 1/ 2012

− 2/ 2013 (((121

- 82

- 4 OR Cyclonite OR RDX OR Cyclotrimethylenetrinitramine OR "cyclotrimethylene trinitramine" OR "Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine" OR "Hexahydro

- 1,3,5

- trinitro

- s

- triazine" OR Hexogen OR "1,3,5

- trinitro

1,3,5

- triazine" OR "1,3,5

- Triaza

- 1,3,5

- trinitrocyclohexane" OR "1,3,5

- Trinitro

1,3,5

- triazacyclohexane" OR "1,3,5

- Trinitrohexahydro

- 1,3,5

- triazine" OR "1,3,5

- Trinitrohexahydro

- s

- triazine" OR "1,3,5

- Trinitroperhydro

- 1,3,5

- triazine" OR "Esaidro

- 1,3,5

- trinitro

- 1,3 ,5

- triazina" OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazin" OR "Perhydro

- 1,3,5

- trinitro

- 1,3,5

- triazine" OR 26 titles screened (6 selected for full records and added to HERO)

This document is a draft for review purposes only and does not constitute Agency policy. 

### B

- 7 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Database Terms Hits Cyclotrimethylenenitramine OR Trimethylenetrinitramine OR "Trimethylene trinitramine" OR Trimethyleentrinitramine OR "Trinitrocyclotrimethylene triamine " OR Trinitrotrimethylenetriamine OR "CX 84A" OR Cyklonit OR Geksogen OR Heksogen OR Hexogeen OR Hexolite OR "KHP 281" OR "PBX (af) 108" OR "PBXW 108(E)" OR "Pbx(AF) 108") NOT (patent/dt OR tscats/fs)) AND (py> 2012 OR ed> 2012 0101)) AND (chronic OR immunoto x? OR neurotox? OR toxicokin? OR biomarker? OR neurolog? OR pharmacokin? OR subchronic OR pbpk OR epidemiology/st,ct, it) OR acute OR subacute OR ld50# OR lc50# OR (toxicity OR adverse OR poisoning)/st,ct,it OR inhal? OR pulmon? OR nasal? OR lung? OR respi r? OR occupation? OR workplace? OR worker? OR oral OR orally OR ingest? OR gavage? OR diet OR diets OR dietary OR drinking(w)water OR (maximum and concentration? and (allowable OR permissible)) OR (abort? OR abnormalit? OR embryo? OR cleft? OR fetus? OR fo etus? OR fetal? OR foetal? OR fertil? OR malform? OR ovum OR ova OR ovary OR placenta? OR pregnan? OR prenatal OR perinatal? OR postnatal? OR reproduc? OR steril? OR teratogen? OR sperm OR spermac? OR spermag? OR spermati? OR spermas? OR spermatob? OR sper matoc? OR spermatog? OR spermatoi? OR spermatol? OR spermator? OR spermatox? OR spermatoz? OR spermatu? OR spermi? OR spermo? OR neonat? OR newborn OR development OR developmental? OR zygote? OR child OR children OR adolescen? OR infant OR wean? OR offspri ng OR age(w)factor? OR dermal? OR dermis OR skin OR epiderm? OR cutaneous? OR carcinog? OR cocarcinog? OR cancer? OR precancer? OR neoplas? OR tumor? OR tumour? OR oncogen? OR lymphoma? OR carcinom? OR genetox? OR genotox? OR mutagen? OR genetic(w)toxic? O R nephrotox? OR hepatotox? OR endocrin? OR estrogen? OR androgen? OR hormon? OR rat OR rats OR mouse OR mice OR muridae OR dog OR dogs OR rabbit? OR hamster? OR pig OR pigs OR swine OR porcine OR goat OR goats OR sheep OR monkey? OR macaque? OR marmoset? O R primate? OR mammal? OR ferret? OR gerbil? OR rodent? OR lagomorpha OR baboon? OR bovine OR canine OR cat OR cats OR feline OR pigeon? OR occupation? OR worker? OR epidem?) AND (biosis/fs OR caplus/fs))

Notes: Duplicates were removed

. Toxcenter

Date limit: 11/ 2012

− 1/ 2014 (((121

- 82

- 4 OR Cyclonite OR RDX OR Cyclotrimethylenetrinitramine OR "cyclotrimethylene trinitramine" OR "Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine" OR "Hexahydro

- 1,3,5

- trinitro

- s

- triazine" OR Hexogen OR "1,3,5

- trinitro

1,3,5

- triazine" OR "1,3,5

- Triaza

- 1,3,5

- trinitrocyclohexane" OR "1,3,5

- Trinitro

1,3,5

- triazacyclohexane" OR "1,3,5

- Trinitrohexahydro

- 1,3,5

- triazine" OR "1,3,5

- Trinitrohexahydro

- s

- triazine" OR "1,3,5

- Trinitroperhydro

- 1,3,5

- triazine" OR "Esaidro

- 1,3,5

- trinitro

- 1,3,5

- triazina" OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazin" OR "Perhydro

- 1,3,5

- trinitro

- 1,3,5

- triazine" OR Cyclotrimethylenenitramine OR Trimethylenetrinitramine OR "Trimethylene trinitr amine" OR Trimethyleentrinitramine OR "Trinitrocyclotrimethylene triamine" OR Trinitrotrimethylenetriamine OR "CX 84A" OR Cyklonit OR Geksogen OR Heksogen OR Hexogeen OR Hexolite OR "KHP 281" OR "PBX (af) 108" OR "PBXW 108(E)" OR "Pbx(AF) 108") NOT (patent /dt OR tscats/fs)) AND (py> 2012 OR ed> 2012 1101)) AND (chronic OR immunotox? OR neurotox? OR toxicokin? OR biomarker? OR neurolog? OR pharmacokin? OR subchronic OR pbpk OR epidemiology/st,ct, it) OR acute OR subacute OR ld50# OR lc50# OR (toxicity OR advers e OR poisoning)/st,ct,it OR inhal? OR pulmon? OR nasal? OR 20 titles screened (0 selected for full records; none added to HERO )

This document is a draft for review purposes only and does not constitute Agency policy. 

### B

- 8 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Database Terms Hits lung? OR respir? OR occupation? OR workplace? OR worker? OR oral OR orally OR ingest? OR gavage? OR diet OR diets OR dietary OR drinking(w)water OR (maximum and concentration? and (allowable OR per missible)) OR (abort? OR abnormalit? OR embryo? OR cleft? OR fetus? OR foetus? OR fetal? OR foetal? OR fertil? OR malform? OR ovum OR ova OR ovary OR placenta? OR pregnan? OR prenatal OR perinatal? OR postnatal? OR reproduc? OR steril? OR teratogen? OR spe rm OR spermac? OR spermag? OR spermati? OR spermas? OR spermatob? OR spermatoc? OR spermatog? OR spermatoi? OR spermatol? OR spermator? OR spermatox? OR spermatoz? OR spermatu? OR spermi? OR spermo? OR neonat? OR newborn OR development OR developmental? OR zygote? OR child OR children OR adolescen? OR infant OR wean? OR offspring OR age(w)factor? OR dermal? OR dermis OR skin OR epiderm? OR cutaneous? OR carcinog? OR cocarcinog? OR cancer? OR precancer? OR neoplas? OR tumor? OR tumour? OR oncogen? OR lymphom a? OR carcinom? OR genetox? OR genotox? OR mutagen? OR genetic(w)toxic? OR nephrotox? OR hepatotox? OR endocrin? OR estrogen? OR androgen? OR hormon? OR rat OR rats OR mouse OR mice OR muridae OR dog OR dogs OR rabbit? OR hamster? OR pig OR pigs OR swine O R porcine OR goat OR goats OR sheep OR monkey? OR macaque? OR marmoset? OR primate? OR mammal? OR ferret? OR gerbil? OR rodent? OR lagomorpha OR baboon? OR bovine OR canine OR cat OR cats OR feline OR pigeon? OR occupation? OR worker? OR epidem?) AND (bios is/fs OR caplus/fs))

Notes: Duplicates were removed

. Toxcenter

Date limit: 11/ 2013

− 1/ 2015 (((121

- 82

- 4 OR Cyclonite OR RDX OR Cyclotrimethylenetrinitramine OR "cyclotrimethylene trinitramine" OR "Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine" OR "Hexahydro

- 1,3,5

- trinitro

- s

- triazine" OR Hexogen OR "1,3,5

- trinitro

1,3,5

- triazine" OR "1,3,5

- Triaza

- 1,3,5

- trinitrocyclohexane" OR "1,3,5

- Trinitro

1,3,5

- triazacyclohexane" OR "1,3,5

- Trinitrohexahydro

- 1,3,5

- triazine" OR "1,3,5

- Trinitrohexahydro

- s

- triazine" OR "1,3,5

- Trinitroperhydro

- 1,3,5

- triazine" OR "Esaidro

- 1,3,5

- trinitro

- 1,3,5

- triazina" OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazin" OR "Perhydro

- 1,3,5

- trinitro

- 1,3,5

- triazine" O R Cyclotrimethylenenitramine OR Trimethylenetrinitramine OR "Trimethylene trinitramine" OR Trimethyleentrinitramine OR "Trinitrocyclotrimethylene triamine" OR Trinitrotrimethylenetriamine OR "CX 84A" OR Cyklonit OR Geksogen OR Heksogen OR Hexogeen OR Hexol ite OR "KHP 281" OR "PBX (af) 108" OR "PBXW 108(E)" OR "Pbx(AF) 108") NOT (patent/dt OR tscats/fs)) AND (py > 2013 OR ed> 2013 1101)) AND (chronic OR immunotox? OR neurotox? OR toxicokin? OR biomarker? OR neurolog? OR pharmacokin? OR subchronic OR pbpk OR epi demiology/st,ct, it) OR acute OR subacute OR ld50# OR lc50# OR (toxicity OR adverse OR poisoning)/st,ct,it OR inhal? OR pulmon? OR nasal? OR lung? OR respir? OR occupation? OR workplace? OR worker? OR oral OR orally OR ingest? OR gavage? OR diet OR diets OR dietary OR drinking(w)water OR (maximum and concentration? and (allowable OR permissible)) OR (abort? OR abnormalit? OR embryo? OR cleft? OR fetus? OR foetus? OR fetal? OR foetal? OR fertil? OR malform? OR ovum OR ova OR ovary OR placenta? OR pregnan? O R prenatal OR perinatal? OR postnatal? OR reproduc? OR steril? OR teratogen? OR sperm OR spermac? OR spermag? OR spermati? OR spermas? OR spermatob? OR spermatoc? OR spermatog? OR spermatoi? OR spermatol? OR spermator? OR spermatox? OR spermatoz? OR sperma tu? OR spermi? OR 80 titles screened (3 selected for full records and added to HERO)

This document is a draft for review purposes only and does not constitute Agency policy. 

### B

- 9 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Database Terms Hits spermo? OR neonat? OR newborn OR development OR developmental? OR zygote? OR child OR children OR adolescen? OR infant OR wean? OR offspring OR age(w)factor? OR dermal? OR dermis OR skin OR epiderm? OR cutaneous? OR carcinog? OR cocarcinog? OR cancer? OR precancer? OR neoplas? OR tumor? OR tumour? OR oncogen? OR lymphoma? OR carcinom? OR genetox? OR genotox? OR mutagen? OR genetic(w)toxic? OR nephrotox? OR hepatotox? OR endocrin? OR estrogen? OR androgen? OR hormon? OR rat OR rats OR

mouse OR mice OR muridae OR dog OR dogs OR rabbit? OR hamster? OR pig OR pigs OR swine OR porcine OR goat OR goats OR sheep OR monkey? OR macaque? OR marmoset? OR primate? OR mammal? OR ferret? OR gerbil? OR rodent? OR lagomorpha OR baboon? OR bovine OR canine OR cat OR cats OR feline OR pigeon? OR occupation? OR worker? OR epidem?) AND (biosis/fs OR caplus/fs))

Note: Duplicates were removed

. 

### 1

2

3 **Ta ble B** 

**- 2. Summary of detailed search strategies for RDX (DTIC)** 

Database Terms Hits

DTIC Online Access Controlled

Date searched: 1/2015 Synonyms in all fields search box ("121

- 82

- 4" OR "RDX" OR "Cyclotrimethylenetrinitramine" OR "Cyclonite" OR "cyclotrimethylene trinitramine" OR "Hexogen" OR "Hexahydro

- 1,3,5

- trinitro

1,3,5

- triazine" OR "Hexahydro

- 1,3,5

- trinitro -s- triazine" OR "Trimethylene trinitramine" OR "Trimethylenetrinitramine" OR "Hexolite" OR "Trinitrotrimethylenetriamine") Keywords in citation box ("toxicity" OR "toxicology" OR "poisoning" OR "cancer" OR "carcinogens" OR "carcinogen" OR "neoplasms" OR "neoplasm" OR "oncogenesis" OR "teratogenic compounds" OR "lethality" OR "death" OR "body weight" OR "immunology" OR "genotoxicity" OR "mutagenicity" OR "mutagens" OR "mutations" OR "oral " OR "gavage" OR "inhalation" OR "dermal" OR "metabolism" OR "pharmacokinetics" OR "pharmacokinetic" OR "PBPK" OR "pharmacology" OR "organs" OR "skin" OR "tissues" OR "body fluids" OR "toxic agents" OR "rats" OR "mice" OR "mouse" OR "rat") Limited to Con tent type: Documents Distribution: Approved for Public Release 826 (85 selected and added to HERO)
Distribution: U.S. Gov't and Contractors 239 (0 selected and added to HERO)
Distribution: U.S. Gov't Only 199 ( 0 selected and added to HERO)

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 10 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**T able B** 

**- 3. Processes used to augment the search of core databases for RDX**  1

Selected k ey r eference(s) or s ources Date Additional r eferences i dentified
“Forward” and “backward” Web of Science searches a Sweeney et al. (2012a)

. Assessing the non

- cancer risk for RDX (hexahydro

1,3,5

- trinitro

- 1,3,5

- triazine) using physiologically based pharmacokinetic (PBPK) modeling. Regul Toxicol Pharmacol 62(1):107−114. ( forward search ) 1 search result 3/ 2013 0 citations added

Sweeney et al. (2012b)

. Cancer mode of action, weight of evidence, and proposed cancer reference value for hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine (RDX). Regul Toxicol Pharmacol 64(2):205−224 ( backwards search ) 0 search results 3/ 2013 0 citations added

Sweeney et al. (2012a)

. Assessing the non

- cancer risk for RDX (hexahydro

1,3,5

- trinitro

- 1,3,5

- triazine) using physiologically based pharmacokinetic (PBPK) modeling. Regul Toxicol Pharmacol 62(1):107−114. ( review of 35 references cited in this paper ) 3/ 2013 0 citations added

Sweeney et al. (2012b)

. Cancer mode of action, weight of evidence, and proposed cancer reference value for hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine (RDX). Regul Toxicol Pharmacol 64(2):205−224 ( review of 69 references cited in this paper ) 3/ 2013 3 citations added

Online regulatory sources
Combination of Chemical Abstracts Registry Number ( CASRN ) and synonyms searched on the following websites:

Agency for Toxic Substances and Disease Registry ( ATSDR ) http://www.atsdr.cdc.gov/substances/index.asp (Note: the reference list for the ATSDR toxicological profile for RDX was compared to the search results and relevant references were added) California Environmental Protection Agency (Office of Environmental Health Hazard Assessment) ( http://www.oehha.ca.gov/risk.html ) eChemPortal ( http://www.echemportal.org/echemportal/participant/page.action? pageID=9 ) EPA Acute Exposure Guideline Levels ( http://www.epa.gov/oppt/aegl/pubs/chemlist.htm ) EPA

— IRISTrack/New Assessments and Reviews ( http://cfpub.epa.gov/ncea/iristrac/ ) to find dates ( http://www.epa.gov/ncea/iris/index.html ) to find data EPA National Serv ice Center for Environmental Pu blications ( NSCEP ) ( http://www.epa.gov/ncepihom/ ) EPA Science Inventory ( http://cfpub.epa.gov/si/ ) Federal Docket www.regulations.gov Health Canada First Priority List Assessments ( htt p://www.hc

- sc.gc.ca/ewh

- semt/pubs/contaminants/psl1

- lsp1/index

eng.php ) 4/11/ 2012
1/27/2014

1/22/2015 15 citations added

1 citation added

0 citations added

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 11 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Selected k ey r eference(s) or s ources Date Additional r eferences i dentified Health Canada Second Priority List Assessments ( http://www.hc

- sc.gc.ca/ewh

- semt/pubs/contaminants/psl2

- lsp2/index

eng.php ) Internatio n al Agency for Research on Cancer ( IARC ) ( http://monographs.iarc.fr/htdig/search.html ) International Programme on Chemical Safety ( IPCS ) INCHEM ( http://www.inchem.org/ ) National Academy of Science ( NAS) via the National Academies Press ( http://www.nap.edu/ ) National Cancer Institute ( NCI ) ( http://www.cancer.gov ) National Center for Toxicological Research ( NCTR ) ( http://www.fda.gov/AboutFDA/CentersOffices/OC/OfficeofScientific and MedicalPrograms/NCTR/default.htm ) National Institute o f Environmental Health Sciences (NIEHS) ( http://www.niehs.nih.gov/ ) National Ins t itute for Occupational Safety and Health (NIOSH) NIOSHTIC 2 ( http://www2a.cdc.gov/nioshtic

- 2/ ) National Toxicology Program ( NTP )— RoC, status, results, and management reports ( http://ntpsearch.niehs.nih.gov/query.html ) World Health Organization ( WHO ) assessments

— Concise International Chemical Assessment Documents ( CICAD s), Environmental Health Criteria ( EHC ) ( http://www.who. int/ipcs/assessment/en/ ) 

### 1 2 3 4 a Sweeney et al. (2012a) and Sweeney et al. (2012b) were selected for forward and backward searching in the Web of Science as the two more recent reviews of the health effects of RDX toxicity in the published literature.

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 12 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine



# APPENDIX C. INFORMATION IN SUPPORT OF HAZARD IDENTIFICATION AND DOSE

- RE S PONSE ANALYSIS

**C.1. TOXICOKINETICS**  1
2 3
4
5 6
7

8
9
10
11
12
13
14
15 16
17 18 19

20
21
22
23
24 25
26
27 28 Hexahydro- 1,3,5

- trinitro

- 1,3,5

- triazine ( RDX ) is absorbed following exposure by inhalation and oral routes . The rate and extent of absorption are dependent upon the dosing preparation. RDX is systemically distributed, can be transfer re d from mother to fetus
, and can transfer in maternal milk . Metabolism of RDX is extensive and includes denitration, ring cleavage, and generation of CO2 possibly through cytochrome P450 (CYP450) . RDX metabolites are eliminated primarily via urinary excre tion and exhalation of CO2.

**C.1.1. Absorption**  Absorption of RDX following oral exposure has been demonstrated in humans and laboratory animals (rats, mice, swine, and voles) through measurement of radiolabeled RDX and/or metabolites in excreta (urine and expired air) and tissues (including blood)

. Quantitative estimates of oral absorption (e.g., oral bioavailability or fractional absorption) are not available in humans. Results of animals studies indicate that oral bioavailability ranges from approximately 50 to 90% and may vary based on the physical form of RDX and the matrix (e.g., soil, plants) in which it is administered

. Studies investigating absorption of RDX following inhalation exposure were not identified . Results of an intratracheal administration stu dy in rats provide limited evidence of absorption of RDX from the respiratory tract . The only data evaluating dermal absorption of RDX is provided by in vitro studies showing that RDX can be absorbed through excised skin of humans and animals.

Oral Absor ption Quantitative information on blood levels following accidental exposure to RDX is limited to two studies of accidental oral exposures ( Küçükardali et al., 2003 ; Woody et al., 1986 ) and one

s tudy of mixed derm al and inhalation exposure ( Özhan et al., 2003 ) . A number of qualitative case
st udies of accidental exposures with similar toxic effects provide additional support that RDX is absor bed into the body ( Hett and Fichtner, 2002 ; Harrell- Bruder and Hutchins, 1995 ; Goldberg et al., 1 992 ; Ketel and Hughes, 1972 ; Hollander and Colbach, 196 9 ; Stone et al., 1969 )

. The oral ab sorption of RDX in humans was demonstrated in a case report of a 3

- year
- old male child wh o ingested plasticized RDX material that adhered to his mother’s work boots and clothing ( Woody et

This document is a draft for review purposes only and does not constitute Agency policy. 

### B-1 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

al., 1986 )

. RDX was measured in serum, urine, cerebrospinal fluid, and feces

. Based on a kinetic 1
2 3
4
5 6
7 8
9 10
11
12 13
14
15 16
17 18
19 20
21
22
23
24
25
26
27
28
29
30 31
32
33 34
35
36 an alysis of the serum RDX concentrations, the dose was estimated to be 85 mg/kg and the first

order absorption rate constants were estimated to be 0.34

− 2.20 hour

− 1 

### ( Woody et al., 1986 ) 2



### . Sweeney et al. (2012a) estimated the absorption rate constant for this same subject to be 0 .060 hour

− 1 

### . The large range in the calculated absorption rate constants re sulted from uncertainty in the dose and time to peak serum RDX levels, and the models that were used to simulate the RDX toxicokinetics. Özhan et al. (2003) su mmarized plasma RDX levels in five military personnel who

w ere accidentally exposed to toxic levels of RDX. Although Özhan et al. (2003) reported that
p ersonnel were exposed through dermal and inhaled routes, secondary oral exposure may have occurred

. Based on physiologically based pharmacokinetic ( PBPK ) model fits to the plasma RDX concentration data, Sweeney et al. (2012a) estimated a first

- order absorption rate constant of 0 .033 hour

− 1



### . Küçükardali et al. (2003) summarized plasma RDX levels in five military personnel

w ho ingested toxic levels of RDX (doses were not reported). RDX was detected in plasma of all patients within 3 hours after ingestion. Quantitative data to directly support estimates of oral bioavailability are available from studies in rats and mice ( Guo et al., 1985 ; Schneider et al., 1978 , 1977 )

. Results of single and

r epeated oral dose studies in adult Sprague

- Dawley rats indicate that approximately 83

− 87% of the administered dose i s absorbed from the gastrointestinal (GI) tract

. Following gavage administration of 50 mg/kg [ 14 

### C]

- RDX dissolved in dimethylsulfoxide (DMSO), approximately 90% of the administered carbon

- 14 was recovered 4 days after dosing, with ~3% in feces, 34% in urin e, 43% in expired air, and 10% in the carcass ( Schneider et al., 1977 )

. It is unclear if the carcass
in cludes the GI tract, which may include unabsorbed RDX . Assuming that all of the carbon

- 14 in feces represents unabsorbed RDX (rather than RDX that was absorbed and subsequently secreted into the intestine), results of this study indicate that at least 87% of the administered dose was absorbed from the GI tract

. Similar results were observed following repeated daily oral exposure of Sprague

- Dawley rats t o [ 14 

### C]

- RDX by gavage (in DMSO) or drinking water for 1 week

. Based on recovery of carbon

- 14 in urine and expired air and the carbon

- 14 retained in carcass, approximately 83% (drinking water) to 85 % (gavage) of the administered dose was absorbed ( Schn eider et al., 1978 ). An estimate of oral bioavailability in rats can also be obtained from data on blood RDX concentrations reported in Krishnan et al. (2009) . Male Sprague

- Dawley rats received a single
i ntravenous (i.v.) (0.77 or 1.04 mg/kg) or oral (1.53 or 2.07 mg/kg, dissolved in water) dose of RDX

. Estimates of bioavailability were obtained based on the reported blood RDX concentrations, terminal elimination rate constants (estimated for this review by fitting the serum RDX data with a first- order exponential function, see Table C -5 in Section C.1.4, Excretion ) and the blood area under the curve ( AUC) values (calculated for this review using the trapezoid rule extrapolated to infinite

2 Woody et al. (1986) repo rted the absorption rate constants in units of L/hour; however, this appears to have been a typographical error for 1/hour or hour

− 1

. This document is a draft for review purposes only and does not constitute Agency policy. 

### B-2 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

t ime)

. Calculated dose

- adjusted AUC values were 9.6 and 8.4 hours

- kg/L for the i.v. studies and 1
2 3
4
5 6
7 8
9 10
11
12 13
14
15 16
17 18
19 20
21
22
23
24
25
26
27
28
29
30 31
32
33 34
35
36 37
38 4.7 and 6.0 hours

- kg/L for the oral dosing studies . These AUC values correspond to estimated oral bioavailability ranging from 50 to 70% . Recovery of administered radiolabel was incomplete (~90% of the administered carbon

- 14) in the studies ( S chneider et al., 1978
, 1977 ) ; therefore, it is
p ossible that oral bioavailability is actually higher than 83

− 87%. Guo et al. (1985) reported data on b lood tritium kinetics in mice that received i.v. (0.055 mg RDX or ~2.5 mg/kg body weight) or oral (50 mg/kg) doses of [ 3 

### H]

- RDX

. Based on the reported blood tritium concentratio ns (% of dose/g) and terminal t 1/2 

### values for concentrations of tritium in blood (1.1 days for i.v. and 2.2 days for oral), the corresponding AUCs of the blood concentration versus time curves were calculated (calculated for this review using the trapezoid rule extrapolated to infinite time) to be 30 and 16 hours

- % dose/g for i.v. and oral dosing, respectively

. This corresponds to an oral bioavailability of RDX

- derived tritium concentration of approximately 50% (i.e., 16/30). In Yucatan miniature swine ad ministered a single dose of [ 14 

### C]

- RDX (43

− 45 mg/kg as a suspension in carboxymethylcellulose), approximately 0.8

− 6% of the administered carbon

- 14 was eliminated in feces 24 hours after dosing ( Musick et al., 2010 ; Major et al., 2007 )

. Although results

o f swine studies suggest that GI absorption of RDX was nearly complete, data cannot be used to determine a quantitative estimate of oral bioavailability because it is unlikely that fecal excretion of unabsorbed RDX was complete 24 hours after dosing ( Snoeck et al., 2004 ). Oral bioavailability of RDX appears to vary depending upon the physical form of RDX and the matrix (e.g., soil, vegetation) in which it is administered. Schneider et al. (1977) compared the

o ral absorption of a single 100 mg/kg gavage dose of coarse granular [ 14 

### C]

- RDX as a slurry in isotonic saline with a single 50 mg/kg gavage dose of a finely powdered [ 14 

### C]

- RDX solution in saline in Sprague

- Dawley rats. Plasma carbon

- 14 levels were measured for 24 hours after dosing. For both [ 14 

### C]

- RDX preparations, peak plasma levels of carbon

- 14 were observed 24 ho urs after oral administration, with a higher 24

- hour plasma concentration for the 50 mg/kg dose (~4.7 μg/mL) compared to the 100 mg/kg dose (3.12 μg/mL)

. Results of this study indicate that the oral bioavailability of RDX may be greater for the finely pow d ered preparation than for the coarse granular preparation consistent with a greater surface area available for absorption with finely powdered RDX . However, blood levels were only measured 24 hours after dosing, and the lower 24

- hour carbon

- 14 plasma concentration for the coarse granular preparation could be due to slower absorption of coarse RDX granules compared with fine RDX powder, rather than lower overall bioavailability. Oral bioavailability of RDX is lower when administered as RDX

- contaminated so il or when RDX is in plant materials that were grown on RDX

- contaminated soils. Crouse et al. (2008)
in vestigated the oral bioavailability of RDX in contaminated soils relative to pure RDX by comparison of the A UC for the RDX blood concentration versus time curves. Adult male Sprague

- Dawley rats were administered oral doses (in gelatin capsules) of pure RDX (99.9% purity; neat) or an equivalent amount of RDX in contaminated soils from the Louisiana Army Ammuniti on

This document is a draft for review purposes only and does not constitute Agency policy. 

### B-3 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Pl ant (AAP) or Fort Meade

. Blood concentrations for rats dosed with Louisiana AAP soil 1
2 3
4
5 6
7 8
9 10
11
12 13
14
15 16
17 18
19 20
21
22
23
24
25
26
27
28
29
30 31 32
33
34
35
36 37
38 (1.24 mg/kg) and neat RDX (1.24 mg/kg) peaked at approximately 6 hours . The AUC and 6

- hour RDX blood concentration were both approximately 25% lower for Louisiana A AP soil than for neat RDX ( p ≤ 0.003 for AUC), suggesting that oral bioavailability of RDX from Louisiana AAP soil was 25% lower than neat RDX. For Fort Meade soil (0.2 mg/kg), RDX blood concentrations peaked at 6 hours compared to 4 hours for neat RDX (0 .2 mg/kg) . The 4

- hour blood concentration for Fort Meade soil was approximately 15% lower than for neat RDX, although the AUC for Fort Meade soil was only 5% lower than for neat RDX (not statistically significant). Collectively, these results suggest tha t RDX in soil is absorbed following oral exposure and that it has a lower bioavailability than neat RDX. F ellows et al. (2006) showed that plants (alfalfa shoots and corn leaves) incorporated [ 14 

### C]

- RDX grown on [ 14 

### C]

- RDX

- amended soils

. [ 14 

### C]

- RDX and plant metabolites of [ 14 

### C]

- RDX were absorbed by voles following oral administration ( Fellows et al., 2006 )

. In adult male prairie voles ( M icrotus ochrogaster ) fe d diets containing RDX incorporated in plants for 5 or 7 days (average RDX dose per animal of 2.3 mg/kg

- day), 94.8 and 96.6% (respectively) of the administered carbon

- 14 was eliminated in excreta (combined feces, urine, and CO 2 

### ) and 3

− 5% was retained in th e carcass

. Feces, urine, and CO 2 

### contained 74

− 79, 13

− 14, and 8

− 12% of the total carbon

- 14 in excreta, respectively

. Based on carbon

- 14 elimination in urine and CO 2 

### plus that retained by the carcass, the study authors estimated the oral bioavailability of plant

- derived RDX to be >20%. However, if biliary excretion of RDX and/or RDX metabolites is a major excretory pathway in voles (as is the case with mice), estimat es of bioavailability of plant

- derived RDX could be substantially higher. In Yorkshire piglets administered single doses of 5 or 10 mg/kg in gelatin capsules, peak plasma concentrations were proportional to the administered dose ( Bannon, 2006 ) . However, the
p otential for dose

- dependence has not been evaluated over a wide range of doses. RDX appears in blood within 1 hour following oral dosing; however, the rate of absorption may depend upon the physical form or dose of RDX ( Bannon et al., 2009a ; Crouse et al., 2008 ; Bannon, 2006 ; Guo et al., 1985 ; MacPhail et al., 1985 ; Schneider et al., 1977 )

. Oral absorption of R DX was rapid in LACA mice following stomach perfusion with [ 3 

### H]

- RDX (50mg/kg in methyl cellulose) ( Guo et al., 1985 ) . The tritium radiolabel was detected in blood 15 minutes following d osing and the highest concentrations in blood were observed 30 minutes after dosing. Based on

a n analysis of the blood tritium concentration kinetics, the authors estimated an absorption rate constant of 8.7 hour −1

. In Sprague

- Dawley rats administered single doses (0.2

− 18.0 mg/kg) of RDX in gelatin capsules, peak blood RDX concentrations were obser ved between 2.5 and 6 hours ( Bannon et al., 2009a ; Krishnan et al., 2009 ; Crouse et al., 2008 )

. Peak blood concentrations

o ccurred at 24 hours after Sprague

- Dawley rats were administered a single oral dose (100 mg/kg) of coarse granular RDX in saline ( Schneider et al., 1977 )

. Similarly, peak RDX blood concentrations
in swine administered single doses (5

− 10 mg/kg) of finally powdered (>98% pure) RDX in gelatin capsules occurred at 3

− 8 hours after dosing ( Bannon et al., 2009a ) , compared to 24 hours after a

This document is a draft for review purposes only and does not constitute Agency policy. 

### B-4 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

single dose (100 mg/kg) of RDX administered as a finely powdered in saline ( Bannon et al., 2009a ; 1
2 3
4
5 6
7 8
9
10
11

12
13
14
15 16
17 18 19
20
21

22
23
24 25
26
27 28
29
30
31 32
33
34 35
36
37 Schneider et al., 1977 )

. Peak plasma concentrations in Yucatan miniature swine administered a

s ingle dose of [ 14 

### C]

- RDX (45 mg/kg as a suspension in carboxymethylcellulose) were reached within 6

− 12 hours after dosing ( Musick et al., 2010 ). Krishnan et al. (2009) and Sweeney et al. ( 2012a) estimated absorption rates in rats dosed with higher doses of coarse granular RDX to be
ap proximately 100 times slower than absorption rates in rats dosed with lower doses of finely powdered neat RDX preparations or neat RDX dissolved in aqueous vehicles

. For example, Krishnan et al. (2009) estimated the absorption rate constant to be 0.75 hour −1 for rats dosed with

n eat RDX dissolved in water (1.53 or 2.07 mg/kg) or neat RDX in a gelatin capsule (0.2 or 1.24 mg/kg) ( Crouse et al., 2008 ) , compared to 0.0075 hour

− 1 

### for rats dosed with coarse granular RD X (100 mg/kg) ( Schneider et al., 1977 )

. Inhalation Absorption Studies evaluating absorption of RDX in humans following inhalation expo sure were not identified

. Several case reports have documented seizures and other neurological effects in individuals exposed to RDX either in a manufacturing setting or in the course of using RDX as a cooking fuel ( Testud et al., 1996a ; T estud et al., 1996b ; Ketel and Hughes, 1972 ; Hollander and C olbach, 1969 ; Kaplan et al., 1965 ; Barsotti and Crotti, 1949 ) . These reports suggest that RDX may be absorbed by the respiratory system . However, in several cases, the study authors were unable to clearly identify the primary route of exposure. In some cases, incidental oral exposure was suggested

. Studies in laboratory animals have not investigated the absorption of RDX following inhalation exposure.

Dermal Absorption In vitro studies have demonstrated the dermal absorption of RDX in human and pig skin ( Reddy et al., 2008 ; Reifenrath et al., 2008 ). Reddy et al. (2008) reported that 5.7% of the applied R D X dose (in acetone) was absorbed into excised human skin in 24 hours

. Dermal absorption of [ 14 

### C]

- RDX from both a low

- carbon (1.9%) and a high

- carbon (9.5%) soil was also assessed in this system . Approximately 2.6% of the RDX applied in the low

- carbon so il and 1.4 % applied in the high

- carbon soil was absorbed after 24 hours

. Thus, the dermal absorption of RDX from soils was reduced when compared with absorption from acetone, and absorption was lower in the high

- carbon soil than in the low

- carbon soil. Reifenrath et al. (2008) investigated the influence of skin surface moisture conditions, soil

c arbon content, and soil aging on the in vitro percutaneous penetration of [ 14 

### C]

- labeled RDX in excised pig skin

. Mean skin absorption of RDX was higher for low

- carbon soil (1. 2%), regardless of soil age and skin surface moisture . Absorption and evaporation were <1% for RDX regardless of soil type and age. While dermal absorption of certain munitions (e.g., 2,6

- dinitrotoluene) is greatly enhanced by hydration of the skin surfa ce, hydration had minimal effect on RDX
, mostly due to the lack of RDX volatility (e.g., <0.5% evaporation).

This document is a draft for review purposes only and does not constitute Agency policy. 

### B-5 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**C .1.2.**  **Distribution** 1
2
3
4
5 6
7 8 9
10
11
12 13
14
15 16
17 18 19
20
21
22 23
24
25 26
27
28
29 30
31
32 33
34
35
36 37 I nformation on the distribution of absorbed RDX in humans is limited to a few cases of accidental exposures to RDX that provide data o n the kinetics of RDX in blood and cerebrospinal fluid ( Küçükardali et al., 2003 ; Özhan et al., 2003 ; Woody et al., 1986 )

. Concentrations of RDX in

s erum and cerebrospinal fluid were similar (11 and 9 mg/L, respectively) in a child 24 hours after ingesting an estimated dose of 85 mg/kg RDX ( Woody et al., 1986 )

. More extensive information on
t issue distribution is available for animals, including mice, rats, and swine ( Musick et al., 2010 ; Banno n, 2006 ; Reddy et al., 1989 ; Guo et al., 1985 ; MacPhail et al., 1985 ; Schneider et al., 1977 )

. In
t hese studies, RDX or radiolabeled RDX ([ 14 

### C] or [ 3 

### H) was administered by the oral, intraperitoneal (i.p.), i.v., or intratracheal route and the distribution of the RDX or radiolabel was measured. Since metabolism of RDX can result in loss of carbon

- 14 or triti um from the parent compound, the distribution of radiolabel will not necessarily reflect the distribution of RDX ( Schneider et al., 1977 )

. To compare tissue distributions in studies in which animals received different doses by different routes of administ ration, distribution data are expressed as ratios of tissue RDX or radiolabel to that of either whole blood or plasma, whichever was reported. RDX in blood distributes into red blood cells (RBCs) and plasma to achieve concentration ratios that are close t o unity

. The plasma:whole blood carbon

- 14 ratio in swine that received a single oral dose of [ 14 

### C]

- RDX (4 5 mg/kg) was approximately 1.3 ( Musick et al., 2010 ) , and whole rat blood incubated in vitro

w ith RDX had a plasma: RBC RDX ratio of approximately 1.0 ( Krishnan et al., 2009 )

. As a result of
t he similarity between plasma and whole blood concentrations, tissue distribution is approximately equivalent when expressed as ratios of blood or plasma. Studies co nducted in rats, mice, and swine indicate that absorbed RDX distributes to many different tissues. Schneider et al. (1977) estimated the volume of distribution of RDX to be
ap proximately 2.18 L/kg in rats, based on plasma RDX kinetics in rat s that received a single i.p. dose of RDX (5

− 6 mg/kg)

. Consistent with this estimate are observations of tissue:blood (or plasma) concentration ratios that exceed 1 in various tissues, including brain (showing that RDX can cross the blood:brain barrier), heart, kidney, and liver ( Musick et al., 2010 ; Bannon et al., 2006 ; MacPhail et al., 1985 ; Schneider et al., 1977 )

. Distribution within the brain may not be uniform

. H owever, Bannon et al. (2006) observed tissue:blood concentrations for RDX of approximately 4 in b rain hippocampus and 3.5 in brain cortex of swine that received a single oral dose of 10 mg/kg [ 14 

### C]

- RDX, although this is the only study that reported distribution for brain regions

. Reported tissue:blood (or plasma) concentration ratios of RDX 24 hours following a single dose (oral or i.p.) were 1

− 9 for kidney, 1

− 7 for liver, and 1

− 3 for heart ( Table C

- 1) ( Bannon, 2006 ; Schneider et al., 1 977 ) . With repeated oral dosing (e.g., 30

− 90 days), tissue:blood ratios of RDX for these tis sues are

c onsistently greater than unity ( Schneider et al., 1978 )

. There is no consistent evidence that RDX

ac cumulates in fat, although estimates of the fat:blood partition coefficient range from 6 to 8 and exceed that of other tissues ( Sweeney et al., 2012a ; Krishnan et al., 2009 )

.

This document is a draft for review purposes only and does not constitute Agency policy. 

### B-6 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**T able**  **C-1. Distribution of RDX or radiolabel from administered RDX**  **a** 

### 1

2 3 4 5 6 7 8 9 10 11
12
13
14
15 16

17
18
19
20
21
22
23
24
25
26
27 28 Animal Route Dose (mg/kg) Time (hrs) Brain Heart Kidney Liver Fat Source

Swine Oral 45 b 24 0.6 c 0.7 2.4 7.3 0.4 Musick et al. (2010)

Swine Oral 10 d 3 3.5

− 4.0 d 2 ≤1 <1 NA g Bannon et al. (2006)

Swine Oral 100 d 24 1.5 c 1.1 1.2

− 1.9 0.9 1.8 Schneider et al. (1977)

Rat Oral 100 d 24 3.4 c 2.9 6.6 0.7 NA Schneider et al. (1977)

Rat i.p. 50 d 2 3.4 c 2.6 8.8 5.7 NA Schneider et al. (1977)

Rat i.p. 500 d ≤6.5 2.5 c 2.1 4.8 3.3 NA Schneider et al. (1977)

Mouse Oral 50 e 24 1 c 0.8 1 1.4 0.8 Guo et al. (1985)

Mouse i.v. 2.5 e 24 0.6 f 0.8 0.7 1.6 0.4 Guo et al. (1985)

a Values are tissue:blood or tissue:plasma ratios following a single dose of either RDX, [ 14 C]- RDX, or [ 3 H]

- RDX.
b Carbon

- 14

. c Tissue:plasma

. d RDX

. e Tritium

. f Tissue:blood

. g Not available

.


### I n rats, RDX can cross the placental:blood barrier resulting in exposure to the fetus, and can also be transported into maternal milk. Hess

- Ruth et al. (2007) detected RDX in the brain tissue of
p ostnatal day ( P ND ) 1 rat pups (concentrations ranged from 0.64 to 7.6 μg/g brain tissue, with no differences between males and females) after maternal exposure to 6 mg/kg RDX via gavage from gestation al day (GD) 6 to PND 10

. RDX was also detected in maternal milk (conce ntrations ranged from 3 to 5.7 μg/mL on PND 1 and from 0.7 to 3.1 μg/mL on PND 10).

