EPA/635/R

- 13/107 Preliminary Materials www.epa.gov/iris



## Preliminary Materials **for the Integrated Risk Information System (IRIS)**  **Toxicological R**  **eview of**  tert

**- Butyl Alcohol (**  tert

**- Butanol)** 

[CASRN 75

- 65

- 0]

Ju ly 2013

**NOTICE**

This document is comprised of preliminary materials, consisting of a literature search strategy, evidence tables, and exposure

- response arrays. This in formation is distributed solely for the purpose of pre

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

National Center for Environmental Assessment Office of Research and Development U.S. Environmental Pro tection Agency Washington, DCPreliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

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

— DO NOT CITE OR QUOTE **DISCLAIMER**

This document is comprised of preliminary materials for review purposes only. This information is distributed solely for the purpose of pre

- dissemination 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 or recommendation for use.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

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

— DO NOT CITE OR QUOTE 

# CONTENTS

PREFACE

................................................................................................ ........................................................ v

1. DRAFT LITERATURE SEARCH STRATEGY

.............................................................................................. 1-1

1.1. Literature Search and Screening Strategy for tert

- Butanol

.......................................................... 1-1

1.2. List of References Based on Sea rch Strategy for tert

- Butanol

...................................................... 1-5

1.2.1. Primary Sources of Health Effects Data

............................................................................. 1-5

1.2.2. Not Primary Source of Health Effects Data, but Kept as Additional Resources

.............. 1- 13

1.2.3. Kept for Possible Further Review

.................................................................................... 1- 17

2. PRELIMINARY EVIDENCE TABLES AND EXPOSURE

- RESPONSE ARRAYS

............................................... 2-1

2.1. Data Extraction: Preparation of Preliminary Evidence Tables and Exposure -Response Arrays

................................................................................................ ............................................ 2-1

2.2. Kidney Effects

................................................................................................ ................................ 2-3

2.3. Thyroid Effects

............................................................................................................................ 2- 13

2.4. Reproductive, Developmental, and Neurodevelopmental Effects

............................................. 2- 16

2.5. Central Nervous System (CNS) Effects

........................................................................................ 2- 25

2.6. Other Systemic Effects (Body Weight, Liver, and Urinary Bladder)

............................................ 2- 28

2.7. Genotoxic Effects

........................................................................................................................ 2- 37Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

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

— DO NOT CITE OR QUOTE 

# TABLES

Table 1- 1. Database search strategy for tert

- butanol

............................................................................... 1-3 Table 1

- 2. Summary of additional search strategies for tert

- butanol

...................................................... 1-4 Table 2

- 1. Evidence pertaining to kidney effects in animals following oral exposure to tert

butanol

............................................................................................................................. 2-3 Table 2

- 2. Evidence pertaining to kidney effects in animals following inhalation exposure to tert

butanol

............................................................................................................................. 2-9 Table 2

- 3. Evidence pertaining to thyroid effects in animals following oral exposure to tert

butanol

........................................................................................................................... 2- 13 Table 2

- 4. Evidence pertaining to reproductive effects in animals following exposure to tert

butanol

........................................................................................................................... 2- 16 Table 2

- 5. Evidence pertaining to developmental effects in animals following exposure to tert

butanol

........................................................................................................................... 2- 18 Table 2

- 6. Evidence pertaining to neurodevelopmental effects in animals following exposure to
tert

- butanol

.................................................................................................................... 2- 21 Table 2

- 7. Evidence pertaining to CNS effects in animals following exposure to tert

- butanol

.............. 2- 25 Tabl e 2

- 8. Evidence pertaining to effects on body weight in animals following oral exposure to
tert

- butanol

.................................................................................................................... 2- 28 Table 2

- 9. Evidence p ertaining to liver effects in animals following oral exposure to tert

- butanol

...... 2- 30 Table 2

- 10. Evidence pertaining to liver effects in animals following inhalation exposure to tert

butanol

........................................................................................................................... 2- 33 Table 2

- 11. Evidence pertaining to urinary bladder effects in animals following oral exposure to
tert

- butanol

.................................................................................................................... 2- 34 Table 2

- 12. Evidence pertaining to genotoxic effects of tert

- butanol in vitro and in vivo studies

......... 2- 37



# FIGURES

Figure 1

- 1. Literature search approach for tert

- butanol.

......................................................................... 1-2 Figure 2

- 1. Exposure response array of kidney effects in rats and mice following oral exposure
to tert

- butanol.

.............................................................................................................. 2- 11 Figure 2

- 2. Exposure

- response array of kidney effects in rats and mice following subchronic inhalation exposure to tert

- butanol.

............................................................................. 2- 12 Figure 2

- 3. Exposure

- response array of thyroid follicular cell effects in rats and mice following chronic oral exposure to tert

- butanol.

.......................................................................... 2- 15 Figure 2

- 4. Exposure

- response array of reproductive, developmental and neurodevelopmental effects following oral exposure to tert

- butanol.

........................................................... 2- 23 Fi g ure 2

- 5. Exposure

- response array of reproductive and developmental effects following inhalation exposure to tert

- butanol.

............................................................................. 2- 24 Figure 2

- 6. Exposure

- response array of body weight, liver effects, and urinary bladder effects following oral exposure to tert

- butanol.

....................................................................... 2- 36Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

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

v DRAFT

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# PREFACE 1

2

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33 This document presents the draft literature search strategy, preliminary evidence tables, and preliminary exposure

- res p onse arrays for tert

- butyl alcohol (henceforth referred to a s tert

b utanol ) p repared under the auspices of EPA’s Integrated Risk Information System (IRIS) Program.
Th is material is being released for public viewing and comment prior to a public meeting, providing an opportunity for the IRIS Program to engage in early discussions w it h stakeholders and the public on data that may be used to identify adverse health effects and characterize exposure

- resp o nse relationships. The draft literature search strategy, preliminary evidence tables, and preliminary exposure

response arrays are r esp o nsive to the National Research Council (NRC) 2011 report Review of the Environmental Protection Agency’s Draft IRIS Assessment of Formaldehyde

. The li t erature search strategy, which describes the processes for identifying scientific literature, screenin g studies for consideration, and selecting studies for inclusion in evidence tables, is responsive to NRC recommendations regarding systematic review of the scientific literature. In a d dition, NRC recommendations for standardized presentation of key stud y data are addressed in the preliminary evidence tables and preliminary exposure

- resp o nse arrays. EPA welcomes all comments on the draft literature search strate g y , preliminary evidence tables, and preliminary exposure

- resp o nse arrays, such as remark s on the following:

• the clarity and transparency of the materials;

• the approach for identifying pertinent studies;

• the selection of studies for data extraction to preliminary evidence tables and exposure

res p onse arrays;

• any methodological considerations th at could affect the interpretation of or confidence in study results; and

• any additional studies published or nearing publication that may provide data for the
evaluat ion of human health hazard or exposure
- res p onse relationships.
The preliminary evidence tables and exposure

- re s ponse arrays should be regarded solely as representing the data on each endpoint that have been identified as a result of the draft literature search strategy. They do not reflect any conclusions as to hazard identification or dos e- resp o nse assessment. After obtaining public input and conducting additional study evaluation and data integration, EPA will revise these materials to support the hazard identification and dose

- resp o nse assessment in a draft Toxicological Review.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

Thi s document is a draft for review purposes only and does not constitute Agency policy. 1-1 DRAFT

— DO NOT CITE OR QUOTE 

# 1. DRAFT LITERATURE SEARCH STRATEGY

**1.1. L** 

Th



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## a

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li 

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## ch

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## a

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## n

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## d

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## S

p 

## c

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## r

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## e

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## en
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## i

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## n

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## g

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## St

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## r

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## a

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## t

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## eg

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## y cally



## fo

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## r

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## -

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in itial
chemical

- specific search was conducted in four online scientific databases in December, 2012
, and January, 2013
, using the keywords and limits described in Table 1 -1 . After electr onically eliminating duplicates from the citations retri eved through these databases, 2,907 unique citations

were identified

. An additional seven citations were obtained using additional search strategies
described in Table 1- 2. The resulting 2,4 10 cita tions were scr
t een
ert ed using the title, abstract, and/or full text for pertinence to examining the health effects of -b utanol exposure. A total of 2,226 references

were identified as not being pertinent and were excluded from further consideration (see F igure 1

- 1 for the exclusion categories)

. A total of 106 references were identified as primary sources of health effects data and were considered for data extraction to evidence tables and exposure

response arrays (see Section 1.2.1)

. A total of 60 r efere nces were considered pertinent, but not as primary sources of health effects data (e.g., ADME studies)
, and kept as additional resources for development of the T oxicological R eview (see Section 1.2.2 ). A total of 18 references did not provide enough material to evaluate pertinence ( e.g., foreign language )
, and were reserved for further possible review ( see Section 1.2.3 ). 