**C.1.3. Metabolism** 
The metabolism of RDX is not well characterized

. No studies investigating the metabolism of RDX in humans were identified. Studies in animals indicate that RDX undergoes extensive metabolism, including denitration, ring cleavage, and generation of CO 2 

### . Predominant metabolic pathways and major organs involved in RDX metabolism have not been identified, although results of in vitro studies suggest a role for CYP4 50. RDX undergoes metabolism through processes that generate CO 2 

### . In Sprague

- Dawley rats administered a single 50 mg/kg gavage dose of [ 14 

### C]

- RDX, 43% was recovered as exhaled [ 14 

### CO 2 

### ] after 4 days ( Schneider et al., 1977 )

. Similarly, approximately 30−50% of the radioactivity was

r ecovered as exhaled [ 14 

### CO 2 

### ] in rats administered [ 14 

### C]

- RDX in saturated drinking water or daily gavage for up to 3 months ( Schneider et al., 1978 )

. Metabolism of RDX to CO 2 

### was also observed in
p rairie voles following dietary exposure (average RDX dose per animal of 2.3 mg/kg

- day) to

This document is a draft for review purposes only and does not constitute Agency policy. 

### B-7 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

[ 14 

### C]

- RDX incorporated plant materials for 5

− 7 days, with approximately 9% of the administered 1
2 3
4
5 6
7 8
9 10
11
12 13
14
15 16
17 18
19 20
21
22
23
24
25
26
27
28
29
30 31
32
33 34
35
36 37
38 [ 14 

### C]

- RDX dose eliminated as exhaled [ 14 

### CO 2 

### ] ( F ellows et al., 2006 )

. Terminal metabolites of RDX have been identified in the urine of rats and swine, with very little urinary excretion of parent compound, indicating extensive metabolism of RDX

. Following oral a dministration of a single 50 mg/kg gavage dose of [ 14 

### C]

- RDX, 3.6% of the urinary radioactivity was identified as unmetabolized RDX ( Schneider et al., 1977 ) . Total urinary radiolabel accounted f or about one

- third of the administered label and unmetabolized RDX contributed 3−5% of total urinary radioactivity in rats exposed to [ 14 

### C]

- RDX

- saturated drinking water for 1 or 13 weeks ( Schneider et al., 1978 )

. Similar results were observed in Yucatan swine administered a single 4 5 mg/kg oral dose of [ 14 

### C] -R DX, with approximately 1

− 3.5% of the urinary radioactivity as parent RDX ( Major et al., 2007 )

. Urinary metabolites were not characterized in these studies ( Schneider et al., 1 978 , 1977 )

. However, Schneider et al. (1978) cited unpublished findings in their laboratory
t hat, in addition to carbon dioxide, other one

- carbon intermediates were produced
, including bicarbonate and formic acid. In the environment, the predominant breakdown products of RDX are methylenedinitramie and 4

- nitro -2- diazbutanal ( Sweeney et al., 2012b ; Paquet et al., 2011 )

. RDX metabolism in animals
i s less well understood

. N-N itroso RDX metabolites have been identified as derived through anaerobic metabolism ( ATSDR, 2012 ; Pan et al., 20 07b )

. Based on characterization of RDX

me tabolites in urine and plasma of miniature swine, metabolism of RDX appears to involve loss of nitro groups and ring cleavage ( Musick et al., 2010 ; Major et al., 2007 )

. The two principal urinary

me tabolites identified in miniature swine following a single oral dose of 43 or 45 mg/kg were 4- nitro

- 2,4

- diazabutanal and 4

- nitro

- 2,4

- diaza

- butanamide (see Table C -2)

. Bhushan et al. (2003)

s uggested that the formation of the 4

- nitro

- 2,4

- diazabutanal metabolite occurred via denitration followed by hydroxylation and spontaneous hydrolytic decomposition resulting in ring cleavage and aldehyde formation . In the miniature swine gavage studies, only trace amounts of the nitrosamine RDX metabolites
, hexahydro -1- nitroso

- 3,5

- dinitro

- 1,3,5

- triazine (MNX) and 1

- nitro

3,5

- dinitroso

- 1,3,5

- triazacyclohexane (DNX)
, were found in urine ( Musick et al., 2010 ; Major et al., 2 007 ) . In plasma, most of the radioactivity existed as RDX, with trace levels of MNX, DNX, and 1 ,3,5

- trinitroso

- 1,3,5

- triazacyclohexane (TNX) . The study authors suggested that the trace levels of MNX, DNX, and TNX in plasma may have been formed within the GI tract via sequential nitrogen reduction by intestinal bacteria ( Major et al., 2007 )

. The low levels of these compounds in urine

an d plasma were attributed to the nearly complete absorption of RDX from the GI tract, leaving little parent compound available for bacterial metabolism within the GI tract

. In a study of female deer mice ( Peromyscus maniculatus ) fed diets containing RDX at concent r ations of 12 and 120 mg/kg for 9 days
, MNX and DNX were identified in the stomach
, but TNX was not detected ( Pan
et

al., 2007b ) . MNX and DNX were also measured in various organs of female B6C3F 1 

### mice provided RD X in feed at doses of 0.38

− 522 mg/kg; TNX was also detected in some organ compartments, but not in the liver

. T he authors concluded that RDX can be metabolized into its N

- nitroso compounds

This document is a draft for review purposes only and does not constitute Agency policy. 

### B-8 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

i n mice, but did not identify a mechanism for the formation of the metabolites. Comparing RDX 1
2 3
4

5 6

7 8 9 10
11
12
13
14
15 16
17 18
19
20
21

22
23
24
25
26
27
28
29
30 with MNX and TNX, RDX was the most potent compound at causing overt signs of toxici ty (seizures and mortality) as determined through identification of the median lethal dose using the EPA up

and

- down procedure in deer mice of varying ages ( S mith et al., 2009 ; Rispin et al., 2002 ).

**Table C-2.**  **Principal urinary metabolites of RDX in miniature swine 24 hours**  **after dosing with RDX**

Sample origin Metabolite name Metabolite structure Urine peak 1 M1 4

- Nitro

- 2,4

- diazabutanal

Urine peak 2 M2 4

- Nitro

- 2,4

- diaza

- butanamide

Sources: M ajor et al. (2007) ; Musick et al. (2010)

.


### A lthough the metabolic pathways and major tissues involved in RDX metabolism have not been identified , the re is some evidence for the in volvement of the liver and CYP450 enzymes

. Comparison of hepatic radioactivity to liver concentrations of RDX after a single gavage dose to rats suggested the presence of RDX metabolites and a possible role for hepatic metabolism of RDX ( Schneider et al., 1977 ) . In vitro data indicated that CYP450 may be involved in the metabolism of RD X ( Bhushan et al., 2003 )

. Incubation of RDX with nicotinamide adenine dinucleotide phosphate ( N ADPH ) and rabbit liver CYP450 2B4 under anaerobic conditions produced nitrite, 4

- nitro

2,4

- diazabutanal, formaldehyde, and ammonium ion ( Bhushan et al., 2003 ) . The reaction rate

u nder aerobic conditions was approximatel y one
- third of that observed under anaerobic conditions

. Several CYP450 inhibitors (ellipticine, metyrapone, phenylhydrazine, 1

- aminobenzo

triazole, and carbon monoxide) decreased the formation of RDX metabolites (55−82% inhibition), providing support for the role of CYP450 in RDX metabolism.

**C.1.4. Excretion**  The primary routes of elimination of absorbed RDX are excretion of RDX and metabolites in urine, and exhalation of CO 2 

### liberated from metabolism of RDX ( Sweeney et al., 2012a ; Musick et al., 2 010 ; Krishnan et al., 2009 ; Major et al., 2007 ; Schneider et al., 1977 )

. Tritium derived from

a dministered [ 3 

### H]

- RDX has been detected in mouse gall bladder contents, suggesting biliary secretion in this species ( Guo et al., 1985 ) ; however, biliary secretion of RDX or metabolites has not b een confirmed in other animal species

. Studies conducted in the rat and swine sugge st that metabolism is the dominant mechanism of elimination of absorbed RDX

. In both species, metabolites dominated the carbon

- 14 distribution in urine of animals that received doses of

This document is a draft for review purposes only and does not constitute Agency policy. 

### B-9 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

[ 14 

### C]

- RDX, with RDX accounting for <5% of the urinary carbon

- 14 ( Musick et al., 2010 ; Schneider et 1
2 3
4
5 6
7 8
9 10
11
12 13

14

15 16 17 18 19 20 21 22 23
24
25
26
27
28
29 30 al., 1 977 ). Data on kinetics of elimination of absorbed RDX from blood are available from reports of accidental exposures of humans to RDX ( Table C -3 ). Woody et al. (1986) estimated the elimination
t 1/2 

### to be approximately 15 hours in a child who ingested approximately 85 mg of RDX per kg of body weight

. The t 1/2 

### estimate was based on measured serum concentrations of RDX made between 24 and 120 hours following ingestion for RDX

. Based on plasma RDX concentration data from five adults exposed to RDX (m easurements made between 24 and 96 hours following
exposure ) ( Özhan et al., 2003 )
, a first

- order elimination t 1/2 

### of 20

− 30 hours was derived (calculated f or this review by fitting the serum RDX data to a first

- order exponential function). It needs to be noted that it is not possible to draw reliabl e inferences from these values since they are based on accid ental, acute exposures and, in particular, the data for the child are based on a single set of

mea surements for one individual.

**Table C-3.**  **Elimination t 1/2**  

### values for RDX or radiolabeled RDX

Animal Route Dose (mg/kg) Time a 

### t 1/2 

### (hrs) Source Human (child) Oral 85 b 24

− 120 hrs 15.0 c Woody et al. (1986) Human (adult) Oral NA 24

− 96 hrs 21

− 29 c,d Özhan et al. (2003)

Rat i.v. 5

− 6 0.5 min

− 6 hrs 10 b Schneider et al. (1977)

Rat i.v. 0.8

− 1.0 30 min

− 10 hrs 4.6 c,d Krishnan et al. (2009)

Rat Oral 1.53

− 2.07 1

− 10 hrs 6.9 c,d Krishnan et al. (2009)

Mouse Oral 35, 60, 80 4 5 min

− 4 hrs 1.2 d Sweeney et al. (2012b)

a Observation period follow ing exposure on which the t 1/2 values were based

. b Reported estimate of dose based on blood kinetics

. c Value for blood RDX

. d Calculated for this review based on reported blood RDX concentrations

.



### T he kinetics of elimination of absorbed RDX from blood has been evaluated in rats and mice . I n rats elimination kinetics were biphasic ( Krishnan et al., 2009 ; Guo et al., 1985 ; Schneider et al., 1 977 )

. As shown in Table C -3 , estimated t 1/2 

### values for the terminal elimination phase in rats

r ange from 5 to 10 hours ( Krishnan et al., 2009 ; Schneider et al., 1977 ). Blood concentration time

c ourse measurements of RDX can be used to estimate an apparent metabolism and elimi nation of RDX from blood. The RDX bl ood concentrations reported in Sweeney et al. (2012b) after gavage d osing of 35, 60
, and 80 mg/kg RDX found a consistent terminal elimination t 1/2 

### of approximately 1.2 hours. The elimination t 1/2 

### estimated for rats ( Krishnan et al., 2009 ; Schneider et al., 1977 ) is as

m uch as an order of magnitude longer than mice ( Sweeney et al., 2012b ).

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 10 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**C.1.5. Physiologically**  **B ased**  **P harmacokinetic**  **(PBPK) M**  **odels** 1

2
3
4
5 6
7 8
9
10
11
12
13
14

15

16 17 18

19

20 Ov erview of Available PBPK Models A PBPK model to simulate the pharmacokinetics of RDX in rats was first developed by Krishnan et al. (2009) and improved upon to extend the model to humans and mice ( Sweeney et al., 20 12a ; Sweeney et a l., 2012b )

. The Sweeney et al. (2012a) model consists of six main

co mpartments: blood, brain, fat, liver, and lumped compartments for rapidly perfused tissues and slowly perfused tissues ( Figure C -1) . The model can simulate RDX exposures via the i.v. or oral
route

. Distribution of RDX to tissues is assumed to be flow

- limited . Oral absorption is represented in this model as first

- order uptake from the GI tract into the liver, with 100% of the dose absorbed. RDX is assumed to be cleared by first -order metabolism in the liver. However, there is no representation of the kinetics of any RDX metabolites

. The acslX model code (Advanced Continuous Simulation Language, Aegis, Inc., Huntsville, Alabama) was obtained from the authors of Sweeney et al. (2 012a)

.

Exposure to RDX is by the i.v. or oral route, an d clearance occurs by metabolism in the liver. See Table C

- 4 for definitions of parameter abbreviations. The GI tract is represented as one compartment in Krishnan et al. (2009) (on the left) and two compartments in Sweeney et al. (2012a) (on the right).


### Figure **C-1. PBPK model structure for RDX in rats and humans** 

**.**

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 11 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

The parameter values used in the Sweeney et al. (2012a) rat model are listed in Table C -4

. 1
2 3
4
5 6
7 8
9 10
11
12 13
14

15 16 The physiological model parameter values for cardiac output, tissue volumes
, and blood perfusion of tissues were obtained from the literature ( Timchalk et al., 2002 ; B r own et al., 1997 )

. RDX tissu e:blood partition coefficien ts for liver (PL), brain (PB), and richly perfused tissues (PR) were estimated with an algorithm that relates the measured n

- octanol:water partition coefficient for RDX to reported compositions of water and lipids in specific rat tissues ( Poulin and Theil, 2000 ; Poulin and Kr ishnan, 1995 )

. Tissue:blood partition coefficients for fat (PF) and slowly perfus ed tissues (PS ), as well as the metabolic rate constant (KfC)
, were simultaneously optimized to fit rat blood RDX concentrations following i.v. doses of 0.77 or 1.04 mg/kg RDX ( Krishnan et al., 2009 )
pro ducing values of 5.57, 0.15, and 2.6 kg 0.33 

### /h ou r for PF, PS, and KfC
, respectively

. While the optimized value for PS is much smaller than that used by Krishnan et al. (2009) ( 1.0 kg 0.33 

### /hour)
, the optimized values for PF and KfC were fairly similar to those used by Krishnan et al. (2009) (7 .5 5, and 2.2 kg 0.33 

### /h ou r) . The rat model with these parameter values also had good agreement with blood RDX concentrations after a 5

− 6 mg/kg i.v. exposure ( Schneider et al., 1977 ).

**Table C-4**  **. Parameter values used in the Sweeney et al. (2012a)**  **and Sweeney**  **et a l. (2012b)**  **PBPK models for RDX in rats, humans, and mice**

P a r a m e t e r ( a bbr e v i a t i o n; uni t s ) R a t H um a n Mo us e S ou r c e

Bo d y w e i g h t (BW ; k g ) 0. 3 70 0. 0206 D efa u l t v a l u es ; s t u d y

- s p e ci f i c va l u e s u s e d i f

a v a i l a b l e
C a r d i a c o u t p u t ( K Q C , L /h r /k g 0. 74 ) 15 14 15 T i m ch a l k e t a l

. ( 2002 ) ; B r o w n e t a l

. ( 1997 )

Ti s su e v o l u me s ( f ra c t i o n o f B W )

L i v e r ( K VL ) 0. 04 0. 026 0. 04 T i m ch a l k e t a l

. ( 2002 ) ; B r o w n e t a l

. ( 1997 )

B r a i n ( K VB ) 0. 012 0. 02 0. 012 T i m ch a l k e t a l

. ( 2002 ) ; B r o w n e t a l

. ( 1997 )

F a t (KV F ) 0. 07 0. 21 0. 07 T i m ch a l k e t a l

. ( 2002 ) ; B r o w n e t a l

. ( 1997 )

Ri ch l y p e r f u s e d t i s s u e s ( K VR ) 0. 04 0. 052 0. 04 T i m ch a l k e t a l

. ( 2002 ) ; B r o w n e t a l

. ( 1997 )

B l o o d ( K VV) 0. 06 0. 079 0. 06 T i m ch a l k e t a l

. ( 2002 ) ; B r o w n e t a l

. ( 1997 )

S l o wl y p er fu s ed t i s s u e s ( K V S ) 0. 688 0. 523 0. 688 0. 91 − ( K VL + K VB + K VF + K VR + K VV)

B l o o d f l o w s ( f ra c t i o n o f c a rd i a c o u t p u t )

L i v e r ( K Q L ) 0. 25 0. 175 0. 25 T i m ch a l k e t a l

. ( 2002 ) ; B r o w n e t a l

. ( 1997 )

Br a i n (KQ B) 0. 03 0. 114 0. 03 T i m ch a l k e t a l

. ( 2002 ) ; B r o w n e t a l

. ( 1997 )

F a t (KQ F ) 0. 09 0. 085 0. 09 T i m ch a l k e t a l

. ( 2002 ) ; B r o w n e t a l

. ( 1997 )

S l o wl y p er fu s ed t i s s u e s ( K Q S ) 0. 2 0. 2449 0. 2 T i m ch a l k e t a l

. ( 2002 ) ; B r o w n e t a l

. ( 1997 )

Ri ch l y p e r f u s e d t i s s u e s ( K Q R) 0. 43 0. 3811 0. 43 1 − (KQ L + K Q B + K QF + KQ S )

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 12 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Parameter (abbreviation; units) Rat Human Mouse Source Tissue:blood partition coefficients Liver (PL) 1.2 1.3 1.3 Krishnan et al. (2009) a Brain (PB) 1.4 1.6 1.6 Krishnan et al. (2009) a Richly perfused tissues (PR) 1.4 1.6 1.6 Krishnan et al. (2009) a Fat:blood (PF) 5.57 5.57 5.57 Sweeney et al. (2012a) b Slowly perfused tissues (PS) 0.15 0.15 0.15 Sweeney et al. (2012a) b

Metabolism

First

- order metabolic rate constant (KfC; kg 0.33 /hr) 2.6 9.87 (child); 11.2 (adult) 102 Sweeney et al. (2012a) b,c ; Sweeney et al. (2012b) d GI absorption Dosing via gavage

Absorption from compartment 1 (KAS, /hr) 0.83 0.033 0.51 Sweeney et al. (2012a) c,d,e

Transfer from compartment 1 to compartment 2 (KT, /hr) 1.37 0 0 Sweeney et al. (2012a) c,d

Absorption from compartment 2 (KAD, /hr) 0.0258 0 0 Sweeney et al. (2012a) c,d

Dosing via capsule (KAS, /h r) 0.12 NA NA Sweeney et al. (2012a) e

“ coarse” RDX formulation (KAS, /hr) 0.005 NA NA Sweeney et al. (2012a) e 

### 1 2 3 4 5 6 7 8 9 10 11
12 13
14
15 16
17 18
19 20 a Predicted from n

- octanol:water partition coefficient.
b Optimized from rat i.v. data.

c Optimized from human data of Özhan et al. (2003) an d Woody et al. (1986)

. d Optimized from mouse oral data.

e Optimized from rat oral data of Bannon et al. (2009a) , Crouse et al. (2008) , Krishnan et al. (2009) , and Schneider

et al. (1977)

.

Note: P arameter v alues used in the Sweeney et al. (2012a) and Sweeney et al. (2012b) PBPK models for RDX in

rats, humans, and mice.



### The GI tract oral absorption rate constant (KAS) was optimized to fit the time

- course concentration data for rat oral dosing studies

. The Krishnan et al. (2009) model used a

one
- co mpartm ent GI tract

. KAS was fit to the RDX blood concentrations in Krishnan et al. (2009)
, and the model with this parameter value had good agreement with the blood RDX concentrations after 0.2 and 1.24 mg/kg oral exposures ( Crouse et al., 2008 ) . The value of KAS was adjusted to fit th e RDX blood concentrations in the Schneider et al. (1977) study. Sweeney et al. (2012a) modified th e GI tract description by adding a second GI compartment and corresponding oral absorption parameters (KAS, KAD, and KT) to fit the bloo d concentrations from Krishnan et al. (2009)

. For the
ot her oral dosing studies
, the two

- compartment GI model did not improve the model fit to the data,

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 13 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

so KT was set equal to zero making the GI submodel equivalent to a one
- compartment model

. The 1
2 3
4
5 6
7 8
9 10
11
12 13
14
15 16
17 18
19 20
21
22
23
24
25
26
27
28
29
30 31
32
33 34
35
36 37
38 v alu e of KAS was adjusted separately to fit the oral studies with RDX in capsules ( Bannon et al., 20 09a ; Crouse et al., 2008 ) and coarse

- grain RDX in a saline slurry ( Schneider et al., 1977 ). The Sweeney et al. (2012a) model fits to blood and brain RDX concentrations for rats were

mo stly within a factor of 1.5 of the experimentally measured values indicating a tightly calibrated model. Human RDX toxicokinetics were modeled with the same model structure as for rats

. Values for the human physiological parameters such as tissue volumes and blood perfusion of tissues were obtained from the literature ( Brown et al., 1997 ) . Human absorption and metabolic clearance rate

con stants were optimized to fit observed RDX blood concentrations from a case study of ingestion by a 3- year
- old boy ( Woody et al., 1986 ) , and a study where five soldiers were intentionally or

acc identally exposed to RDX powder via inhalation or dermal contact ( Özhan et al., 2003 )

. The

am ounts of RDX ingested in both studies were unknown
, so Sweeney et al. (2012a) estimated the do se amount by optimizing this parameter to fit the data ( Table C -4 ). Sweeney et al. (2012a) in itially simulated each individual soldier’s blood level data separately . The resulting parameter values were similar, so data from the five soldiers were combin ed and the rate constants were reestimated using the combined data . For comparison, the rat metabolic rate constant (KfC) was scaled to humans ; the rat KfC (from fitting to in vivo data) was multiplied by the ratio of the human to rat metabolic rate constants measured in vitro and by the ratio of human to rat microsomal protein levels ( Cao et al., 2008 ; Lipscomb and Poet, 2008 ) . The scaling from rats yielded a human in
vi vo metabolic rate constant of 12.4 kg

- BW 0 .33 

### /h ou r, which is similar to the values that Sweeney et al. (2 012a) derived by fitting the combined Özhan et al. (2003) adult data (11.2 kg

- bw 0.33 

### /h ou r) and the Woody et al. (1986) child data (9.87 kg

- bw 0.33 

### /h ou r). Mouse RDX toxicokinetics were also m odeled by Sweeney et al. (2012b) using the same

mo del structure as for rats . Values for the mouse physiological parameters such as tissue volumes and bl ood perfusion of tissues were assumed to be the same as the body weight normalized parameter values in the rat model. RDX tissue:blood partition coefficients for liver (PL), brain (PB), and richly perfused tissues (PR) were estimated with an algorithm tha t relates the measured

n- octanol:water partition coefficient for RDX to reported compositions of water and lipids in specific mouse tissues ( Poulin and Theil, 2000 ; Poulin and Krishnan, 1995 ) . The KfC and KAS were
op timized to fit measured mouse RDX blood concentrations ( Sweeney et al., 2012b ) . The KfC value
est imated for the mouse (102 kg 0.33 

### /h ou r) is much higher than those estimated for rats and humans (2.6 and 11.2 kg0 .33 

### /h ou r, respectively); however, the KAS value (0.51/h ou r) fit to mouse data is similar to the value (0.83/h ou r) used in the RDX rat model for gavage in water. The Sweeney et al. (2 012b) model predictions of blood RDX concentrations were in good a greement with the
exp erimental mouse gavage data reported in the same study. The above PBPK model was evaluated and subsequently modified by EPA for use in dose

response modeling in this assessment

. This is detailed in the following section.

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 14 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

PBPK Model Evaluation and Further Development of the Sweeney et al . (2012a) and Sweeney et 1 2
3
4
5 6

7

8 9 10 11 12

13 14 15

16 17

18 19 20

21
22
23
24
25
26
27 28

29 30 al. (2012b) Models EPA evaluated and performed a quality control check of the PBPK models for RDX in rats, humans
, and mice published by Sweeney and colleagues ( Sweeney et al., 2012a ; Sweeney et al., 20 12b )

. The conclusions from these analyses are summarized below and then discussed in more de tail:

1) The model code and the parameter values matched the published reports.
2) The absorption of RDX from the GI tract did not use a consistent structure; for gavage doses
, the model used a two

- compartment GI submodel and for other oral exposures (e.g., gelatin caps ule), the model used a one
- compartment GI submodel . The model was revised to have

a one
- compartment GI submodel to simulate all oral exposures with a consistent set of absorption parameters for each dosage formulation of administered RDX.
3) Additional oral rat data were identified from single

- dose studies ( MacPhail et al., 1985 ; Schneider et al., 1977 ) and subchronic studies ( Schneider et al., 1978 ) and were used for mod el calibration as well as for independent comparison against model predictions.
4) In addition to the sensitivity analysis conducted by Swee ney et al. (2012b) on the mouse

mo del, a sensitivity analysis in the rat and human models was performed.
5) The Sweeney et al. (2012b) mouse model u sed the same physiological parameters scaled to bo dy weight as the rat model. This mouse model was revised to use mouse

- specific physiological parameters.

The Sweeney et al. (2012a) model for rats was modified by changing the oral absorption

ra te constants (as discussed below) and the partition coefficient s for the fat and slowly perfused tissues (PF and PS) as shown in Table C -5 . The partition coefficients for the fat and slowly perfused tissues were set to the values calculated by Krishnan et al. (2009) relating the measured n

- octanol:
wat er partition coefficient for RDX to reported compositions of water and lipids in those tissues

. The fits to RDX blood time course data after i .v. exposure ( Figure C -2 ) are slightly worse than the Sweeney et al. (2012a) rat model because the Sweeney et al. (2012a) rat model optimized the fat: blood and slowly perfused tissue partition coefficients to fit the dat a.

**Table C-5.**  **Parameters values used in the EPA application of the rat, human ,**  **and mouse models**

Parameter ( a bbreviation; units) Rat Human Mouse Source Body weight (BW; kg) 0.3 70 0.0206 Default values shown; study

- specific values used if available Cardiac output (KQC; L/h r /kg 0.74 ) 15 14 15 Timchalk et al. (2002) ; Brown et al. (1997)

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 15 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Parameter ( a bbreviation; units) Rat Human Mouse Source

Tissue volumes (fraction of BW) Liver (KVL) 0.04 0.026 0.055 Timchalk et al. (2002) ; Brown et al. (1997) Brain (KVB) 0.012 0.02 0.017 Timchalk et al. (2002) ; Brown et al. (1997) Fat (KVF) 0.07 0.21 0.07 Timchalk et al. (2002) ; Brown et al. (1997) Richly perfused tissues (KVR) 0.04 0.052 0.071 Timchalk et al. (2002) ; Brown et al. (1997) Blood (KVV) 0.06 0.079 0.049 Timchalk et al. (200 2) ; Brown et al. (1997) Slowly perfused tissues (KVS) 0.688 0.523 0.648 0.91 − (KVL + KVB + KVF + KVR + KVV) Blood flows (fraction of cardiac output) Liver (KQL) 0.25 0.175 0.25 Timchalk et al. (2002) ; Brown et al. (1997) Brain (KQB) 0.03 0.114 0.03 Timchalk et al. (2002) ; Brown et al. (1997) Fat (KQF) 0.09 0.085 0.09 Timchalk et al. (2002) ; Brown et al. (1997) Slowly perfused tissues (KQS) 0.2 0.2449 0.2 Timchalk et al. (2002) ; Brown et al. (1997) Richly perfused tissues (KQR) 0.43 0.3811 0.43 1 − (KQL + KQB + KQF + KQS) Tissue:blood partition coefficients and metabolism Liver (PL) 1.2 1.3 1.3 Krishnan et al. (2009) a Brain (PB) 1.4 1.6 1.6 Krishnan et al. (2009) a Richly perfused tissues (PR) 1.4 1.6 1.6 Krishnan et al. (2009) a Fat:blood PC (PF) 7.55 7.55 7.55 Krishnan et al. (2009) a Slowly perfused tissues (PS) 1.0 1.0 0.9 Krishnan et al. (2009) a

First

- order metabolic rate constant (KfC; kg 0.33 /hr) 2.6 9.87 (small boy); 11.2 (soldiers) 77 Sweeney et al. (2012a) b,c ; Sweeney et al. (2012b) d

Absorption Absorption from GI to liver (KAS; /hr) Table C

- 6 1.75 0.6 F it to rat, human , and mouse oral data

Absorption from lung to blood (Klung; /hr) 0.75 F it to human data



### 1 2 3 4 5 6 a P redicted from n

- octanol:water partition coefficient

. b Optimized from rat i.v. data.

c Optimized from human data of Özhan et al. (2003) and Woody et al. (1986)

. d Optimized from mouse oral data, and differs from that obtained by Sweeney et al. (2012b)

.

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 16 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1 2 3

4 5 6 7

8
9
10
11
12
13
14
15 16
17 18 19
20
21
22
23
24
25 26
27
28 29 A) data from Krishnan et al. (2009) (0.4 kg rats) and B) data from Schneider et al. (1977) (simulation of 0.25 kg rats and 5.5 mg/kg dose for 0.2−0.25 kg rats and 5−6 mg/kg dose).


### Figure **C-2. EPA rat PBPK model predictions fitted to observed RDX blood**  **concentrations in male and female Sprague** 

**- Dawley**  **rats following i.v.**  **exposure.**

A bs orption of RDX from the GI T ract As discussed above in the oral absorption section under toxicokinetics ( Section C.1 .1)
, the rate of oral absorption depends on the physical form of RDX

. This was demonstrated by comparing the Schneider et al. (1977) studies
, which used gavage do s es of 100 mg/kg of co u rse , granular RDX and 50 mg/ kg finely powdered RDX
, and observing that the 50 mg/kg finely powdered RDX had a higher peak plasma level

. These results are likely explained by the smaller surface area to mass ratio of the coarse

- grain RDX leading to slower dissolution and absorption. To follow the rule of model parsimony (i.e., use no more parameters than needed for the best fit to all of the data), oral absorption was modeled with a one
- compartment GI tract submodel for all simulations

. To account for the differences in absorption due to the physical form of RDX
, separate rate constants for RDX oral absorption were optimized to fit measured blood concentrations of RDX according to the type of dosing formulation ; the model fits obtained with the EPA’s revised parameters for rats are shown in Figures C

- 3 to C -5

. The oral dosing formulations were grouped into four categories : RDX dissolved in water, RDX in capsules, fine

- grain RDX, and

coarse
- grain RDX . The absorp tion rate constant for RDX dissolved in water was optimized to the data in the Krishnan et al. (2009) study ( Figure C -3)

. The absorption rate constant for RDX in
caps ules was optimized to the data in the Crouse e t al. (2008) and Bannon et al. (2009a) studies ( Figur e C -4)

. The absorption rate constant for fine

- grain RDX was optimized to the data described below ( A dditional RDX Time -C ourse D ata ) in the MacPhail et al. (1985) and Schneider et al. (1977) studie s ( Figure C -7)

. The Schneider et al. (1977) study was used to estimate the absorption rate

c onsta nts for coarse

- grain RDX ( Figure C -5 ; as represented by the fit to the data obtained by the solid curve at 100% bioavailability). Overall, the fits of the EPA revised model to the blood time

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 17 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

course data of these studies are similar to the fits of the Sweeney et al. (2012a) rat model

. The fits 1
2 3
4
5

6

7

8 9

10

11 12 13 14

15 16

17 to RDX brain time course data after oral exposure to RDX in capsules ( Figure C -6 A) are similar to the fits of the Sweeney et al. (2012a) rat model

. The absorption rate constants for each dosing fo rmulation are listed in Table C -6.

Male and female Sprague

- Dawley rats (0.4 kg) were dosed by gavage ( Krishnan et al., 2009 ).


### Figure **C-3. EPA rat PBPK model predictions fitted to observed RDX blood**  **concentrations following oral exposure to RDX dissolved in water.**

The ingested RDX doses were: A) 0.2 and 1.24 mg/kg RDX in male Sprague

- Dawley rats (0.4 kg, data from Crouse et al. (2008) ) and B) 3 and 18 mg/kg RDX in male and female Sprague

- Dawley rats (0.35 kg, data from Bannon et al. (2009a) ) for KAS = 0.35/hour.



### F **igure C-4.**  **EPA rat model predictions fitted to observed RDX blood concentrations following oral exposure to**  **RDX in dry capsules.**

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 18 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1

2 3 4 5 6 7 8 9

10 11 12

13 14 15 16

17 18 Symbols denote observed RDX blood concentrations measured in male Sprague

- Dawley rats (0.225 k g) resulting from oral doses of 100 m g/kg RDX ( Schneider et al., 1977 ) . The KAS fit to these data assuming 100% bioavailability re sulted in the same estimate (KAS = 0.00497/hour) as obtained by Sweeney et al. (2012a) . Alternatively, for KAS fixed at the value fit to fine

- grain RDX in a saline slurry (KAS = 0.019/hour fit to data from Schneider et al. (1977) and MacPhail et al. (1985) ; Figure C

- 7), the estimate of oral bioavailability fit to the RDX blood concentrations was 35%. A bioavailability of 40% and KAS = 0.019/hour is also shown for comparison.



### F **ig ure**  **C-5. Effect of varying oral absorption parameters on EPA rat model**  **predictions fitted to observed RDX blood concentrations following oral exposure to coarse** 

**- grain RDX** 

**.**

A) 3 and 18 mg/kg RDX in dry capsules (0.35 kg male and female rat data from Bannon et al. (2009a) ; best fit KAS = 0.35/hour. B) 50 mg/kg fine

- grain RDX in a saline slurry (0.25 kg male and female rats data from MacPhail et al. (1985) ; best fit KAS = 0.019/hour.



### F **igure C-6.**  **EPA rat model predictions fitted to observed RDX brain tissue**  **concentrations following oral exposure to RDX.**

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 19 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table C-6**  **. Doses, dosing formulations, and absorption rate constants in** 1 2

3 4 5 6 7 8 9
10
11

12
13
14
15 16
17 18
19
20 **animal and human studies**

Formulation Study Dose Estimated KA (/hr)

RDX dissolved in water Krishnan et al. (2009) 1.53, 2.07 mg/kg
, single gavage 1.75 Schneider et al. (1978) ~5−8 mg/kg

- d , drinking water 90 d Dry RDX in capsules a Crouse et al. (2008) 0.2, 1.24 mg/kg
, single dose 0.35 Bannon et al. (2009a) 3, 18 mg/kg
, single dose

Fine

- grain RDX in saline slurry Schneider et al. (1977) 50 mg/kg
, single gavage 0.19 MacPhail et al. (1985) b 50 mg/kg
, single gavage

Coarse

- grain RDX in saline slurry Schneider et al. (1977) 100 mg/kg
, single gavage 0.00497

a Capsules were filled with dry RDX from stock solution of acetone , and acetone was evaporated off.
b RDX particle size was ≤66 μm in diameter suspended in a 2% solution of carboxymethylcellulose.



### An alternative to varying the KAS for each RDX formulation would be to vary the oral bioavailab ility, in effect modifying the administered exposure concentration. Therefore, the sensitivity of the model fit to variations in oral bioavailability was examined in Figure C

- 5 and an analysis of model sensitivity to oral bioavailability was conducted as discussed further in the section, S ensitivity A nalysis of the R at PBPK M odel

.

Additional RDX T ime -C ourse D ata The EPA revised models were simultaneously fit ted against additional RDX time

- course data (not used in the original Sweeney et al. (2012a) model calibration) . These data came from (1 ) two studies in which animals received oral dos es of fine

- grain RDX ( MacPhail et al., 1985 ; Schneider et al., 1977 ) ( Figure C-7) and (2) RDX brain time

- course data from a study in which

an imals received oral dose s of fine

- grain RDX ( MacPhail et al., 1985 ) ( Figure C -6 B)

. Overall
, the cali brated EPA rat model predictions are within a factor of 1.5 of the measured values from different data sets, and are therefore likely to provide a more robust estimated parameter.

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 20 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1 2 3 4

5 6

7 8
9
10
11
12
13
14
15 16
17 Oral doses of 50 mg/kg RDX were administered to: A) male Sprague

- Dawley rats (0.225 kg) ( Sch neider et al., 1977 ) and B) male and female Sprague

- Dawley rats (0.25 kg) data ( MacPhail et al., 1985 ) . Best fit KAS = 0.019/hour.


### Figure **C-7. EPA rat model predictions**  **fitted to observed RDX blood concentrations following oral exposure to fine** 

**- grain RDX in a saline slurry** 

**.**

F ollowing calibration, the EPA model was further tested by comparison with results from two other subchronic oral studies in male and female rats ( Schneider et al., 1978 ) . These were a
g avage study where 20 mg/kg RDX was administered in saline slurry and a drinking water study where rats were provided with RDX

- saturated drinking water (50

− 70 μg /mL) ad libitum for which the study authors estim ated a daily dose between 5 and 8 mg RDX/kg body weight

. It is striking that the observed RDX blood concentrations in the gavage study ( Figure C -8, symbols) were virtually the same
, or only slightly elevated
, as compared to the blood concentrations report ed in the drinking water exposures
, with a n approximately threefold lower daily administered dose in the drinking water study ( Figure C -9 , symbols)

. This is counter to the expectation that higher doses cause higher blood levels and is discussed further be low.

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 21 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1

2 3 4 5

6 7

8

9

10 11 12 13 14

15 16 Model fits and mean observed RDX blood concentrations resulting from daily gavage doses of 20 m g/kg RDX for 90 days to male and female Sprague

- Dawley rats (0.225 kg). The RDX in saline slurry was assumed to be coarse

- g rained with an oral absorptio n rate constant KAS = 0. 00497/hour.



### F **igure C-8** 

**. Comparison of EPA rat model predictions with data from**  **Schneider et al. (1978) for the subchronic gavage study.** 

Model fits and mean observed RDX blood concentrations resulting from a daily estimated dose of 6.5 m g RDX/kg

- day for 90 days to male and female Sprague

- Dawley rats (0.225 kg). The large peak to trough change in the simulation results from model representation of the daily o r al ingestion of drinking water primarily during the waking state. The oral absorption rate constant for RDX dissolved in water was used (KAS = 1. 75/hour).



### F **igure C-9** 

**. Comparison of EPA rat model predictions with data from**  **Schneider et al. (1978) for the subchronic drinking water study.** 

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 2 2 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

EPA’s modified PBPK model was set up to simulate drinking water exposures with a 1
2 3
4
5 6
7 8
9 10
11
12 13
14
15
16
17 18 19
20
21
22
23
24 25
26

27
28
29
30
31 32
33
34 non continuous sipping pattern based on Sp iteri (1982)
, which assumed 80% of the consumption to

oc cur episodically at night when the rats were awake 3



### . The model predict s blood concentrations to increase in proportion to the total dose; for the gavage study
, the model predictions yielded an RDX blood concentration approximately three fold higher than the reported mean blood concentrations ( Figure C -8), while for the subchr onic drinking water study
, the model fit the data reasonably well ( Figure C -9 ). It is possible that multiple mechanisms such as elimination of unabsorbed RDX or metabolic induction may explain why the observed RDX blood concentrations did not increase in proportion to the higher administered dose in the gavage studies compared to the drinking water study

. Elimination of unmetabolized RDX may be an insignificant factor in the single

- dose studies used for calibration of the absorption constant for the RDX in saline slurry, but for repeated, higher doses this elimination route could be significant

. Schneider et al. (1978) found similar RDX

co ncentrations in the feces of rats in the gavage and drinking water studies (3.1 ± 2.0 and
2.7 ± 1.3 μg RDX per g dry weight feces, respectively)

. The total recovery of radioactivity in feces was also similar in the gavage study (4.8 ± 0.8%, week 1 only) and drinking water study (4.4 ± 0.6%, measured over the course of the study). Thus
, the differen ce in fecal elimination for the two routes does not appear significant. It is also possible that metabolic induction occurred during the repeated dosing of RDX in the gavage study leading to the lower observed RDX blood concentrations . The reasonably goo d fits of the model to the drinking water data set demonstrated achievement of regular periodic levels, and indicate a lack or much lower extent of metabolic induction over time from those repeated doses , possibly because the dose rate was lower: 5

− 8 v ersus 20 mg/kg

- day in the gavage study. Overall, the reasonable agreement of the modified EPA RDX rat model with the subchronic drinking water data support the use of the model in estimating and extrapolating blood levels following chronic exposure at or below this exposure range (5

− 8 mg/kg

- day), particularly in drinking water

. Simulating E xposures in H umans The Sweeney et al. (2012a) model for humans was modified in the same ways as the rats, by cha nging the partition coefficients for the fat and slowly perfused tissues (PF and PS) as shown in Table C

- 5 and fitting the rate constants for oral absorption and metabolism to RDX blood concentration data

. In the studies of humans with measured RDX blood concentrations by Woody et al. ( 1986) and Özhan et al. (2003)

, the RDX doses were unknown and the doses were therefore al so optimized to fit the data

. The model predictions for the Woody et al. (1986) data using the best fi t values of dose = 58.9 mg/kg, KAS = 1.75/h ou r, and KfC = 9.87 kg 0.33 

### /h ou r are shown in

3 A constant drinking water ingestion rate interspaced between episodes of no ingestion was assumed. Each 12

- hour awake period consisted of eight cycles that alternated between 1.5

- hour cycles of frequent sipping (continuous ingestion) and zero ingestion fo r 45 minutes each. Each 12

- hour sleeping period consisted of four cycles with regular sipping periods of 30 min utes followed by 2.5 hours of no ingestion.
This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 23 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Figure C- 10. The model predictions for the Özhan et al. (2003) data using the best fit values o f an 1
2 3

4

5

6 7 8

9

10 11 12

13 14 15 o ral dose = 3.5 mg/kg, KAS = 1.75/h ou r, and KfC = 9.87 kg 0.33 

### /h ou r are shown in Figure C

- 11.

The best fit values were KAS = 1.75/hour, dose = 58.9 mg/kg, and KfC = 9.87 kg 0.33 /hour.



### F **igure C-**  **10**

**. EPA human model predictions fitted to observed RDX blood**  **concentrations resulting from an accidental ingestion of RDX by a 14.5**

**- kg boy**  **( Woody et al., 1986**  **)**

**.**

For an assumed oral exposure, the best fit values were KAS = 1.75/hour, dose = 3.5 mg/kg, and KfC = 9.87 kg 0.33 /hour. For the same 3.5 mg/kg dose and metabolism rate constant, an inhalation exposure found a best fit value for Klung of 0.75/hour.



### F **igure C-**  **11**

**. EPA human model predictions fitted t**  **o observed RDX blood concentrations resulting from accidental exposure to adults assumed to be**  **70 kg**  **( Özhan et al., 2003**  **)**

**.**

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 24 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

E PA’s calibration of the model differed in another import ant respect from that carried out 1
2 3
4
5 6
7 8
9 10
11
12 13
14
15 16
17 18
19 20

21
22 23
24
25 26
27
28
29 30
31
32 33
34
35
36 37 by S weeney et al. (2012a)

. As previously mentioned, Sweeney et al. (2 012a) simulated the soldiers’

e xposure from the Özhan et al. (2003) study as an oral exposure, although the study report states
t hat the exposure was via inhalation and dermal routes. An inhalation or dermal exposure could change the amount of RDX reaching the blood compared with an oral exposure due to first

- pass metabolism in the liver after oral absorption. Dermal absorption was not considered by EPA to be a sign ificant route of RDX exposure and was therefore not modeled. This decision is supported by a study that used excised human skin and reported that only 5.7% of the applied dose was absorbed into the skin by 24 hours post dosing ( Reddy et al., 2008 )

. The model was modified to simulate an
i nha lation exposure and compared with the data from Özhan et al. (2003)

. There are insufficient d ata on blood:air partitioning to modify the Sweeney et al. (2012a) model with a lung

co mpartment; therefore, inhalation exposure was modeled in an approximate manner as a direct input to the blood with an optimized absorption rate to represent absorption from air containing RDX into the blood

. The soldiers’ inhalation exposure was simulated as a conti nuous 8

- hour exposure (i.e., assuming that the soldiers were exposed occupationally during an 8

- hour workday), and for the same dose of 3.5 mg RDX/kg that was estimated by Sweeney et al. (2012a)

. The model

as sumed that 100% of the inhaled dose was absorbed and that the absorption rate constant was optimize d to fit the measured blood concentrations of RDX . The model predictions were in good agreement with the RDX blood concentrations reported by Özhan et al. (2003) as sh own in F igure C- 11.

Sensitivity A nalysis of the R at and H uman PBPK M odel s A sensitivity analysis was performed to see how each model parameter affects the model output

. A sensitivity coefficient, defined as the change in a specified dose metric due to a 1% increase in the value of a parameter, was calculated for each parameter in the rat and human models

. This analysis was carried out for both short

- term (24 hours following a single oral do se of 1.5 mg/kg RDX) and longer

- term (90 days of repeated oral do sing with 1.5 mg/kg RDX) exposures for the dose

- metric of blood AUC

. Parameters with sensitivity coefficients > 0.1 in absolute value (i.e., considered sensitive) are presented in Table C-7

. For the blood AUC dose

- metric , the only sensitive RDX

- specific parameter is the KfC. This sensitivity is likely because bioavailability was assumed to be 100% and metabolism is the only route of elimination in the model

. These assumptions mean that all administered RDX will be absorbed and metabolized ; in other words
, the blood AUC is proportional to the dose and inversely proportional to the metabolic clearance rate
constant

. For the parameter values in this mode l, the rate of metabolism is relatively slow compared to the transport of RDX between other tissues and the site of metabolism in the liver, so that the blood AUC is not sensitive to parameters that impact transport such as KQC and KQL

. Because the metabolic clearance rate constant is scaled to body weight and by liver volume, the blood AUC is also sensitive to these parameters

. The sensitivity analysis by Sweeney et al. (2012b)

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 25 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

for the AUC of RDX in the liver found the model was sensitive to the liver:blood partition coefficient 1
2

3

4 5 6 7 8 9 10 11
12
13
14
15 16
17 18
19
20

21
22
23
24
25
26
27
28
29
30 31
32 (PL) in addition to the same parameters (KfC, KVL
, and BW) found for the blood AUC.

**Ta ble**  **C-7. Sensitivity c**  **oefficients for rat and human RDX PBPK models**

Parameter Rat sensitivity coefficient Human sensitivity coefficient

F ractional liver volume (KVL)

− 1

− 1

B ody weight (BW) 0.3 0.3

M etabolic rate constant (KfC)

− 1

− 1

Parameters with sensitivity coefficients < 0 .1 in absolute value are considered not sensitive, and are listed below:

• ca rdiac output (KQC);

• f ractional blood flow to all tissues (liver, KQL; fat , KQF; slowly perfused tissues , KQS; brain , KQB)

• f ractional tissue volume of fat (KVF), brain (KVB), and blood volume (KVV)

• b lood partition coefficients to all tissues (liver , P L ; fat , PF ; rapidly perfused , PR ; slowly perfused , PS ; brain , PB)

• a bsorption rates from GI (KAS, KT, KAD)



### The model is also very sensitive to oral bioavailability, with a sensitivity coefficient of 0.8 in the case of the rat model. As discussed above in the oral absorption section of toxicokinetics (Section C. 1 .1), estimates of the bioavailability of RDX range from 50 to 87% or greater and may depend upon the physical form of RDX ( Krishnan et al., 2009 ; Schneider et al., 1978
, 1977 )

. Ho wever, as seen in Figure C

- 5, it was not possible to identify the bioavailability and the absorption rate (KAS) as separate parameters by fitting to the available RDX blood concentration time course. Introducing oral bioavailability as an additional unknown parameter and recalibrating the model did not appear to provide an advantage. Therefore, 100% bioavailability was assumed in the model and was acknowledged as an unce rtainty.

Simulating E xposures in M ice Physiological parameters specific to mice were obtained from the literature ( Brown et al., 19 97 ) and are shown in Table C -5 . The partition coefficients calculated for mi ce by Sweeney et al. (2 012b) were used, and include the liver, brain
, and richly perfused tissues

. The partition

co efficients for the fat and slowly per fused tissues from the Sweeney et al. (2012b) mouse model

we re not used because they were estimated via optimization of fits to rat i .v. data

. Inste ad
, the partition coefficient for fat tissues was set equal to the value calculated by Krishnan et al. (2009) for
rat fat tissue, 7.55. The partition coefficient for slowly perfused tissues (0.9) was calculated for mouse tissues using the same metho dology as Krishnan et al. (2009)

. The rate constants for oral ab sorption and metabolism were optimized to fit the data from Sweeney et al. (2012b) for mouse bl ood RDX concentrations

. The model predictions were in good agreement with the RDX blood concentrations reported by Sweeney et al. (2012b)
, as shown in Figure C

- 12

.

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 26 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1

2

3

4

5 6 7

8 9

10 11
12
13
14
15 16
17 Model fits and mean and standard deviation of observed RDX blood concentrations in female B6C3F 1 mice (0.0205 kg) for doses of 35, 60, and 80 mg/kg with KAS = 0.6/hour and KfC = 77 kg 0.33 /hour . Experimental data from Sweeney et al. (2012b)

. 

### Figure **C- 12** 

**. Comparison of EPA mouse PBPK model predictions with data**  **from oral exposure to RDX dissolved in water** 

**.**

Th e only information on RDX metabolism in the mouse comes from a study by Pan et al. (2 013)

. Pan et al. (2013) measured nitrosamine RDX metabolites of RDX (MNX, DNX, and TNX, the la tter repres enting a minor metabolic pathway) in mice at the end of a 28

- day exposure to RDX in feed (ad libitum). These measurements were a single time point without controlling the time between the last RDX ingestion and measurement, and were therefore judged not t o be sufficient for use in parameterizing a PBPK model of the nitrosamine metabolites.

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 27 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

R at to Human Extrapolations 1
2
3
4
5 6
7 8 9
10
11
12 13
14
15 16
17 18 19
20
21
22 23
24
25 26

27
28 29
30
31 32
33
34
35 36
37 T he rat and human PBPK models as described above were applied to derive human equivalent doses ( HEDs ) for candidate points of departure ( PODs ) for endpoints selected from rat bioassays. The rat and human PBPK models were used to estimate two dose metrics

— the AUC and the peak concentration (C max 

### ) for RDX concentration in arterial blood. The relationships between administered dose and both internal metrics (AUC and C max 

### ) were evaluated with the rat PBPK model over the range of 1 μg/kg

- day to 100 mg/kg

- day and with the human PBPK model over the
range of 0.05 μg/kg

- day to 200 mg/kg

- day, ranges that encompas s the PODs. The times to reach steady state for the dose metrics were shorter than the duration of the toxicity studies, so the steady state values were considered representative of the study and were used. To calculate steady

- state values for daily exposure, the simulations were run until the daily average had a <1% change between consecutive days. For both the rat and human PBPK models, both dose metrics correlated linearly with the administered dose. For rats dosed via gavage, the slope of administered dose versus AUC was 6.800 mg/ L- day / mg/kg

- day and that for C max 

### was 0.4718 mg/L / mg/kg

- day. For a continuous dose, the slope of dose versus AUC was the same ( 6.800 mg/L

- day / mg/kg -day) and for C max 

### was 0.3951 mg/L / mg/kg

- day. For humans, assuming a drinking water dose sipping pa ttern, the slope of administered dose versus AUC was 13.95 mg/L

- day / mg/kg

- day and that for C max 

### was 0.7316 mg/L / mg/kg

- day. Given this linearity in internal metrics and assuming that equal internal metrics in rats and humans are associated with the sam e degree of response, the HEDs could then be directly determined b y multiplying the lower bound on the benchmark dose (BMDL) in rats by the ratio of these slopes. For a gavage dose in rats converted to a human drinking water dose
, the ratio for AUC was 6.800 / 13.95 = 0.487 and C max 

### was 0.4718 / 0.7316 = 0.645. For a continuous dose in rats converted to a human drinking water dose, the ratio for AUC was 6.800 / 13.95 = 0.487 and for C max 

### was 0.3951 / 0.7316 = 0.540. These ratios were applied in Table 2 -2 to calculate the POD HED 

### from the rat benchmark dose lower confidence limits ( BMDLs ) and no

- observed

- adverse

- effect levels ( NOAELs ) for each endpoint.

Mouse to Human Extrapolations The mouse and human PBPK models as described above were applied to derive HEDs for candidate PODs for endpoints selected from mouse bioassays. The mouse and human PBPK models were used to estimate two dose metrics

— the area under the curve (AUC) and peak concentration (C max 

### ) for RDX concentration in arterial blood. The relation ships between administered dose and both internal metrics (AUC and C max 

### ) were evaluated with the mouse PBPK model over the range 10 μg/kg

- day to 100 mg/kg

- day and with the human PBPK model over the range 0.05 μg/kg

- day to 200 mg/kg

- day, ranges that encompa ss the PODs. The times to reach steady state for the dose metrics were shorter than the duration of the toxicity studies, so the steady state values were considered representative of the study and were used. To calculate steady state values for daily
exp osure, the simulations were run until the daily average had a <1% change between consecutive

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 28 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

d ays. For both the mouse and human PBPK models, both dose metrics correlated linearly with the 1
2 3
4
5 6
7 8
9 10
11
12 13
14

15
16
17 18 19
20
21
22 23
24
25 26
27
28
29 30
31
32 33
34
35

36 37 administered dose. For mouse dosed via gavage, the slope of administered dose versus AUC was 0.0656 mg/L

- day / mg/kg

- day and that for C max 

### was 0.0273 mg/L / mg/kg

- day. For a continuous dose, the slope of dose versus AUC was the same 0.0656 mg/L

- day / mg/kg

- day and for C max 

### was 0.0081 mg/L / mg/kg

- day. For humans, assum ing a drinking water dose sipping pattern, the slope of administered dose versus AUC was 13.95 mg/L

- day / mg/kg

- day and that for C max 

### was 0.7316 mg/L / mg/kg

- day. Given this linearity in internal metrics and assuming that equal internal
metrics in mice an d humans are associated with the same degree of response, the HEDs could then be directly determined by multiplying the BMDL in mice by the ratio of these slopes. For a gavage dose in mice converted to a human drinking water dose, the ratio for AUC was 0. 0656 / 13.95 = 0.0047 and C max 

### was 0.0273 / 0.7316 = 0.373, respectively. For a continuous dose in mice converted to a human drinking water dose, the ratio for AUC was 0.0656 / 13.95 = 0.0047 and for C max 

### was 0.0081 / 0.7316 = 0.011. These ratios were applied in Table 2

- 2 to calculate the POD HED 

### from the mouse BMDLs and NOAELs for each endpoint.

S ummary of C onfidence in PBPK M odels for RDX Overall, good fits to the rat, mouse, and human time

- course data for RDX internal concentrations were obtained . For the rat and human models, calibration was based generally on fitting to more than one data set obtained from different studies originating in different laboratories or accidental exposure settings. Predi ctions from the rat model compared well with data from a subchronic drinking water study that was not used in model calibration.
T he metabolic rate constant used in the human model was fit to limited data from accidentally exposed humans ; however
, the value of the metabolic rate constant has additional support from in vitro experimental data . The rat meta bolic rate constant, fit to mult iple experimental data sets, was scaled to humans using the ratio of human to rat rate constants measured with in vitro m ethods . This scaled value of the human metabolic rate constant was very similar to the rate constant estimated by fitting the model to the human data

. The congruence in values increases the confidence in using the EPA

- modified PBPK model for predicting h uman blood RDX concentrations. There are several uncertainties in these models ( listed below ), the most significant of which pertain to the mouse PBPK model. The mouse model was based on a single data set, which used high RDX doses to obtain detectable RD X blood concentrations, and the type s of additional data that increased the confidence in the rat and human models are not available for mice. The additional data not available for mice are in vitro measurements of RDX metabolism by mouse cells and quanti fication of potential routes of RDX elimination in mice. Overall
, these uncertainties result in lower confidence in the mouse model than in the rat an d human models.

1) R DX is readily metabolized in several species, yet there are no data on the toxicokineti cs of RDX metabolites in the rat and human. Some data are available for the n

- nitrosoamine

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 29 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

m etabolites (a minor metaboli c pathway) in mice
, but the data are too sparse to help better 1 2 3

4 5 6

7 8

9 10 11 12 13 14 15

16 17 18 19

20 21 22 23 24 25

26 27 28 29 30 31 32 33 34 35 36

37
38
39
40 paramet e rize a PBPK model . Consequently, the PBPK models used in this a ssessment do not inco r porate the kinetics of RDX metabolites.

2 ) The available toxicokinetic data are not sufficient to uniquely identify a parameter value for RDX oral bioavailability

. Consequently
, the model assumes 100% bioavailability even though some studies in rats suggest that a lower bioavailability is likely.
3) The human model is based on single accidental exposures, and the exposure concentrations are not known.
4) The only route of clearance of RDX used in the models is that of total metabolism, which appears reasonable for the rat for which only roughly 5% of the RDX was detected unmetabolized in urine and feces . However, no data on the excretion of RDX are available for the mouse

. This inabil ity to properly characterize the fraction of RDX that is metabolized in the mouse is problematic considering some evidence to indicate that the role of metabolism in RDX toxicity may be different across species

. This uncertainty decreases the confidence in the mouse PBPK model.
5) The PBPK model for the mouse is based on a single data set. This single data set is used to fit both the absorption and metabolic rate constants. There are no in vitro data to independently estimate the metabolic rate constant fo r the mouse . Consequently, the confidence in the mouse model parameter values is low.
6) T he analytical detection limit in the mouse pharmacokinetic study is too high to enable detection at the lower doses. The lowest dose at which RDX was detected was 35 m g/kg; this concentration was high enough to manifest some toxicity in the chronic mouse bioassay

. The measured blood concentration at the 35 mg/kg dose is based on the level measured from one single animal (others were non

- detects or excluded as outliers)

. This decreases the confidence in the calibration of the mouse PBPK model.
7) T he metabolic rate constant as estimated by the PBPK model for mice was 30

- fold higher than the rat (after accounting for body weight differences)
, suggesting that the toxicokinet ics of RDX could be significantly different in the mouse than in the rat

. Mice may have more efficient or higher expression of the CYP450 enzymes

. Alternatively, mice may have other unknown metabolic pathways responsible for metabolizing RDX

. Identifyin g the specific CYP 450 enzymes and measuring expression levels and in vitro metabolic rate constants in mice would allow for in vitro scaling from rats to mice
, which could be used to independently evaluate the mouse metabolic rate constant. Given the high sensitivity of the model to the metabolic rate constant
, this uncertainty in the mouse toxicokinetics significantly decreases the confidence in using the mouse PBPK model for predicting mouse blood RDX concentrations.

Model C ode for RDX PBPK M odel U sed in the A ssessment The PBPK acslX model code is made available electronically through the Health and Environmental Research Online ( HERO ) database

. All mod el files may be downloaded in a zipped workspace from HERO ( U.S. EPA, 2014 ).

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 30 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine



## C.2. **HUMAN STUDIES** 1
2
3
4

5 Table C-8 presents a summary of case reports of humans acutely exposed to RDX. Table C-9 provides a chronological summary of the methodologic features of the available epidemiology studies of RDX.

**Table C-8.**  **Summary of case reports of exposure to RDX** 

Reference,

n umber of c ases,
e xposure setting Exposure Effects observed Comments

Barsotti and Crotti (1949)

17 males among 20 male Italian workers ( 1939

− 1942 )

Manufacturing Inhalation of RDX powder during the drying, cooling, sieving, and packing processes of its manufacture Generalized convulsions of a tonic

- clonic (epileptic) type followed by postictal coma; loss of consciousness without convulsions; vertigo; vomiting and confusion; transient arterial hypertension

Symptoms occurred e ither without prodromal symptoms or were preceded by several days of insomnia, restlessness, irritability, or anxiety Tobacco and alcohol use were considered by the study authors to be aggravating factors

Kaplan et al. (1965 )

5 males among 26 workers (April

− July 1962 )

Manufacturing Inhalation, ingestion, and possible skin absorption as a result of the release of RDX dust into the workroom air during the dumping of dried RDX powder, screening and blending, and cleanup of spilled material without adequate ventilation Sudden convulsions or loss of consciousness without convulsions; few or no premonitory symptoms (e.g., headache, dizziness, nausea, vomiting); stupor, disorientation, nausea, vomiting, and weakness; no change s in complete blood counts or urinalysis Mild cases of RDX intoxication may have been masked by viral illness with nonspecific symptoms (e.g., headache, weakness, upset GI tract) ; n o method was available for determining RDX concentrations in air; recovery was complete without sequelae Merrill (1968)

2 males

Wartime, Vietnam Ingestion of unknown quantity of C

- 4 with moderate amounts of alcohol Coma, vomiting, hyperirritability, muscle twitching, convulsions, mental confusion, and amnesia; kidney damage (oliguria, gross hematuria, proteinuria, elevated BUN); li ver or muscle damage (high AST) ; leukocytosis Confounding factors included ingestion of C

- 4 while intoxicated with ethanol (vodka), which may have caused GI symptoms, and smoking (1−1.5 packs of cigarettes per day)

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 31 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference,

n umber of c ases,
e xposure setting Exposure Effects observed Comments Stone et al. (1969)

4 mal es (March

− December 1968 )

Wartime, Vietnam Ingestion of 180 g (patient 1), or 25 g (patients 2, 3) of C

- 4 (91% RDX) Generalized seizures, lethargy, nausea, vomiting, fever, muscle soreness, headaches, twitching, (semi)comatose, headaches, hematuria, abnor mal laboratory findings, muscle injury, elevated AST; no kidney damage

For the patient who ingested the highest dose, anemia and loss of recent memory present after 30 d Troops ingested small quantities of RDX to get a feeling of inebriation similar to that induced by ethanol

Hollander and Colbach (1969)

5 males (June 1968

− January 1969 )

Wartime, Vietnam Inhalation (all five cases) and ingestion of unknown quantity of C

- 4 ( two cases) Tonic

- clonic seizures; nausea and vomiting occurred before and after admission; hyperirritability, muscle twitching, convuls ions, mental confusion, and amnesia; kidney damage (oliguria, gross hematuria, proteinuria, elevated BUN); liver or muscle damage (high AST) ; leukocytosis; symptoms cleared by the next day except for amnesia (in case 2), oliguria (lasted for 4 d), and gros s hematuria (decreased by 9 th hospital day) Knepshield and Stone (1972)

6 males

Wartime, Vietnam Ingestion of C

- 4, range 25−180 g, average 77 g Generalized seizures, coma, lethargy, severe neuromuscular irritability with twitching and hyperactive reflexes, myalgi a, headache, nausea, vomiting, oliguria, gross hematuria, low

grade fever; abnormal laboratory findings (neutrophilic leukocytosis, azotemia, elevated AST) Includes data on two patients from Merrill (1968)

Ketel and Hughes (1972)

18 males (December 1968

− December 1969 )

Wartime, Vietn am Inhalation while cooking with C

- 4 and possible ingestion CNS signs ( confusion, marked hyperirritability, involuntary twitching of the extremities, severe prolonged generalized seizures, prolonged postictal mental confusion , amnesia ); renal effects (oliguria and proteinuria , one case of acute renal failure requiring hemodialysis) ; GI toxicity (nausea, vomiting) C

- 4 was cut with the same knife used to stir/prep are food

This document is a draft for review purposes only and does not constitute Agency policy. 

### B

- 32 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference,

n umber of c ases,
e xposure setting Exposure Effects observed Comments Woody et al. (1986)

1 male child (August 1984 )

Manufacturing Ingestion of plasticized RDX from mother's clothing and/or boots; estimated ingested dose of 1.23 g RDX was normalized to the patient's body weight (84.82 mg/kg) Seizures before and after admission; EEG revealed prominent diffuse slowing that was greatest in the occipital regions with no evidence of epileptiform activity; elevated AST on admission and after 24 hr s ; within 24 hr s , the child was extubated and intensive care withdrawn; normal mental status and normal neurological examination at discharge Mothe r worked at an explosive plant in which RDX was manufactured in a plasticized form

Goldberg et al. (1992)

1 male

Nonwartime Ingestion after chewing a piece (unknown size) of “Semtex” plastic explosive 4 h r s before first seizure Frontal headache and two tonic

clonic seizures; progressively disseminating petechial rash suggestive of meningococcal infection apyrexial; normotensive; no photophobia; no neurological abnormalities; florid petechial rash over the face and trunk; lacerated tongue

Initial results included leukocyte count of 10.8 × 10 9 /d L (87% neutrophils); hemoglobin, platelet count, coagulation screen, serum and CSF biochemistry all within normal limits; CSF and blood bacteriologically unremarkable

Shortly following admission, headache and rash disappeared ; no further seizures
Harrell

- Bruder and Hutchins (1995)

1 male

Nonwartime Ingestion of C

- 4 (chewing on a piece of undetermined size) Tonic

- clonic seizures ; postictal state ; EEGs were normal; brisk deep tendon reflexes

Testud et al. (1996a)

1 male

Manufacturing Inhalation and possible dermal exposure during the RDX manufacturing process Malaise with dizziness, he adache, and nausea progressing to unconsciousness and generalized seizures without involuntary urination or biting of the tongue ; b lood chemistries were in the normal range and blood alcohol content was null

This document is a draft for review purposes only and does not constitute Agency policy. 

### B

- 33 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference,

n umber of c ases,
e xposure setting Exposure Effects observed Comments

Testud et al. (1996b)

2 males

Manufacturing Inhalation and possible dermal exposure during the RDX manufacturing process Sudden loss of consciousness and generalized s eizures; blood serum level of 2 mg/L RDX measured Smoker and alcohol drinker

Hett and Fichtner (2002)

1 male

Nonwartime Ingestio n of a cube (1 cm across) of C

- 4 Nausea and vomiting ; tonic

- clonic seizure lasting 2 min, followed by two seizures of about 30 sec each; myoclonic jerks in all limbs; petechial hemorrhages around face and trunk after seizures Küçükardali et al. (2003)

5 males

Nonwartime Ingestio n (accidental) of 37−250 mg/kg body weight RDX during military training via food contaminated with RDX Abdominal pain, nausea, vomiting, myalgia, headache, generalized weakness, repetitive tonic

- clonic convulsions, lethargic or comatose between seizures, hyperactive deep tendon reflexes, sinusal tachycardia ; e levated serum levels of AST and ALT ; k idney damage ; p lasma RDX levels 3 hr s after ingestion ranged from 268 to 969 pg/m L Davies et al. (2007)

17 males

Nonwartime Ingestion of unknown quantity C

- 4 under unclear circumstances, but unrelated to recreational abuse Seizures, headache, nausea , and vomiting; hypokalemia and elevated creatine kinase, lactate dehydrogenase, and phosphate noted in all but two patients ;
m etabolic acidosis only occurred in two patients directly following seizures Patient histories may have been affect ed by the fact that the incident was the focus of a military police investigation

Kasuske et al. (2009)

2 males

Nonwartime Ingestion of C

- 4 after handling explosive ordnance Seizures, postictal state, confusion, drowsiness, headache,

nausea , and vomiting ; b lood work revealed high WBC count and elevated creatine phosphokinase ; p roteinuria and gross hematuria observed 

### 1 2 3 ALT = alanine aminotransferase; AST = aspartate transaminase; BUN = blood urea nitrogen; CNS = central nervous system; C SF = cerebrospinal fluid; EEG = electroencephalogram; WBC = white blood cell

This document is a draft for review purposes only and does not constitute Agency policy. 

### B- 34 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**T able**  **C-9. Occupational e**  **pidemiologic s**  **tudies of**  **RDX : summary of methodologic features**  1

Reference,

s etting and d esign Participants,

s election, f ollow

- up Consideration of l ikely

s election b ias Exposure m easure and

r ange Outcome

m easure Consideration of l ikely

c onfounding Analysis and

r esults d etails Sample s ize

Ma and Li (1993) (China) a Industrial workers (t ranslated article ) Details of industrial process and subject selection framework not reported ;
r eferents chosen from same plant; age, employment duration, and education similar across groups ; p articipation rate not reported Sparse reporting of details on subject recruitment and participation Details of exposure monitoring not reported

. Two groups of exposed subjects: Group A, intensity, 0.407 (0.332) mg/m 3 [mean ( standard deviation )], daily cumulative, 2.66 (1.89 ) mg/m 3

. Group B, intensity, 0. 672 (0.556) mg/m 3 ; daily cumulative, 2.53 (8.40 ) mg/m 3

. Neurobehavioral battery administered by trained personnel : memory retention, simple reaction time, choice reaction time, letter cancellation, and block design No adjustment for other risk factors ( e.g., alcohol consumption );

n o consideration or attempt to distinguish TNT Comparisons of mean scaled score on memory retention, letter cancellation, or block design test; mean time on reaction tests; and total behavioral score ;
v ariance (F test), linear a nd multiple regression, and correlation analysis 60 exposed ; Group A (n = 30; 26 males, 4 females) ; Group B (n = 30); 32 referents (27 males, 5 females)

Hathaway and Buck (1977) (United States) Ammunition workers 2 , 022 workers (1, 017 exposed to open explosives (TNT, RDX, HM X), 1,005 referents) at five U.S. Army ammunition plants (I owa , I llinois , T ennessee ) ; p articipation rate : 76% exposed, 71% referents Potential h ealthy worker effect Atmospheric samples of all operations with potential exposure to open explosives taken in 1975

. Range: not detected to 1.57 mg/m 3

. Seventy exposed workers with RDX at >0.01 mg/m 3 [the LOD]; mean: 0.28 mg/m 3 [ standard deviation not presented]

. Job title used to initially identify exposed or unexposed status and reassigned to one of two Liver function, renal function, and hematology tests [blood] Workers with TNT exposure excluded from exposed groups Comparison of mean value; prevalence of elevated value on an individual test 69 RDX exposed (43 male s, 26 females), 24 RDX/HMX exposed (all males), 338 referents (237 males, 101 females)

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 35 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference,

s etting and d esign Participants,

s election, f ollow

- up Consideration of l ikely

s election b ias Exposure m easure and

r ange Outcome

m easure Consideration of l ikely

c onfounding Analysis and

r esults d etails Sample s ize exposed groups (nondetected, >0. 01 mg/m 3 ) based on subject’s industrial hygiene monitoring results.
West and Stafford (1997) (United Kingdom) Ammunition workers (c ase

control study) 378 of 404 subjects (excluded 3 deaths and 23 subjects with unknown addresses) previously studied in 1991 , 32 cases with

a bnormal hematology test and 322 controls with normal hematology test ; p articipation rate among eligible subjects : 97% cases, 93% controls Former employees who were unable to work due to adverse health outcome were not included in the 1991 prevalence study Semiquantitative assessment; source of industrial hygiene data not reported

. Interviews with current and past employees and job title analysis were conducted to identify potential exposure to 100 agents, including RDX . Exposure surrogate was >50 hrs in duration and intensity was low (1

− 10 ppm) , moderate ( 10

− 100 ppm) , or high ( 100

− 1, 000 ppm)

. RDX exposure prevalence (males) was 83%. Abnormal hematology value in 1991 survey indicating possible mye lodysplasia: neutropenia (2.0 x 10 9 /L ), low platelet count (<150 x 109/ L ), or macrocytosis (mean corpuscular volume = 99 f L or >6% macrocytes) Cases and controls were not matched and statisti cal analyses were not adjusted for other risk factor or occupational exposures ; n o consideration or attempt to distinguish TNT Unadjusted prevalence odds ratios and 95% CIs ; a nalyses limited to males because of low frequency of exposure to females 32 cases (29 males, 3 females) and 322 controls (282 males, 12 females)



### 1 2 3 4 5 a Ma and Li (1993) d escribe symptoms reported by subjects during a physical examination , but these are not included in the evidence table because responses for individual symptoms were not identified.

CI = confidence interval; HMX = o ctahydro

- 1,3,5,7

- tetranitro

- 1,3,5,7

- tetr azocine ; LOD = limit of detection; TNT = trinitrotoluene

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 36 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine



## C.3. **OTHER PERTINENT TOXICITY INFORMATION**  1

2
3
4
5 6
7 8
9
10
11
12
13
14
15 16
17 18
19
20
21
22
23
24 25
26
27 28
29
30
31 32 **C.3.1. Mortality in Animals**  Evaluations of the evidence for specific health effects associated with RDX exposure are provided in Sections 1.2.1 to 1.2.6. In addition to these specific organ/system health effects, reduced survival associated with RDX exposure has been observed in experimental animals across multiple studies of varying exposure duration and study design (Table C

- 10). Evidence pertai n ing to mortality in experimental animals exposed to RDX is summarized in Table C

- 10; studies are ordered in the evidence table by duration of exposure and then species. Following chronic dietary exposure, an increased rate of mortality in F344 rats, and in particular male rats, at 40 mg/kg

- day was largely attributed to RDX

- related effects on the kidneys ( Levine et al., 1983b ) 4 

### ; see further discussion in Section 1. 2 .2. In a comparable chronic study, mice

w ere less sensitive than rats with respect to mortality following RDX exposure. After the high dose was red uced from 175 to 100 mg/kg

- day at week 11 in a 2

- year dietary study in B6C3F 1 

### mice because of high mortality, the mortality curve at 100 mg/kg

- day in surviving mice was not significantly different from the control group for the duration of the 2

- year study ( Lish et al., 1984 )

. T he investigators did not identify the probable cause of death at 175 mg/kg

- day. Increased rates of mortality were also observed in experimental animals that ingested RDX for durations up to 6 months ( Li sh et al., 1984 ; Levine et al., 1983b ; Levine et al., 1981a ; Cholakis et al., 1 980 ; von Oettingen et al., 1949 ) . The most detailed data on RDX

- related mortality come from a 9 0

- day gavage study in F344 rats by Crouse et al. (2006) . Across groups of rats exposed to 8

− 15 mg/kg

- day RDX, pre

- term deaths occurred in male rats as early as day 26

− 78 and in female rats as early as day 8

− 16 ( Johnson, 2015 ; Crouse et al., 2006 ) . Treatment

- related mortality was als o observed in the dams of rats exposed gestationally by gavage at doses ranging from 20 to 120 mg/kg

- day ( Angerhofer et al., 1986 ; Cholakis et al., 1980 ) . Deaths were additional ly reported i n one of 40 pregnant dams in both 2 and 6 mg/kg

- day groups in the rat developmental toxicity study by Angerhofer et al. (1986) In general, the evidence suggests that mortality occurs at lower doses in rats than in mice ( e.g., compar ison of rates from the 2

- year dietary studies in mice by Lish et al. (1984) and in rats by Levine et al. (1983b) ) , and at lower doses following gavage administration than dietary

a dministration (e.g., comparison of rates from the 13

- week rat studies using gavage ( Crouse et al., 2 006 ) and dietary ( Levine et al., 1981a ) administration ) . An RDX formulation with a larger particle siz e (e.g., ~200 μm) ( Cholakis et al., 1980 ) , which could potentially reduce the ability of RDX to

4 Deaths in high

- dose (40 mg/kg

- day) male rats were reported as early as week 4 (estimated from Figu re 4 in Levine et al. (1983a) ); the cause of death in rats that died prior to 6 months on study was generally not determined ( Levine et al., 19 83c ). Survival rates in both male and female rats at doses ≤8 mg/kg

- day RDX were similar to the control.

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C- 37 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

en ter the bloodstream, appears to produce less mortality than formulations with finer particle sizes 1
2 3
4
5 6
7 8
9 10
11
12

13 (e.g., median particle diameter of 20 μm) ( L evine et al., 1981a ) . There is evidence that mortality

ma y be associated with nervous system effects; several investigators reported that unscheduled deaths were frequently preceded by convulsions or seizures ( Crouse et al., 2006 ; Levine et al., 1 983b ; Cholakis et al., 1980 ) . In a number of studies, treatment

- related mortality was observed at d oses as low as doses associated with nervous system effects ( Crouse et al., 2006 ; Ange rhofer et al., 1 986 ; Levine et al., 1983b ; Levine et al., 1981a ; Cholakis et al., 1980 ; von Oettingen et al., 1949 )

. Th e evidence for an association between nervous system effects and mortality is discussed in more detail in Section 1. 2 .1, Nervous System Effects. In humans, there were no reports of mortality in case reports involving workers exposed to RDX during manufacture or in individuals exposed acutely as a result of accidental or intentional ingestion (see Appendix C, Section C.2).

**Table C**

**- 10. Evidence per**  **taining to mortality in animals a** 



### Reference and study design Results Lish et al. (1984) Mice, B6C3F 1 , 85/sex/group; interim sacrifices (10/sex/group) at 6 and 12 mo 89.2

− 98.7% pure, with 3

− 10% HMX as contaminant; 83

− 89% of particles <66 μm 0, 1.5, 7.0, 35, or 175/100 mg/kg

- d (high dose reduced to 100 mg/kg

- d in wk 11 due to excessive mortality) Diet 2 yrs ( mortality incidence also provided for mice through week 11 when the high dose was dropped because of high mort ality at that dose, and from the report of the 6

- month interim sacrifice) Doses 0 1.5 7.0 35 175/100 Mortality at 11 wks (incidence)

M 1 /85 0 /85 0 /85 0 /85 30 /85

F 0 /85 0 /85 0 /85 0 /85 36 /85 Mortality at 6 mo (incidence)

M 1/85 2/85 3/85 3/85 34/85

F 0/85 1/85 0/85 0/85 36/85 Mortality at 2 yrs (incidence)

M 20/65 23/65 25/65 29/65 41/65

F 16/65 21/65 14/65 21/65 42/65 After the high dose was reduced to 100 mg/kg

- d, survival was similar to controls. Levine et al. (1983b) Rats, F344, 75/sex/group; interim sacrifices (10/sex/group) at 6 and 12 mo 89.2−98.7% pure, with 3

− 10% HMX as contaminant: 83−89% of particles <66 μm 0, 0.3, 1.5, 8.0, or 40 mg/kg -d Diet 2 yrs ( m ortality incidence also provided for mice through week 1 3, and from the report of the 6

- month scheduled sacrifice) Doses 0 0.3 1.5 8.0 40 Mortality at 13 wks (incidence)

M 0 / 75 0 / 75 0 / 75 0 / 75 0 / 75

F 0 / 75 0 / 75 0 / 75 0 / 75 0 / 75 Mortality in 6

- mo interim sacrifice animals (incidence) #

# includes spontaneous death and moribund sacrifice

M 0/ 75 0/ 75 0/ 75 0/ 75 5/ 75
F 0/ 75 0/ 75 0/ 75 0/ 75 0/ 75 Mortality at 2 yrs (incidence)

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C- 38 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results

M 17/55 19/55 30/55* 26/55 51/55*

F 12/55 10/55 13/55 14/55 27/55* Cholakis et al. (1980) Mice, B6C3F 1 , 10−12/sex/group 88.6% pure, with 9% HMX and 2.2% water as contaminants; ~200 μm particle size 0, 40, 60, or 80 mg/kg

- d for 2 wks followed by 0, 320, 160, or 80 mg/kg

- d (TWA doses of 0, 79.6, 147.8, or 256.7 mg/kg

- d for males and 0, 82.4, 136.3, or 276.4 mg/kg

- d for females) b Diet 13 wks Doses 0 79.6 147.8 256.7 Mortality (incidence)

M 0/10 0/10 0/10 4/10*

F 0/11 0/12 0/10 2/12

Cholakis et al. (1980) Rats, F344, 10/sex/group 88.6% pure, with 9% HMX and 2.2% water as contaminants; ~200 μm particle size 0, 10, 14, 20, 28, or 40 mg/kg

- d Diet 13 wks Doses 0 10 14 20 28 40 Mortality (incidence)

M 0/10 0/10 0/10 0/10 0/10 0/10

F 1/10 (accidental

death) 0/10 0/10 0/10 0/10 0/10

Cholakis et al. (1980) Rats, CD, two

- generation study; F0: 22/sex/group; F1: 26/sex/group; F2: 10/sex/group 88.6% pure, with 9% HMX and 2.2% water as contaminants; ~200 μm particle size F0 and F1 parental animals: 0, 5, 16, or 50 mg/kg

- d Diet F0 exposure: 13 wks pre

- mating, and during mating, gestation, and lactation of F1; F1 exposure: 13 wks after weaning, and during mating, gestation, and lactation of F2; F2 exposure: until weaning Doses 0 5 16 50 Mortality in F0 adults (incidence) c M (F0) 0/22 0/22 0/22 2/22 F (F0) 0/22 0/22 0/22 6/22

M + F (F0) 0/44 0/44 0/44 8/44*

Crouse et al. (2006) Rats, F344, 10/sex/group 99.99% pure 0, 4, 8, 10, 12, or 15 mg/kg

- d Gavage 13 wks Doses 0 4 8 10 12 15 Mortality (incidence)

M 0/10 0/10 1/10 3/10 2/10 3/10

F 0/10 0/10 1/10 2/10 5/10 4/10

( Levine et al. (1990) ; Levine et al. (1981a) , 1981b) ) d Rats, F344, 10/sex/group; 30/sex for control Doses 0 10 30 100 300 600 Mortality (incidence) e

M 0/30 0/10 0/10 8/10 10/10 10/10

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 39 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results

84.7 ± 4.7% purity, ~10% HMX, median particle diameter 20 μm, ~90% of particles ≤66 μm 0, 10, 30, 100, 300, or 600 mg/kg

- d Diet 13 wks F 0/30 1/10 0/10 5/10 10/10 10/10

von Oettingen et al. (1949) Rats, sex/strain not specified, 20/group 90−97% pure, with 3−10% H MX; particle size not specified 0, 15, 25, or 50 mg/kg

- d Diet 13 wks Doses 0 15 25 50 Mortality (incidence) 0/20 1/20 (probably not related

to RDX) 8/20 8/20

Hart (1974) Dogs, Beagle, 3/sex/group Pre

- mix with ground dog chow containing 20 mg RDX/g

- chow, 60 g dog food ; p urity and particle size not specified 0, 0.1, 1, or 10 mg/kg

- d Diet 13 wk s Doses 0 0.1 1 10 Mortality (incidence)

M 0/3 0/3 1/3 (not related to RDX) 0/3

F 0/3 0/3 0/3 0/3

Martin and Hart (1974) Monkeys, Cynomolgus or Rhesus f
, 3/sex/group Purity and particle size not specified 0, 0.1, 1, or 10 mg/kg

- d Gavage 13 wks Doses 0 0.1 1 10 Mortality (incidence)

M 0/3 0/3 0/3 0/3

F 0/3 0/3 0/3 1/3 (animal exhibited neurological effects; euthanized)

von Oettin gen et al. (1949) Dogs, breed not specified, 5 females/group (control); 7 females/group (exposed) 90−97% pure, with 3−10% HMX; particle not specified 0 or 50 mg/kg

- d Diet 6 d/wk for 6 wks Doses 0 50 Mortality (incidence)

F 0/5 1/7

MacPhail et al. (1985) Rats, Sprague

- Dawley derived CD, 8−10 males or females/group Purity 84 ± 4.7%; ≤ 66 μm particle size 0, 1, 3, or 10 mg/kg

- d Gavage 30 d No mortality was rep orted (incidence data were not provided).