### 1

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9
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17Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

Thi s document is a draft for review purposes only and does not constitute Age ncy policy. 1-2 DRAFT

— DO NOT CITE OR QUOTE 

### 1

2 Database Searches (see Table 1 for keywords and limits)

(After duplicates removed electronically by HERO) n = 2,907 Pubmed n = 1,512 Web of Science n = 462 Toxline (incl. TSCATS) n = 931 TSCATS 2 n = 2

Combined Dataset (After all duplicates removed) n = 2,410 Additional Search Strategies (see Table 2

- 1 for methods and results)

n = 7

Excluded (not pertinent) 80 Biodegradation/environmental fate 75 Chemical analysis/fuel chemistry 1,143 Other chemical/ non

- tert

- butanol 86 Method of detection/exposure and biological monitoring 624 Methodology/solvent 62 Not relevant exposure paradigms (e.g., dermal, eye irritation, etc.) 39 Not relevant species/matrix (e.g., amphibians, fish, etc.) 40 Abstract only/comment 25 Book chapter/section 15 General review papers 22 Society abstracts 14 QSAR 1 Unpublished data subsequently published Kept for Possible Further Review
18 Foreign language

Primary Sources of Health Effects Data
4 Human health effects studies (including one case report) 37 Animal toxicology studies 22 Genotoxicity studies 43 Other studies involving direct administration of chemical, including mechanistic studies Manual Screening For Pertinence (Title/Abstract/Full Text)

Not Primary Source of Health Effects Data, but Kept As Additional Resource 17 Reviews, editorials 8 Health effects assessments by others 29 Toxicokinetic studies (not including PBTK) 6 PBTK modeling studies

**Figure 1-1** 

**. Literature**  **s earch**  **a pproach for**  tert **-b utanol** 

.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

Thi s document is a draft for review purposes only and does not constitute Agency policy. 1-3 DRAFT

— DO NOT CITE OR QUOTE **Table 1-1.**  **Database s**  **earch s**  **trategy for** tert **-b uta**  **nol** 

### 1

2
3 Database (Search Date) Keywords Limits PubMed (12/20/2012) t

- b utanol OR 75

- 65

- 0[rn] OR “t

b utyl hydroxide” OR “2 -m ethyl -2- propanol” OR “ t rimethyl carbinol” OR “t -b utyl alcohol” OR tert

b utanol OR "tert

- butyl alcohol" OR tert

- butyl alcohol[mesh] None

Web of Science (12/20/2012) Topic = (t

- b utanol OR 75

- 65

- 0 OR "t -b utyl hydroxide" OR "2 -m ethyl

2- propanol" OR " t rimethyl carbinol" OR "t -b utyl alcohol" OR "tert -b utanol" OR "tert

- butyl alcohol") Refined by: Research Areas = ( cell biology OR respiratory system OR microscopy OR biochemistry molecular biology OR gastroenterology hepatology OR public environmental occupational health OR oncology OR physiology OR cardiovascular system cardiology or toxicology OR life sciences biomedicine other topics OR hematology OR pathology OR neurosciences neurology OR developmental biology) Toxline (includes TSCATS) (1/11/2013) t

- butanol OR 75

- 65

- 0 [rn] OR t

butyl hydroxide OR 2

- methyl -2- propanol OR trimethyl carbinol OR
t- butyl alcohol OR tert

- butanol OR
tert

- butyl alcohol OR tert

- butyl alcohol Not PubMed

TSCATS2 (1/4/2013) 75

- 65

- 0 NonePreliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

Thi s document is a draft for review purposes only and does not constitute Agency policy. 1-4 DRAFT

— DO NOT CITE OR QUOTE **Table 1-2** 

**. Summary of**  **a dditional**  **s earch**  **s trategies for**  tert **-b utanol**  

### 1

Approach u sed Source(s) Date performed Number of a dditional citations i dentified Manual search of citations from reviews Review article : McGregor, D. ( 2010 )

. Tertiary -b utanol : A toxicological review. Crit Rev Toxicol 40(8): 697

- 727. 1/2013 5 citations

Review article: Chen
, M

. ( 2005 )

. Amended final re port of the safety assessment of t -b utyl a lcohol as used in cosmetics.” Int J Toxicol 24(2): 1

- 20. 1/2013 2 citations

Manual search of citations from r eviews conducted by other international and federal agencies IPCS (International Programme on Chemical Safety) ( 1987a )

. Butanols: Four isomers: 1

- butanol, 2- but anol, tert

- butanol, isobutanol [WHO EHC]. Geneva, Switzerland: World Health Organization

. 1/2013 None

OSHA (Occupational Safety & Health Administration). ( 1992 ). Occupational safety and health guideline for tert

- butyl alcohol. Cincinnati, OH: National Institute for Occupational Safety and Health. 1/2013 None



### 2
3Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

Thi s document is a draft for review purposes only and does not constitute Agency policy. 1-5 DRAFT

— DO NOT CITE OR QUOTE 

## 1.2. **List of References Based on Search Strategy for**  tert

**- Butanol**  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 Citations for excluded references are not listed here, but can be found on the Health and Environmental Research Online (HERO) Web site ( http://hero.epa.gov/ tert

- Butanol ).