Cholakis et al. (1980) Doses 0 0.2 2.0 20 Mortality (incidence)

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 40 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results Rabbits, New Zealand White (NZW), 11−12 pregnant females/group 88.6% pure, with 9% HMX and 2.2% water as contaminants; ~200 μm particle size 0, 0.2, 2.0, or 20 mg/kg -d Diet GDs 7−29 F 0/11 0/11 0/11 0/12

Cholakis et al. (1980) Rats, F344, 24−25 females/group 88.6% pure, with 9% HMX and 2.2% water as contaminants 0, 0.2, 2.0, or 20 mg/kg -d Gavage GDs 6−19 Doses 0 0.2 2.0 20 Mortality (incidence)

F 0/24 0/24 0/24 5/24 (1 rat accidentally killed; removed from analysis) Angerhofer et al. (1986) (range

- finding study) Rats, Sprague

- Dawley, 6 pregnant females/group Purity 90%; 10% HMX and 0.3% acetic acid occurred as contaminants 0, 10, 20, 40, 80, or 120 mg/kg -d Gavage GDs 6−15 Doses 0 10 20 40 80 120 Mortality (incidence)

F 0/6 0/6 0/6 6/6 6/6 6/6

Angerhofer et al. (1986) Rats, Sprague

- Dawley, 39−51 mated females/group Purity 90%; 10% HMX and 0.3% acetic acid occurred as contaminants 0, 2, 6, or 20 mg/kg -d Gavage GDs 6−15 Doses 0 2 6 20 Mortality (incidence)

F 0/39 1/40 1/40 16/51 Not reported whether deaths at 2 and 6 mg/kg

- d related or likely related to RDX exposure 

### 1 2 3 4 5 6

c 

### 7 8 9 10 11
f 

### 12 13 14 15 *Statistically significant ( p < 0.05) based on analysis by the study authors.

a The 2

- year rat study by Hart (1976) was not included in this evidence table because a malfunctioning heating system incident resulted in the premature deaths of 59 animals on study days 75−76 across groups, thereby confounding mortality findings.
b Doses were calculated by the study authors. Data for male and female rats were combined for statistical analysis.
d Levine et al. (1981a) is a laboratory report of a 13

- week study of RDX in F344 rats; two subsequently published papers ( Levine et al., 1990 ; Levine et al., 1981b ) present subsets of t he data provided in the full laboratory report.

e Animals receiving 300 mg/kg

- day died by week 3 of the study; animals receiving 600 mg/kg

- day died by week 1 of the study. The species of monkey used in this study was inconsistently reported in the study as either Cynomolgus (in the methods section) or Rhesus (in the summary).

TWA = time

- w eighted average

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C- 41 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**C .3.2.**  **Other Noncancer Effects** 1
2
3
4
5 6
7 8

9
10
11
12
13
14
15 16
17 18
19
20
21
22
23
24
25
26
27 28
29
30

31
32 33
34
35 T here are isolated reports of RDX inducing systemic effects in several organs/systems, including the eyes and the musculoskeletal, cardiovascular, immune, and GI systems. However, there is less evidence for these effects compared to organ systems describe d in Section 1.2. Overall, at the present time, the evidence does not support identifying these other systemic effects as human hazards of RDX exposure. S ummaries of the evidence for other systemic effects in humans

an d animals are shown in Tables C- 11 a nd C- 12 , respectively. Experimental animal studies are ordered in the evidence table by type of effect, and then by duration of exposure and by species.

Ocular Effects
There are no reports of ocular effects in human case reports or epidemiological studies . In experimental animals, evidence of ocular effects comes from cataract findings in one 2

- year bioassay. Specifically, the incidence of cataracts was 73% in female F344 rats exposed to 40 mg/kg

- day RDX in the diet for 2 years, compared with 32% in the control group ( Levine et al., 1 983b ) . After 76 weeks of exposure, the incidence of cataracts in female rats at 40 mg/kg

- day ( 23%) was also elevated compared to controls (6%). The incidence of cataracts was not increased in RDX

- exposed male rats in the same study ( Levine et al., 1983b ) , and other studies have not

o bserved ocular effects associated with RDX exposure. Only two rats (do se groups not reported) were observed to have mild cataracts in a 90

- day study of male and female F344 rats exposed to RDX at doses up to 15 mg/kg

- day by gavage; however, the authors noted that these observations are common in F344 rats at 4 months of age and should not be attributed to treatment ( Crouse et al., 2 006 ) . Furthermore, cataracts were not observed in male or female F344 rats exposed to 4 0 mg/kg

- day RDX by diet for 90 days ( Cholakis et al., 1980 ) or in male or female B6C3F 1 

### mice

ex posed to RDX in the diet for 2 years at doses up to approximately 100 mg/kg

- day ( Lish et al., 1 984 ) . A statistically significant increase in the incidence of cataracts in male mice was initially

no ted by Lish et al. (1984) , but was not confirmed when mice used for orbital bleedings were

ex cluded from the analysis, suggesting that the effect was not treatment related. No ocular effects were observed in monkeys exposed by gavage for 90 days at doses up to 10 mg/kg

- day ( Martin and H art, 1974 )

. In summary, the incidence of cataracts w as statistically significantly increased in high

- dose female rats in one chronic oral study; however, this finding was not reproduced in other subchronic and chronic studies in rats or mice.

Cardiovascular Effects
Human evidence for cardiovascular effects is limited to case reports that include observations of transient arterial hypertension in male workers following inhalation of RDX during manufacturing ( Barsotti and Crotti, 1949 ) , sinus tachycardia, and in one instance
, premature

v entricular beats in five men following accidental ingesti on of RDX at 37

− 250 mg/kg body weight ( Küçükardali et al., 2003 ) (s ee Appendix C, Section C.2).
This document is a draft for review purposes only and does not constitute Agency polic y. 

### C- 42 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

I nconsistent observations of cardiovascular effects have been reported in animal studies. 1
2 3
4
5 6
7 8
9 10
11
12 13
14
15 16
17 18
19 20
21
22

23
24 25
26
27
28 29
30

31
32
33 34
35 An increase in the relative heart

- to

- body weight ratio was observed at the highest dose tested in B6C3F 1 

### mice (male: 13%; female : 17%) and F344 rats (male: 22%; female : 15%) following chronic dietary administration of RDX ( L ish et al., 1984 ; Levine et al., 1983b ) ; however, these do ses also

r esulted in reductions in body weight in both males and females. Dose

- related decreases in absolute heart weight in rats were reported in some subchronic (dietary) studies ( Levine et al., 1 990 ; Levine et al., 1981a
, b ; Cholakis et al., 1980 ) , whereas little change or modest increases in

a bsolute heart weight were observed in other subchronic studies in rats or mice ( Crouse et al., 2 006 ; Cholakis et al., 1980 ) . A subchronic study in male dogs reported a 31% increase in absolute h eart weight at the highest dose tested (10 mg/kg

- day) ( Hart, 1974 ). Evidence for changes in histopathology associated with RDX exposure is limited to findings of an increased incidence of focal myocardial degeneration in female rats (6/10 versus 2/10, respectively ) and male mice (5/10 versus 0/10, respectively) compared with controls following exposure to RDX in the diet for 90 days ( Cholakis et al., 1980 ) . With the exception of one male

m ouse, the severity of the lesion was characterized as minimal. In each study, the finding of myocardial degeneration was limited to one sex and to the high

- dose group only; the high dose in the male mouse study caused 40% mortality. Other studies in monkeys ( Martin and Hart, 1974 )

a nd rats ( von Oettingen et al., 1949 ) reported no observable cardiovascular effects. In summary, evidence for cardiovascular effects associated with RDX exposure consists of
two case reports of cardiovascular effects following acute exposure, inconsistent findings of changes in heart weight in experimental animals, and one report of minimal histopathological changes in a 90

- day rat study t hat was not confirmed in other toxicity studies.

Musculoskeletal Effects
Evidence of musculoskeletal effects in humans consists of case reports that include observations of muscle twitching, myalgia/muscle soreness, and muscle injury as indicated by elevated levels of aspartate aminotransferase ( AST ) or myoglobinuria ( Küçükardali et al., 2003 ; Hett and Fichtner, 2002 ; Hollander and Colbach, 1969 ; Stone et al., 1969 ; Merrill, 1968 ) (see A ppendix C, Section C.2). Histological evaluations of musculature or skeletal tissue did not reveal any alterations in mice ( Lish et al., 1984 ) or rats ( Levine et al., 1983b ; Hart, 1976 ) following chronic

o ral exposure to RDX, in mice and rats followi ng subchronic exposure ( Cholakis et al., 1980 ) , or in d ogs following a 90

- day dietary exposure ( Hart, 1974 )

. Immune System Effects
RDX is struct urally similar to various drugs known to induce the autoimmune disorder systemic lupus erythematosus (SLE). Based on the initial identification of three cases of SLE at

one U.S. Army munitions plant, further investigation was conducted on a population of 69 employees at five U.S. Army munitions plants with potential exposure to RDX ( Hathaway and B uck, 1977 ) ; no additional cases of SLE were identified. Increased WBC counts have been reported

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C- 43 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

in some case reports of individuals (troops during the Vietnam war) who ingested or inha led RDX 1
2 3
4
5 6
7 8
9 10
11
12 13
14
15 16
17 18
19 20
21
22
23
24
25
26

27
28 29
30
31 32
33
34
35 36 or C

- 4 (91% RDX) ( Kn epshield and Stone, 1972 ; Hollander and Colbach, 1969 ; Stone et al., 1969 ; Merrill, 1968 )

. In animal studies , increased WBC count in female rats following subchronic dietary exposure to RDX was the only dose

- related immune effect reporte d ( Levine et al., 1990 ; Levine et al., 1 981a , b ) ; WBC counts in male rats were unaffected. Conversely, decreased WBC counts were

r eported in male and female rats in a 2

- year study ( Hart, 1976 ) . Changes in spleen weights were

o bserved across studies, but the responses were not consistent and did not appear to be dose

related. For example, in 90

- day studies, Cholakis et al. (1980) identified a statistically significant d ecrease in absolute spleen weight in female F344 rats at 40 mg/kg

- day, while Crouse et al. (2006)

o bserved a statistically significant increase in spleen weight at >10 mg/kg

- day. Across studies, there was no significant or dose

- dependent pattern of response to suggest that the WBC changes reflect RDX

- induced immunotoxicity. No dose

- related immune effects from oral exposure to RDX were observed in other animal studies, including a 90

- day study in F344 rats specifically designed to evaluate immunotoxicity (parameters included evaluation of red blood cell [RBC] and WBC populations, proportion of cell surface markers, cellularity in proportion to organ weight, B and T cells in the spleen, and CD4/CD8 antigens of maturing lymphocytes in the thymus) ( Crouse et al., 2 006 ) . Routine clinical and histopathology evaluations of immune -related organs in a two

g eneration study in rats ( Cholakis et al., 1980 ) and chronic studies in rats ( Levine et al., 1983b ) and
mi ce ( Lish et al., 1984 ) provide no evidence of immunotoxicity associated with oral (diet ary)

ex posure to RDX. In summary , evidence for immunotoxicity associated with RDX exposure is limited to findings from one study of increased WBC counts in female rats ( Levine et al., 1981a
, b ) . Evidence
t hat RDX is not immunotoxic comes from several animal studies, including other repeat

- dose oral studies in mice and rats ( Crouse et al., 2006 ; Lish et al., 1984 ; Levine et al., 1983b ; Cholakis et al., 1 980 )

. Gastrointestinal Effects
Clinical signs of nausea and/or vomiting have been frequently identified in case reports of accidental or intentional RDX poisonings, and have generally been concurrent with severe neurotoxicity ( Kasuske et al., 2009 ; Davies et al., 2007 ; Küçükardali et al., 2003 ; Hett and Fichtner, 2 002 ; Ketel and Hughes, 1972 ; Knepshield and Stone, 1972 ; Hollander and Colbach, 1969 ; Stone et al., 1 969 ; Merrill, 1968 ; Kaplan et al., 1965 ; Barsotti and Crotti, 1949 ) (see Appendix C, Section C.2). I n animal studies, vomiting has also been observed following oral exposure in swine (single

- dose study) ( Musick et al., 2 010 ) , dogs ( Hart, 1974 ) , and monkeys ( Martin and Hart, 1974 ) . One

s ubchronic oral (diet) rat study from the early literature reported congestion of the GI tract at doses also associated with elevated mortality ( von Oettingen et al., 1949 ) ; however, none of the

su bsequent subchronic or chronic animal studies reported histological changes of the GI tract

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C- 44 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

related to RDX administered via gavage or the diet ( Crouse et al., 2006 ; Lish et al., 1984 ; Levine et 1
2 3
4
5 6

7 8
9
10
11
12
13
14
15 16
17 18
19
20
21
22
23
24 25
26
27
28 29
30
31 32
33
34
35 36 al., 1 983b ; Hart, 1974 ; Martin and Hart, 1974 )

. In summary , evidence for GI tract effects associated with RDX exposure consists largely of reports of nausea and vomiting in humans acutely exposed to RDX and similar reports of vomiting in swine, dogs, and monkeys. In general, histopathological changes have not be reported in experimental animals exposed to RDX in the diet.

Hematological Effects
Elevated prevalence odds ratios (ORs) for hematological abnormalities (i.e., neutropenia, low platelet count, or macrocytosis; see Table C-11 for criteria used to define abnormal) were observed in a case

- control study of men (24 cases, 199 controls) exposed to RDX in ordnance factories ( West and Stafford, 1997 ) (see Table C

- 11 ). The prevalence OR for an association between R DX exposure and hematological abnormalities was 1.7 (95% confidence interval [CI] 0.7−4.2) for men with >50 hours of low

- intensity exposure (based on 22 cases), while the prevalence OR was 1.2 (95% CI 0.3−5.3) for men with >50 hours of high

- intensity exposure (based on 2 cases). The ORs from this study must be interpreted with caution given the small sample size, wide CIs, and lack of identification of co

- exposures. No changes in hematological parameters (including hemoglobin, hematocrit, and reticulocyte count) were observed in a cross

- sectional epidemiologic study of 69 workers exposed to RDX by inhalation (RDX exposure range: undetectable [ <0.01 mg/m 3 

### ] to 1.6 mg/m 3 

### ) ( Hathaway and Buck, 1977 ) . Humans who ingested or inhaled

u nknown amounts of RDX or C

- 4 (~91% RDX) for an acute duration di splayed temporary hematological alterations, including anemia, decreased hematocrit, hematuria, and methemoglobinemia ( Kasuske et al., 2009 ; Küçükardali et al., 2003 ; Knepshield and Stone, 1972 ; Hollander and Colbach, 1969 ; Stone et al., 1969 ; Merrill, 1968 ) . In other case reports, normal blood

c ounts were observed in accidentally exposed individuals ( Testud et al., 1996a ; Goldberg et al., 1 992 ; Woody et al., 1986 ; Ketel and Hughes, 1972 ; Kaplan et al., 1965 ) (see Appendix C, S ection C.2). In animals , hematological alterations were obse rved following oral exposure in chronic and subchronic studies in both sexes of rats (F344 or Sprague

- Dawley) and B6C3F 1 

### mice (see Table C- 12 ). Increases in platelet count were observed in male and female mice and rats in some subchronic and chronic studi es at doses ranging from 0.3 to 320 mg/kg

- day ( Lish et al., 1984 ; Levine et al., 1983b ; Cholakis et al., 1980 ) ; however, changes were generally inconsistent across

s tudies and were not generally dose

- dependent. Similarly, decreased hemoglobin levels/anemia were observed in some chronic and subchronic studies ( Levine et al., 1983b ; Cholakis et al., 1980 ; von Oettingen et al., 1949 ) , particularly at doses ≥15 mg/kg

- day, but trends in hemoglobin levels

a cross studies did not show a consi stent relationship with dose. Other hematological parameters, including WBC counts, reticulocyte counts, and hematocrit, showed conflicting results between studies, marginal responses, or inconsistent changes with increasing dose. Other subchronic

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C- 45 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

studie s in rats and dogs ( Crouse et al., 2006 ; Hart, 1974 ; von Oettingen et al., 1949 ) did not identify 1
2 3
4
5 6
7 8
9

10 11 an y changes in hematological parameters. In summary , evidence for hematological effects associated with RDX exposure in humans comes from several case reports that found transient fluctuations in hematological endpoints after acute exposures. Hematological findings from the case

- control study and the cros s- sectional study were not consistent. The small number of cases or exposed individuals, respectively, from the case

control and cross

- sectional study may contribute to the difficulty in interpreting the results across studies (Table C -11 ). In general, a nimal studies of chronic and subchronic durations showed no consistent, dose

- related pattern of increase or decrease in hematological parameters.

**Table C**

**- 11**  **. Evidence pertaining to other noncancer**  **effects (hematological) in humans** 

Reference and study d esign Results Hematological effects West and Stafford (1997) (United Kingdom) Case

- control study of ordnance factory workers, 32 cases with abnormal and 322 controls with normal hematology test drawn from 1991 study of 404 workers at ammunitions plant; participation rate 97% of cases, 93% of controls. Analysis limited to men (29 cases, 282 controls). Anal ysis specific to RDX: 22 low - and 2 high

- intensity cases; 182 low - and 17 high

- intensity controls. Exposure measure s: Exposure determination based on employee interviews and job title analysis; data included frequency (hrs/d, d/yr), duration (yrs), and int ensity (low [1−10 ppm], moderate [10−100 ppm], and high [100−1,000 ppm], based on ventilation considerations). Effect measures : Hematology tests; blood disorder defined as neutropenia (2.0 × 10 9 /L), low platelet count (<150 × 10 9 /L), or macrocytosis (mean corpuscular volume = 99 fl or >6% macrocytes). Analysis : Unadjusted OR. Hematological abnormality (neutropenia, low platelet count, or macrocytosis) (OR; 95% CI [number of exposed cases]) Low intensity, 50

- hr

- duration 1.7; 0.7,4.2 [22]
Medium intensity, 50

- hr duration 1.6; not reported [not reported] High intensity, 50

- hr duration 1.2; 0.3, 5.3 [2]

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C- 46 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study d esign Results Hathaway and Buck (1977) (United States) Cross

- sectional study, 2,022 workers, 1,491 participated (74% response rate). Analysis limited to whites; 69 exposed to RDX alone and 24 exposed to RDX and HMX; 338 not exposed to RDX, HMX, or TNT. Exposure measures : Exposure determination based on job title and industrial hygiene evaluation. Exposed subjects assigned to two groups: <LOD or ≥0.01 mg/m 3 (mean for employees with exposures ≥LOD: 0.28 mg/m 3 ). Effect measures : Hematology tests. Analysis : Types of statistical tests were not reported (assumed to be t

- tests for comparison of means and χ 2 tests for comparison of proportions). Hematology tests in men (mean; standard deviation not reported)

Test Referent (n = 237) RDX exposed* Undetected

(<LOD) (n = 22) >0.01 mg/m 3 (n = 45) Hemoglobin 15.2 14.7 15.2

Hematocrit 42 45.6 47 Reticulocyte count 0.7 0.9 0.7

*Includes both workers exposed to RDX alone and RDX and HMX. No differences were statistically significant. Similar results in

women. Hematology tests in men (prevalence of abnormal values)

Test (abnormal

range) Referent RDX exposed*
Undetected

(<LOD) >0.01 mg/m 3 Hemoglobin (< 14) 15/237 3/22 4/45

Hematocrit (<40) 1/237 1/22 1/45

Reticulocyte count (>1.5) 18/237 3/22 2/45

*Includes both workers exposed to RDX alone and RDX and HMX. No differences were statistically significant. Similar results in

women.

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C- 47 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**T able**  **C-12 . Evidence pertaining to**  **other noncancer effects in animals**  **a** 

### 1

Reference and study design Results Ocular effects Lish et al. (1984) Mice, B6C3F 1 , 85/sex/group; interim sacrifices (10/sex/group) at 6 and 12 mo 89.2−98.7% pure, with 3−10% HMX as contaminant; 83−89% of particles <66 μm 0, 1.5, 7.0, 35, or 175/100 mg/kg

- d (high dose reduced to 100 mg/kg

- d in wk 11 due to excessive mortality) Diet 2 yrs Doses 0 1.5 7.0 35 175/100 Cataracts; 103 wks (incidence) b

M 2/47 2/41 0/41 2/37 2/16

F 2/50 1/37 6/52 0/46 1/26

Levine et al. (1983b) Rats, F344, 75/sex/group; interim sacrifices (10/sex/group) at 6 and 12 mo 89.2−98.7% pure, with 3−10% HMX as cont aminant; 83−89% of particles <66 μm 0, 0.3, 1.5, 8.0, or 40 mg/kg -d Diet 2 yrs Doses 0 0.3 1.5 8.0 40 Cataracts; 103 wks (incidence)

M 8/40 6/39 6/31 8/35 2/6

F 14/44 4/48 11/44 8/43 22/30*

Cholakis et al. (1980) Rats, F344, 10/sex/group 88.6% pure, with 9% HMX and 2.2% water as contaminants; ~200 μm particle size 0, 10, 14, 20, 28, or 40 mg/kg -d Diet 13 wks No ocular effects were observed (gross examination of eye was performed in all animals, and microscopic examination was performed in control and 40 mg/kg

- d animals).

Crouse et al. (2006) Rats, F344, 10/sex/group 99.99% pure 0, 4, 8, 10, 12, or 15 mg/kg -d Gavage 13 wks No ocular effects were observed (ophthalmic examinations were performed in all animals within 1 wk of sacrifice, and microscopic examination of the eye was performed in control and 15 mg/kg -d animals).

Martin and Hart (1974) Monkeys, Cynomolgus or Rhesus c , 3/sex/group Purity of test material not specified 0, 0.1, 1, or 10 mg/kg -d Gavage 13 wks No ocular effects were observed (ophthalmoscopic examination was performed at the end of exposure).

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C- 48 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results Cardiovascular effects Lish et al. (1984) Mice, B6C3F 1 , 85/sex/group; interim sacrifices (10/sex/group) at 6 and 12 mo 89.2−98.7% pure, with 3−10% HMX as contaminant; 83−89% of particles <66 μm 0, 1.5, 7.0, 35, or 175/100 mg/kg

- d (high dose reduced to 100 mg/kg

- d in wk 11 due to excessive mortality) Diet 2 yrs Doses 0 1.5 7.0 35 175/100 Absolute heart weight; 104 wks (percent change compared to control)

M 0% 4% 4% 5% 7%

F 0% 1% 5% 2%

− 5%
Relative heart

- to

- body weight; 104 wks (percent change compared to control)

M 0% 7% 5% 5% 13%*

F 0% 0% 6% 4% 17%* Body weight was significantly lower at termination in males and females exposed to 175/100 mg/kg

- d (−5 and −19%, respectively). Hart (1976) Rats, Sprague

- Dawley, 100/sex /group Purity and particle size not specified 0, 1.0, 3.1, or 10 mg/kg

- d Diet 2 yrs Doses 0 1.0 3.1 10 Myocardial fibrosis (percent incidence; number not reported)

M 20%

− − 5%

F 5%

− − 1%
Endocardial disease (percent incidence; number not reported)

M 1%

− − 3%

F 0%

− − 0% Absolute heart weight; 104 wks (percent change compared to control)

M 0%

− 6%

− 2%

− 5%

F 0% 13% 3% 15%
Relative heart

- to

- body weight; 104 wks (percent change compared to control)

M 0%

− 2% 4% 1%

F 0% 23% 13% 27% Levine et al. (1983b) Rats, F344, 75/sex/group; interim sacrifices (10/sex/group) at 6 and 12 mo 89.2−98.7% pure, with 3−10% HMX as contaminant; 83−89% of particles <66 μm 0, 0.3, 1.5, 8.0 , or 40 mg/kg

- d Diet 2 yrs Doses 0 0.3 1.5 8.0 40 Absolute heart weight; 104 wks (percent change compared to control)

M 0% 3%

− 2%

− 2% 1%

F 0%

− 1% 0%

− 4%

− 3%
Relative heart

- to

- body weight; 104 wks (percent change compared to control)

M 0% 2% 6% 0% 22%

F 0%

− 2% 3%

− 1% 15%

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 49 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results Cholakis et al. (1980) Mice, B6C3F 1 , 10−12/sex/group 88.6% pure, with 9% HMX and 2.2% water as contaminants; ~200 μm particle size Experiment 1 : 0, 10, 14, 20, 28, or 40 mg/kg

- d Diet 13 wks Doses 0 10 14 20 28 40 Absolute heart weight (percent change compared to control)

M 0%

− − − 7% 7%

F 0%

− − − 0% 0% Relative heart weight (percent change compared to control)

M 0%

− − − 6% 0%

F 0%

− − − − 4% 0% Experiment 2 : 0, 40, 60, or 80 mg/kg

- d for 2 wks followed by 0, 320, 160, or 80 mg/kg

- d (TWA doses of 0, 79.6, 147.8, or 256.7 mg/kg

- d for males and 0, 82.4, 136.3, or 276.4 mg/kg

- d for females) d Diet 13 wks Doses 0 80 160 320 Focal myocardial degeneration (incidence)

M # 0/10

− − 5/10* , ‡

F † 0/11

− − 2/11 Absolute heart weight (percent change compared to control)

M 0% 0% 0% 8%

F 0% 0% 0% 8%
Relative heart

- to

- body weight (percent change compared to control)

M 0% 0%

− 2% 6%

F 0% 0%

− 2% 2%

# Includes one affected and three unaffected animals that died prematurely.

† Includes one unaffected animal that died prematurely.
‡ Minimal severity in four rats, moderate in one. Cholakis et al. (1980) Rats, F344, 10/sex/group 88.6% pure, with 9% HMX and 2.2% water as contaminants; ~200 μm particle size 0, 10, 14, 20, 28, or 40 mg/kg

- d Diet 13 wks Doses 0 10 14 20 28 40 Focal myocardial degeneration, minimal severity (incidence)

M 3/10

− − − − 1/10

F 2/10

− − − − 6/10 Absolute heart weight (percent change compared to control)

M 0%

− − − 0%

− 8%*

F 0%

− − − − 6%

− 11%*
Relative heart

- to

- body weight (percent change compared to control)

M 0%

− − − 3% 0%

F 0%

− − − − 3%

− 8%
Relative heart

- to

- brain weight (percent change compared to control)

M 0%

− − − − 4%

− 10%*

F 0%

− − − − 5%

− 11%*

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 50 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results Cholakis et al. (1980) Rats, CD, two

- generation study; F0: 22/sex/group; F1: 26/sex/group; F2: 10/sex/group 88.6% pure, with 9% HMX and 2.2% water as contaminants; ~200 μm particle size F0 and F1 parental animals: 0, 5, 16, or 50 mg/kg

- d Diet F0 exposure: 13 wks pre

- mating, and during mating, gestation, and lactation of F1; F1 exposure: 13 wks after weaning, and during mating, gestation, and lactation of F2; F2 exposure: until weaning No cardiac effects were observed (microscopic examination of heart was performed in randomly selected F2 animals).

Doses 0 5 16 50 Absolute heart weight (percent change compared to control)

F2 M 0% 3.2%

− 6.5%

−
F2 F 0% 15%

− 3.7%

−

Crouse et al. (2006) Rats, F344, 10/sex/group 99.99% pure 0, 4, 8, 10 , 12, or 15 mg/kg

- d Gavage 13 wks Doses 0 4 8 10 12 15 Cardiomyopathy (incidence)

M 2/10

− − − − 3/8

F 0/10

− − − − 1/6 Absolute heart weight (percent change compared to control)

M 0%

− 2%

− 7%

− 1% 1% 11%

F 0%

− 2% 0% 8% 7% 6%
Relative heart

- to

- body weight (percent change compared to control)

M 0% 4% 2% 1%

− 1% 8%

F 0%

− 2%

− 7%

− 6%

− 9%

− 16%* ( Levine et al. (1990) ; Levine et al. (1981a) , 1981b) ) e Rats, F344, 10/sex/group; 30/sex for control 84.7 ± 4.7% purity, ~10% HMX, median particle diameter 20 μm, ~90 % of particles ≤66 μm 0, 10, 30, 100, 300, or 600 mg/kg

- d Diet 13 wks All animals in the 300 and 600 mg/kg

- d groups died prior to study termination.

Doses 0 10 30 100 300 600 Chronic focal myocarditis (incidence)

M 8/30 8/10 6/10 1/10 1/10 0/10

F 8/30 3/10 1/10 1/10 1/10 1/9 Absolute heart weight (percent change compared to control)

M 0%

− 2%

− 10%

− 15%

− −
F 0%

− 3% 0%

− 5%

− − Relative heart

- to

- body weight (percent change compared to control)

M 0% 2%

− 4% 3%

− −
F 0%

− 2% 0%

− 3%

− −

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 51 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results von Oettingen et al. (1949) Rats (sex/strain not specified); 20/group Purity and particle size not specified 0, 15, 25, or 50 mg/kg

- d Diet 13 wks The study authors reported that there were no cardiac effects (microscopic examination of the heart was performed in all rats; data were not shown).

Hart (1974) Dogs, Beagle, 3/sex/group Pre

- mix with ground dog chow containing 20 mg RDX/g

- chow, 60 g dog food ; p urity and particle size not specified 0, 0 .1, 1, or 10 mg/kg

- d Diet 13 wks Doses 0 0.1 1 10 Focal hyalinization of the heart (incidence)

M 0/3

− − 0/3

F 0/3

− − 1/3 Absolute heart weight (percent change compared to control)

M 0%

− − 31%

F 0%

− − 5.7% Martin and Hart (1974) Monkeys, Cynomolgus or Rhesus c , 3/sex/group Purity of test material not specified 0, 0.1, 1, or 10 mg/kg

- d Gavage 13 wks Doses 0 0.1 1 10 Myocarditis (percent change compared to control)

M 1/3

− − 1/3

F 0/3

− − 0/3 Absolute heart weight (percent change compared to control)

M 0% 7%

− 1% 5%

F 0% 10% 12%

− 12% Immune effects Lish et al. (1984) Mice, B6C3F 1 , 85/sex/group; interim sacrifices (10/sex/group) at 6 and 12 mo 89.2−98.7% pure, with 3−10% HMX as contaminant; 83−89% of particles

< 66 μm 0, 1.5, 7.0, 35, or 175/100 mg/kg

- d (high dose reduced to 100 mg/kg

- d in wk 11 due to excessive mortality) Diet 2 yrs No immune effects were observed with routine hematology, clinical chemistry, or histopathology evaluations.

Doses 0 1.5 7.0 35 175/100 WBC count; 105 wks (percent change compared to control)

M 0%

− 13%

− 8%

− 16%

− 30%

F 0% 12% 39%* 28% 0% Absolute spleen weight; 105 wks (percent change compared to control)

M 0% 24% 31%

− 10%

− 28%

F 0% 4% 15%

− 17% 16% Relative spleen weight; 105 wks (percent change compared to control)

M 0% 26% 32%

− 11%

− 21%

F 0% 4% 15%

− 17% 44%

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 52 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results Hart (1976) Rats, Sprague

- Dawley, 100/sex/group Purity and particle size not specified 0, 1.0, 3.1, or 10 mg/kg

- d Diet 2 yrs Doses 0 1.0 3.1 10 WBC count; 104 wks (percent change compared to control)

M 0%

− 13%

− 22%*

− 34%*

F 0% 5%

− 32%*

− 12% Absolute spleen weight; 104 wks (percent change compared to control)

M 0%

− 11%

− 16%

− 4%

F 0% 58% 8% 37% Relative spleen weight; 104 wks (percent change compared to control)

M 0%

− 11%

− 14% 1%

F 0% 77% 19% 55% Levine et al. (1983b) Rats, F344, 75/sex/group; interim sacrifices (10/sex/group) at 6 and 12 mo 89.2−98.7% pure, with 3−10% HMX as contaminant; 83−89% of particles <66 μm 0, 0.3, 1.5, 8.0, or 40 mg/kg

- d Diet 2 yrs No immune effects were observed with routine hematology, clinical chemistry, or histopathology evaluations.

Doses 0 0.3 1.5 8.0 40 WBC count; 105 wks (percent change compared to control)

M 0%

− 11% 103% f 184% f 15%

F 0% 7% 12% 354% f 251% f Absolute spleen weight; 105 wks (percent change compared to control)

M 0% 5%

− 10%

− 32%

− 49%

F 0%

− 28%

− 44%

− 35% 17% Relative spleen weight; 105 wks (percent change compared to control)

M 0% 9% 4%

− 29%

− 38%

F 0%

− 34%

− 45%

− 36% 9% Cholakis et al. (1980) Mice, B6C3F 1 , 10−12/sex/group 88.6% pure, with 9% HMX and 2.2% water as contaminants; ~200 μm particle size Experiment 1: 0, 10, 14, 20, 28, or 40 mg/kg

- d Diet 13 wks Doses 0 10 14 20 28 40 Absolute spleen weight (percent change compared to control)

M 0%

− − − 18% 13%

F 0%

− − − − 2%

− 8% Relative spleen weight (percent change compared to control)

M 0%

− − − 24% 14%

F 0%

− − − − 3%

− 5%

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 53 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results
Experiment 2: 0, 40, 60, 80 mg/kg

- d for 2 wks followed by 0, 320, 160, or 80 mg/kg

- d (TWA doses of 0, 79.6, 147.8, or 256.7 mg/kg

- d for males and 0, 82.4, 136.3, or 276.4 mg/kg

- d for females) d Diet 13 wks Doses 0 80 160 320

WBC count (percent change compared to control)

M 0%

− 27%

− 12% 30%

F 0%

− 17% 3%

− 3% Absolute spleen weight (percent change compared to control)

M 0% 17% 0%

− 17%

F 0%

− 22% 0% 0% Relative spleen weight (percent change compared to control)

M 0% 25% 5% 0%

F 0%

− 12% 0%

− 3% Cholakis et al. (1980) Rats, F344, 10/sex/group 88.6% pure, with 9% HMX and 2.2% water as contaminants; ~200 μm particle size 0, 10, 14, 20, 28, or 40 mg/kg

- d Diet 13 wks Doses 0 10 14 20 28 40

WBC count (percent change compared to control)

M 0%

− − − − 12% 7%

F 0%

− − − 17% 30% Absolute spleen weight (percent change compared to control)

M 0%

− − − 2%

− 4%

F 0%

− − − − 10%

− 12%* Relative spleen weight (percent change compared to control)

M 0%

− − − 5% 5%

F 0%

− − − − 8%

− 8% Cholakis et al. (1980) Rats, CD, two

- generation study; F0: 22/sex/group; F1: 26/sex/group; F2: 10/sex/group 88.6% pure, with 9% HMX and 2.2% water as conta minants; ~200 μm particle size F0 and F1 parental animals: 0, 5, 16, or 50 mg/kg

- d Diet F0 exposure: 13 wks pre

- mating, and during mating, gestation, and lactation of F1; F1 exposure: 13 wks after weaning, and during mating, gestation, and lactation of F2; F2 exposure: until weaning No immune effects were observed upon routine histopathology evaluation.

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 54 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results Crouse et al. (2006) Rats, F344, 10/sex/group 99.99% pure 0, 4, 8, 10, 12, or 15 mg/kg

- d Gavage 13 wks No effects were observed on thymus or spleen histology, RBC or WBC populations, or lymphocyte populations.

Doses 0 4 8 10 12 15

WBC count (percent change compared to control)

M 0%

− 5%

− 12%

− 7% 1%

− 3%

F 0% 22% 45% 12% 52% 29% Absolute spleen weight (percent change compared to control)

M 0%

− 3%

− 6% 3% 1% 5%

F 0% 1% 8% 23%* 17%* 24%* Relative spleen weight (percent change compared to control)

M 0% 3% 4% 7%

− 1% 2%

F 0% 1% 0% 6%

− 1%

− 2% Absolute thymus weight (percent change compared to control)

M 0%

− 1% 3%

− 10%

− 12%

− 25%

F 0%

− 7% 12% 19% 32% 19% Relative thymus weight (percent change compared to control)

M 0%

− 1% 3%

− 10%

− 12%

− 25%

F 0%

− 7% 4% 4% 12%

− 6% ( Levine et al. (1990) ; Levine et al. (1981a) , 1981b) ) e Rats, F344, 10/sex/group; 30/sex for control 84.7 ± 4.7% purity, ~10% HMX, median particle diameter 20 μm, ~90% of particles ≤66 μm 0, 10, 30, 100, 300, or 600 mg/kg

- d Diet 13 wks Data were not reported for rats in the 300 or 600 mg/kg

- d dose groups because all of the rats died before the 13

- wk necropsy.

Doses 0 10 30 100 300 600

WBC count (percent change compared to control)

M 0% 4% 7% 15%

− −
F 0% 23%* 24%* 62%*

− − Absolute spleen weight (percent change compared to control)

M 0%

− 11%

− 16%

− 34%

− −
F 0% 2% 12% 0%

− − Relative spleen weight (percent change compared to control)

M 0%

− 9%

− 12%

− 21%

− −
F 0% 2% 12% 3%

− −

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 55 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results von Oettingen et al. (1949) Rats, sex/strain not specified , 20/group 90−97% pure, with 3−10% HMX; particle size not specified 0, 15, 25, or 50 mg/kg

- d Diet 13 wks Doses 0 15 25 50

WBC count (percent change compared to control)

M 0%

− 30% 7%

− 6%

Hart (1974) Dogs, Beagle, 3/sex/group Pre

- mix with ground dog chow containing 20 mg RDX/g

- chow, 60 g d og food ; p urity and particle size not specified 0, 0.1, 1, or 10 mg/kg

- d Diet 13 wks Doses 0 0.1 1 10

WBC count (percent change compared to control)

M 0% 5% 2%

− 19%

F 0%

− 2% 24% 6% Absolute spleen weight (percent change compared to control)

M 0%

− − 123%

F 0%

− − − 11% Martin and Hart (1974) Monkeys, Cynomolgus or Rhesus c , 3/sex/group Purity of test material not specified 0, 0.1, 1, or 10 mg/kg

- d Gavage 13 wks Doses 0 0.1 1 10

WBC count (percent change compared to control)

M 0%

− 32% 0%

− 3%

F 0%

− 38%

− 1%

− 41%

Gastrointestinal effects Lish et al. (1984) Mice, B6C3F 1 , 85/sex/group; interim sacrifices (10/sex/group) at 6 and 12 mo 89.2−98.7% pure, with 3−10% HMX as contaminant; 83−89% of particles <66 μm 0, 1.5, 7.0, 35, or 175/100 mg/kg

- d (high dose r educed to 100 mg/kg

- d in wk 11 due to excessive mortality) Diet 2 yrs No GI tract effects were observed as clinical signs or on gross pathology or histopathology examination.

Levine et al. (1983b ) Rats, F344, 75/sex/group; interim sacrifices (10/sex/group) at 6 and 12 mo 89.2−98.7% pure, with 3−10% HMX as contaminant; 83−89% of particles <66 μm 0, 0.3, 1.5, 8.0, or 40 mg/kg

- d Diet 2 yrs No GI tract effects were observed as clinical signs or on gross pathology or histopathology examination.

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 56 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results Crouse et al. (2006) Rats, F344, 10/sex/group 99.99% pure 0, 4, 8, 10, 12, or 15 mg/kg

- d Gavage 13 wks No GI tract effects were observed on gross pathology or histopathology

ex amination. Increased salivation and blood stains around the mouth were noted (affected doses and incidences were not reported); it is not clear whether these effects occurred in animals also experiencing convulsions.

von Oettingen et al. (1949) Rats (sex/strain not specified); 20/group 90−97% pure, with 3−10% HMX; particle size not specified 0, 15, 25, or 50 mg/kg

- d Diet 13 wks Congestion of the GI tract was observed in 50 and 100 mg/kg

- d rats that also exhibited mortality (40%) and severe neurotoxicity.

Martin and Hart (1974) Monkeys, Cynomolgus or Rhesus c , 3/sex/group Purity of test material not specified 0, 0.1, 1, or 10 mg/kg

- d Gavage 13 wks Vomiting was observed more frequently in the 1 and 10 mg/kg

- d groups compared to the control or 0.1 mg/kg

- d groups, although some episodes occurred during the intubation procedure.

Hart (1974) Dogs, Beagle, 3/sex/group Pre

- mix with ground dog chow containing 20 mg RDX/g

- chow, 60 g d og food ; p urity and particle size not specified 0, 0.1, 1, or 10 mg/kg

- d Diet 13 wks Some nausea and vomiting were reported (incidences and affected dose groups were not reported).

Hematological effects Lish et al. (1984) Mice, B6C3F 1 , 85/sex/group; interim sacrifices (10/sex/group) at 6 and 12 mo 89.2−98.7% pure, with 3−10% HMX as contaminant; 83−89% of particles <66 μm 0, 1.5, 7.0, 35, or 175/100 mg/kg

- d (high dose re duced to 100 mg/kg

- d in wk 11 due to excessive mortality) Diet 2 yrs Doses 0 1.5 7.0 35 175/100 RBC count; 105 wks (percent change compared to control)

M 0%

− 4% 3%

− 3% 14%

F 0% 4%

− 7% 5% 3% Hemoglobin; 105 wks (percent change compared to control)

M 0%

− 6% 3%

− 5% 9%

F 0% 2%

− 7% 3% 1% Hematocrit; 105 wks (percent change compared to control)

M 0%

− 4% 3%

− 4% 9%

F 0% 3%

− 6% 3% 1% Platelets; 105 wks (percent change compared to control)

M 0% 33% 9% 21% 27%

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 57 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results

F 0%

− 14%

− 7% 1% 5% Hart (1976) Rats, Sprague

- Dawley, 100/sex/group Purity and particle size not specified 0, 1.0, 3.1, or 10 mg /kg

- d Diet 2 yrs Doses 0 1.0 3.1 10 RBC count; 104 wks (percent change compared to control)

M 0% 3% 7%

− 2%

F 0%

− 14% 7% 2% Reticulocyte count; 104 wks (percent change compared to control)

M 0% 250% 500%* 850%*

F 0% 180%*

− 40% 20% Hemoglobin; 104 wks (percent change compared to control)

M 0% 3% 4% 0%

F 0%

− 1% 1%

− 2% Levine et al. (1983b) Rats, F344, 75/sex/group; interim sacrifices (10/sex/group) at 6 and 12 mo 89.2

− 98.7% pure, with 3−10% HMX as contaminant; 83−89% of particles <66 μm 0, 0.3, 1.5, 8.0, or 40 mg/kg

- d Diet 2 yrs Doses 0 0.3 1.5 8.0 40 Hemoglobin levels; 105 wks (percent change compared to control)

M 0% 6% 6% 3%

− 13%

F 0%

− 5% 1%

− 9%

− 14% RBC count; 105 wks (percent change compared to control)

M 0% 5% 2%

− 1%

− 9%

F 0%

− 2% 2%

− 9%

− 13% Platelet count; 105 wks (percent change compared to control)

M 0% 6%

− 4%

− 10%

− 7%

F 0% 14%

− 4% 5% 22% Hematocrit; 105 wks (percent change compared to control)

M 0% 5% 5% 2%

− 7%

F 0%

− 5% 0%

− 8%

− 12% Cholakis et al. (1980) Mice, B6C3F 1 , 10−12/sex/group 88.6% pure, with 9% HMX and 2.2% water as contaminants; ~200 μm particle size 0, 80, 60, or 40 mg/kg

- d for 2 wks followed by 0, 80, 160, or 320 mg/kg

- d (TWA doses of 0, 79.6, 147.8, or 256.7 mg/kg

- d for males and 0, 82.4, 136.3, or 276.4 mg/kg

- d for females) d Diet 13 wks Doses 0 80 160 320

RBC count (percent change compared to control)

M 0%

− 5%

− 12%*

− 2%

F 0%

− 10%

− 1% 1% Reticulocytes (percent change compared to control)

M 0%

− 36%

− 13% 15%

F 0% 21% 25%

− 19%
Hematocrit (percent change compared to control)

M 0%

− 1%

− 6% 0%

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 58 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results

F 0%

− 8% 2% 1%
Hemoglobin (percent change compared to control)

M 0%

− 2%

− 7%*

− 3%

F 0%

− 5% 4% 1%
Platelets (percent change compared to control)

M 0% 33% 28% 22%

F 0% 3% 9% 39% Cholakis et al. (1980) Rats, F344, 10/sex/group 88.6% pure, with 9% HMX and 2.2% water as contaminants; ~200 μm particle size 0, 10, 14, 20, 28, or 40 mg/kg

- d Diet 13 wks Doses 0 10 14 20 28 40

RBC count (percent change compared to control)

M 0%

− − − 3%

− 1%

F 0%

− − − − 1%

− 7% Hemoglobin (percent change compared to control)

M 0%

− − − 2%

− 1%

F 0%

− − − − 1%

− 1%
Platelet (percent change compared to control)

M 0%

− − − 11% 16%*

F 0%

− − − − 23%

− 13% Reticulocytes (percent change compared to control)

M 0%

− − − 26% 76%*

F 0%

− − − − 2% 17%
Hematocrit (percent change compared to control)

M 0%

− − − 3% 0%

F 0%

− − − 0%

− 2% Crouse et al. (2006) Rats, F344, 10/sex/group 99.99% pure 0, 4, 8, 10, 12, or 15 mg/kg

- d Gavage 13 wks Doses 0 4 8 10 12 15

RBC count (percent change compared to control)

M 0% 1%

− 7%

− 2%

− 4%

− 5%

F 0% 3% 3%

− 1% 2%

− 2% Hemoglobin (percent change compared to control)

M 0%

− 1%

− 5% 0%

− 1%

− 6%

F 0% 2% 4%

− 1 4%

− 4%
Platelet count (percent change compared to control)

M 0% 21% 11% 13%

− 8% 34%

F 0% 6% 40% 47% 34%

− 36%

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 59 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results

Hematocrit (percent change compared to control)

M 0% 2%

− 5% 0%

− 1%

− 4%

F 0% 3% 4% 0% 4%

− 2% ( Levine et al. (1990) ; Levine et al. (1981a) , 1981b) ) e Rats, F344, 10/sex/group; 30/sex for control 84.7 ± 4.7% purity, ~10% HMX, median particle diameter 20 μm, ~90% of particles ≤66 μm 0, 10, 30, 100, 300, or 600 mg/kg

- d Diet 13 wks Data were not reported for rats in the 300 or 600 mg/kg

- d dose groups because all of the rats died before the 13

- wk necropsy.

Doses 0 10 30 100 300 600
Hematocrit (percent change compared to control)

M 0%

− 2%

− 1%

− 5%

− −
F 0% 0%

− 4%

− 7%

− − Hemoglobin (percent change compared to control)

M 0%

− 3%

− 1%

− 6%

− −
F 0% 0%

− 4%

− 8%*

− −
RBC count (percent change compared to control)

M 0%

− 2%

− 2%

− 5%

− −
F 0%

− 1%

− 4%

− 5%

− − Reticulocytes (percent change compared to control)

M 0%

− 4% 10% 28%

− −
F 0% 9% 73% 71%

− − von Oettingen et al. (1949) Rats, sex/strain not specified, 20/group 90−97% pure, with 3−10% HMX; particle size not specified 0, 15, 25, or 50 mg/kg

- d Diet 13 wks Doses 0 15 25 50

RBC count (percent change compared to control)

M + F 0%

− 23%

− 12%

− 14% Hemoglobin (percent change compared to control)

M + F 0%

− 25%

− 7%

− 11% Hart (1974) Dogs, Beagle, 3/sex/group Pre

- mix with ground dog chow containing 20 mg RDX/g

- chow, 60 g dog food ; p urity and particle size not specified 0, 0.1, 1, or 10 mg/kg

- d Diet 13 wks Doses 0 0.1 1 10

RBC count (percent change compared to control)

M 0%

− 3% 3% 2%

F 0% 13% 7% 11% Reticulocyte count (percent change compared to control)

M 0%

− 66% 0%

− 50%

F 0%

− 17%

− 50% 0%
Hematocrit (percent change compared to control)

M 0%

− 4% 2% 0%

F 0% 6% 1% 7%

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C

- 60 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference and study design Results Hemoglobin (percent change compared to control)

M 0% 5%

− 2% 0%

F 0% 8%

− 2% 8% Martin and Hart (1974) Monkeys, Cynomolgus or Rhesus c , 3/sex/group Purity of test material not specified 0, 0.1, 1, or 10 mg/kg -d Gavage 13 wks Histopathological examination revealed increased numbers of degenerate or necrotic megakaryocytes in all bone marrow sections.

Doses 0 0.1 1 10

RBC count (percent change compared to control)

M 0%

− 3% 2%

− 3%

F 0% 0%

− 1% 2% Reticulocyte count (percent change compared to control)

M 0%

− 33%

− 50%

− 50%

F 0%

− 18%

− 36% 45%
Hematocrit (percent change compared to control)

M 0%

− 7%

− 4%

− 1%

F 0% 10% 7% 3% Hemoglobin (percent change compared to control)

M 0%

− 10%

− 8%

− 6%

F 0% 6% 6% 3% 

### 1 2 3 4 5 6 7 8 9 10 11 12 13 14 *Statistically significantly different compared to the control, as determined by study authors ( p < 0.05).

a No musculoskeletal evidence is presented in this table as no animal study reported effects on the musculoskeletal

system and all human effects were in case reports (see summary in Appendix C, Section C.2).
b Incidence counts exclude individuals from which blood was obtained via the orbital sinus.

c The species of monkey used in this study was inconsistently reported in the study as either C ynomolgus (in the

Methods section) or R he sus (in the Summary).
d Doses were calculated by the study authors.

e Levine et al. (1981a) is a laboratory report of a 13

- week study of RDX in F344 rats; two subsequently published

papers ( Levine et al., 1990 ; Levine et al., 1981b ) present subsets of the data provided in the full laboratory report.
f Standard deviations accompanying the mean response in a given dose group were high, suggesting uncertainty in

the accuracy of the reported percent change compared to control.

Note: A dash (“



### − ”) indicates that the study authors did not measure or report a value for that dose group.

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C- 61 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**C .3.3.**  **Genotoxicity** 1

2
3
4
5 6
7 8
9
10
11
12
13
14
15 16
17 18 19
20
21
22
23
24 25
26
27
28 29
30
31 32
33
34
35 R DX RDX has tested negative in a variety of in vitro tests for g enotoxicity, including mutation assays in multiple strains of Salmonella typhimurium (with or without metabolic activation), recombination in Saccharomyces cerevisiae strain D3, and forward mutations in both V79 Chinese hamster lung cells and mouse lymphoma L5178Y cells. However, in genotoxicity assays designed to be more sensitive, RDX did show some positive results . For example, when the concentration of S9 was doubled, the mutagenicity of RDX was about twice that of background

. RDX also showed positiv e mutagenic results with metabolic activation in a chemiluminescent assay (Mutatox assay)

. In some cases, the interpretation of testing data for RDX was complicated by the tendency of the compound to precipitate out of DMSO solution (the usual vehicle) at concentrations ≥250 μg/mL ( Reddy et al., 2005 )

. As with other studies of RDX, the purity of the test compound was unknown in

s everal (particularly older) studies

. A summary of the results of in vitro genotoxicity studies of RDX is pr esented in Table C- 13

. RDX has produced negative results in all reverse mutation assays in S. typhimurium that use standard levels of the metabolic activation system (S9). No evidence of reverse mutation was observed in S. typhimurium (strains TA98, TA100 , TA1535, TA1537, and TA1538), either with or without the addition of S9 metabolic activating mixture ( Neuwoehner et al., 2007 ; George et al., 2 001 ; Lachance et al., 1999 ; Tan et al., 1992 ; Minor et al., 1982 ; Cholakis et al., 1980 ; Whong et al., 1 980 ; Simmon et al., 1977b )

. One exception is a finding of weak mutagenic activity of RDX to S. typhimurium strain TA97a (mutagenicity index = 1.5

− 2.0) ( Pan et al., 2007a ) . However, this assay

u sed a high percentage of S9 fraction (9% instead of 4%), indicating that extensive metabolic activation is needed to elicit a mutagenic response. R DX did not cause gene recombination in S. cerevisiae strai n D3 at concentrations up to 23 μg/mL, with or without metabolic activation ( Simmon et al., 1977b )

. The study authors not ed
t hat the negative findings should be considered in the context of the low concentrations tested. Similarly, RDX did not induce forward mutations (HGPRT locus) in V79 Chinese hamster lung cells, with or without metabolic activation, although minimal cyt otoxicity was observed at 180 μM ( Lachance et al., 1999 ) . However, RDX produced revertants in two of three trials in the Mutatox

a ssay with the bacterium V ibrio fisheri when tested at doses up to 2.5 μg/tube, with and without S9 ( Arfsten et al., 1994 ) . In the presence of S9, a dose

- response was observed; in the absence of S9, no d ose- response relationship was detected ( Arfsten et al., 1994 )

. RDX did not induce forward

mu tations in mouse lymphoma L5178Y cells with or without metabolic activation ( Reddy et al., 2 005 )

. During an accompanying range

- finding study, precipitates occurred at doses ≥250 μg/mL,

s uggesting that concentrations of RDX in DMSO reported beyond 250 μg/mL may not be accurate.

This document is a draft for review purposes only and does not constitute Agency polic y. 

### C- 62 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**T able**  **C- 13** 

**. Summary of in vitro studies of the genotoxicity of RDX**  1

Endpoint Test system Dose/ concentration a 

### Results b



### Comments Reference Without activation With activation Genotoxicity studies in prokaryotic organisms

Reverse mutation Salmonella typhimurium TA1535, TA1537, TA1538, TA98, TA100 1,000 μg/plate

− − Metabolic activation with S9 Cholakis et al. (1980)

Reverse mutation S. typhimurium TA1535, TA1537, TA1538 TA100, TA98 14 μg/plate

− − Effect of disinfection treatments on mutagenicity tested: RDX was not mutagenic in any strain before or after disinfection treatment with chlorine or ozone Simmon et al. (1977a)

Reverse mutation S. typhimurium TA98, TA100 250 μg/plate

− − Study authors noted that results were consistent with literature George et al. (2001)

Reverse mutation S. typhimurium TA98, TA100 1 mg/plate

− − Metabolic activation with S9 Tan et al. (1992)

Reverse mutation S. typhimurium TA98, TA100 1,090 μ g/plate

− − High S9 activation (9%) used Pan et al. (2007a)

Reverse mutation S. typhimurium TA97a 32.7 μg/plate

− ± High S9 activation (9%) used; study authors concluded that RDX “required intensive metabolic activation” to exhibit mutagenicity in this strain Pan et al. (2007a)

Reverse mutation Vibrio fischeri 0.004 μg/tube ± + Mutatox assay with metabolic activation (S9) Arfsten et al. (1994)

Reverse mutation ( u mu test) Salmonella choleraesius subsp. chol. (prior S. typhimurium ) TA1535/pSK1002 20.6 μg/mL

− − No observed effect concentration; tested at highest concentration where the induction rate was below 1.5 for the first time and the growth factor was below 0.5 Neuwoehner et al. (2007)

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 63 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Endpoint Test system Dose/ concentration a 

### Results b



### Comments Reference Without activation With activation

Reverse mutation ( NM 2009 test) S. choleraesius subsp

. chol

. NM 2009 , TA1535/pSK1002/pNM12 20.6 μg/mL

− − No observed effect concentration; tested at highest concentration where the induction rate was below 1.5 for the first time and the growth factor was below 0.5 Neuwoehner et al. (2007)

Induction of the sfiA gene (SOS chromotest) Escherichia coli PQ37 20.6 μg/mL

− − No observed effect concentration; tested at highest concentration where the induction rate was below 1.5 for the first time and the growth factor was below 0.5 Neuwoehner et al. (2007)

Reverse mutation S. typhimurium , TA98, TA1 00 24.8 μg/mL

− − No observed effect concentration; metabolic activation with S9 Neuwoehner et al. (2007)

Reverse mutation S. t yphimurium TA98, TA100 2.6 μg/mL

− − No observed effect concentration; metabolic activation with S9; minimal cytotoxicity was observed at 180 μM Lachance et al. (1999)

Reverse mutation S. typhimurium TA1535, TA1536, TA1537, TA1538 TA100, TA98 30.8 μg/mL

− − Metabolic activation with S9 Cotruvo et al. (1977)

Genotoxicity studies in nonmammalian eukaryotic organisms
Recombination induction S accharomyces cerevisiae D3 23 μg/mL

− − Study authors concluded that this microorganism did not appear to be useful for detecting mutagenicity in several compounds tested Simmon et al. (1977a)

Recombination induction S. cerevisiae D3 30.8 μg/mL

− − Metabolic activation with S9 Cotruvo et al. (197 7)

This document is a draft for review purposes only and does not constitute Agency policy. 

### C

- 64 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Endpoint Test system Dose/ concentration a 

### Results b



### Comments Reference Without activation With activation Genotoxicity studies in mammalian cells
Forward mutation Chinese hamster lung fibroblasts V79 cells 40 μg/mL

− − Minimal cytotoxicity observed at 40 μg/mL (limit of solubility) Lac hance et al. (1999)

Mutation L5178Y mouse lymphoma cells 500 μg/mL

− − No or low cytotoxicity seen at these concentrations; however, precipitate was observed at >250 μg/mL Reddy et al. (2005)

Unscheduled DNA synthesis; DNA repair WI

- 38 cells, human diploid fibroblasts 4,000 μg/mL

− − Precipitates were observed at concentrations of RDX ≥40 μg/mL Dilley et al. (1979)



### 1 2 3 a Lowest effective dose for positive results; highest dose tested for negative results.
b + = positive; ± = equivocal or weakly positive; − = negative

.

This d ocument is a draft for review purposes only and does not constitute Agency policy. 

### C- 65 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

R DX did not induce unscheduled DNA synthesis, with or without metabolic activation, using 1
2 3
4
5 6
7 8
9 10
11
12

13
14
15 16
17 18 19
20
21
22 23
24
25 26
27
28
29 30
31
32 33 34 human diploid fibroblasts (WI

- 38 cells) when tested in DMSO at concentrations up to 4,000 μg/mg; however, precipitation of RDX at high concentrations in cell culture media makes interpretation of these results difficult ( D illey et al., 1979 ) . Only two in vivo studies are available for the genotoxicity

o f RDX; these are summarized in Table C

- 14 . RDX di d not decrease the ratio of polychromatic erythrocytes (PCEs) to normochromatic erythrocytes (NCEs), nor did it induce micronucleated PCEs in an in vivo mouse bone marrow micronucleus assay in young adult male CD

- 1 mice ( Reddy
et

al., 2005 ) . RDX was considered negative for the induction of dominant lethal mutations in male C D rats fed RDX at nominal doses from 0 to 50 mg/kg

- day for 15 weeks prior to mating with untreated virgin females ( Cholakis et al., 1980 ) . Females sacrificed at midgestation showed no
st atistically significant effec ts on number of corpora lutea, implants, or live or dead embryos ( Cholakis et al., 1980 ).

Metabolites of RDX Several metabolites of RDX, N

- nitroso derivatives of the parent compound (mononitroso, dinitroso, and trinitroso compounds, abbreviated MNX, DNX, and TNX, respectively) ( Musick et al., 2 010 ; Major et al., 2007 ) have been tested directly for genotoxicity ( Pan et al., 2007a ; George et al., 2 001 ; Snodgrass, 1984 ) . Miniature pigs were used to detect these trace metabolites because the

sw ine model of the GI tract more closely resembles that of humans ( Major et al., 2007 ) ; an i dentification and quantification of RDX metabolites in humans has not been conducted. A summary of the results of in vitro and in vivo genotoxicity studies of metabolites of RDX is presented in Table C- 15

. Pan et al. (2007a) studied the mutagenicity of two metabolites, MNX and TNX. These

m etabolites were no t mutagenic in S. typhimurium strain TA97a at normal levels of S9, but were clearly mutagenic at enhanced concentrations of S9 (4% versus 9% S9). The observation that these metabolites were positive in S. typhimurium strain TA97a is likely due to this str ain’s higher sensitivity for frameshift mutations that occur at a cluster of cytosine residues in the mutated gene for histidine synthesis in this strain ( Pan et al., 2007a ) . These metabolites were also weakly

mu tagenic in S. typhimurium strain TA102, again with high levels of S9. Strain TA102 was developed with an A:T base pair at the site of mutation and its sensitivity was increased by the addition of some 30 copies of a plasmid containing the mutant gene that is available for back mutation. This strain is sensitive to many oxidative mutagenic compounds ( Levin et al., 1982 )

. O ther metabolites with potential human relevance identified in the urine of miniature pigs have not been assessed for their genotoxicity ( Major et al., 2007 ).

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 66 DRAFT―DO NOT CI TE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table C-**  **14**

**. Summary of in vivo studies of the genotoxicity of RDX**  1

2 Endpoint Test system Dose/ concentration Results Comments Reference In vivo genotoxicity studies in mammalian systems
Micronucleus formation CD

- 1 mouse bone marrow Single dose of 62.5, 125, or 250 mg/kg No significant decrease in PCE:NCE ratios; no induction of micronucleated PCE at any dose 250 mg/kg was maximum tolerated dose determined in dose range

- finding study Reddy et al. (2005)

Dominant lethal mutations Male CD rats dosed and mated with untreated female rats 0, 5, 16, or 50 mg/kg

- d for 15 wk No statistically or biologically significant effects on fertility; determined to be negative for the induc tion of lethal mutations Males in the high

- dose group experienced lower food consumption and weight gain compared with all other groups Cholakis et al. (1980)

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 67 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**T able**  **C- 15** 

**. Summary of in vitro and in vivo studies of the genotoxicity of RDX metabolites**  1

2 3 4 Endpoint Te st system Dose/ concentration a 

### Results b



### Comments Reference Without activation With activation Genotoxicity studies in prokaryotic organisms

Reverse mutation Salmonella typhimurium TA97a, TA102 22 μg/plate

− + Mono and trinitroso metabolites (MNX and TNX); high S9 activation (9%) used Pan et al. (2007a)

Reverse mutation S. typhimurium TA1535, TA1537, TA1538, TA98, TA100 500 μ g/plate + + Positive only for TNX; MNX and DNX were negative George et al. (2001 )

Reverse mutation S. typhimurium TA1535, TA1537, TA1538, TA98, TA100 NR

− − Mononitroso metabolite, MNX; metabolic activation with S9 Snodgrass (19 84)

Genotoxicity studies in mammalian cells

— in vitro
Forward mutation Mouse lymphoma thymidine kinase NR + + Mononitroso metabolite, MNX; metabolic activation with S9 Snodgrass (1984)

Chromosomal aberrations Chinese hamster ovary cells NR

− + Mononitroso metabolite, MNX; metabolic activation with S9 Snodgrass (1984)

Unscheduled DNA synthesis; DNA repair Primary rat hepatocytes NR + ND Mononitroso metabolite, MNX; additional metabolic activation not required with S9 Snodgrass (1984)

In vivo genotoxicity studies in mammalian systems

Dominant lethal mutations Male mice dosed and mated with untreated female mice NR

− ND Mononitroso metabolite, MNX; additional metabolic activation not required with S9 Snodgrass (1984)

a Lowest effective dose for positive results; highest dose tested for negative results; NR = not reported

. b + = positive; ± = equivocal or weakly positive; − = negative ; ND = not determined

.

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 68 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Th e genotoxicity of MNX was positive in three out of five assays conducted for the U.S. Army 1
2 3
4
5 6
7 8
9 10
11
12 13 ( S nodgr ass, 1984 ) . MNX was positive with or without metabolic activation in the mouse lymphoma f orward mutation assay at the thymidine kinase locus, for chromosomal aberrations in Chinese hamster ovary cells, and in the primary rat hepatocyte unscheduled DNA synthesis assay. MNX was not considered positive in S. typhimurium (strains TA98, TA100, TA1535, TA1537, and TA1538), either with or without the addition of S9 metabolic activating mixture or in an in vivo dominant lethal mutation assay in mice. However, this study i s of limited use due to a significant lack of details including information on dosing, raw data, and statistical reporting. In summary, RDX is not mutagenic or genotoxic in vitro or in vivo in typical assays used to detect genotoxicity. In two in vitro s tudies using more sensitive assays and conditions for detecting mutagenicity, RDX was found to be positive. Several studies showed that the N

- nitroso metabolites are genotoxic, but the formation and quantification of these metabolites in humans is not kno wn.

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 69 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine



# APPENDIX D. DOSE

- RESPONSE MODELING FOR THE DERIVATION OF REFERENCE VALUES FOR EFFECTS OTHER THAN CANCER AND THE DERIVATION OF CANCER RISK ESTIMATES

This appendix provides technical detail on dose

- response evaluation and determination of 1
2 3
4
5 6
7 8

9 10

11
12

13 points of departure (POD) for r elevant toxicological endpoints. The endpoints were modeled using the U.S. Environmental Protection Agency ( EPA ) Benchmark Dose Software (BMDS, Versions 2.4). Sections D.1 (noncancer) and D.2 (cancer) describe the common practices used in evaluating the model fit and selecting the appropriate model for determining the POD, as outlined in the Benchmark Dose Technical Guidance Document ( U .S. EPA, 2012b ). In some cases , it may be
ap propriate to use alternative methods, based on statistical judgement; exceptions are noted as necessary in the summary of the modeling results.



## D.1. **BENCHMARK DOSE MODELING SUMMARY FOR N ONCANCER**  **E NDPOINTS** 

The noncancer endpoints that were selected for dose

- response modeling are presented in Table D-1. For each endpoint, the doses and re sponse data used for the modeling are presented.

**Table D-1.**  **Noncancer endpoints selected for dose**

**- response modeling for RDX** 

Endpoint and

reference Species/sex Dose (mg/kg

- d) Incidence/ t otal (%)

Convulsions Crouse et al. (2006) a Female F344 rat 0 4 8 10 12 15 0/10 (0%) 0/10 (0%) 2/10 (20%) 3/10 (30%) 5/10 (50%) 5/10 (50%)
Male F344 rat 0 4 8 10 12 15 0/10 (0%) 0/10 (0%) 1/10 (10%) 3/10 (30%) 8/10 (80%) 7/10 (70%)

This document is a draft for review purposes only and does not constitute Agency policy. 

### C-1 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Endpoint and

reference Species/sex Dose (mg/kg

- d) Incidence/ t otal (%)

Male and female F344 rat, combined 0 0/20 (0%) 4 0/20 (0%) 8 3/20 (15%) 10 6/20 (30%) 12 13/20 (65%) 15 12/20 (60%)
Convulsions Cholakis et al. (1980) Female F344 rat (gestational exposure) 0 0/24 (0%) 0.2 0/24 (0%)

2 1/24 (4%) 20 18/24 (75%)
Testicular degeneration Lish et al. (1984) Male B6C3F 1 mouse 0 0/63 (0%) 1.5 2/60 (3%) 7 2/62 (3%) 35 6/59 (10%) 107 3/27 (11%) Prostate suppurative inflammation Levine et al. (1983b) Male F344 rat 0 2/54 (4%) 0.3 4/55 (7%) 1.5 9/52 (17%) 8 12/55 (22%) 40 19/31 (61%) 

### 1 2 3 4 5 6
7 8

9 10 a For convulsions in Crouse et al. (2006) , the incidence rates across doses were determined to be not statistically significantly different between the males and females using an exact Cochran

- Mantel

- Haenszel test ( p ≥ 0.10). The data were combin ed acr o ss sex for this endpoint prior to modeling.


### In addition to the endpoints presented in Table D

- 1, the combined incidence of seizure and mortality was modeled for Crouse et al. (2006) to determine the effect of possible under estimation of seizur es , as discussed in Section 2.1.6. Table D

- 2 presents the data on this combined incidence.

**Table D-**  **2. Convulsion or mortality**  **e ndpoints**  **from Crouse et al. (2006)**  **selected for dose**

**- res**  **ponse modeling for RDX**

Endpoint and

reference Species/sex D os e (mg/kg

- d) Incidence/t otal (%)

Convulsion or mortality Johnson (2015) Female F344 rat 0 4 8 10 12 15 0/10 (0%) 0/10 (0%) 3/10 (30%) 5/10 (50%) 9/10 (90%) 8/10 (80%)

This document is a draft for review purposes only and does not constitute Agency policy. 

### C-2 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Endpoint and

reference Species/sex Dose (mg/kg

- d) Incidence/t otal (%)

Male F344 rat 0 4 8 10 12 15 0/10 (0%) 0/10 (0%) 2/10 (20%) 4/10 (40%) 8/10 (80%) 7/10 (70%)
Male and female F344 rat, combined 0 4 8 10 12 15 0/20 (0%) 0/20 (0%) 3/20 (15%) 6/20 (30%) 13/20 (65%) 12/20 (60%)


### 1 2 3 4 5 6

7
8 9
10
11
12 13
14
15 16
17 18 19
20
21
22 23
24 a Incidence was defined for each animal as the presence of convulsion or mortality . T he incidence rates across doses f or this endpoint were determined to be not statistically significantly different between the males and females using an exact Cochran

- M antel

- Haenszel test ( p ≥ 0.10). The data were combined across sex for this endpoint prior to modeling.



### D **.1.1. Evaluation of Model Fit**  **and Model Selection**
For each dicho tomous endpoint, BMDS dichotomous models 5 

### were fitted to the data using the maximum likelihood method. Each model was tested for goodness

- of

- fit using a chi

- square goodness

- of

- fit test (χ 2 

### p
- value < 0.10 indicates lack of fit). Other factors were also us ed to assess model fit, such as scaled residuals, visual fit, and adequacy of fit in the low

- dose region and in the vicinity of the benchmark response ( BMR ). F rom among the models exhibiting adequate fit , the best

- fit model was selected for estimation of the BMD. This model selection was conducted in two stage, first from among only the multistage models to determine a representative multistage model, and second from among the representative multistage m odel and the non

- multistage models. In each stage, t he BMDL estimate s (95% lower confidence limit on the BMD, as estimated by the profile likelihood method) and AIC value s of the models considered in that stage were used to make the selection, as follows

. If the BMDL estimates were “sufficiently close,” that is, differed by three fold or less , the model selected was the one that yielded the lowest AIC value. If the BMDL estimates were not sufficiently close, the model with the low est BMDL was selected

. T he model selected in the second stage was considered the best

- fit model. The BMDL estimate (95% lower confidence limit on the benchmark dose [ BMD ] , as estimated by the profile likelihood method) and Akaike’s information criterion ( AIC ) value were

5 Unless otherwise specified, all available BMDS dichotomous models besides the alternative and nested dichotomous models were fitted. The follo wing parameter restrictions were applied: f or the L og -L ogistic model, restrict slope ≥1; for the G amma and Weibull models, restrict power ≥1.
This document is a draft for review purposes only and does not constitute Agency policy. 

### C-3 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

u sed to select a best

- fit model from among the models exhibiting adequate fit. If the BMDL 1
2 3
4

5
6
7

8
9
10
11
12
13
14
15

16 17

18 19 20 estimates were “sufficiently close” (i.e. , differed by at most threefold ) , the model selected was the one that yielded the lowest AIC value. If the BMDL estimates were no t sufficiently close, the lowest BMDL was selected as the POD.

**D .1.2.**  **Modeling Results**
The tables that follow summariz e the modeling results for the noncancer endpoints modeled.

Nervous System Effects
Tables D

- 3 to D

- 11 (and Figures D

- 1 to D

- 9) present the BMD model ing results for incidence of convulsions for female, male, and male and female F344 rats combined based on data from Crouse et al. (2006) , using BMRs of 10, 5, and 1% extra risk (ER) . Tables D

- 12 to D -14 (and Figures D- 1 0 to D

- 12) present the BMD model ing results for incidence of convulsions for female F344 rats based on data from Cholakis et al. (1980) , using BMRs of 10, 5, and 1% ER

. Table D

- 15 (and Figure D- 1 3) presents the BMD modeling results for combined incidence of convulsions a nd mortality for male and female rats combined based on data from Crouse et al. (2006)

.

**Table D-3** 

**. Model predictions for convulsions in female F344 rats exposed to**  **RDX by gavage for 90 days (**  **Crouse et al., 2006 )**  **; BMR = 10% ER** 

Model a 

### Goodness of fit BMD 10Pct 

### (mg/kg

- d) BMDL 10Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma 0.923 55.085 6.46 2.92 The Q uantal

- L inear model was selected based on lowest BMDL (BMDLs differed by more than threefold). Logistic 0.733 56.607 6.76 4.75 LogLogistic 0.929 55.076 6.42 3.04
Probit 0.793 56.086 6.64 4.54 LogProbit 0.952 54.798 6.54 3.39
Weibull 0.892 55.420 6.16 2.62 Multistage 2° 0.954 53.595 5.46 2.47 Quantal

- Linear 0.733 56.131 2.76 1.84 Multistage 5° b 0.885 55.525 5.98 2.47 Multistage 4° 0.885 55.525 5.98 2.47 Multistage 3° 0.885 55.525 5.98 2.49
Dichotomous

- Hill 0.964 56.265 7.10 5.75

a Selected model in bold; scaled residuals for selected model for doses 0, 4, 8, 10, 12, and 15 mg/kg -d ay were 0.00,

− 1. 29,

− 0.46,

− 0.12, 0.87, and 0.41, respectively.

This document is a draft for review purposes only and does not constitute Agency policy. 

### C-4 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

b The Multistage 5° model may appear equivalent to the Multistage 4° model ; however , differences exist in digits 

### 1 2 3

4

5

6 7 8

9
10
11
12
13
14
15 16
17 18

19 20 not displayed in the table.

BMR = 10% ER; dose shown in mg/kg

- day.



### F **igure D-1** 

**. Plot of incidence rate by dose, with fitted curve for selected model,**  **for convulsions in female F344 rats exposed to RDX by gavage for 90 days (**  **Crouse et al., 2006 ).** 
**Quantal Linear Model using Weibull Model** (Version: 2.16; Date: 2/28/ 2013 ) The form of the probability function is: P[response] = background + (1

- background)*[1

-- EXP(

- slope*dose)]

**Benchmark Dose Computation** BMR = 10% ER BMD = 2.75544 BMDL at the 95% confidence level = 1.84489

Parameter Estimates Variable Estimate Default i nitial p arameter v alues Background 0 0.0833333 Slope 0.0382372 0.0404091

Power Not applicable 1



### A nalysis of Deviance Table

Model Log (likelihood) Number of p aram eter s Deviance Test degrees of freedom (d.f.) p - value

Full model

− 24.9756 6 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8

0 2 4 6 8 10 12 14Fraction Affected

dose Quantal Linear Model, with BMR of 10% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the

14:08 02/27 2014 BMDL BMD Quantal Linear

This document is a draft for review purposes only and does not constitute Agency policy. 

### C-5 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Fitted model

− 27.0654 1 4.17949 5 0.5239
Reduced model

− 33.7401 1 17.529 5 0.003598 

### 1 2 3 4

5

a 

### 6 7 8 9 AIC : = 56.1307

Goodness

- of

- Fit Table
Dose Est. p rob. a 

### Expected Observed Size Scaled residuals

0 0 0 0 10 0

4 0.1418 1.418 0 10

− 1.286

8 0.2635 2.635 2 10

− 0.456

10 0.3178 3.178 3 10

− 0.121

12 0.368 3.68 5 10 0.866

15 0.4365 4.365 5 10 0.405



### Estimated probab i lity

Chi^2 = 2.79 d.f

. = 5 p

- value = 0.7325

This document is a draft for review purposes only and does not constitute Agency policy. 

### C-6 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-4** 

**. Model predictions for convulsions in female F344 rats exposed to**  1 2

3 4 5 6 7 8

9

10

11 12 13 **RDX by gavage for 90 days (**  **Cr ouse et al., 2006**  **) ; BMR= 5%**  **ER**

Model a 

### Goodness of fit BMD 5Pct 

### (mg/kg

- d) BMDL 5Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma 0.923 55.085 5.09 1.54 The Q uantal

- L inear model was selected based on lowest BMDL (BMDLs differed by more than threefold). Logistic 0.733 56.607 4.76 2.83 LogLogistic 0.929 55.076 4.99 1.71
Probit 0.793 56.086 4.80 2.69 LogProbit 0.952 54.798 5.33 2.17
Weibull 0.892 55.420 4.55 1.30 Multistage 2° 0.954 53.595 3.81 1.21 Quantal

- Linear 0.733 56.131 1.34 0.898 Multistage 3° 0.885 55.525 4.30 1.21 Multistage 4° b 0.885 55.525 4.30 1.21 Multistage 5° 0.885 55.525 4.30 1.20
Dichotomous

- Hill 0.964 56.265 6.25 2.30

a Selected model in bold; scaled residuals for selected model for doses 0, 4, 8, 10, 12, and 15 mg/kg -d ay were 0.00,

− 1. 29,

− 0. 46,

− 0.12, 0.87, and 0.41, respectively. The BMD 10 and BMDL 10 f or the selected model were 2.76 and 1.84 mg/kg -d ay , r espectively.
b The Multistage 4° model may appear equivalent to the Multistage 3° model ; however , differences exist in digits

not displayed in the table.

BMR = 5% ER; dose shown in mg/kg

- day.


### Fig **ure D-2** 

**. Plot of incidence rate by dose, with fitted curve for selected model,**  **for convulsions in female F344 rats exposed to RDX by gavage for 90 days (**  **Crouse et al., 2006 ).**  0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8

0 2 4 6 8 10 12 14Fraction Affected

dose Quantal Linear Model, with BMR of 5% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the B

14:07 02/27 2014 BMDL BMD Quantal Linear

This document is a draft for review purposes only and does not constitute Agency policy. 

### C-7 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Q uantal Linear**  **Model using Weibull Model** (Version: 2.16; Date: 2/28/ 2013 ) 1
2 3
4
5 6
7 8 9 10

11 12

13 14 15 16

17 18 19 T he form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- slope*dose)]

**Benchmark Dose Computation** BMR = 5% ER BMD = 1.34145 BMDL at the 95% confidence level = 0.89 816

Parameter Estimates Variable Estimate Default i nitial p arameter v alues Background 0 0.0833333 Slope 0.0382372 0.0404091

Power Not applicable 1



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 24.9756 6
Fitted model

− 27.0654 1 4.17949 5 0.5239
Reduced model

− 33.7401 1 17.529 5 0.003598



### AIC: = 56.1307

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0 0 0 10 0

4 0.1418 1.418 0 10

− 1.286

8 0.2635 2.635 2 10

− 0.456

10 0.3178 3.178 3 10

− 0.121

12 0.368 3.68 5 10 0.866

15 0.4365 4.365 5 10 0.405



### Chi^2 = 2.79 d.f

. = 5 p

- value = 0.7325

This document is a draft for review purposes only and does not constitute Agency policy. 

### C-8 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-5** 

**. Model predictions for convulsions in female F344 rats exposed to**  1 2

3 4 5 6

7

8

9 10 11 **RDX by gavage for 90 days (**  **Cr ouse et al., 2006**  **) ; BMR= 1%**  **ER**

Model a 

### Goodness of fit BMD 1Pct 

### (mg/kg

- d) BMDL 1Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma 0.923 55.085 3.10 0.355 The Q uantal

- L inear model had a BMD more than 10 times lower than the lowest dose, so this model was excluded from consideration. Of the remaining models, the M ultistage 5° model was selected based on lowest BMDL (BMDLs differed by more than threefold). Logistic 0.733 56.607 1.60 0.681 LogLogistic 0.929 55.076 2.87 0.468
Probit 0.793 56.086 1.86 0.649 LogProbit 0.952 54.798 3.63 0.919
Weibull 0.892 55.420 2.30 0.259 Multistage 2° 0.954 53.595 1.69 0.236
Quantal

- Linear 0.733 56.131 0.263 0.176 Multistage 3° 0.885 55.525 1.99 0.238 Multistage 4° 0.885 55.525 1.99 0.236 Multistage 5° 0.885 55.525 1.99 0.235
Dichotomous

- Hill 0.964 56.265 4.77 0.778

a Selected model in bold; scaled residuals for selected model for doses 0, 4, 8, 10, 12, and 15 mg/kg -d ay were 0.00 ,

− 0. 67, 0.14, 0.11, 0.64, and

− 0 .51, respectively. The BMD 10 and BMDL 10 f or the selected model were 5.98 and 2.47 mg /kg

- d ay , respectively.

BMR= 1% ER; dose shown in mg/kg

- day.


### Fig **ure D-3** 

**. Plot of incidence rate by dose, with fitted curve for selected model,**  **for convulsions in female F344 rats exposed to RDX by gavage for 90 days (**  **Crouse et al., 2006 ).**  0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8

0 2 4 6 8 10 12 14Fraction Affected

dose Multistage Model, with BMR of 1% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BM

14:06 02/27 2014 BMD BMDL Multistage

This document is a draft for review purposes only and does not constitute Agency policy. 