**1.2.1. Pri**  **mary Source s**  **of Health Effects Data**
Data from citations in **bold** ar e d isplayed in Section 2. See Section 2. 1 for a description of the process of selecting these studies for evidence tables and exposure

- r es ponse array s.

Human health effects studies

1) Ed wards, EK, Jr; Edwards, EK. ( 1982 ) . Allergic reaction to tertiary butyl alcohol in a sunscreen. Cutis 29: 476

- 4 7 8.

2) Johanson, G; Nihlen, A; Lof, A. ( 1995 ) . Toxicokinetics and acute effects of MTBE and ETBE in male volunteers. Toxicol Lett 82/83: 713

- 71 8.

3) L aire, G; Viaene, MK; Veulemans, H; Masschelein, R; Nemery, B. ( 1997 ) . Noct urnal oxygen desaturation, as assessed by home oximetry, in long

- te rm solvent

- exposed workers. Am J Ind Med 32: 656

- 6 6 4.
4) Prah, JD; Goldstein, GM; Devlin, R; Otto, D; Ashley, D; House, D; Cohen, KL; Gerrity, T. ( 1994 )

. Sensory, symptomatic, inflammatory, and ocular responses to and the metabolism of methyl tertiary butyl ether in a controlled human exposure experiment. Inhal Toxicol 6: 521

- 53 8.

Animal toxicology studies

**1) A**  **charya, S; Mehta, K; Rodrigues, S; Pereira, J; Krishnan, S; Rao, CV.**  **( 1995**  **)**

**. Administration of subt**  **o x**  **ic doses of t**

**- butyl alcohol and trichloroacetic acid to male**  **Wistar rats to study the interactive toxicity. Toxicol Lett 80: 97**

**- 104.** 

**2) A**  **charya, S; Mehta, K; Rodriguez, S; Pereira, J; Krishnan, S; Rao, CV. (**  **1997 )**  **. A histopathological study of liver and kidney in male Wistar rats treated with subtoxic**  **doses of t**

**- bu**  **tyl alcohol and trichloroacetic acid. Exp Toxicol Pathol 4 9: 369** 

**- 373.** 

3) ARCO (ARCO Chemical Company). ( 1992 ) . Initial submission: letter submitting preliminary results from subchronic toxicity studies of te rt iary butyl alcohol in rats and mice dated 10 / 1 4 / 9 2 and attachments. (88930000018). Newton Square, PA: Arco Chemical Company.

4) Atrens, D; van der Reest, A; Balleine, B; Menéndez, J; Siviy, S. ( 1989 ) . Effects of ethanol and tertiary butanol on blood glucose levels and body temperature of rats. Alcohol 6: 183

- 1 8 7.
5) Belknap, JK; Deutsch, CK. ( 1982 ) . Differential neurosensitivity to three alcohols and phenobarbital in C57BL/6J and DBA/2J mice. Behav Genet 12: 309

- 3 1 7.
6) Bellin, SI; Edmonds, HL, Jr. ( 1976 ) . The use of tert

- butanol in alcohol dependence studies. Proc West Pharmacol Soc 19: 351

- 3 5 4.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

Thi s document is a draft for review purposes only and does not constitute Agency policy. 1-6 DRAFT

— DO NOT CITE OR QUOTE **7) Billitti, JE; Faul**  **kner, BC; Wilson, BW. (**  **2005 )**  **. Absence of acute testicular toxicity of** 

### 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 **methyl**

**- tert butyl ether and breakdown products in mice. Bull Environ Contam**  **Toxicol 75: 228**

**- 235.** 

8) C irvello, JD; Radovsky, A; Heath, JE; Farnell, DR; III, LC. ( 1995 ) . Toxicity and carcinogenicity of t

- b utyl alcohol in rats and mice following chronic exposure in drinking water. Toxicol Ind H ealth 11: 151

- 16 5.

**9) D**  **aniel, MA; Evans, MA. (**  **1982 )**  **. Quantitative comparison of maternal ethanol and maternal tertiary butanol diet on postnatal development. J Pharmacol Exp Ther 222:**  **294**

**- 300.** 

10) Dow Chemical Co (Dow Chemical Company). ( 1992 ) . Letter submitting multiple studies on multiple chemicals required for docket opts

- 8 2 036 with attachments (sanitized).

11) Dow Chemical Co (Dow Chemical Company). ( 1994 ) . Animal toxicity experiments with ethyl alcohol, tertiary butyl alcohol, and stoddard's solvent with cover letter dated 03/28/94. (sanitized). (8694000270S). Midland, MI.