### C-9 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Multistage Model** (Version: 3.3; Date: 02/28/ 2013 ) The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)]

**Benchmark Dose Computation** BMR = 1% ER BMD = 1.98616 BMDL at the 95% confidence level = 0.235433 1
2 3
4
5 6
7 8 9 10

11 12

13 14 15 16

17 18 Parameter Estimates Variable Estimate Default i nitial p arameter v alues Background 0 0 Beta(1) 0 0.0172961 Beta(2) 0.00234798 0.002476 Beta(3) 0.000100566 0 Beta(4) 0 0 Beta(5) 0 0



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 24.9756 6
Fitted model

− 25.7624 2 1.57351 4 0.8135
Reduced model

− 33.7401 1 17.529 5 0.003598



### AIC: = 55.5247

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0 0 0 10 0

4 0.043 0.43 0 10

− 0.671

8 0.1827 1.827 2 10 0.141

10 0.2849 2.849 3 10 0.106

12 0.4006 4.006 5 10 0.641

15 0.5801 5.801 5 10

− 0.513



### Chi^2 = 1.16 d.f

. = 4 p

- value = 0.8854

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 10 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-6** 

**. Model predictions for convulsions in male F344 rats exposed to**  1 2

3 4 5 6 7

8

9

10 11 12 **RDX by gavage for 90 days (**  **Cr ouse et al., 2006**  **) ;**  **BMR = 10% ER** 

Model a 

### Goodness of fit BMD 10Pct 

### (mg/kg

- d) BMDL 10Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma 0.482 48.534 7.40 5.11 Of the models that provided an adequate fit, the M ultistage 3° model was selected based on lowest AIC. Logistic 0.335 49.692 7.18 5.03 LogLogistic 0.522 48.248 7.48 5.29
Probit 0.363 49.460 7.17 4.86 LogProbit 0.530 48.224 7.53 5.38
Weibull 0.376 49.496 6.84 4.47 Multistage 2° 0.307 50.335 4.54 2.95
Quantal

- Linear 0.0553 56.530 1.98 1.38 Multistage 5° b Multistage 4° 0.361 49.607 6.85 3.91

Multistage 3° 0.515 47.803 6.17 3.95
Dichotomous

- Hill 0.701 48.408 8.34 6.32

a Selected model in bold; scaled residuals for selected model for doses 0, 4, 8, 10, 12, and 15 mg/kg -d ay were 0.00,

− 0. 54,

− 0. 82,

− 0.40, 1.66, and

− 0.61, respectively.
b For the Multistage 5° model, the b4 coefficient estimates was 0 (boundary of parameters space). The models in this row reduced to the Multistage 4° model.

BMR = 10% ER; dose shown in mg/kg

- day.


### Fig **ure D-4.**  **Plot of incidence rate by dose, with fitted curve for selected model, for convulsions in male F344 rats exposed to RDX by gavage for 90 days**  **( Crouse et al., 2006**  **)**

**.** 0 0.2 0.4 0.6 0.8

1

0 2 4 6 8 10 12 14Fraction Affected

dose Multistage Model, with BMR of 10% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BM

14:11 02/27 2014 BMD BMDL Multistage

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 11 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1 2
3
4
5 6
7 8
9 10 11

12 13

14 15 16 17

18 19 **M ulti**  **stage Model** (Version: 3.3; Date: 02/28/ 2013 ) The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)]

**Benchmark Dose Computation** BMR = 10% ER BMD = 6.17392 BMDL at the 95% confidence level = 3.94501

Parameter Estimates Variable Estimate Default i nitial p arameter v alues Background 0 0 Beta(1) 0 0 Beta(2) 0 0.00691555 Beta(3) 0.000447707 0



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 20.4721 6
Fitted model

− 22.9013 1 4.85838 5 0.4334
Reduced model

− 37.4599 1 33.9755 5 < 0 .0001



### AIC: = 47.8027

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0 0 0 10 0

4 0.0282 0.282 0 10

− 0.539

8 0. 2049 2.049 1 10

− 0.822

10 0.3609 3.609 3 10

− 0.401

12 0.5387 5.387 8 10 1.658

15 0.7793 7.793 7 10

− 0.605



### Chi^2 = 4.24 d.f

. = 5 p

- value = 0.5153

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 12 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-7** 

**. Model predictions for convulsions in male F344 rats exposed to**  **RDX by gavage for 90 days (**  **Crouse et al., 2006 )**  **; BMR = 5% ER**  1 2

3 4 5 6 7 8

9

10

11 12 13 Model a 

### Goodness of fit BMD 5Pct 

### (mg/kg

- d) BMDL 5Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma 0.482 48.534 6.47 3.96 Of the models that provided an adequate fit, the M ultistage 3° model was selected based on lowest AIC. Logistic 0.335 49.692 5.74 3.34 LogLogistic 0.522 48.248 6.51 4.14
Probit 0.363 49.460 5.92 3.26 LogProbit 0.530 48.224 6.71 4.40
Weibull 0.376 49.496 5.58 3.16 Multistage 2° 0.307 50.335 3.17 1.63
Quantal

- Linear 0.0553 56.530 0.964 0.670 Multistage 5° b Multistage 4° 0.361 49.607 5.56 1.99

Multistage 3° 0.515 47.803 4.86 2.19
Dichotomous

- Hill 0.701 48.408 7.76 5.27

a Selected model in bold; scaled residuals for selected model for doses 0, 4, 8, 10, 12, and 15 mg/kg -d ay were

0.000,

− 0.54,

− 0.82,

− 0.40, 1.66, and

− 0.61, respectively. The BMD 10 and BMDL 10 f or the selected model were 6.17 and 3.95 mg/kg -d ay , respectively.
b For the Multistage 5° model, the beta coefficient estimates were 0 (boundary of parameters space). The models

in this row reduced to the Multistage 4° model.

BMR = 5% ER; dose shown in mg/kg

- day. 0 0.2 0.4 0.6 0.8

1

0 2 4 6 8 10 12 14Fraction Affected

dose Multistage Model, with BMR of 5% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BM

14:10 02/27 2014 BMD BMDL Multistage



### Figure **D-5**

**. Plot of incidence rate by dose, with fitted curve for selected model,**  **for convulsions in male F344 rats exposed to RDX by gavage for 90 days (**  **Crouse et al., 2006 ).** 
This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 13 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1 2
3
4
5 6
7 8
9 10 11

12 13

14 15 16 17

18 19 **Multi stage Model**  (Version: 3.3; Date: 02/28/ 2013 ) The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)]

**Benchmark Dose Computation** BMR = 5% ER BMD = 4.85686 BMDL at the 95% confidence level = 2. 1924 4

Parameter Estimates Variable Estimate Default i nitia l p arameter v alues Background 0 0 Beta(1) 0 0 Beta(2) 0 0.00691555 Beta(3) 0.000447707 0



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 20.4721 6
Fitted model

− 22.9013 1 4.85838 5 0.4334
Reduced model

− 37.4599 1 33.9755 5 < 0 .0001



### AIC: = 47.8027

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0 0 0 10 0

4 0.0282 0.282 0 10

− 0.539

8 0. 2049 2.049 1 10

− 0.822

10 0.3609 3.609 3 10

− 0.401

12 0.5387 5.387 8 10 1.658

15 0.7793 7.793 7 10

− 0.605



### Chi^2 = 4.24 d.f

. = 5 p

- value = 0.5153

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 14 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-8** 

**. Model predictions for convulsions in male F344 rats exposed to**  1 2

3 4 5 6 7 8

9

10

11 12 13 **RDX by gavage for 90 days (**  **Crou se et al., 2006**  **) ; BMR = 1%**  **ER**

Model a 

### Goodness of fit BMD 1Pct 

### (mg/kg

- d) BMDL 1Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma 0.482 48.534 4.96 2.32 The M ultistage 2 ° model was selected based on lowest BMDL (BMDLs differed by more than threefold). Logistic 0.335 49.692 2.86 0.975 LogLogistic 0.522 48.248 4.79 2.38
Probit 0.363 49.460 3.60 1.01 LogProbit 0.530 48.224 5.41 3.00
Weibull 0.376 49.496 3.52 1.43 Multistage 2° 0.307 50.335 1.40 0.363
Quantal

- Linear 0.0553 56.530 0.189 0.131 Multistage 5° b 0.361 49.607 3.42 0.392 Multistage 4° c 0.361 49.607 3.42 0.392 Multistage 3° 0.515 47.803 2.82 0.457
Dichotomous

- Hill 0.701 48.408 6.64 3.47

a Selected model in bold; scaled residuals for selected model for doses 0, 4, 8, 10, 12, and 15 mg/kg -d ay were 0.00 ,

− 0.92,

− 1. 26,

− 0. 65, 1.76, and 0.11, respectively. The BMD 10 and BMDL 10 for the selected model were 4.54 and 2.95 mg/kg

- d ay , respectively.
b The Multistage 5° model may appear equivalent to the Multistage 4° model ; however , differences exist in digits

not displayed in the table.

BMR = 1% ER; dose shown in mg/kg

- day.


### Figur **e D-6.**  **Plot of incidence rate by dose, with fitted curve for selected model, for convulsions in male F344 rats exposed to RDX by gavage for 90 days**  **( Crouse et al., 2006**  **).** 0 0.2 0.4 0.6 0.8

1

0 2 4 6 8 10 12 14Fraction Affected

dose Multistage Model, with BMR of 1% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BM

14:09 02/27 2014 BMD BMDL Multistage

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 15 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Multistage Model** (Version: 3.3; Date: 02/28/ 2013 ) 1
2 3
4
5 6
7 8
9 10

11 12

13 14 15 16

17 18 19 The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)]

**Benchmark Dose Computation** BMR = 1% ER BMD = 1.40125 BMDL at the 95% confidence level = 0.363499

Parameter Estimates Variable Estimate Default i nitial p arameter v alues Background 0 0 Beta(1) 0 0 Beta(2) 0.00511858 0.00691555



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 20.4721 6
Fitted model

− 24.1672 1 7.39017 5 0. 1932
Reduced model

− 37.4599 1 33.9755 5 < 0 .0001



### AIC: = 50.3345

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0 0 0 10 0

4 0.0786 0.786 0 10

− 0.924

8 0.2793 2.793 1 10

− 1.264

10 0.4006 4.006 3 10

− 0.649

12 0.5215 5.215 8 10 1.763

15 0.6839 6.839 7 10 0.11



### Chi^2 = 5.99 d.f

. = 5 p

- value = 0.3069

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 16 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-9** 

**. Model predictions for convulsions in male and female F344 rats**  1 2

3 4 5 6 7

8

9

10 11 12 **exposed to RDX by gavage for 90 days (**  **Crou se et al., 2006**  **) ; BMR = 10%**  **ER**

Model a 

### Goodness of fit BMD 10Pct 

### (mg/kg

- d) BMDL 10Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma 0.484 101.79 6.92 5.09 Of the models that provided an adequate fit, the M ultistage 3° model was selected based on lowest AIC. Logistic 0.231 104.55 6.86 5.34 LogLogistic 0.512 101.66 6.93 5.15
Probit 0.291 103.61 6.83 5.19 LogProbit 0.557 101.25 7.01 5.31
Weibull 0.369 102.91 6.52 4.62 Multistage 2° 0.364 103.03 4.97 3.75
Quantal

- Linear 0.0369 111.56 2.32 1.77 Multistage 3° b Multistage 4° Multistage 5° 0.502 100.91 6.60 4.59

Dichotomous

- Hill 0.696 101.64 7.73 5.98

a Selected model in bold; scaled residuals for selected model for doses 0, 4, 8, 10, 12, and 15 mg/kg -d ay were 0.00,

− 0.69,

− 0. 25,

− 0. 06, 1.62, and

− 1.08, respectively.
b For the Multistage 4° and 5° models, the b4 and b5 coefficient estimates were 0 (boundary of para meters space). The models in this row reduced to the Multistage 3° model.

BMR = 10% ER; dose shown in mg/kg

- day.


### Figur **e D-7** 

**. Plot of incidence rate by dose, with fitted curve for selected model,**  **for convulsions in male and female F344 rats exposed to RDX by gavage for**  **90 days**  **( Crouse et al., 2006**  **).** 0 0.2 0.4 0.6 0.8

0 2 4 6 8 10 12 14Fraction Affected

dose Multistage Model, with BMR of 10% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BM

14:05 02/27 2014 BMD BMDL Multistage

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 17 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1 2
3
4
5 6
7 8
9 10 11

12 13

14 15 16 17

18 19 **Mult istage Model**  (Version: 3.3; Date: 02/28/ 2013 ) The form o f the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)]

**Benchmark Dose Computation** BMR = 10% ER BMD = 6.60247 BMDL at the 95% confidence level = 4.59346

Parameter Estimates Variable Estimate Default i nitial p arameter v alues Background 0 0 Beta(1) 0 0.00163806 Beta(2) 0 0.00485933 Beta(3) 0.000366065 0



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 47.0806 6
Fitted model

− 49.4567 1 4.75213 5 0.4469
Reduced model

− 71.5289 1 48.8965 5 < 0 .0001



### AIC: = 100.913

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0 0 0 20 0

4 0.0232 0.463 0 20

− 0.689

8 0.1709 3.418 3 20

− 0.248

10 0.3065 6.131 6 20

− 0.063

12 0.4688 9.375 13 20 1.624

15 0.7093 14.186 12 20

− 1.076



### Chi^2 = 4.34 d.f

. = 5 p

- value = 0.5021

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 18 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-**  **10**

**. Model predictions for convulsions in male and female F344 rats**  1 2

3 4 5 6 7 8

9

10

11 12 13 **exposed to RDX by gavage for 90 days (**  **Crou se et al., 2006**  **) ; BMR = 5%**  **ER**

Model a 

### Goodness of fit BMD 5Pct 

### (mg/kg

- d) BMDL 5Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma 0.484 101.79 5.78 3.80 Of the models that provided an adequate fit, the M ultistage 3° model was selected based on lowest AIC. Logistic 0.231 104.55 5.13 3.49 LogLogistic 0.512 101.66 5.74 3.85
Probit 0.291 103.61 5.29 3.43 LogProbit 0.557 101.25 6.01 4.20
Weibull 0.369 102.91 5.11 3.18 Multistage 2° 0.364 103.03 3.47 2.20
Quantal

- Linear 0.0369 111.56 1.13 0.860 Multistage 3° 0.502 100.91 5.19 2.66 Multistage 4° Multistage 5° b 0.502 100.91 5.19 2.65

Dichotomous

- Hill 0.696 101.64 6.98 4.80

a Selected model in bold; scaled residuals for selected model for doses 0, 4, 8, 10, 12, and 15 mg/kg -d ay were 0.00,

− 0.69,

− 0. 25,

− 0. 06, 1.62, and

− 1.08, respectively. The BMD 10 and BMDL 10 f or the selected model were 6.60 and 4.59 mg/kg -d ay , r es pectively.
b For the Multista ge 5° model, the b5 coefficient estimate was 0 (boundary of parameters space). The models in this row reduced to the Multistage 4° model.

BMR = 5% ER; dose shown in mg/kg

- day.


### Figur **e D-8** 

**. Plot of incidence rate by dose, with the fitted curve of the selected**  **model, for convulsions in male and female F344 rats exposed to RDX by gavage for 90 days**  **( Crouse et al., 2006**  **).** 0 0.2 0.4 0.6 0.8

0 2 4 6 8 10 12 14Fraction Affected

dose Multistage Model, with BMR of 5% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BM

14:04 02/27 2014 BMD BMDL

Multistage

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 19 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1
2 3
4
5 6
7 8
9 10 11

12 13

14 15 16 17

18 19 20 **Mult istage Model**  (Version: 3.3; Date: 02/28/ 2013 ) The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)]

**Benchmark Dose Computation** BMR = 5% ER BMD = 5. 1939 9 BMDL at the 95% confidence level = 2.65815

Parameter Estimates Variable Estimate Default i nitial p arameter v alues Background 0 0 Beta(1) 0 0.00163806 Beta(2) 0 0.00485933 Beta(3) 0.000366065 0



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 47.0806 6
Fitted model

− 49.4567 1 4.75213 5 0.4469
Reduced model

− 71.5289 1 48.8965 5 < 0 .0001



### AIC: = 100.913

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0 0 0 20 0

4 0.0232 0.463 0 20

− 0.689

8 0.1709 3.418 3 20

− 0.248

10 0.3065 6.131 6 20

− 0.063

12 0.4688 9.375 13 20 1.624

15 0.7093 14.186 12 20

− 1.076



### Chi^2 = 4.34 d.f

. = 5 p

- value = 0.5021

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 20 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-**  **11**

**. Model predictions for convulsions in male and female F344 rats**  1 2

3 4 5 6 7 8 9

10

11 12 13 14 **exposed to RDX by gavage for 90 days (**  **Cr ouse et**  **al., 2006 )**  **; BMR = 1% ER** 

Model a 

### Goodness of fit BMD 1Pct 

### (mg/kg

- d) BMDL 1Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma 0.484 101.79 4.02 2.03 The Quantal

- Linear model did not fit the data adequately ( p

- value
< 0.10), so it was excluded from consideration. Of the remaining multistage models, the Multistage 3° model was selected based on lowest AIC. From among the multistage 3° and

non

- multistage models, the multistage 3° model was selected based on lowest BMDL ( BMDLs differed by more than threefold). Logistic 0.231 104.55 2.04 0.987 LogLogistic 0.512 101.66 3.79 2.00
Probit 0.291 103.61 2.57 1.03 LogProbit 0.557 101.25 4.50 2.69
Weibull 0.369 102.91 2.94 1.35 Multistage 2° 0.364 103.03 1.53 0.544
Quantal

- Linear 0.0369 111.56 0.222 0.169 Multistage 5° b Multistage 4° 0.502 100.91 3.02 0.549

Multistage 3° 0.502 100.91 3.02 0.569
Dichotomous

- Hill 0.696 101.64 5.62 2.90

a Selected model in bold; scaled residuals for selected model for doses 0, 4, 8, 10, 12, and 15 mg/kg -d ay were 0.00 ,

− 0. 69 , − 0. 25 , − 0.06 , 1. 62 , and

− 1.08 , respectively. The BMD 10 and BMDL 10 for the selected model were 6.60 and 4.59 mg/kg

- d a y, respectively.
b For the Multistage 5° model, the beta coefficient estimates were 0 (boundary of parameters space). The models

in this row reduced to the Multistage 4° model.



### Fig **ure D**

**- 9.**  **Plot of incidence rate by dose, with the fitted curve of the multistage 3°**  **model, for convulsions in male and female F344 rats exposed to RDX by gavage for 90 days**  **( Crouse et al., 2006**  **)**

**.** BMR = 1% ER; dose shown in mg/ kg

- day. 0 0.2 0.4 0.6 0.8

0 2 4 6 8 10 12 14Fraction Affected

dose Multistage Model, with BMR of 1% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BM

14:03 02/27 2014 BMD BMDL Multistage

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 21 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Multistage Model. (Version: 3.3; Date: 02/28/2013) 1 2 3 4 5 6 7 8 9 10 11 12

13 14

15 16 17 18

19 20 2 2
2 1

2
3

This document is a draft for review purposes only and doe 

### C- 22 The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)]

Benchmark Dose Computation. BMR = 1% Extra risk BMD = 3.01676 BMDL at the 95% confidence level = 0.569284

Parameter Estimates
Variable Estimate Default Initial Parameter Values Background 0 0 Beta(1) 0 0.00163806 Beta(2) 0 0.00485933 Beta(3) 0.000366065 0



### Analysis of Deviance Table
Model Log(likelihood ) # Param's Deviance Test d.f. p - value

Full model

- 47.0806 6
Fitted model

- 49.4567 1 4.75213 5 0.4469
Reduced model - 71.5289 1 48.8965 5 <.0001



### AIC: = 100.913

Goodness of Fit Table
Dose Est. Prob. Expected Observed Size Scaled Resid

0 0 0 0 20 0

4 0.0232 0.463 0 20

- 0.689

8 0.1709 3.418 3 20

- 0.248

10 0.3065 6.131 6 20

- 0.063

12 0.4688 9.375 13 20 1.624

15 0.7093 14.186 12 20

- 1.076



### Chi^2 = 4.34 d.f = 5 P

- value = 0.5021

s not constitute Agency policy. 

### DRA FT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-**  **12**

**. Model predictions for convulsions in female F344 rats exposed to**  1 2 3

4

a 

### 5 6
b 

### 7 8

9

10

11 12 13 14 **RDX by gavage on gestation al**  **days (GDs)**  **6**

**− 19**  **( Chola**  **kis et al., 1980 )**  **; BMR**  **= 10%**  **ER**

Model a 

### Goodness of fit BMD 10Pct 

### (mg/kg

- d) BMDL 10Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma 0.989 42.003 3.62 1.56 The Q uantal

- L inear model is selected based on lowest BMDL (BMDLs differed by more than threefold). Logistic 0.526 43.556 8.92 6.14 LogLogistic 0.991 41.996 3.45 1.53
Probit 0.577 43.348 7.64 5.39 LogProbit 1.000 41.963 3.13 1.51
Weibull 0.983 42.026 3.81 1.55 Multistage 3° b 0.960 42.113 4.26 1.54 Multistage 2° 0.960 42.113 4.26 1.54 Quantal

- Linear 0.669 42.077 1.88 1.29

Selected model in bold; scaled residuals for selected model for doses 0, 0.2, 2, and 20 mg/kg -d ay were 0.00,

− 0.52,

− 1. 03, and 0.49, respectively. T he Multistage 3° model may appear equivalent to the Multistage 2° model ; ho we ver , differences exist in digits not displayed in t he table.

BMR = 10% ER; dose shown in mg/kg

- day.



### Figur **e D-**  **10**

**. Plot of incidence rate by dose, with the fitted curve of the**  **selected model, for convulsions in female F344 rats exposed to RDX by gavage**
**on GDs**  **6**

**− 19**  **( Cholakis et al., 1980**  **).** 0 0.2 0.4 0.6 0.8

0 5 10 15 20Fraction Affected

dose Quantal Linear Model, with BMR of 10% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the

09:29 06/06 2014 BMDL BMD Quantal Linear

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 23 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Quantal Linear Model using Weibull Model**  (Version: 2.16; Date: 2/28/ 2013 ) 1
2 3
4
5 6
7 8
9 10

11 12

13 14 15 16

17 18 19 The f orm of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- slope*dose)]

**Benchmark Dose Computation** BMR = 10% ER BMD = 1.87886 BMDL at the 95% confidence level = 1.28909

Parameter Estimates Variable Estimate Default i nitial p arameter v alues Background 0 0.0384615 Slope 0.056077 0.0588587

Power Not applicable 1



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 18.9808 4
Fitted model

− 20.0384 1 2.11537 3 0.5488
Reduced model

− 47.9793 1 57.9972 3 < 0 .0001



### AIC: = 42.0769

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0 0 0 24 0

0.2 0.0112 0.268 0 24

− 0.52

2 0.1061 2.546 1 24

− 1.025

20 0.6742 16.856 18 25 0.488



### Chi^2 = 1.56 d.f

. = 3 p

- value = 0.6686

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 24 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-**  **13**

**. Model predictions for convulsions in female F344 rats exposed to**  **RDX by gavage on GDs**  **6**

**− 19**  **( Cholakis et al., 1980**  **) ; BMR = 5%**  **ER** 1 2

9

10

11 12 13

3 4 5 6 7 8 Model a 

### Goodness of fit BMD 5Pct 

### (mg/kg

- d) BMDL 5Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma 0.989 42.003 2.31 0.759 The Q uantal -L inear model is selected based on lowest BMDL (BMDLs differed by more than threefold). Logistic 0.526 43.556 6.53 3.90 LogLogistic 0.991 41.996 2.27 0.823
Probit 0.577 43.348 5.41 3.34 LogProbit 1.000 41.963 2.18 0.902
Weibull 0.983 42.026 2.36 0.756 Multistage 2° 0.960 42.113 2.51 0.747 Quantal- Linear 0.669 42.077 0.915 0.628 Multistage 3° b 0.960 42.113 2.51 0.747

a Selected model in bold; scaled residuals for selected model for doses 0, 0.2, 2, and 20 mg/kg -d ay were 0.00,

− 0.52,

− 1.03, and 0.49, respectively. The BMD 10 and BMDL 10 for the selected mod el were 1.88 and 1.29 mg/kg -d ay , respectively.
b The Multistage 3° model may appear equivalent to the Multistage 2° model ; however , differences exist in digits

not displayed in the table.

Quantal Linear Model, with BMR of 5% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the B

0.6 0.8Affected Quantal Linear

BMR = 5% ER; dose shown in mg/kg

- day. 0 0.2 0.4

0 5 10 15 20Fraction

dose 15:24 05/23 2014 BMDL BMD



### Figure **D- 11** 

**. Plot of incidence rate by dose, with the fitted curve of the**  **selected model, for convulsions in female F344 rats exposed to RDX by gavage** 
**on GDs**  **6**

**− 19**  **( Cholakis et al., 1980**  **).**

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 25 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1 2
3
4
5 6
7 8
9 10 11

12 13

14 15 16 17

18 19 20 **Qua ntal Linear Model using Weibull Model**  (Version: 2.16; Date: 2/28/ 2013 ) The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- slope*dose)]

**Benchmark Dose Computation** BMR = 5% ER BMD = 0.914694 BMDL at the 95% confidence level = 0.627577

Parameter Estimates Variable Estimate Default i nitial p arameter v alues Background 0 0.0384615 Slope 0.056077 0.0588587

Power Not applicable 1



### Analysis of Deviance Table Model Log likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 18.9808 4
Fitted model

− 20.0384 1 2.11537 3 0.5488
Reduced model

− 47.9793 1 57.9972 3 < 0 .0001



### AIC: = 42.0769

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0 0 0 24 0

0.2 0.0112 0.268 0 24

− 0.52

2 0.1061 2.546 1 24

− 1.025

20 0.6742 16.856 18 25 0.488



### Chi^2 = 1.56 d.f

. = 3 p

- value = 0.6686

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 26 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-**  **14**

**. Model predictions for convulsions in female F344 rats exposed to**  1 2

3 4 5 6 7 8 9

10

11

12 13 14 **RDX by gavage on GDs**  **6**

**− 19**  **( C**  **hola kis et al., 1980**  **) ; BMR =**  **1 %**  **ER**

Model a 

### Goodness of fit BMD 1Pct 

### (mg/kg

- d) BMDL 1Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma 0.989 42.003 0.866 0.149 The Q uantal

- L inear model is selected based on lowest BMDL (BMDLs differed by more than threefold). Logistic 0.526 43.556 2.46 1.05 LogLogistic 0.991 41.996 0.902 0.201
Probit 0.577 43.348 1.96 0.871 LogProbit 1.000 41.963 1.11 0.335
Weibull 0.983 42.026 0.798 0.148 Multistage 3° b 0.960 42.113 0.638 0.146 Multistage 2° c 0.960 42.113 0.638 0.146 Quantal

- Linear 0.669 42.077 0.179 0.123

a Selected model in bold; scaled residuals for selected model for doses 0, 0.2, 2, and 20 mg/kg -d ay were 0.00 ,

− 0.52,

− 1. 03, and 0.49, respectively. The BMD 10 and BMDL 10 for the selected model were 1.88 and 1.29 mg/kg

- d ay , respectively.
b The Multistage 3° model may appear equivalent to the Multistage 2° model ; however , differences exist in digits

not displayed in the table.

BMR = 1% ER; dose shown in mg/kg

- day.


### Figur **e D-**  **12**

**. Plot of incidence rate by dose, with the fitted curve of the**  **selected model, for convulsions in female F344 rats exposed to RDX by gavage**
**on GDs**  **6**

**− 19**  **( Cholakis et al., 1980**  **).** 0 0.2 0.4 0.6 0.8

0 5 10 15 20Fraction Affected

dose Quantal Linear Model, with BMR of 1% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the B

14:19 07/28 2014 BMDLBMD Quantal Linear

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 27 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Quantal Linear Model using Weibull Model** (Version: 2.16; Date: 2/28/ 2013 ) 1
2 3
4
5 6
7 8
9 10

11 12

13 14 15 16

17 18 19 The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- slope*dose)]

**Benchmark Dose Computation** BMR = 1% ER BMD = 0.179224 BMDL at the 95% confidence level = 0.122966

Parameter Estimat es Variable Estimate Default i nitial p arameter v alues Background 0 0.0384615 Slope 0.056077 0.0588587

Power Not applicable 1



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 18.9808 4
Fitted model

− 20.0384 1 2.11537 3 0.5488
Reduced model

− 47.9793 1 57.9972 3 < 0 .0001



### AIC: = 42.0769

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0 0 0 24 0

0.2 0.0112 0.268 0 24

− 0.52

2 0.1061 2.546 1 24

− 1.025

20 0.6742 16.856 18 25 0.488



### Chi^2 = 1.56 d.f

. = 3 p

- value = 0.6686

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 28 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D**

**- 15** 

**. M**  **odel predictions for combined incidence of convulsion and mortality in male and female F344 rats exposed to RDX by gavage for 90 days**  **( Crouse et al., 2006**  **) ; BMR = 1% ER**  1 2 3

4 5 6 7 8 9 10 11

12 13

14 15 16 17 Model a 

### Goodness of fit BMD 1Pct 

### (mg/kg

- d) BMDL 1Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma 0.245 99.260 3.73 2.10 The log

- logistic and quantal

- linear models did not achieve an adequate fit (goodness

- of

- fit p

- value < 0.10). The multistage 2° model was excluded from model selection because the residual in the lowest dose group, near the BMD, was above 1.5 in absolute value. Of the remaining models, the multistage 3° model was selected based on lowest BMDL (BMDLs diffe red by more than threefold). Dichotomous

- Hill 0.436 98.317 5.22 3.04 Logistic 0.0859 102.17 1.81 0.846 LogLogistic 0.305 98.593 3.70 2.20
Probit 0.101 101.85 2.16 0.853 LogProbit 0.316 98.465 4.22 2.75
Weibull 0.152 101.16 2.45 1.24 Multistage 4° b 0.229 99.182 2.56 0.486 Multistage 3° c 0.229 99.182 2.56 0.486 Multistage 2° 0.165 102.01 1.22 0.470
Quantal

- Linear 0.0052 113.90 0.144 0.113

a Selected model in bold; scaled residuals for selected model for doses 0, 4, 8, 10, 12, and 15 mg/kg

- day were 0,

− 0.88, −0.14, −0.01, 1.92, −1.55, respectively.
b The Multistage 4° model may appear equivalent to the Multistage 3° model; however, differences exist in digits

not displayed in the table.

c The Multistage 3° model may appear equivalent to the Multistage 4° model; however, differences exist in digits

not displayed in the table.

BMR = 1% ER; dose shown in mg/kg

- day. 0 0.2 0.4 0.6 0.8

1

0 2 4 6 8 10 12 14Fraction Affected

dose Multistage Model, with BMR of 1% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BMDL

13:21 01/06 2016 BMD BMDL Multistage



### Figure **D-13**

**. Plot of incidence rate by dose with fitted curve for Multistage 3°**  **model for Model predictions for combin**  **e d incidence of convulsion and**  **mortality in male and female F344 rats exposed to RDX by gavage for 90 days (**  **Crouse et al., 2006 ).** 

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 29 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1
2 3
4
5 6
7 8
9 10 11

12 13

14 15 16 17

18 19 20 **Mul tistage Model.**  (Version: 3.4; Date: 05/02/ 2014 ) The form of the probabilit y function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)]

**Benchmark Dose Computation.** BMR = 1% Extra risk BMD = 2.56012 BMDL at the 95% confidence level = 0.486284

Parameter Estimates Variable Estimate Default i niti al p arameter v alues Background 0 0 Beta(1) 0 0.027 2036 Beta(2) 0 0.00626035 Beta(3) 0.000598962 0



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 44.71 6
Fitted model

− 48.59 1 7.76102 5 0.17
Reduced model

− 9.88 1 70.3406 5 < 0 .0001



### AIC: = 99.1817

Goodness

- of

- Fit Table
Dose Est. Prob. Expected Observed Size Scaled r esid uals

0 0 0 0 20 0

4 0.0376 0.752 0 20

− 0.88

8 0.2641 5.282 5 20

− 0.14

10 0.4506 9.012 9 20

− 0.01

12 0.6448 12.896 17 20 1.92

15 0.8675 17.351 15 20

− 1.55



### Chi^2 = 6.88 d.f

. = 5 p

- value = 0.2294

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 30 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

M

ale Reproductive Effects 1
2
3

4 5

6 7 8 9 10 11 12 13 14 T able D -16 (and Figure D

- 14) presents the BMD model ing results for incidence of testicular degeneration for male B6C3F 1 

### mice based on data from Lish et al. (1984) , using a BMR of 10% ER

.

**Table D-16** 

**. Model predictions for testicular degeneration in male B6C3F**  **1** 

### mice exposed to RDX by diet for 24 months **( Lish et al., 1984**  **) ; BMR = 10%**  **ER**

Model a 

### Goodness of fit BMD 10Pct 

### (mg/kg

- d) BMDL 10Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma b Weibull Quantal

- Linear 0.357 101.10 66.5 35.4 The L og

- P robit model was selected based on lowest BMDL (BMDLs differed by more than threefold). Logistic 0.159 103.40 97.1 66.1 LogLogistic 0.377 100.91 63.6 32.3
Probit 0.178 103.12 93.1 61.4 LogProbit 0.876 97.564 56.0 16.3 Multistage 2° c Multistage 3° Multistage 4° 0.357 101.10 66.5 35.4

a Selected model in bold; scaled residuals for selected model for doses 0, 1.5, 7, 35, and 107 mg/kg

- ay were 0.00,

0.32,

− 0. 61, 0.43, and

− 0.17, respectively.
b For the Gamma and Weibull models, the power parameter estimates were 1 (boundary of parameter spac e). The

models in this row are equivalent to the Quantal

- L inear model.

c The Multistage 3° and 4° model had b3 and b4 coefficient estimates of 0 (boundary of parameters space). The

models in this row reduced to the Multistage 2° model. The models in this r ow may appear equivalent to the Gamma model ; h owever , differences exist in digits not displayed in the table.

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 31 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1

2

3 4 5

6 7 8 9
10
11
12 13
14
15 16
17 18

1 9 BMR = 10% ER; dose shown in mg/kg

- day.


### Fig **ure D-**  **14**

**. Plot of incidence rate by dose, with fitted curve for selected**  **model, for testicular degeneration in male B6C3F**  **1** 

### mice exposed to RDX by **diet for 24 months (**  **Lish et al., 1984 ).** 

**P robit Model**  (Version: 3.3; Date: 2/28/ 2013 ) 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 Slope parameter is not restricted

**Benchmark Dose Computation** BMR = 10% ER BMD = 55.9784 BMDL at the 95% confidence level = 16.2787

Parame ter Estimates Variable Estimate Default i nitial p arameter v alues

B ackground 0 0

I ntercept

− 2.0054E+00

− 1.9976E+00

S lope 0.179828 0.172286 0 0.05

0.1 0.15

0.2 0.25

0.3

0 20 40 60 80 100Fraction Affected

dose LogProbit Model, with BMR of 10% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the B

13:42 02/14 2014 BMDL BMD LogProbit

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 32 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

A nalysis of Deviance Table 1

2 3 4 5

6 7 Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 46.4212 5
Fitted model

− 46.7817 2 0.721088 3 0.8682
Reduced model

− 52.1663 1 11.4902 4 0.02157



### AIC: = 97.5635

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0 0 0 63 0

1.5 0.0267 1.599 2 60 0.321

7 0.0489 3.033 2 62

− 0.608

35 0.086 5.072 6 59 0.431

107 0.122 3.294 3 27

− 0.173



### Chi^2 = 0.69 d.f

. = 3 p

- value = 0.8759

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 33 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

K

idney Effects 1
2
3
4

5 6 7

8 9 10 11 12 13 14 15 16 T able D

- 17 (and Figure D

- 15) presents the BMD model results for incidence of suppurative inflammation of the prostate in male F344 rats based on data from Levine et al. (1983b) , using a B MR of 10% ER

.

**Table D-**  **17**

**. Model predictions for prostate suppurative inflammation in male**  **F344 rats exposed to RDX by diet for 24 months (**  **Levine et al., 1983b )**  **; BM**  **R =**  **10% ER** 

Model a 

### Goodness of fit BMD 10Pct 

### (mg/kg

- d) BMDL 10Pct 

### (mg/kg

- d) Basis for model selection p - value AIC

Gamma b Multistage 2° Quantal

- Linear Multistage 3° Multistage 4° 0.288 200.37 4.61 3.24 The L og

- P robit model is selected based on lowest BMDL (BMDLs differ by more than threefold).

Logistic 0.102 203.50 10.8 8.58 LogLogistic 0.328 200.05 3.33 2.09
Probit 0.116 203.10 9.91 7.96 LogProbit 0.204 202.03 1.67 0.469
Weibull g 0.288 200.37 4.61 3.24

a Selected model in bold; scaled residuals for selected model for doses 0, 0.3, 1.5, 8, and 40 mg/kg -d ay were

− 0. 289, 0.172, 0.846,

− 1.298, and 0.819, respectively.
b The Gamma model had a power parameter estimate of 1 (boundary of parameter space). The M ultistage 2 ° , 3 ° ,

and 4 ° m odels had b2, b3, and b4 coefficients of 0 (boundary of parameter spac e). The models in this row are equivalent to the Quantal

- L inear model.

c The Weibull model may appear equivalent to the Q uantal -L inear model ; however , differences exist in digits not

displayed in the table.

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 34 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1

2

3 4 5

6 7 8 9
10
11
12 13
14
15 16
17 18

19 BMR = 10% ER; dose shown in mg/kg

- day.


### Fig **ure D-**  **15**

**. Plot of incidence rate by dose, with fitted curve for selected**  **model, for prostate suppurative inflammation in male F344 rats exposed to RDX by diet for 24 months**  **( Levine et al., 19**  **83b )** 

**.**

**P robit Model**  (Version: 3.3; Date: 2/28/ 2013 ) 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 Slope paramete r is not restricted

**Benchmark Dose Computation** BMR = 10% ER BMD = 1.67454 BMDL at the 95% confidence level = 0.468688

Parameter Estimates Variable Estimate Default i nitial p arameter v alues

B ackground 0.0452202 0.037037

I ntercept

− 1.4970E+00

− 1.3564E+00

S lope 0.417872 0.36341 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8

0 5 10 15 20 25 30 35 40Fraction Affected

dose LogProbit Model, with BMR of 10% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BM

13:39 02/14 2014 BMDL BMD LogProbit

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 35 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Analysis of Deviance Table 1

2 3 4 5

6 7 8

9
10
11

12 Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 96.3905 5
Fitted model

− 98.0147 3 3.24837 2 0. 1971
Reduced model

− 118.737 1 44.6933 4 < 0 .0001



### AIC: = 202.029

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0.0452 2.442 2 54

− 0.289

0.3 0.0669 3.682 4 55 0.172

1.5 0.1332 6.927 9 52 0.846

8 0.2982 16.402 12 55

− 1.298

40 0.5396 16.726 19 31 0.819



### Chi^2 = 3.18 d.f

. = 2 p

- value = 0. 2035

**D.1.3. Mortality: Dose** 

**- Response Analysis and BMD Modeling Documentation**  This appendix also presents a quantitative dose

- response analysis of mortality incidence from studies identified in Section 2.1.6 (see Table D

- 18).

**Table D-**  **1 8. Mortality data selected for dose** 

**- response modeling for RDX** 

Reference Species/sex D ose Incidence / Total (%) or Mean ± SD (no. of animals)
Lish et al. (1984) (mortality at 11 wks) Male B6C3F 1 mouse 0 mg/kg -d 1.5 7 35 175/100 1 / 85 (1%) 0 / 85 (0%) 0 / 85 (0%) 0 / 85 (0%) 30 / 85 (35%) Lish et al. (1984) (mortality at 11 wks) Female B6C3F 1 mouse 0 mg/kg -d 1.5 7 35 175/100 0 / 85 (1%) 0 / 85 (0%) 0 / 85 (0%) 0 / 85 (0%) 36 / 85 (42%)

Levine et al. (1981b) a Female F344 rat 0 mg/kg -d 10 30 100 300 600 0 / 30 (0%) 1 / 10 (10%) 0 / 10 (0%) 5 / 10 (50%) 10 / 10 (100%) 10 / 10 (100%)

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 36 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference Species/sex Dose Incidence / Total (%) or Mean ± SD (no. of animals)

Male F344 rat 0 mg/kg

- d 10 30 100 300 600 0 / 30 (0%) 0 / 10 (0%) 0 / 10 (0%) 8 / 10 (80%) 10 / 10 (100%) 10 / 10 (100%)

Male and female F344 rat, combined 0 mg/kg

- d 10 30 100 300 600 0 / 60 (0%) 1 / 20 (5%) 0 / 20 (0%) 13 / 20 (65%) 20 / 20 (100%) 20 / 20 (100%) von Oettingen et al. (1949) Rats, sex/strain not specified 0 mg/kg

- d 15 25 50 0 / 20 (0%) 0 / 19 (0%) b 8 / 20 (40%) 8 / 20 (40%) Cholakis et al. (1980) (2

- generation study) Female CD rat 0 mg/kg

- d 5 16 50 0 / 22 (0%) 0 / 22 (0%) 0 / 22 (0%) 6 / 22 (27%) Levine et al. (1983b) (mortality at 13 wks) Male F344 rat 0 mg/kg

- d 0.3 1.5 8 40 0 / 75 (0%) 0 / 75 (0%) 0 / 75 (0%) 0 / 75 (0%) 0 / 75 (0%)

Female F344 rat 0 mg/kg

- d 0.3 1.5 8 40 0 / 75 (0%) 0 / 75 (0%) 0 / 75 (0%) 0 / 75 (0%) 0 / 75 (0%) Cholakis et al. (1980) (13

- week study) Male F344 rat 0 mg/kg

- d 10 14 20 28 40 0 / 10 (0%) 0 / 10 (0%) 0 / 10 (0%) 0 / 10 (0%) 0 / 10 (0%) 0 / 10 (0%)

Female F344 rat 0 mg/kg

- d 10 14 20 28 40 0 / 9 (0%) c 0 / 10 (0%) 0 / 10 (0%) 0 / 10 (0%) 0 / 10 (0%) 0 / 10 (0%) ( Crouse et al. (2006) ) Female F344 rat 0 mg/kg

- d 4 8 0 / 10 (0%) 0 / 10 (0%) 1 / 10 (20%)

This document is a draft for review purposes only and does not constitute Agency policy. 