12) Eas tman Kodak Company. ( 1963 ) . Toxicity report (summary) of t

- butyl alcohol with cover letter dated 02 / 15 / 94. Rochester, New York: Eastman Kodak Company.

**13) Faulkner, TP; Wiechart, JD; Hartma**  **n , DM; Hussain, AS.**  **( 1989**  **) . The effects of prenatal**  **tertiary butanol administration in CBA/J and C57BL/6J mice. Life Sci 45: 1989**

**- 1995** 

**.** 14) Feller, DJ; Crabbe, JC. ( 1991 ) . Effect of alcohols and other hypnotics in mice selected for differential sensitivity to hypothermic actions of ethanol. J Pharmacol Exp Ther 256: 947

- 953.

**15) G**  **rant, KA; Samson, HH. (**  **1981 )**  **. Development of physical dependence on t**

**- butanol in**  **rats: An examination using schedule**

**- i**  **nduced drinking. Pharmacol Biochem Behav 14: 633** 

**- 637.** 

**16) G**  **rant, KA; Samson, HH. (**  **1982 )**  **. Ethanol and tertiary butanol induced microcephaly in the neonatal rat: Comparison of brain growth parameters. Neurobehav Toxicol**  **Teratol 4: 315**

**- 321.** 

17) G S RI (Gulf South Research Institute). ( 1979a ) . Repeated dose test of t

- butanol (c55367) in B6C3F 1 mice and F ischer 344 rats with cover letter dated 031594. (86940000173). Orleans Parrish, LA.

18) G S RI (Gulf South Research Institute). ( 1979b ) . Subchronic test of t

- butanol (c55367) in B6C3F 1 mice and F ischer 344 rats in drinking water with cover letter dated 031594. (86940000172). Orleans Parish, LA.

**19) H**  **ard, GC; Bruner, RH; Cohen, SM; Pletcher, JM; Regan, KS. (**  **2011 )**  **. Renal histopathology in toxicity and carcinogenicity studies with tert** 

**- bu**  **tyl alcohol administered in drinking water to F344 rats: A pathol**  **ogy w orking group review and** 

**re -**  **e valuation. Regul Toxicol Pharmacol 59: 430** 

**- 436.** Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

Thi s document is a draft for review purposes only and does not constitute Agency policy. 1-7 DRAFT

— DO NOT CITE OR QUOTE 20) Lindamood, C; Farnell, D; Giles, H; Prejean, J; Collins, J; Takahashi, K; Maronpot, R. ( 1992 )

. 

### 1 2 3

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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 Subchronic toxic ity studies of t

- butyl alcohol in rats and mice. Fundam Appl Toxicol 19: 91

- 100.

21) L yondell Chemical Co. (Lyondell Chemical Company). ( 2003 ) . An oral gavage reproductive and developmental toxicity screening in rats. (Document Control Number: 88030000222).

**22) L**  **yondell Chemical Co. (Lyondell Chemical Company). (**  **2004 )**  **. Reproductive and developmental toxicity screening test in rats by oral gavage. (Document Control**  **Number: 89**

**- 040000106).** 

**23) M**  **ccomb, J; Goldstein, D. (**  **1979a )**  **. Additive physical dependence: evidence for a common mechanism in alcohol dependence. J Pharmacol Exp Ther 210: 87** 

**- 90** 

**. 24)**  **M ccomb, J; Goldstein, D.**  **( 1979b**  **) . Quantitative comparison of physical dependence on**  **tertiary butanol and ethanol in mice: Correlation with lipid solubility. J Pharmacol Exp Ther 208: 113** 

**- 117.** 

25) Nelson, BK. ( 1986 ) . Developmental neurotoxicology of in utero exposure to industrial solvents in experimental animals. Neurotoxicology 7: 441

- 4 4 7.
**26) Nelson, BK; Brightwell, WS; Khan, A; Burg, JR; Goad, PT.**  **( 1989**  **) . Lack of selective**  **developmental toxicity of three butanol isomers administered by inhalation to rats. Fundam Appl Toxicol 12: 469** 

**- 4**  **79.**

**27) Nelson, BK; Brightwell, WS; Khan, A; Shaw, PB; Krieg, EF, Jr; Massari,**  **VJ. (**  **1991 )** 

**. Behavioral teratology investigation of tertiary** 

**- butanol administered by inhalation to**  **rats. Pharmacopsychoecologia 4: 1**

**- 7.** 

28) N elson, BK; Brightwell, WS; Krieg, EF, Jr. ( 1990 ) . Developmental to xicology of industrial alcohols: A summary of 13 alcohols administered by inhalation to rats. Toxicol Ind Health 6: 373

- 387.