### C

- 37 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reference Species/sex Dose Incidence / Total (%) or Mean ± SD (no. of animals)

10 12 15 2 / 10 (20%) 5 / 10 (50%) 4 / 10 (40%)

Male F344 rat 0 mg/kg

- d 4 8 10 12 15 0 / 10 (0%) 0 / 10 (0%) 1 / 10 (10%) 3 / 10 (30%) 2 / 10 (20%) 3 / 10 (30%)

Male and female F344 rat, combined 0 mg/kg

- d 4 8 10 12 15 0 / 20 (0%) 0 / 20 (0%) 2 / 20 (10%) 5 / 20 (25%) 7 / 20 (35%) 7 / 20 (35%) Cholakis et al. (1980) (gestational exposure) Female F344 rats (gestational exposure) 0 mg/kg

- d 0.2

2 20 0 / 24 (0%) 0 / 24 (0%) 0 / 24 (0%) 5 / 24 (21%) Angerhofer et al. (1986) Female SD rate (gestational exposure) 0 mg/kg

- d 2 6 20 0 / 39 (0%) 1 / 40 (3%) 1 / 40 (3%) 16 / 51 (31%) Cholakis et al. (1980) Female New Zealand white rabbit (gestational exposure) 0 mg/kg

- d 0.2

2 20 0 / 11 (0%) 0 / 11 (0%) 0 / 11 (0%) 0 / 12 (0%)

a For Levine e t al. (1981a) and Crouse et al. (2006) , the incidence rates across doses were determined to be not

statistically significantly different between the males and females using an exact Cochran

- Mantel

- Haenszel test (p ≥ 0.10) . The data were combined across sex for each of these endpoints prior to modeling.
b For von O ettingen et al. (1949) , one mortality was reported in the 15 mg/kg

- d dose group. However, this

mortality was most likely not related to RDX, so the animal that died was excluded.

c For Cholakis et al. (1980) , one accidental death was reported in the 0 mg/kg

- d dose group. The animal that died

was excluded. 

### 1 2
3
4
5 6
7 8 Ta bles D

- 19 to D

- 22 present the BMD modeling results for incidence of mortality from Crouse et al. (2006)
, von Oettingen et al. (1949)
, Levine et al. (1983b) , and Angerhofer et al. (1986)

. T he following datasets were not modeled because each had either no response or a p ositive response only in the highest dose group: 11

- week mortality from Lish et al. (1984) , both male (one d eath in control group) and female; 13

- week mortality data from Levine et al. (1983b) , both male

an d female; mortality in female CD rats and male and female F344 rats from Cholakis et al. (1980) ;

an d mortality in female F344 rats during gestational exposure from Cholakis et al. (1980)

. This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 38 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**T able**  **D- 19. Model predictions for**  **combined mortality in male and female** 1 2 3

4

5

6 7 8

9 **F344 rats exposed to RDX by diet for 13 weeks (Levine et al., 1981); BMR = 1% ER** 

Model a Goodness of fit BMD 1Pct (mg/kg

- day) BMDL 1Pct (mg/kg

- day) Basis for model selection p - value AIC

Gamma 0.401 41.100 49.8 12.7 The quantal

- linear model was excluded because the fit has a residual below

- 2 at 4 mg/kg

- d. The multistage 4° model was selected as the representative multistage model based on lowest AIC. From among the multistage 4° and non

- multistage models, the multistage 4° model was selected based on lowest BMDL (BMDLs differed by more than threefold). Logistic 0.346 41.429 18.3 8.25 LogLogistic 0.257 43.098 73.2 16.9
Probit 0.328 41.727 15.0 6.82 LogProbit 0.257 43.098 58.6 19.5
Weibull 0.257 43.101 56.6 b 6.51 b
Multistage 2° 0.424 42.942 7.72 2.01
Quantal

- Linear 0.139 50.257 1.12 0.818 Multistage 3° 0.503 41.520 7.71 2.08 Multistage 4° 0.535 40.935 7.85 2.15 Multistage 5° 0.371 42.928 7.86 2.15

a Selected model in bold; scaled residuals for selected model for doses 0, 10, 30, 100, 300, and 600 mg/kg/day

were 0.00, 1.48,

- 0.97, 0.05, 0.00, and 0.00, respectively. The BMD 10 and BMDL 10 estimates for the selected model were 47.2 and 22.2 mg/kg

- d, respectively.
b The parameter convergence parameter was increased to 2e

- 08 to obtain convergence.



### F **igure D**

**- 16. Plot of incidence rate by dose, with the fitted curve of the**  **multistage 2° model, for combined mortality in male and female F344 rats exposed to RDX by diet for 13 weeks (Levine et al**  **., 1981); BMR = 1% E R.** 

0 0.2 0.4 0.6 0.8

1

0 100 200 300 400 500 600Fraction Affected

dose Multistage Model, with BMR of 1% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BM

13:54 02/27 2014 BMD BMDL Multistage

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 39 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Mul tistage Model. (Version: 3.3; Date: 02/28/2013) 1 2 3 4 5 6 7 8 9 10 11 12

13 14

15 16 17 18

19 20 21 T he form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)]

Benchmark Dose Computation. BMR = 1% Extra risk BMD = 7.85287 BMDL at the 95% confidence level = 2.15059

Parameter Estimates
Variable Estimate Default Initial Parameter Values Background 0 0 Beta(1) 0.00127544 1.9710E+17 Beta(2) 0 0 Beta(3) 0 0 Beta(4) 9.0721E

- 09 0



### Analysis of Deviance Table
Model Log(likelihood ) # Param's Deviance Test d.f. p - value

Full model

- 16.9192 6
Fitted model

- 18.4677 2 3.09685 4 0.5418
Reduced model - 102.298 1 170.758 5 <.0001



### AIC: = 40.9353

Goodness of Fit Table
Dose Est. Prob. Expected Observed Size Scaled Resid

0 0 0 0 60 0

10 0.0128 0.255 1 20 1.484

30 0.0446 0.892 0 20

- 0.966

100 0.6447 12.894 13 20 0.05

300 1 20 20 20 0

600 1 20 20 20 0



### Chi^2 = 3.14 d.f = 4 P

- value = 0.5352

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 40 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D**

**- 20. Summary of BMD**  **m odeling**  **r esults for**  **m odel predictions for**  1 2 3

4

5 6

7 8 9 10

11 **mortality (number found dead) in rats exposed to RDX in the diet for 13 weeks (von Oettingen et al., 1949)** 

Model a Goodness of fit BMD 1Pct (mg/kg

- day) BMDL 1Pct (mg/kg

- day) Basis for model selection

p - value AIC

Gamma 0.0341 66.088 3.14 0.648 All the models besides dichotomous

- Hill had goodness

of

- fit p

- values less than 0.10 and thus did not provide an adequate fit to the data. For the dichotomous

- Hill model, the slope parameter acheived the BMDS internal upper bound (18), so the results from t his model were not reliable. No model was selected. Dichotomous

- Hill 0.984 57.888 17.2 10.9 Logistic 0.0044 70.074 3.40 2.08 LogLogistic 0.0397 65.853 3.39 0.529
Probit 0.0056 69.283 3.28 1.94 LogProbit 0.0426 65.464 5.67 0.409
Weibull 0.0349 66.233 2.30 0.641 Multistage 3° a 0.0351 66.517 1.22 0.628 Multistage 2° b 0.0351 66.517 1.22 0.628
Quantal

- Linear 0.0995 64.639 0.919 0.623

a The Multistage 3° model may appear equivalent to the Multistage 2° model, however differences exist in digits

not displayed in the table.
b The Multistage 2° model may appear equivalent to the Multistage 3° model, however differences exist in digits

not d isplayed in the table.



### Fig **ure D**

**- 17. Plot of incidence rate by dose with fitted curve for Dichotomous** 

**** **Hill model for Model predictions for mortality (number found dead) in rats exposed to RDX in the diet for 13 weeks (von Oettingen et al., 1949); dose**  **shown in mg/kg**

**- day.** 

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 41 DRAFT―DO NOT CITE OR QU 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7

0 10 20 30 40 50Fraction Affected

dose Dichotomous-Hill Model, with BMR of 1% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BMDL

14:56 02/04 2016 BMDL BMD Dichotomous-Hill



### OTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**T able**  **D- 21. Model predictions for combined mortality (number found dead)**  1 2 3 **in male and female F344 rats exposed to RDX by gavage for 90 days (Crouse et al., 2006); BMR= 1% E**  **R**

Model a Goodness of fit BMD 1Pct (mg/kg

- day) BMDL 1Pct (mg/kg

- day) Basis for model selection

p - value AIC

Gamma 0.794 93.263 3.46 0.840 The multistage 2° model was selected as the representative multistage model based on lowest AIC. From among the multistage 2° and non

- multistage models, the multistage 2° model was selected based on lowest BMDL (BMDLs differed by more than threefold). Logistic 0.474 95.709 2.11 1.11 LogLogistic 0.794 93.332 3.17 0.872
Probit 0.574 94.797 2.40 1.07 LogProbit 0.854 92.832 3.96 1.48
Weibull 0.743 93.698 2.76 0.641 Multistage 2° 0.858 91.926 2.11 0.463
Quantal

- Linear 0.535 95.345 0.405 0.288 Multistage 5° b 0.731 93.851 2.42 0.433 Multistage 4° c 0.731 93.851 2.42 0.433 Multistage 3° 0.731 93.851 2.42 0.439
Dichotomous

- Hill 0.998 93.343 5.96 1.95

a Selected model in bold; scaled residuals for selected model for doses 0, 4, 8, 10, 12, and 15 mg/kg/day were

0.00,

- 0.86,

- 0.46, 0.53, 0.72, and 0.45, respectively. The BMD 10 and BMDL 10 for the selected model were 6.82 and 4.41 mg/kg

- d, respectively.
b The Multistage 5° model may appear equivalent to the Multistage 4° model, however differences exist in digits

not displayed in the table.

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 42 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1

2

3 4 5 6

7 8 9 10 11 12 13 14 15 16 17 18

19 20

21 **Figur e D** 

**- 18. Plot of incidence rate by dose, with the fitted curve of the**  **multistage 2° model, for mortality in male and female F344 rats exposed to RDX by gavage for 90 days**  **( Crouse et al.,**  **2006 )**  **. BMR = 1% ER; dose shown in mg/kg** 

**- day.** 
Multistage Model. (Version: 3.3; Date: 02/28/2013) The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)] 0 0.1 0.2 0.3 0.4 0.5 0.6

0 2 4 6 8 10 12 14Fraction Affected

dose Multistage Model, with BMR of 1% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BM

14:12 02/27 2014 BMD BMDL Multistage



### Be nchmark Dose Computation. BMR = 1% Extra risk BMD = 2.10625 BMDL at the 95% confidence level = 0.462994

Parameter Estimates
Variable Estimate Default Initial Parameter Values Background 0 0 Beta(1) 0 0.0134587 Beta(2) 0.00226548 0.00141278



### Analysis of Deviance Table
Model Log(likelihood ) # Param's Deviance Test d.f. p - value

Full model

- 43.6462 6
Fitted model

- 44.963 1 2.63354 5 0.7563
Reduced model - 55.6472 1 24.0019 5 0.0002169

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 43 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

AIC: = 91.926 1 2 3

4 5 6

7 8 9

10 Goo dness of Fit Table
Dose Est. Prob. Expected Observed Size Scaled Resid

0 0 0 0 20 0

4 0.0356 0.712 0 20

- 0.859

8 0.135 2.699 2 20

- 0.458

10 0.2027 4.054 5 20 0.526

12 0.2784 5.567 7 20 0.715

15 0.3993 7.987 7 20

- 0.451



### Chi^2 = 1.94 d.f = 5 P

- value = 0.8576

**Table D**

**- 22. Model predictions for mortality in**  **female Sprague**

**- Dawley rats**  **exposed to RDX by gavage on gestation days 6**

**– 15 (Angerhofer, 1986); BMR =**  **1% E R** 

Model a Goodness of fit BMD 1Pct (mg/kg

- day) BMDL 1Pct (mg/kg

- day) Basis for model selection

p - value AIC

Gamma 0.314 89.496 5.15 0.538 The multistage 3 ° model was selected based on lowest AIC. From among the multistage 3° and non

- multistage models, the multistage 3° model was selected based on lowest BMDL (BMDLs differed by more than threefold). Logistic 0.667 87.213 3.88 2.16 LogLogistic 0.312 89.473 4.88 0.560
Probit 0.643 87.196 3.37 1.87 LogProbit 0.319 89.522 5.58 0.885
Weibull 0.309 89.458 4.62 0.541
Quantal

- Linear 0.450 87.502 0.652 0.452 Multistage 3° 0.655 86.906 1.68 0.588 Multistage 2° 0.554 87.291 1.78 0.555

a Selected model in bold; scaled residuals for selected model for doses 0, 2, 6, and 20 mg/kg

- d were 0.00, 0.76,

-

0.52, and 0.04, respectively. The BMD 10 and BMDL 10 for the selected model were 10.9 and 6.09 mg/kg

- d, respectively.

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 44 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1

2 3 4

5 6 7 8 9 10 11 12 13 14 15 16 17

18 19 **Figure D** 

**- 19. Plot of**  **incidence rate by dose, with the fitted curve of the multistage 3° model, for mortality in female Sprague** 

**- Dawley rats exposed to**  **RDX by gavage on gestation days 6**

**– 15 (Angerhofer, 1986); BMR = 1% E**  **R.**

0 0.1 0.2 0.3 0.4 0.5

0 5 10 15 20Fraction Affected

dose Multistage Model, with BMR of 1% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BM

10:18 05/22 2014 BMD BMDL Multistage



### Mult istage Model. (Version: 3.3; Date: 02/28/2013) The f orm of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)]

Benchmark Dose Computation. BMR = 1% Extra risk BMD = 1.68097 BMDL at the 95% confidence level = 0.587568

Parameter Estimates
Variable Estimate Default Initial Parameter Values Background 0 0.00807857 Beta(1) 0.00588873 0.00216407 Beta(2) 0 0 Beta(3) 0.0000319123 0.0000406218



### Analysis of Deviance Table
Model Log(likelihood ) # Param's Deviance Test d.f. p - value

Full model

- 41.0771 4
Fitted model

- 41.4531 2 0.752152 2 0.6866

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 45 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Reduced model - 57.4292 1 32.7043 3 <.0001



### 1 2 3 4

5 6 7 8

9
10
11
12

13

14 15 16

17
18
19 20
21 AIC: = 86.9063

Goodness of Fit Table
Dose Est. Prob. Expected Observed Size Scaled Resid

0 0 0 0 39 0

2 0.012 0.478 1 40 0.759

6 0.0413 1.654 1 40

- 0.519

20 0.3114 15.881 16 51 0.036



### Chi^2 = 0.85 d.f = 2 P

- value = 0.6549



## D.2. **BENCHMARK DOSE MODELING SUMMARY FOR C**  **ANCER E**  **NDPOINTS** The cancer endpoints in the mouse that were selected for dose

- response modeling are presented in Table D- 23

. For each endpoint, the doses and tumor incidence data used for the modeling are presented.

**Table D-**  **23**

**. Cancer endpoints selected for dose** 

**- response modeling for RDX** 

Endpoint and reference Species/sex Dose (mg/kg

- d) Incidence/total
Hepatocellular adenomas or carcinomas Parker et al. (2006) Female B6C3F 1
m ouse 0 1.5 7 35 107 1/67 (1%) a 4/62 (6%) 5/63 (8%) 10 /64 (16%) 4/31 (13%)
Alveol ar/bronchiolar adenomas or carcinomas Lish et al. (1984) Female B6C3F 1
m ouse 0 1.5 7 35 107 7/65 (11%) 3/62 (5%) 8/64 (13%) 12/64 (19%) 7/31 (23%)

a For female mouse hepatocellular tumors from Lish et al. (1984) , tumor incidence and totals are those reported in the Pathology Working Group ( PWG ) reevaluation ( Parker et al., 2006 ).



### D.2.1. **Evaluation of Model Fit and Model Selection for Mouse Tumor Da**  **ta** First, to determine whether a time

- to

- tumor analysis was warranted, the survival curves were compared across dose groups for female mice in Lish et al. (1984) in the study to determine wheth er time of death should be incorporated in the dose

- response analysis of tumors. A log

- rank test on the Kaplan

- Meier survival curves per dose was used to do the comparison, excluding

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 46 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

animals that died prior to week 11 when the dose was reduced in the high

- dose group to 1
2 3
4
5 6
7 8
9 10
11
12 13
14
15 16
17

18
19
20

21

22 23 24

25 26 27 28 29 100 m g/k g- day. The test yielded a nonsignificant result ( p = 0.51), so a time

- to

- tumor analysis was not necessary for this study. Th erefore, non

- time

- dependent dose

- response analyses were conducted using standard BMDS models. For each tumor type , BMDS multistag e- cancer models 6 

### were fitted to the data using the maximum likelihood method. Each model was tested for goodness

- of

- fit using a chi

- square goodness

- of

- fit test (χ 2 

### p
- value <0.05 7 

### indicates lack of fit). Other factors were used to assess model fit, including scaled residuals, visual fit, and adequacy of fit in the low

- dose region and in the vicinity of the BMR. T he BMDL estimate and AIC value were used to select a best

- fit model from among the models exhibiting adequate fit. If the BMDL estimates w ere “sufficiently close” (i.e., differed by threefold or less )
, then the model selected was the one that yielded the lowest AIC value. If the BMDL estimates were not sufficiently close, then the lowest BMDL was selected as the POD. After selecting models for the two endpoints, the results were combined using MS

- COMBO in BMDS. This procedure analyzes the incidence of a tumor (adenoma or carcinoma) defined as present if either the hepatocellular or alveolar/bronchiolar tumor (or both) was present, and not present otherwise. The two endpoints were assumed to be independent.

**D.2.2. Modeling Results**  **for Mouse Tumor Data**
T he BMD modeling results for mouse tumor data sets are provided in Tables D

- 24 to D

- 28 (and Figures D

- 20 to D

- 24)

.

Mouse Tumor Data

— BMD Modeling Results

**Table D-**  **24**

**. Model predictions for combined alveolar/bronchiolar adenoma**  **and carcinoma in female B6C3F 1**  

### mice exposed to RDX by diet for 24 months **( Lish et al., 1984**  **) ; BMR = 10%**  **ER**

Model a 

### Goodness of fit BMD 10Pct 

### (mg/kg

- d) BMDL 10Pct 

### (mg/kg

- d) Basis for model selection p - value AIC Multistage 1° b Multistage 2° Multistage 3° Multistage 4° 0.417 218.68 52.8 27.7 All of the models reduced to the M ultistage 1° model, so this model was selected.

a Selected model in bold. Scaled residuals for the selected model for doses 0, 1.5, 7, 35, and 107 mg/kg -d ay were

0.40,

− 1. 27, 0.50, 0.73, and

− 0 .52, respectively.
b For the M ultistage 2°, 3°, and 4° models, the b2, b3 , and b4 coefficient estimates were 0 ( boundary of parameter

space). The models in this row reduced to the M ultistage 1° model.

6 The coefficients of the multistage

- cancer models were restricted to be nonnegative (beta values ≥0).
7 A significance level of 0.05 from U.S. EPA (2012b) is used for selecting cancer models because the model family (multistage) is selected a priori.
This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 47 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1

2

3

4 5 6 7

8
9
10
11
12
13
14
15 16
17 18
19 20 21

22 BMR = 10% ER; dose shown in mg/kg

- day.


### Figur **e D-**  **20**

**. Plot of incidence rate by dose, with the fitted curve for the**  **selected model, for combined alveolar/bronchiolar adenoma and carcinoma in female B6C3F**  **1** 

### mice exposed to RDX by diet for 24 months **( Lish et al.,**  **1984 )** 

**. Multistage Cancer Model**  (Version: 1.10; Date: 02/28/ 2013 ) The form of the probability function is: P[respon se] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)] The parameter betas are restricted to be positive

**Benchmark Dose Computation** BMR = 10% ER BMD = 52.8078 BMDL at the 95% confidence level = 27.748 Benchmark dose upper bound ( BMDU ) at the 95% confidence level = 194.806 Taken together, (27.748, 194.806) is a 90% two

- sided confidence interval for the BMD Multistage Cancer Slope Factor = 0.00360387 0 0.05

0.1 0.15

0.2 0.25

0.3 0.35

0.4 0.45

0 20 40 60 80 100Fraction Affected

dose Multistage Cancer Model, with BMR of 10% Extra Risk for the BMD and 0.95 Lower Confidence Limit for t

16:09 02/14 2014 BMD BMDL Multistage Cancer Linear extrapolation



### Pa rameter Estimates Variable Estimate Default i nitial p arameter v alues Background 0.093168 0.0998927 Beta(1) 0.00 1995 17 0.00155773

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 48 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Analysis of Deviance Table 1

2 3 4 5

6 7 8 Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 105.777 5
Fitted model

− 107.341 2 3.12764 3 0.3724
Reduced model

− 110.164 1 8.77367 4 0.06701



### AIC: = 218.682

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0.0932 6.056 7 65 0.403

1.5 0.0959 5.944 3 62

− 1.27

7 0.1057 6.768 8 64 0.501

35 0.1543 9.877 12 64 0.734

107 0.2675 8.292 7 31

− 0.524



### Chi^2 = 2.84 d.f

. = 3 p

- value = 0.4168

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 49 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-**  **25**

**. Model predictions for combined alveolar/bronchiolar adenoma**  1 2 3

4 5 6 7 8

9

10

11 12 13

14 **and carcinoma in female B6C3F 1**  

### mice exposed to RDX by diet for 24 months **( Lish et al., 1984**  **) ; BMR = 5%**  **ER**

Model a 

### Goodness of fit BMD 5Pct 

### (mg/kg

- d) BMDL 5Pct 

### (mg/kg

- d) Basis for model selection p - value AIC Multistage 1° b Multistage 2° Multistage 3° Multistage 4° 0.417 218.68 25.7 13.5 All of the models reduced to the M ultistage 1° model, so this model

was selected.

a Selected model in bold. Scaled residuals for the selected model for doses 0, 1.5, 7, 35, and 107 mg/kg -d ay were

0.40,

− 0.40,

− 1. 2 7, 0.50, 0.73, and

− 0.52 , respectively. The BMD 10 and BMDL 10 for the selected model were 52.8 and 27.7 mg/kg -d ay , r espectively.

BMR = 5% ER; dose shown in mg/kg

- day.


### Figur **e D-**  **21**

**. Plot of incidence rate by dose, with fitted curve for selected**  **model, for combined alveolar/bronchiolar adenoma and carcinoma in female B6C3F**  **1** 

### mice exposed to RDX by diet for 24 months **( Lish et al., 1984**  **).** 0 0.05

0.1 0.15

0.2 0.25

0.3 0.35

0.4 0.45

0 20 40 60 80 100Fraction Affected

dose Multistage Cancer Model, with BMR of 5% Extra Risk for the BMD and 0.95 Lower Confidence Limit for th

16:11 02/14 2014 BMD BMDL Multistage Cancer Linear extrapolation

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 50 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Multistage Cancer Model** (Version: 1.10; Date: 02/28/ 2013 ) 1
2 3
4
5 6
7 8
9 10
11
12 13 14

15 16

17 18 19 20

21 22 23 The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- be ta2*dose^2...)] The parameter betas are restricted to be positive

**Benchmark Dose Computation** BMR = 5% ER BMD = 25.7088 BMDL at the 95% confidence level = 13.5087 BMDU at the 95% confidence level = 94.8384 Taken together, (13.5087, 94.8384) is a 90% two

- sided confidence interval for the BMD Multistage Cancer Slope Factor = 0.00370131

Parameter Estimates Variable Estimate Default i nitial p arameter v alues Background 0.093168 0.0998927 Beta(1) 0.00 1995 17 0.00155773



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 105.777 5
Fitted model

− 107.341 2 3.12764 3 0.3724
Reduced model

− 110.164 1 8.77367 4 0.06701



### AIC: = 218.682

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0.0932 6.056 7 65 0.403

1.5 0.0959 5.944 3 62

− 1.27

7 0.1057 6.768 8 64 0.501

35 0.1543 9.877 12 64 0.734

107 0.2675 8.292 7 31

− 0.524



### Chi^2 = 2.84 d.f

. = 3 p

- value = 0.4168

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 51 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-**  **26**

**. Model predictions for combined hepatocellular adenoma and**  1 2 3

4 5 6 7 8 9

10

11

12 13 14

15 **carcinoma in female B6C3F 1**  

### mice exposed to RDX by diet for 24 months **( Parker et al., 2006**  **) ; BMR = 10%**  **ER**

Model a 

### Goodness of fit BMD 10Pct 

### (mg/kg

- d) BMDL 10Pct 

### (mg/kg

- d) Basis for model selection p - value AIC Multistage 1° b Multistage 2° Multistage 3° Multistage 4° 0.160 164.06 64.2 32.6 All of the models reduced to the M ultistage 1° model, so this model was selected.

a Selected model in bold. Scaled residuals for the selected model for doses 0, 1.5, 7, 35, and 107 mg/kg -d ay were

− 1.37, 0.35, 0.54, 1.34, and

− 1

. 05 , respectively.
b For the M ultistage 2°, 3°, and 4° models, the b2, b3 , and b4 coefficient estimates were 0 (boundary of parameter

space). The models in this row reduced to the M ultistage 1° model.

BMR = 10% ER; dose shown in mg/kg

- day.


### Figur **e D-**  **22**

**. Plot of incidence rate by dose, with fitted curve for**  **selected model, for combined hepatocellular adenoma and carcinoma in female B6C3F**  **1** 

### mice exposed to RDX by diet for 24 months **( Parker et al., 2006**  **).** 0 0.05

0.1 0.15

0.2 0.25

0.3

0 20 40 60 80 100Fraction Affected

dose Multistage Cancer Model, with BMR of 10% Extra Risk for the BMD and 0.95 Lower Confidence Limit for t

16:21 02/14 2014 BMD BMDL Multistage Cancer Linear extrapolation

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 52 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Multistage Cancer Model** (Version: 1.10; Date: 02/28/ 2013 ) 1
2 3
4
5 6
7 8
9 10
11
12 13 14

15 16

17 18 19 20

21 22 23 The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)] The parameter betas are restricted to be positive

**Benchmark Dose Computation** BMR = 10% ER BMD = 64.203 BMDL at the 95% confidence level = 32.6282 BMDU at the 95% confidence level = 281.385 Taken together, (32.6282, 281.385) is a 90% two

- sided confidence interval for the BMD Multistage Cancer Slope Factor = 0.00306483

Parameter Estimates Variable Estimate Default i nitial p arameter v alues Background 0.05 2075 5 0.0658334 Beta(1) 0.00164105 0.000876864



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 77.1516 5
Fitted model

− 80.0315 2 5.75967 3 0.1239
Reduced model

− 82.5216 1 10.74 4 0.02965



### AIC: = 164.063

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0.0521 3.489 1 67

− 1.369

1.5 0.0544 3.373 4 62 0.351

7 0.0629 3.963 5 63 0.538

35 0.105 6.719 10 64 1.338

107 0. 2047 6.347 4 31

− 1.045



### Chi^2 = 5.17 d.f

. = 3 p

- value = 0.16

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 53 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-**  **27**

**. Model predictions for B6C3F**  **1** 

### female mouse combined 1 2 3

4 5 6 7 8 9 10

11

12

13 14 15

16 **hepatocellular adenoma and carcinoma in mice exposed to RDX by diet for 24**  **months (**  **Parker e t al., 2006**  **) ; BMR = 5%**  **ER**

Model a 

### Goodness of fit BMD 5Pct 

### (mg/ kg

- d) BMDL 5Pct 

### (mg/kg

- d) Basis for model selection p - value AIC Multistage 1° b Multistage 2° Multistage 3° Multistage 4° 0.160 164.06 31.3 15.9 All of the models reduced to the M ultistage 1° model, so this model was selected.

a Selected model in bold; scaled residuals for selected model for doses 0, 1.5, 7, 35, and 107 mg/kg -d ay were − 1.37,

0.35, 0.54, 1.34, and

− 1.05, r espectively. The BMD 10 and BMDL 10 for the selected model were 64.2 and 32. 6 mg/kg

- d ay , respectively.
b For the M ultistage 2°, 3°, and 4° models, the b2, b3 , and b4 coefficient estimates were 0 (boundary of parameter space). The models in this row reduced to the M ultistage 1° model.

BMR = 5% ER; dose shown in mg/kg

- day.


### Figur **e D-**  **23**

**. Plot of incidence rate by dose, with fitted curve for selected**  **model, for B6C3F 1**  

### female mouse combined hepatocellular adenoma and **carcinoma in mice exposed to RDX by diet for 24 months (**  **Parker et al., 2006 ).**  0 0.05

0.1 0.15

0.2 0.25

0.3

0 20 40 60 80 100Fraction Affected

dose Multistage Cancer Model, with BMR of 5% Extra Risk for the BMD and 0.95 Lower Confidence Limit for th

16:21 02/14 2014 BMD BMDL Multistage Cancer Linear extrapolation

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 54 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Multistage Cancer Model** (V ersion: 1.10; Date: 02/28/ 2013 ) The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)] The parameter betas are restricted to be positive

**Benchmark Dose Computation** BMR = 5% ER BMD = 31.2563 BMDL at the 95% confidence level = 15.8846 BMDU at the 95% confidence level = 136.989 Taken together, (15.8846, 136.989) is a 90% two

- sided confidence interval for the BMD Multistage Cancer Slope Factor = 0.0031477 1
2 3
4
5 6
7 8
9 10
11
12 13 14

15 16

17 18 19 20

21 22 23 Parameter Estimates Variable Est imate Default i nitial p arameter v alues Background 0.052075 5 0.0658334 Beta(1) 0.00164105 0.000876864



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 77.1516 5
Fitted model

− 80.0315 2 5.75967 3 0.1239
Reduced model

− 82.5216 1 10.74 4 0.02965



### AIC: = 164.063

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0.0521 3.489 1 67

− 1.369

1.5 0.0544 3.373 4 62 0.351

7 0.0629 3.963 5 63 0.538

35 0.105 6.719 10 64 1.338

107 0.2047 6.347 4 31

− 1.045



### Chi^2 = 5.17 d.f

. = 3 p

- value = 0.16

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 55 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Combined results for presence of hepatocellular or alveolar/bronchiolar adenoma or** 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 **carcinoma in B6C3F 1**  

### female mice exposed to RDX by diet for 24 months; BMR = 10% **ER**

BMD = 29.0 mg/kg

- day; BMDL = 17.7 mg/kg

- day

MSCOMBO results
BMR of 10% Extra Risk

  Start of combined BMD and BMDL Calculations.** **

Combined Log

- L ikelihood

- 187.3723596892213
Combined Log

- li kelihood Constant 166.01737626058841

Benchmark Dose Computation

Spe cified effect = 0 .1
Risk Type = Extra risk

Confidence level = 0.95

BMD = 28.9753

BMDL = 17 .6574
Multistage Cancer Slope Factor = 0.00 566334

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 56 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Combined results for presence of hepatocellular or alveolar/bronchiolar adenoma or** 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 **carcinoma in B6C3F 1**  

### female mice exposed to RDX by diet for 24 months; BMR = 5% **ER**
BMD = 29.0 mg/kg

- day; BMDL = 17.7 mg/kg

- day

MSCOMBO results
BMR of 5% Extra Risk

  Start of combined BMD and BMDL Calculations.** **

Combined Log

- L ikelihood

- 187.3723596892213
Combined Log

- li kelihood Constant 166.01737626058841

B enchmark Dose Computation

Spe cified effect = 0.05

Ris k Type = Extra risk

Confidence level = 0.95

BM D = 14.1062

BMDL = 8. 59627
Multistage Cancer Slope Factor = 0.00 581647

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 57 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D-**  **28 . Model predictions for combined hepatocellular adenoma and**  1 2 3 4

5 6 7 8 9 10

11 12

13 14 15 16 17 **carcinoma in female B6C3F 1**  

### mice exposed to RDX by diet for 24 months, using **incidence frequencies from Parker et al. (2006)**  **and sample sizes from Lish et**  **al. (1984)**  **; BMR = 10% ER**

Model a 

### Goodness of fit BMD 10Pct 

### (mg/kg

- d) BMDL 10Pct 

### (mg/kg

- d) Basis for model selection p - value AIC Multistage 1° b Multistage 2° Multistage 3° Multistage 4° 0.171 163.98 64.8 32.8 All of the models reduced to the Multistage 1° model, so this model was selected.

a Selected model in bold. Scaled residuals for the selected model for doses 0, 1.5, 7, 35, and 107 mg/kg

- day were

-

1.34, 0.34, 0.49, 1.34,

- 1.03, respectively.
b For the Multistage 2°, 3°, and 4° models, the b2, b3 and b4 coefficient estimates were 0 (bound ary of parameter

space). The models in this row reduced to the Multistage 1° model.

BMR = 10% ER, dose shown in mg/kg

- day.


### Figur **e D-**  **24**

**. Plot of incidence rate by dose with fitted curve for Multistage** 

**** **Cancer 1° model for model predictions for combined hepatocellular adenoma**  **and carcinoma in female B6C3F1 mice exposed to RDX by diet for 24 months ,**  **using incidence frequencies from** Parker et al. (2006) **and sample sizes from** ( Parker et al., 2006 )

**.** 0 0.05 0.1 0.15 0.2 0.25 0.3

0 20 40 60 80 100Fraction Affected

dose Multistage Cancer Model, with BMR of 10% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BMDL

10:00 07/20 2015 BMD BMDL Multistage Cancer Linear extrapolation

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 58 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Multistage Model.** (Version: 3.4; Date: 05/02/ 2014 ) The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)] The parameter betas are restricted to be positive

**Benchmark Dose Computation.** BMR = 10% Extra risk BMD = 64.7853 BMDL at the 95% confidence level = 32.7981 BMDU at the 95% confidence level = 291.495 Taken together, (32.7981, 291.495) is a 90% two

- sided confidence interval for the BMD Multistage Cancer Slope Factor = 0.00304896 1
2 3
4
5 6
7 8
9 10
11
12 13 14

15 16

17 18
19 20

21 22
23 Paramet er Estimates Variable Estimate Default i nitial p arameter v alues Background 0.0525105 0.0656105 Beta(1) 0.0016263 0.000878945



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 77.2 5
Fitted model

− 79.99 2 5.57217 3 0.13
Reduced model

− 82.43 1 10.462 4 0.03



### AIC: = 163.978

Goodness

- of

- Fit Table
Dose Est. Prob. Expected Observed Size Scaled r esid uals

0 0.0525 3.413 1 65

− 1.34

1.5 0.0548 3.399 4 62 0.34

7 0.0632 4.047 5 64 0.49

35 0.1049 6.716 10 64 1.34

107 0.2038 6.319 4 31

− 1.03



### Chi^2 = 5.02 d.f

. = 3 p

- value = 0.1706

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 59 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Combined results for presence of hepatocellular or alveolar/bronchiolar adenoma or** 1 2 3 4 5 6 7 8 9 10 11

12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 **carcinoma in B6C3F 1**  

### female mice exposed to RDX by diet for 24 months; **for hepatocellular adenoma or carcinoma, the incidence frequencies from**  **Parker et al. (2006) and the**  **sample si**  **zes from Lish et al. (1984)**  **were used ;**  **BMR = 10% ER**
BMD = 29.0 mg/kg

- day; BMDL = 17.7 mg/kg

- day

MSCOMBO results
BMR of 10% Extra Risk

  Start of combined BMD and BMDL Calculations.** **
Combined Log

- Like lihood

- 187.33008597565913
Combined Log

- like lihood Constant 166.068416550547

Be nchmark Dose Computation

Spe cified effect = 0.1
Risk Type = Extra risk

Confidence level = 0.95

BM D = 29.0933

BMDL = 17.7048
Multistage Cancer Slope Factor = 0.00564818

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 60 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**D.2.3. Dose** 

**- response**  **A nalysis**  **and BMD Modeling Documentation for Other Tumor Data**  1 2
3
4
5 6

7

8 9 10
11
12
13
14
15 16
17 18
19
20 **Sets**
T his a ppendix also presents a quantitative dose

- response analysis of incidence of liver carcinomas in male F344 rats ( Levine et a l., 1983b ) and incidence of lung carcinomas in male B3 C6F 1 

### mice (Table D -29)

. The resulting candidate oral slope factor s (OSF s ) are presented for comparison with OSF estimates provided in Section 2.3.3 of the Toxicological Review.

**Table D-**  **29 . Liver carcinoma data from**  **Levine et al. (1983b)**

Endpoint and reference Species/sex Dose (mg/kg

- d) Incidence/total Alveolar/bronchiolar carcinomas Lish et al. (1984) Male B6C3F 1 mouse 0 1.5 7 35 107 3/63 (5%) 6/60 (10%) 3/62 (5%) 7/59 (12%) 5/27 (19%) Hepatocellular carcinomas Levine et al. (1983b) Mal e F344 rat 0 0.3 1.5 8 40 1/55 (2%) 0/55 (0%) 0/52 (0%) 2/55 (4%) 2/31 a (6%)

a The denominators listed in the table represent the number of animals that were alive 1 year after dosing began.



### Fo r male mice in Lish et al. (1984)
, a log

- rank test on the Kaplan

- Meier survival curves,
str atified by dose, yielded a nonsignificant result ( p
- value ≥0.10), indicating that the survival curves were similar across dose groups. Therefore, a time

- to

- tumor analysis was not necessary for hepatocellular carcinomas in Lish et al. (1984)

. A non
- time

- dependent dose

- response analysis was

co nducted using BMDS multistage

- cancer models, and the model selection procedures described in Section D.2.1 were used to select the appropriate models. Subsequently, the administered dose was converted to a human equivalent dose ( HED ) on the basis of (body weight) 3/4 

### ( U.S. EPA, 1992 ) , as des cribed in Section 2.3.2. The POD estimate for male mouse carcinomas and OSF calculated from this POD are provided in Table D

- 30 ; det ailed BMD modeling results are provided in Table D

- 31 (and Figure D

- 25).

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 61 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D**

**- 30**  **. Model predictions and oral slope factor for alveolar/bronchiolar**  1 2 3

4 5 6 7

8 9 10

11 12 13 14 15 16

17 18

19 20 21 **carcinomas in ma**  **le B6C3F 1**  

### mice **exposed to RDX by diet for 2 years (**  **Lish et al., 1984**  **)**

Tumor type Selected model BMR BMD, mg/kg

- d BMDL, mg/kg

- d POD = BMDL 10

- HED a



### mg/kg

- d Candidate OSF b 

### (mg/kg

- d) -1
Alveolar/bronchiolar carcinomas Multistage 1° 10% ER 76.1 36.2 5.41 0.0 18

a Based on allometric scaling of administered RDX dose; BMDL 10

- HED = BMDL 10 × (BW a 1/4 /BW h 1/4 ), BW a = 0.035 kg,

and BW h = 70 kg.
b Slope factor = BMR/BMDL 10

- HED , where BMR = 0. 10 (10 % ER ).



### Table **D- 31** 

**. Summary of BMD m**  **odeling r**  **esults for m**  **odel predictions for alveolar/bronchiolar carcinoma in male B6C3F1 mice exposed to RDX by diet**  **for 2 years**  **( Lish et al., 1984**  **) ; BMR = 10% ER** 

Model a 

### Goodness of fit BMD 10Pct 

### (mg/kg

- d) BMDL 10Pct 

### (mg/kg

- d) Basis for model selection p - value AIC Multistage 1° b Multistage 2° Multistage 3° Multistage 4° 0.561 162.00 76.1 36.2 All of the models reduced to the multistage 1° model, so this model was selected.

a Selected model in bold; scaled residuals for selected model for doses 0, 1.5, 7, 35, and 107 mg/kg

- day were

− 0.52, 1.08, −0.74, 0.27, and −0.1, respectively.
b For the Multistage 2°, 3°, and 4° models, the b2, b3, and b4 coefficient estimates were 0 (boundary of parameter

space). The models in this row reduced to the Multistage 1° model.