**29) NTP (National Toxicology Program).**  **( 1995**  **) . Toxicology and carcinogenesis studies of**  **t- bu**  **tyl alcohol (CAS no 75**

**- 65** 

**- 0) in F344/N rats and B6C3F1 mice (**  **d rinking water**  **studies) (pp. 1**

**- 305)**  **. Research Triangle Park, NC.**

**30) N**  **TP (National Toxicology Program). (**  **1997 )**  **. NTP technical report on toxicity studies of t** 

**- bu**  **tyl alcohol (CAS no 75**

**- 65** 

**- 0) administered by inhalation to F344/N rats and**  **B6C3F1 mice. Research Triangle Park, NC.**

31) Pa lmer, AA; Mckinnon, CS; Bergstrom, HC; Phillips, TJ. ( 2002 ) . Locomotor activity responses to ethanol, other alcohols, and GABA

- A a cting compounds in forward - a nd reverse -selected FAST and SLOW mouse lines. Behav Neurosci 116: 958

- 9 6 7.
32) Siviy, SM; Atrens, DM; Jirasek, M; Holmes, LJ. ( 1987 ) . Effects of ethanol and tertiary

- butanol on energy expenditure and subs t rate utilization in the rat. Alcohol 4: 437

- 442.

**33) Snell, D.**  **( 1980**  **) . Impairment of avoidance behavior following short** 

**- term ingestion of**  **ethanol, tertiary**

**- bu**  **tanol, or pentobarbital in mice. Psychopharmacology 69: 53**

**- 57.** Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

Thi s document is a draft for review purposes only and does not constitute Agency policy. 1-8 DRAFT

— DO NOT CITE OR QUOTE **34) Takahashi, K; Lindamood, C; Maronpot, R.**  **( 1993**  **) . Retrospective study of possible**  

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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 **alpha**

**- 2 mu** 

**- globulin nephropathy and associated cell proliferation in male Fischer**  **344 rats dosed with t**

**- butyl alcohol. Environ Health Perspect 101: 281** 

**- 285.** 

**35) T**  **hurman, RG; Winn, K; Urquhart, B. (**  **1980 )**  **. Rat brain cy clic AMP levels and**  **withdrawal behavior following treatment with t**

**- bu**  **tanol. Adv Exp Med Biol 126: 271**

**- 281.** 

**36) W**  **illiams, TM; Borghoff, SJ. (**  **2001 )**  **. Characterization of tert**

**- butyl alcohol binding to**  **"alpha"2u**

**- g**  **lobulin in F**

**- 344 rats. Toxicol Sci 62: 228** 

**- 235.** 

**37) W**  **ood, J; Laverty, R. (**  **1979 )**  **. Physical dependence following prolonged ethanol or t**

  ** butanol administration** **to rats. Pharmacol Biochem Behav 10: 113**

**- 119.** 

Genotoxicity s t udies

1) Abbondandolo, A; Bonatti, S; Corsi, C; Corti, G; Fiorio, R; Leporini, C; Mazzaccaro, A; Nieri, R; B arale, R; Loprieno, N. ( 1980 ) . The use of organic solvents in mutagenicity testing. DNA Repair 79: 141

- 15 0.

**2) A**  **RCO (ARCO Chemical Company). (**  **1994a )**  **. Evaluation of test article t**

**- butyl alcohol**  **99.9% (MRI #635) and arconol (MRI #636) for mutagenic potential employing the l5178Y TK+/**  **- m**  **u tagenesis assay w/cover letter dated 03/24/94. (TSCATS/451796).** 

**3) AR**  **CO (AR C**  **O Chemical Company). (**  **1994b )**  **. In vitro evaluation of t**

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.Preliminary Materials for the IRIS Toxicological Review of **tert**

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1 9) L ee, CW; Mohr, SN; Weisel, CP. ( 2001 ) . Toxicokinetics of human exposure to methyl tertiary

- butyl ether (MTBE) following short

- te r m controlled exposures. J Expo Sci Environ Epidemiol 11: 67

- 7 8

. 20) Miller, MJ; Ferdinandi, ES; Klan, M; Andrews, LS; Douglas, JF; Kneiss, JJ. ( 1997 )

. Pharmacokinetics and disposition of methyl t

- bu t yl ether in Fischer

- 344 rats. J App l Toxicol 17: S3

- S12.

2 1) N ihlen, A; Lof, A; Johanson, G. ( 1995 ) . Liquid/air partition coefficients of methyl and ethyl tbutyl ethers, t- am yl methyl ether, and t- butyl alcohol. J Expo Anal Environ Epidemiol 5: 573

- 582.

22) N ihlen, A; Lof, A; Johanson, G. ( 1998a ) . Controlled ethyl tert

- butyl ether (ETBE) exposure of male volu nteers : I Toxicokinetics. Toxicol Sci 46: 1

- 10.

23) N ihlen, A; Lof, A; Johanson, G. ( 1998b ) . Experimental exposure to methyl tertiary

- butyl ether: I Toxicokinetics in humans. Toxicol Appl Pharmacol 148: 274

- 28 0.

24) N ihlen, A; Lof, G; Johanson, G. ( 1997 ) . Liquid/air partition coefficients of methyl and ethyl tbutyl ethers, t- am yl methyl ether, and t- butyl

- alcohol. J Clean Technol, Environ Toxicol, Occup Med 6: 205

- 2 1 3.

25) Nihlen, A; Sumner, SCJ; Lof, A; Johanson, G. ( 1999 ) . 13C2

- labeled methyl tert

- butyl ether: toxicokinetics and characterization of urinary metabolites in humans. Chem R es Toxicol 12: 822

- 830

.

2 6) P oet, TS; Valentine, JL; Borghoff, SJ. ( 1997 ) . Pharmacokinetics of tertiary butyl alcohol in male and female Fischer 344 rats. Toxicol Lett 92: 179

- 18 6.

27) P rah, J; Ashley, D; Blount, B; Case, M; Leavens, T; Pleil, J; Cardinali, F. ( 2004 ) . Dermal, oral, and inhalation pharmacokinetics of methyl tertiary butyl ether (MTBE) in human volunteers. Toxicol Sci 77: 195

- 20 5.

28) Turini, A; Amato, G; Longo, V; Gervasi, PG. ( 1998 ) . Oxidation of methyl - and ethyl

- tertiary

butyl ethers in rat liver microsomes: role of the cytochrome P450 isoforms. Arch Toxicol 72: 207

- 214.

2 9) Vainiotalo, S; Riihimäki, V; Pekari, K; Teräväinen, E; Aitio, A. ( 2007 ) . Toxicokinetics of methyl tert

- b u tyl ether (MTBE) and tert

- amyl methyl ether (TAME) in humans, and implications to their biological monitoring. J Occup Environ Hyg 4: 739

- 75 0.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

Thi s document is a draft for review purposes only and does not constitute Agency policy. 1- 17 DRAFT

— DO NOT CITE OR QUOTE PBTK m odeling s tudies 

### 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 1) Borghoff, S; Murphy, J; Medinsky, M. ( 1996 ) . Development of physiologically based pharmacokinetic model for methyl tertiary

- b u tyl ether and tertiary

- butanol in male Fisher

344 rats. Fundam Appl Toxicol 30: 264

- 27 5.

2) B orghoff, S; Parkinson, H; Leavens, T. ( 2010 ) . Physiologically bas ed pharmacokinetic rat model for methyl tertiary

- bu tyl ether; comparison of selected dose metrics following various MTBE exposure scenarios used for toxicity and carcinogenicity evaluation. Toxicology 275: 79

- 91

. 3) Ki m, D; Andersen, ME; Pleil, JD; Nylander

- F rench, LA; Prah, JD. ( 2007 ) . Refined PBPK model of aggregate exposure to methyl tertiary

- bu tyl ether. Toxicol Lett 169: 222

- 23 5.

4) L eavens, T; Borghoff, S. ( 2009 ) . Physiologically based pharmacokinetic model of methyl tertiary butyl ether and tertiary butyl alc oh ol dosimetry in male rats based on binding to alpha2u

- g lobulin. Toxicol Sci 109: 321

- 3 35.

5) Nihlen, A; Johanson, G. ( 1999 ) . Physiologically based toxicokinetic modeling of inhaled ethyl tertiary

- b utyl ether in humans. Toxicol Sci 51: 184

- 1 94.

6) Rao, HV; Ginsberg, GL. ( 1997 ) . A physiologically

- based pharmacokinetic model assessment of methyl t

- bu tyl ether in groundwater for a bathing and showering determination. Risk Anal 17: 583

- 5 9 8.

**1.2.3. Kept for Possible Further Review** 

F

oreign language

1) A non. ( 1987 ). Criteria for proposed 1987 short

- term exposure limit s. Sangyo Igaku 29: 416

- 433.

2) A pel, YA. ( 1968 ) . Information on the t oxicology of some i ntermediate p roducts in the pr oduction o f i soprene r ubber. Sbornik Nauchnykh Trudov. Severo osetinskii Gosudarstvennyi i Meditsinskii Institut 5: 87

- 88

. 3) A strand, I; Ovrum, P; Lindqvist, T; Hultengren, M. ( 1976 ) . Exposure to butyl alcohol: Uptake and distribution in man. Scand J Work Environ Health 2: 165

- 17 5.