BMR = 10% ER; dose shown in mg/kg

- day.


### Figur **e D-**  **25**

**. Plot of incidence rate by dose with fitted curve for Multistage** 

**** **Cancer 1° model for Model predictions for alveolar/ bronchiolar carcinoma in**  **male B6C3F1 mice exposed to RDX by diet for 24 months (**  **Lish et al., 1984 ).**  0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4

0 20 40 60 80 100Fraction Affected

dose Multistage Cancer Model, with BMR of 10% Extra Risk for the BMD and 0.95 Lower Confidence Limit for the BMDL

18:19 01/08 2016 BMD BMDL Multistage Cancer Linear extrapolation

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 62 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1 2
3
4
5 6
7 8
9
10
11
12
13
14 15

16 17

18 19 20 21

22 23 **Mult istage Model.**  (Version: 3.4; Date: 05/02/ 2014 ) The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)] The parameter betas are restricted to be positive

**Benchmark Dose Computation.** BMR = 10% Extra risk BMD = 76.1197 BMDL at the 95% confidence level = 36.2316 BMDU at the 95% confidence level = 27443100 T aken together, (36.2316, 27443100) is a 90% two

- sided confidence interval for the BMD Multistage Cancer Slope Factor = 0.00276003

Parameter Estimates Variable Estimate Default i nitial p arameter v alues Background 0.0635642 0.0652915 Beta(1) 0.00138414 0.00131052



### Analysis of Deviance Table Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 78 5
Fitted model

− 79 2 1.99294 3 0.57
Reduced model

− 81.08 1 6.15622 4 0.19



### AIC: = 162.001

Goodness

- of

- Fit Table
Dose Est. Prob. Expected Observed Size Scaled r esid uals

0 0.0636 4.005 3 63

− 0.52

1.5 0.0655 3.93 6 60 1.08

7 0.0726 4.501 3 62

− 0.74

35 0.1078 6.363 7 59 0.27

107 0. 1925 5.197 5 27

− 0.1



### Chi^2 = 2.06 d.f

. = 3 p

- value = 0.561

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 63 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

For male rats in Levine et al. (1983b)
, the high

- dose group had a markedly lower survival 1
2 3
4
5 6
7 8
9 10
11
12 13
14
15 16
17 18
19 20
21
22
23
24
25
26
27
28
29
30 31
32
33 34
35
36 37
38 curve than the other dose groups, indicating a substantial number of early deaths in the high

- dose
group. A log

- rank test on the Kaplan

- Meier survival curves , stratified by dose , y ielded a significant result ( p
- value <0.001), in which case a time

- to

- tumor analysis is generally preferred. However, such an analysis was not possible because the data were insufficient to allow this analysis. Although tumor incidence was listed for each animal in th e source article, the pathology report used a different animal numbering system than the experimental report where the times of death were listed, and the relationship between the two systems was not documented

. This prohibit ed the matching of the times of death and the tumor incidence of the animals
, thus prohibiting the use of a time

- to

- tumor analysis. Therefore, a no n
- time

- dependent dose

- response analys i s w as conducted using BMDS multistage

- cancer models. The model selection procedures described in Section D.2.1 were used to select the appropriate models. To account for the difference in the survival curves across the groups for rats, the number of animals alive at 12 months was used as the denominator in the analysis (d enominators listed in Table D

- 28 ). Because the maximum liver tumor response in the male rat was 6.4%, a BMR of 5% was used to model male rat liver tumor data in order to obtain a BMD and BMDL in the range of the experimental data, as recommended in Section 3.2 of Guidelines for Carcinogen Risk Assessment ( U.S. EPA, 2005a )

. To estimate the HED at the BMDL, HEDs based on both administered dose scaled by BW 3/4 

### and physiologically based pharmacokinetic ( PBPK ) modeling were considered

. C onfidence in the revised rat PBPK model is relatively high (see Appendix C, Section C. 1 .5) ; however, the choice of an internal dose is not straightforward. First, evidence regarding the involvement of metabolites has been discussed in the literature only in the context of the mouse, and the rate of metabolism (allometrically adjusted) appears to be qualitatively slower for the rat. Second, metabolism in the model represents the total of all pathways, whereas it is only the minor N

- nitroso metabolic route, and not the oxidative route that has been proposed as a factor in RDX

- induced mouse carcinogenicity. Third, while blood concentration of RDX as an internal dose would be m ore proximally relevant to the tissue than administered dose, there are no data to indicate that the parent RDX is directly related to its carcinogenicity. Therefore, given the uncertainties, HEDs based on both administered dose scaled by BW 3/4 

### and area u nder the curve ( AUC ) of RDX arterial blood concentration (calculated using the PBPK model) are presented. Extrapolation based on the internal dose of the parent compound is accomplished by assuming toxicological equivalence when dose is expressed in terms of the AUC of the RDX blood concentration. The POD estimate s for rat liver carcinomas and the O S Fs calculated from these PODs are provided in Table D- 32 ; detailed BMD modeling results are provided in Table D

- 33 (and Figure D

26). Results based on two dose- metrics are presented: administered dose of RDX scaled by BW 3/4 

### (when dose is expressed in terms of mg/kg

- day, this entails scaling by BW −1/4 

### ) and AUC of RDX arterial blood concen tration (using PBPK modeling).

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 64 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

**Table D**

**- 32**  **. Model predictions and oral slope factor for hepatocellular** 1 2 3

4 5 6 7 8

9 10 11

12 13 14 15 16 17 **carcinomas in male F344 rats administered RDX in the diet for 2 yea**  **rs (**  **Levine et** 

**al., 1983b )** 

Tumor type Selected model BMR BMD , mg/kg

- d BMDL , mg/kg

- d POD = BMDL 05

- HED ,



### mg/kg

- d Candidate OSF a 

### (mg/kg

- d) -1
Hepatocellular carcinomas Multistage 1° 5% ER 28.5 11.8 2. 88 b , 5. 75 c 0. 017 b , 0. 009 c

a Slope factor = BMR/BMDL 05

- HED , where BMR = 0.05 (5% ER ).
b Based on allometric scaling of administered RDX dose; BMDL 05

- HED = BMDL 05 × (BW a 1/4 /BW h 1/4 ), BW a = 0.25 kg, and

BW h = 70 kg

. c Based on toxicological equivalence of PBPK model derived AUC of RDX blood concentration

.


### Ta **ble D-**  **33**

**. Model predictions for combined hepatocellular adenoma and**  **carcinoma in F344 rats exposed to RDX by diet for 24 months**  **( Levine et al.,**  **1983b )**  **; BMR = 5% ER** 

Model a 

### Goodness of fit BMD 5 Pct 

### (mg/kg

- d) BMDL 5 Pct 

### (mg/kg

- d) Basis for model selection p - value AIC Multistage 1° b Multistage 2° Multistage 3° Multistage 4° 0.493 49.095 28.5 11.8 All of the models reduced to the M ultistage 1° model, so this model was selected.

a Selected model in bold. Scaled residuals for the selected model for doses 0, 0.3, 1.5, 8, and 40 mg/kg -d ay were

0.89 , − 0. 67,

− 0. 74, 0.74, and

− 0.26 , respectively.
b For the M ultistage 2°, 3°, and 4° models, the b2, b3 , and b4 coefficient estimates were 0 (boundary of parameter

space). The models in this row reduced to the M ultistage 1° model.

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 65 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

1

2

3 4 5

6
7 8 9
10
11
12
13
14
15 16
17 18 19

20 BMR = 5% ER; dose shown in mg/kg

- day.


### Figur **e D-**  **26**

**. Pl**  **ot of incidence rate by dose, with fitted curve for selected model, for combined hepatocellular adenoma and carcinoma in F344 rats**  **exposed to RDX by diet for 24 months (**  **Levine et al., 1983b ).** 

**Multista ge Model**  (Version: 3.4; Date: 05/02/ 2014 ) The form of the probability function is: P[response] = background + (1

- background)*[1

- EXP(

- beta1*dose^1

- beta2*dose^2...)] The parameter betas are restricted to be positive

**Benchmark Dose Computation** BMR = 5% ER BMD = 28.4525 BMDL at the 95% confidence level = 11.8487 BMDU at the 95% confidence level = 235.886 Taken together, (11.8487, 235.886) is a 90% two

- sided confidence interval for the BMD Multistage Cancer Slope Factor = 0.0042 1987 0 0.05

0.1 0.15

0.2 0.25

0 5 10 15 20 25 30 35 40Fraction Affected

dose Multistage Cancer Model, with BMR of 5% Extra Risk for the BMD and 0.95 Lower Confidence Limit for th

17:39 07/29 2014 BMD BMDL Multistage Cancer Linear extrapolation



### Pa rameter Estimates Varia ble Estimate Default i nitial p arameter v alues Background 0.00766363 0.00949438 Beta(1) 0.00180277 0.00149364

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 66 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Analysis of Deviance Table 1

2 3 4 5

6 7 8 Model Log (likelihood) Number of p aram eter s Deviance Test d.f. p - value

Full model

− 21.0055 5
Fitted model

− 22.5473 2 3.08372 3 0.3789
Reduced model

− 24.4692 1 6.92747 4 0.1398



### AIC: = 49.0947

Goodness

- of

- Fit Table
Dose Est. p rob. Expected Observed Size Scaled residuals

0 0.0077 0.421 1 55 0.894

0.3 0.0082 0.451 0 55

− 0.674

1.5 0.0103 0.538 0 52

− 0.737

8 0.0219 1.203 2 55 0.735

40 0.0767 2.378 2 31

− 0.255



### Chi^2 = 2.4 d.f

. = 3 p

- value = 0.493

This document is a draft for review purposes only and does not constitute Agency policy. 

### C- 67 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine



# REFERENCES FOR APPENDICES

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 Ang erhofer, R; Davis, G; Balezewski, L. (1986). Teratological assessment of Trinitro - RDX in rats. (7 5

- 51

- 0573

- 86). Aberdeen Proving Ground, MD: U.S. Army Environmental Hygiene Agency. Arfsten, DP; Davenport, R; Schaeffer, DJ. (1994). Reversion of bioluminescent bacteria (Mutatox TM ) to their luminescent state upon exposure to organic compounds, munitions, and metal salts. Bio med Environ Sci 7: 144

- 149. ATSDR (Agency for Toxic Substances and Disease Registry). (2012). Toxicological profile for RDX (U pdate) [ATSDR Tox Profile]. (NT IS/10590213). Atlanta, GA: U.S. Department of Health and Human Services, Public Health Service. Bannon, DI. (2006). Toxicology study no. 87

- XE

- 059F

- 06: Biomar kers of RDX exposure in breath of swin e, August - September 2006. Aberdeen Proving Ground, MD: U.S. Army Center for Health Promotion and Preventive Medicine. Ban non, DI; Dillman, JF; Hable, MA; Phillips, CS; Perkins, EJ. (2009a). Global gene expression in rat br ain and liver after oral exposure to the explosive hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine (RDX). Chem Res Toxicol 22: 620

- 625. http://dx.doi.org/10.1021/tx800444k Bannon, DI; Dillman, JF; Perkins, EJ; Bao, W; Wolfinger, RD; Chu, T; Phillips, CS. (2006). Acute RDX exp osure and gene expression in the rat brain [Abstract]. Oxford, United Kingdom: Oxford University Press. http://www.toxicology.org/pubs/docs/Tox/2006Tox.pdf Bannon, DI; Johnson, M; Williams, L; Adams, V; Perkins, E; Gust, K; Gong, P. (2009b). RDX and miRNA expression in B6C3F1 mice [Letter]. Environ Health Perspect 117: A98. http://dx.doi.org/10.1289/ehp.0800276 Barsotti, M; Crotti, G. (1949). [Attacchi epileptici come manifestazione di intossicazione pr ofessionale da trimetilen

- trinitroamina (T4)]. Med Lav 40: 107

- 112. Bhushan, B; Trott, S; Spain, JC; Halasz, A; Paquet, L; Hawari, J. (2003). Biotransformation of he xahydro- 1,3,5

- trinitro

- 1,3,5

- triazine (RDX) by a rabbit liver cytochrome P45 0: Insight into the mechanism of RDX biodegradation by Rhodococcus sp. strain DN22. Appl Environ Microbiol 69: 1347

- 1351. http://dx.doi.org/10.1128/AEM.69.3.1347

- 1351.2003 Bingham, E; Cohrssen, B; Powell, CH. (2001). Patty's toxicology. In E Bingham; B Cohrssen; CH Powell (Eds.), Patty's toxicology vol 6: Ketones, alcohols, esters, epoxy compounds, organic peroxides (5 ed.). Hoboken, NJ: John Wiley & Sons. http://dx.doi.org/10.1002/0471125474 Brown, RP; Delp, MD; Lindstedt, SL; Rhomberg, LR; Beliles, RP. (1997). Physiological parameter

va lues for physiologically based pharmacokinetic models [Review]. Toxicol Ind Health 13: 407

- 484. http://dx.doi.org/10.1177/074823379701300401 Cao, CJ; Reddy, G; Bannon, DI; Johnson, MS. (2008). In vitro study of hexahydro

- 1,3,5

- trinitro

- 1,3,5

- tr iazine (RDX) metabolism in human liver. Aberdeen Proving Ground, MD: U.S. Army Center for Health Promotion and Preve ntive Medicine. Cholakis, JM; Wong, LCK; Van Goethem, DL; Minor, J; Short, R; Sprinz, H; Ellis, HV, III. (1980). Mam malian toxicological evaluation of RDX. (D AMD17

- 78 -C- 8027). Kansas City, MO: Midwest Research Institute. Cotruvo, JA; Simmon, VF; Spanggord, RJ. (1977). Investigation of mutagenic effects of products of

oz onation reactions in water. Ann N Y Acad Sci 298: 124

- 140. http://dx.doi.org/10.1111/j.1749

- 6632.1977.tb19259.x

This document is a draft for review purposes only and does not constitute Agency policy. 

### R- 1 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Crouse, LCB; Michie, MW; Major, M; Johnson, MS; Lee, RB; Paulus, HI. (2006). Subchronic oral 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 t oxicity of RDX in rats. (Toxicology Study No. 85

- XC

- 5131

- 03). Aberdeen Proving Ground, MD: U.S. Army Center for Health Promotion and Preventive Medicine. Crouse, LCB; Michie, MW; Major, MA; Leach, GJ; Reddy, G. (2008). Oral bioavailabili ty of cy clotrimethylenetrinitramine (RDX) from contaminated site soils in rats. Int J Toxicol 27: 317

- 322. http://dx.doi.org/10.1080/10915810802366885 Davies, JOJ; Roberts, DM; Hittarage, A; Buckley, NA. (2007). Oral C

- 4 plastic explosive in humans - A

c ase series. Clin Toxicol 45: 454

- 457. htt p://dx.doi.org/10.1080/15563650601118044 Dilley, JV; Tyson, CA; Newell, GW. (1979). Mammalian toxicological evaluation of TNT wastewaters. V olume II. Acute and subacute mammalian toxicity of TNT and the LAP mixture. (ADA080957. DAMD17

- 76 -C- 6050.). Menlo Park, CA: SRI International. http://oai.dtic.mil/oai/oai?verb =getRecord&metadataPrefix=html&identifier=ADA080957 Fellows, RJ; Driver, CR; Cataldo, DA; Harvey, SD. (2006). Bioavailability of hexahydro

- 1,3,5

- trinitro

1 ,3,5

- triazine (RDX) to the prairie vole (Microtus ochrogaster). Environ Toxicol Chem 25: 1881

- 1886. http://dx.doi.org/10.1897/05

- 446R1.1 George, SE; Huggins

- Clark, G; Brooks, LR. (2001). Use of a Salmonella microsuspension bioassay to d etect the mutagenicity of munitions compounds at low concentrations. Mutat Res 490: 45

- 56. http://dx.doi.org/10.1016/S1383

- 5718(00)00150 -9 Goldberg, DJ; Green, ST; Nathwani, D; McMenamin, J; Hamlet, N; Kennedy, DH. (1992). RD X in toxication causing seizures and a widespread petechial rash mimicking meningococcaemia. J R Soc Med 85: 181. Guo, L; Xu, H; Chen, Y; Chang, Y. (1985). Dis tribution and metabolism of tritium

- labeled hexogen in

w hite mice. Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi 3: 335

- 339. Harrell- Bruder, B; Hutchins, KL. (1995). Seizures caused by ingestion of composition C

- 4. Ann Em erg Med 26: 746

- 748. http://dx.doi.org/10.1016/S0196

- 0644(95)70049 -8 Hart, ER. (1974). Subacute toxicity of RDX and TNT in dogs. Final report. (A717530). Kensington, M D: Litton Bionetics, Inc. https://ntrl.ntis.gov/NTRL/dashboard/searchResults.xhtml?searchQuery=ADA035717 Hart, ER. (1976). Two

- year chronic toxicity study in rats. (N00014

- 73 -C- 0162). Kensington, MD: L itton Bionetics, Inc. https://ntrl.ntis.gov/NTRL/dashboard/searchResults. xhtml?searchQuery=ADA040161 Hathaway, JA; Buck, CR. (1977). Absence of health hazards associated with RDX manufacture and

u se. J Occup Med 19: 269

- 272. Hess

- Ruth, A; Crouse, L; Roszell, L. (2007). RDX pilot development neurotoxicity test in rats. ( Toxicology Study No. 85

- XC

- 064Y

- 07). Aberdeen Proving Ground: U.S. Ar my Center for Health Promotion and Preventive Medicine. Hett, DA; Fichtner, K. (2002). A plastic explosive by mouth. J R Soc Med 95: 251

- 252. http://dx.doi.org/10.1258/jrsm.95.5.251 Hollander, AI; Colbach, EM. (1969). Composition C

- 4 induced seizures: A report of five cases. Mil M ed 134: 1529

- 1530. Johnson, MS. (2015). Memorandum for National Center for Environmental Assessment (8101R/Dr. L ouis D'Amico), U.S. EPA - Office of Research and Development, 1200 Pennsylvania Ave., NW, Washington, DC 20460. Subject: Additional data from the oral subchronic toxicity of RDX in rats, 2006. (Toxicology study no. 85

- XC

- 5131

- 03), U.S. Army Center for Health Promotion and Preventa tive Medicine, Aberdeen Proving Ground, Maryland; Incidence of seizure relative to mortality events. (MCHB

- IP

- T). Aberdeen Proving Ground, MD: Department of the Army, US Army Institute of Public Health. Kaplan, AS; Berghout, CF; Peczenik, A. (1965). Human intoxication from RDX. Arch Environ Health 1 0: 877

- 883.

This document is a draft for review purposes only and does not constitute Agency policy. 

### R- 2 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

Kasuske , L; Schofer, JM; Hasegawa, K. (2009). Two marines with generalized seizure activity. J 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 Em erg Nurs 35: 542

- 543. http://dx.doi.org/10.1016/j.jen.2008.05.001 Ketel, WB; Hughes, JR. (1972). Toxic encephalopathy with seizures secondary to ingestion of

co mposition C

- 4. A clinical and electroencephalographic study. Neurology 22: 871

- 876. Knepshield, JH; Stone, WJ. (1972). Toxic effects following ingestion of C

- 4 plastic explosive. In W Keu p (Ed.), Drug Abuse: Current Concepts and Research (pp. 296

- 300). Springf ield, IL: Charles C. Thomas. Krishnan, K; Crouse, LCB; Bazar, MA; Major, MA; Reddy, G. (2009). Physiologically based p harmacokinetic modeling of cyclotrimethy lenetrinitramine in male rats. J Appl Toxicol 29: 629

- 637. http://dx.doi.org/10.1002/jat.1455 Küçükardali, Y; Acar, HV; Özkan, S; Nalbant, S; Yazgan, Y; Atasoyu, EM; Keskin, O; Naz, A; Akyatan, N; G ökben, M; Danaci, M. (2003). Accidental oral poisoning caused by RDX (cyclonite): A report

o f 5 cases. J Intensive Care Med 18: 42

- 46. http://dx.doi.org/10.1177/0885066602239123 Lachance, B; Robidoux, PY; Hawari, J; Ampleman, G; Thiboutot, S; Sunahara, GI. (1999). C ytotoxic

a nd genotoxic effects of energetic compounds on bacterial and mammalian cells in vitro. Mutat Res 444: 25

- 39. http://dx.doi.org/10.1016/S1383

- 5718(99)00073 -X Levin, DE; Hollstein, M; Christman, MF; Schwiers, EA; Ames, BN. (1982). A new Salmonella tester

s train (TA102) with AT base pairs at the site of mutation detects oxidative mutagens. PNAS 79: 7445

- 7449. http://dx.doi.org/10.1073/pnas.79.23.7445 Levine, BS; Furedi, EM; Gordon, DE; Barkley, JJ; Lish, PM. (1990). Toxic interactions of the munitions

c ompounds TNT and RDX in F344 rats. Fundam Appl Toxicol 15: 373

- 380. http://dx.doi.org/10.1016/0272

- 0590(90)90062 -O Levine, BS; Furedi, EM; Gordon, DE; Burns, JM; Lish, PM. (1981a). Thirteen week oral (diet) toxicity

s tudy of trinitrotoluene (TNT), hexahydro

- 1, 3, 5 -trinitro- 1, 3, 5

- triazin e (RDX) and TNT/RDX mixtures in the Fischer 344 rat. Final report. (ADA108447. DAMD17

- 79 -C- 9120. DAMD17

- 79 -C- 9161). Chicago, IL: IIT Research Institute. htt p://oai.dtic.mil/oai/oai?verb=getRecord&metadataPrefix=html&identifier=ADA108447 Levine, BS; Furedi, EM; Gordon, DE; Burns, JM; Lish, PM. (1981b). Thirteen wee k toxicity study of h exahydro- 1,3,5

- trinitro

- 1,3,5

- triazine (RDX) in Fischer 344 rats. Toxicol Lett 8: 241

- 245. http://dx.doi.org/10.1016/0378

- 4274(81)90108 -9 Levine, BS; Furedi, EM; Rac, VS; Gordon, DE; Lish, PM. (1983a). Determination of the chronic

ma mmalian toxicological effects of RDX (twenty

- four month chronic toxicity/c arcinogenicity study of hexahydro

- 1,3,5 -trinitro- 1,3,5

- triazine (RDX) in the Fischer 344 rat): Final report

-- Phase V. Volume 1. (ADA160774. IITRI

- L6121

- 6. DAMD17

- 79 -C- 9161). Chicago, IL: IIT Research Institute. http://oai.dtic.mil/oai/oai?verb=getRecord&metadataPrefix=html&identifier=ADA160774 Levine, BS; Lish, PM; Furedi, EM; Rac, VS; Sagartz, JM. (1983b). Determination of the chronic

ma mmalian toxicological effects of RDX (twenty

- four month chronic toxicity/carcinogenicity study of hexahydro

- 1,3,5 -trinitro- 1,3,5

- triazine (RDX) in the Fischer 344 rat): Final repo rt

-- phase V. Chicago, IL: IIT Research Institute. Levine, BS; Lish, PM; Furedi, EM; Rac, VS; Sagartz, JM. (1983c). Determination of the chronic

m ammalian tox icological effects of RDX (twenty

- four month chronic toxicity/carcinogenicity study of hexahydro

- 1,3,5 -trinitro- 1,3,5

- triazine (RDX) in the Fischer 344 rat): Final report

-- phase V. Volume 3. Chicago, IL: IIT Research Institute. Lipscomb, JC; Poet, TS. (2008). In vitro measurements of metabolism for application in p harmacokinetic modeling [Review]. Pharmacol Ther 118: 82

- 103. http://dx.doi.org/10.1016/j.pharmthera.2008.01.006 Lish, PM; Levine, BS; Furedi, EM; Sagartz, JM; Rac, VS. (1984). Determinatio n of the chronic

ma mmalian toxicological effects of RDX: Twenty

- four month chronic toxicity/carcinogenicity study of hexahydro

- 1,3,5 -trinitro- 1,3,5

- triazine (RDX) in the

This document is a draft for review purposes only and does not constitute Agency policy. 

### R- 3 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

B 6C3F1 hybrid mouse (Volumes1

- 3). (ADA181766. DAMD17

- 79 -C- 9161). Fort Detrick, 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 Frederi ck, MD: U.S. Army Medical Research and Development Command. h ttp://oai.dtic.mil/oai/oai?verb=getRecord&metadataPrefix=html&identifier=ADA181766 Ma, B; Li, H. (1993). Neurobehavioral effects of hexogen on exposed workers. Gongye Weisheng yu Z hiyebin 19: 20

- 23. MacPhail, RC; Walker, QD; Cook, L. (1985). Neurotoxicology of cyclotrimethylenetrinitramine ( RDX). Final report. (ADA168266). Research Triangle Park, NC: U.S. Environmental Protection Agency, Health Effects Resear ch Laboratory, Neurotoxicology Division. https://ntrl.ntis.gov/NTRL/dashboard/searchResults.xhtml?searchQuery=ADA168266 Major, MA; Reddy, G; Berge, MA; Patzer, SS ; Li, AC; Gohdes, M. (2007). Metabolite profiling of [ 14C]hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine (RDX) in Yucatan miniature pigs. J Toxicol Environ Health A 70: 1191

- 1202. http://dx.doi.org/10 .1080/15287390701252717 Martin, D; Hart, E. (1974). Subacute toxicity of RDX and TNT in monkeys (pp. 1

- 216). (ADA044650). K ensington, MD: Litton Bionetics, Inc

. https://ntrl.ntis.gov/NTRL/dashboard/searchResults.xhtml?searchQuery=ADA044650 Merrill, SL. (1968). Ingestion of an explosive material, composition C

- 4: A re port of two cases. U SARV Med Bull 40: 5

- 11. Minor, JL; Short, RD, Jr; Van Goethem, DL; Wong, LCK; Dacre, JC. (1982). Mutagenic and

r eproductive studies of hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine (RDX) in rats and rabbits [Abstract]. Musick, TJ; Berge, MA; Patzer, SS; T ilch, KR. (2010). Absorption, distribution, metabolism, and

ex cretion of 14C

- RDX following oral administration to minipigs (DAAD05

- 02 -P- 2319). (ADA526472). Madison, WI: Covance Laboratories Inc. http://www.dtic.mil/cgi

b in/GetTRDoc?AD=ADA526472 Neuwoehner, J; Schofer, A; Erlenkaemper, B; Steinbach, K; Hund

- Rinke, K; Eisentraeger, A. (2007). T ox icological characterization of 2,4,6

- trinitrotoluene, its transformation products, and two nitramine explosives. Environ Toxicol Chem 26: 1090

- 1099. http://dx.doi.org/10.1897/06

- 4 71R.1 NIOSH (National Institute for Occupational Safety and Health). (2012). NIOSH pocket guide to

c hemical hazards: Cyclonite, CAS 121

- 82

- 4. Cincinnati, OH: U.S. Department of Health and Human Services, Centers for Disease Control and Prevention, National Institute for Occupational Safety and Health. http://www.cdc.gov/niosh/npg/npgd0169.html OSHA (Occupational Safety & Health Administration). (2012a). Air contaminants. Table Z

S hipyards. CFR 29 CFR 1915, Subpt. P, App A. OSHA (Occupational Safety & Health Administration). (2012b). Gases, vapors, fumes, dusts, and

m ists. Appendix A to 1926.55

- 1970 American Conference of Governmental Industrial Hygienists' threshold limit values of airb orne contaminants. CFR 29 CFR 1926.55. Özhan, G; Topuz, S; Alpertunga, B. (2003). Determination of cyclonite (RDX) in human plasma by h igh

- performance liquid chromatography. Farmaco 58: 445

- 448. http://dx.doi.org/10.1016/S0014

- 827X(03)00069 -7 Pan, X; Ocho a, KM; San Francisco, MJ; Cox, SB; Dixon, K; Anderson, TA; Cobb, GP. (2013). A bsorption, distribution, and biotransformation of hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine (RDX) in B6C3F1 mice (Mus musculus). Environ Toxicol Chem 32: 1295

- 1303. http://dx.doi.org/10.1002/etc.2188 Pan, X; San Francisco, MJ; Lee, C; Ochoa, KM; Xu, X; Liu, J; Zhang, B; Cox, SB; Cobb, GP

. (2007a). E xamination of the mutagenicity of RDX and its N

- nitroso metabolites using the Salmonella reverse mutation assay. Mutat Res Genet Toxicol Environ Mutagen 629: 64

- 69. http://dx.d oi.org/10.1016/j.mrgentox.2007.01.006 Pan, X; Zhang, B; Smith, JN; San Francisco, M; Anderson, TA; Cobb, GP. (2007b). N

- Nitroso

c ompounds produced in deer mous e (Peromyscus maniculatus) GI tracts following

This document is a draft for review purposes only and does not constitute Agency policy. 

### R- 4 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

h exahydro- 1,3,5

- trinitro

- 1,3,5

- triazine (RDX) exposure. Chemosphere 67: 1164

- 1170. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 h ttp://dx.doi.org/10.1016/j.chemosphere.2006.10.077 Paquet, L; Monteil

- Rivera, F; Hatzinger, PB; Fuller, ME; Hawari, J. (2011). Analysis of the key i ntermediates of RDX (hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine) in gr oundwater: Occurrence, stability and preservation. J Environ Monit 13: 2304

- 2311. http://dx.doi.org/10.1039/c1em10329f Parker, GA; Reddy, G; Major, MA. (2006). Reevaluation of a twenty

- four

- month chronic t oxicity/carcinogenicity study of hexahydro

- 1,3,5 -trinitro- 1,3,5

- triazine (RDX) in the B6C3F1 hybrid mouse. Int J Toxicol 25: 373

- 378. http://dx.doi.org/10.1080/10915810600846245 Poulin, P; Krishnan, K. (1995). An algorithm for predicti ng tissue : blood partition coefficients of

o rganic chemicals from n

- octanol : water partition coefficient data. J Toxicol Environ Health 46: 117

- 129. http://dx.doi.org/10.1080/15287399509532021 Poulin, P; Theil, F,

- P. (2000). A priori prediction of tissue : plasma partition coefficients of drugs to fac ilitate the use of physiologically

- based pharmacok inetic models in drug discovery. J Pharm Sci 89: 16

- 35. http://dx.doi.org/10.1002/(SICI)1520

- 6017(200001)89:1<16::AID

J PS3>3.0.CO;2 -E Reddy, G; Allen, NA; Major, MA. (2008). Absorption of 14C

- cyclotrimethylenetrinitramine (RDX) f rom soils through excised human skin. Toxicol Mech Meth 18: 575

- 579. http://dx.doi.org/10.1080/15376510701703466 Reddy, G; Eisenhut, K; Morrison, SA; Kelly, JA. (1989). Toxicokine tics of 14C RDX ( cyclotrimethylenetrinitramine) in rats after intratracheal administration [Abstract]. Reddy, G; Erexson, GL; Cifone, MA; Major, MA; Leach, GJ. (2005). Genotoxicity assessment of h exahydro- 1,3,5

- trinitro

- 1,3,5

- triazine (RDX). Int J Toxicol 24: 427

- 434. http://dx.doi.org/10.1080/10915810500366922 Reifenrath, WG; Kammen, HO; Reddy, G; Major, MA; Leach, GJ. (2008). Interaction of hydration,

a ging, and carbon content of soil on the evaporation and skin bioavailability of munition contaminants. J Toxicol Environ Health A 71: 486

- 494. http://dx.doi.org/10.1080/15287390801906956 Rispin, A; Farrar, D; Margosches, E; Gupta, K; Stitzel, K; Carr, G; Greene, M; Meyer, W; McCall, D. ( 2002). Alternative methods for the median lethal dose (LD(50)) test: The up

- and

- down procedure for acute oral toxicity. ILAR J 43: 233

- 243. http://dx.doi.org/10.1093/ilar.43.4.233 Safe Work Australia. (2014). Hazardous substances information system (HSIS). Exposure standar d d ocumentation: Cyclonite [Database]. Retrieved from http://www.hsis.safeworkaustralia.gov.au/ExposureStandards/Document?exposureStanda

r dID=174 Schneider, NR; Bradley, SL; Andersen, ME. (1977). Toxicology of cyclotrimethylenetrinitramine: Di stribution and metabolism in the rat and the miniature swine. Toxi col Appl Pharmacol 39: 531

- 541. http://dx.doi.org/10.1016/0041

- 008X(77)90144 -2 Schneider, NR; Bradley, SL; Andersen, ME. (1978). The distribution and metabolism of cy clotrimethylenetrinitramine (RDX) in the rat after subchronic administration. Toxicol Appl Pharmacol 46: 163

- 171. http://dx .doi.org/10.1016/0041

- 008X(78)90147 -3 Simmon, VF; Eckford, SL; Griffin, AF; Spanggord, R; Newell, GW. (1977a). Munitions wastewater t reatments: Does chlorination or ozonation of individual components produce microbial mutagens? [Abstract]. Toxicol Appl Pharmacol 41: 197. http://dx.doi.org/10.1 016/0041

- 0 08X(77)90062 -X Simmon, VF; Spanggord, RJ; Eckford, SL; McClurg, V. (1977b). Mutagenicity of some munition

w astewater chemicals and chlorine test kit reagents (pp. 1

- 205). (ADA057680. DAMD17

-

This document is a draft for review purposes only and does not constitute Agency policy. 

### R- 5 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

7 6 -C- 6013). Menlo Park, CA: SRI International. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 h ttp://oai.dtic.mil/oai/oai?verb=getRecord&metadataPrefix=html&ident ifier=ADA057680 Smith, JN; Espino, MA; Liu, J; Romero, NA; Cox, SB; Cobb, GP. (2009). Multigenerational effects in d eer mice (Peromyscus maniculatus) exposed t o hexahydro

- 1,3,5

- trinitroso

- 1,3,5

- triazine (TNX). Chemosphere 75: 910

- 914. http://dx.doi.org/10.1016/j.chemosphere.2009.01.010 Snodgrass, HL, Jr. (1984). Preliminary assessment of relative toxicity and mutagenicity potential of 1

- nitroso

- 3,5

- dinitro

- 1,3,5

- triazacylcohexane (mononitroso

- RDX) (Final Report ed.). (ADA149351. USAEHA

- 75

- 51

- 0345

- 85). Aberdeen Proving Ground, MD: U.S. Army Environmental Hygiene Agency. http://oai.dtic.mil/oai/oai?verb=getRecord&metadataPrefix=html&identifier=ADA149351 Snoeck, V; Huyghebaert, N; Cox, E; Vermeire, A; Saunders, J; Remon, JP; Verschooten, F; Goddeeris, B M. (2004). Gastrointestinal transit time of nondisintegratin g radio

- opaque pellets in

s uckling and recently weaned piglets. J Control Release 94: 143

- 153. http://dx.doi.org/10.1016/j.jconrel.2003.09.015 Spiteri, NJ. (1982). Circadian patterning of feeding, drinking and activity during diurnal food access i n rats. Physiol Behav 28: 139

- 147. http:/ /dx.doi.org/10.1016/0031

- 9384(82)90115 -9 Stone, WJ; Paletta, TL; Heiman, EM; Bruce, JI; Knepshield, JH. (1969). Toxic effects following i ngestion of C

- 4 plasti c explosive. Arch Intern Med 124: 726

- 730. http://dx.doi.org/10.1001/archinte.1969.00300220078015 Sweeney, LM; Gut, CP, Jr; Gargas, ML; Reddy, G; Williams, LR; Johnson, MS. (2012a). Assessing the

no n
- cancer risk for RDX (hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine) using physiologically based pharmacokinetic (PBPK) modeling [Review]. Regul Toxic ol Pharmacol 62: 107

- 114. http://dx.doi.org/10.1016/j.yrtph.2011.12.007 Sweeney, LM; Okolica, MR; G ut, CP, Jr; Gargas, ML. (2012b). Cancer mode of action, weight of

ev idence, and proposed cancer reference value for hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine (RDX). Regul Toxicol Pharmacol 64: 205

- 224. http://dx.doi.org/10.1016/j.yrtph.2012.07.005 Tan, EL; Ho, CH; Griest, WH; Tyndall, RL. (1992). Mutagenicity of trinitrotoluene and its metabolites f o rmed during composting. J Toxicol Environ Health A 36: 165

- 175. http://dx.doi.org/10.1080/15287399209531632 Testud, F; Glanclaude, J,

- M; Descotes, J. (1996a). Acute hexogen poisoning after occupational

ex posure. J Toxicol Clin Toxicol 34: 109

- 111. http://dx.doi.org/10.3109/1556365960902024 4 Testud, F; Glanclaude, J; Imperatori, J; Le Meur, B; Descotes, J. (1996b). [Acute hexogen poisoning aft er occupational exposure, report of 2 cases]. Arch Mal Prof Med Trav Secur Soc 57: 342

- 346. Timchalk, C; Nolan, RJ; Mendrala, AL; Dittenber, DA; Brzak, KA; Mattsson, JL. (2002). A p hysiologically based pharmacoki netic and pharmacodynamic (PBPK/PD) model for the organophosphate insecticide chlorpyrifos in rats and humans. Toxicol Sci 66: 34

- 53. http://dx.doi.org/10.1093/toxsci/66.1.34 U.S. EPA (U.S. Environmental Protection Agency). (1992). A cross

- species scaling factor for

c arcinogen risk assessment based on equivalence of mg/kg3/4/day [EPA Report] (pp. 24152

- 241 61). Washington, DC. U.S. EPA (U.S. Environmental Protection Agency). (2005a). Guidelines for carcinogen risk

as sessment [EPA Report]. (EPA/630/P

- 03/001F). Was hington, DC: U.S. Environmental Protection Agency, Risk Assessment Forum. http://www2.epa.gov/osa/guidelines

c arcinogen

- risk

- assessment U.S. EPA (U.S. Environmental Protection Agency). (2005b). Supplemental guidance for assessing susceptibility from early

- life exposure to carcinogens. (EPA/630/R

- 03/003F). Washington,

This document is a draft for review purposes only and does not constitute Agency policy. 

### R- 6 DRAFT―DO NOT CITE OR QUOTE

### Supplemental Information― Hexahydro

- 1,3,5

- trinitro

- 1,3,5

- triazine

D C: U.S. Environmental Protection Agency, Risk Assessment Forum. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 h ttp://www.epa.gov/raf/publications/pdfs/childrens_supplement_final.pdf U.S. EPA (U.S. Environmental Protection Agency). (2012a). Advances in inhalation gas dosimetry for derivation of a reference concentration (RFC) a nd use in risk assessment. (EPA/600/R

12/044). Washington, DC. http://cfpub.epa.gov/ncea/cfm/recordisplay.cfm?deid=244650 U.S. EPA (U.S. Environmental Protection Agency). (2012b). Benchmark dose technical guidance. ( EPA/100/R

- 12/001). Washington, DC: U.S. Environmental Protection Agency, Risk Assessment Forum. http://www.epa.gov/raf/publications/benchmarkdose.htm U.S. EPA (U.S. Environmental Protection Agency). (2012c). EPA Federal facilities forum issue paper: Site characterization for munitions constituents [EPA Repor t] (pp. 1

- 170). (EPA

- 505 -S- 11

- 001). Washington, D.C. http://www2.epa.gov/fedfac/epa

- federal

- facilities

- forum

- issue

pa per
- site -c haracterization

- munitions

- constituents U.S. EPA (U.S. Environmental Protection Agency). (2014). RDX PBPK files in acsIX format [PBPK]. von Oettingen, WF; Donahue, DD; Yagoda, H; Monaco, AR; Harris, MR. (1949). Toxicity and potential d angers of cyclotrimethylenetrinitramine (RDX). J Ind Hyg Toxicol 31: 21

- 31. West, RR; Stafford, DA. (1997). Occupational exposures and haematological abnormalities among

o rdnance factory workers: A case control study. Leuk Res 21: 675

- 680. http://dx.doi.org/10.1016/S0145

- 2126(97)00042 -8 Whong, W,

- Z; Speciner, ND; Edwards, GS. (1980) . Mutagenic activity of tetryl, a nitroaromatic

ex plosive, in three microbial test systems. Toxicol Lett 5: 11

- 17. http://dx.doi.org/10.1016/0378

- 4274(80)90142 -3 Woody, RC; Kearns, GL; Brewster, MA; Turley, CP; Sharp, GB; Lake, RS. (1986). The neurotoxicity of cy clotrimethylenetrinitramine (RDX) in a child: A clinical and pharmac okinetic evaluation. Clin Toxicol 24: 305

- 319. http://dx.doi.org/10.3109/15563658608992595

This document is a draft for review purposes only and does not constitute Agency policy. 

### R- 7 DRAFT―DO NOT CITE OR QUOTE