4) B arthelmess, A; Elkabarity, A. ( 1962 ) . Chemically induced multipolar mitoses. P art 3. Protoplasma 54: 455

- 47 5.

5) B ASF

. ( 1994 ) . Letter from BASF Corp to U.S. EPA r egarding submission of health and safety data subject to 8d reporting with attachments, dated 05/05/94 (sanitized). N ot e: studies are in german.
6) B rusewitz, S; Criteria Group for Occupational Standards; Wennberg, A. ( 1984 ) . Butanol och butylacetat. Arbete och Hälsa 13 : 1

- 45

. 7) B rusewitz, S; Wennberg, A. ( 1984 ) . Criteria document for setting exposure limits for butanol and butyl acetate. Ar bet e och Hälsa 171.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

Thi s document is a draft for review purposes only and does not constitute Agency policy. 1- 18 DRAFT

— DO NOT CITE OR QUOTE 8) Consejo Interamericano de Seguridad . ( 1996 ) . tert

- Butyl alcohol

. 

### 1

2 3

4 5

6 7

8 9

10 11

12

13

14 15

16 17

18 19 20 9) Conso, F; Mignâee, C. ( 1984 ) . Monohydric alcohols other than ethyl alcohol. Encyclopƒedie M ƒedico Ch irurgic al e Intoxications 101.

10) Damgard Nielsen, G ; Kr agh Hansen, M; Molhave, L. ( 1982 ) . Toxicological evaluation of a number of substances that may pollute the workplace air. (pp. 1 6

- 18). Denmark.

11) De Laat, J; Maouala

- M akata, P; Dore, M. ( 1996 ) . Rate constants for reactions of ozone and hydroxyl radicals with several phenyl

- u r eas and acetamides. Environ Technol 17: 707

- 716.

12) DFG (Deutsche Forschungsgemeinschaft). ( 1 995 ) . 2

- Amino -2- methyl -1-propanol. I n MAK value documentations. Weinheim, Germany: Wiley

- V C H

. 13) Henschler, D. ( 1984 ) . Harmful industrial substances - Toxicological and medical basis for the establishment of maximum permissible concentrations. V er lag Chemie.

14) INRS ( I nstitut national de recherche et de sécurité ). ( 1992 ) . [Pentanols].

15) N oda, T. ( 1994 ) . [Teratogenicity of organotin compounds]. Seikatsu Eisei 38: 139

- 156.

16) Rozengart, E. V. ( 1969 ) . The effect of a liphatic alcohols on the anticholinesterase efficacy of certain organophosphorus compounds. Biokhimiya 34: 813

- 8 1 5.

17) Weese, H. ( 1928 ) . Comparative s tudies o f t he e fficacy a nd t oxicity of the vapors of lower aliphatic alcohols. Arch Exp Pathol Phar 135:118

- 13 0.

18) Y amamoto, I; Haseba, T; Kurosu, M; Watanabe, T. ( 1992 ) . [Allosterism of acidic a lcohol dehydrogenase (class III ADH) of mouse liver and its role in alcohol metabolism]. Nippon Ika Daigaku Zasshi 59: 136

- 1 4 4.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2-1 DRAFT

— DO NOT CITE OR QUOTE 

# 2. PRELIMINARY EVIDENCE TABLES AND EXPOSURE

- RESPONSE ARRAYS 1

2
3

4
5
6
7
8
9

10

11 12 13

14

15

16
17
18
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20
21
22
23
24
25
26
27
28
29
30 

## 2.1. **Data Extraction: Preparation of Preliminary**  **Evidence Table s**  **and Exposure** 

**- Response Array**  **s**
The 1 06 references identified as primary sources of health effects data were considered for data extraction to evidence tables and exposure

- res p onse arrays. References were first collated with respect to exposure route, exposur e d uration, and type of endpoint, to identify those most pertinent for evaluating the human health effects from chronic oral or inhalation exposure to tert

b utanol . A s a result, data from 73 st udies with one or more of the following characteristics were not extracted into evidence tables or exposure

- res p onse arrays:

• The study involved dermal exposure;

• The study only involved acute or short

- t er m exposures (less than 90 days/13 weeks), and it was not conducted in the context of immune, ne u rological, developmental
, or reproductive toxicity;

• The data in the study only included endpoints related to possible mechanisms of toxicity;

• The study’s endpoints did not e x hibit responses in any of the 106 available references

.
Data from t he 33 r emaining references wer e prepared in preliminary evidence tables. No studies were excluded based on study quality considerations, so as to allow for public input on methodological considerations that could affect the interpretation of or confidence in each study’s
results. Wit h r eg ard to noncancer effects, health effect endpoints that were consistently affected in chronic or subchronic studies were included in the evidence tables. All data demonstrating carcinogenic effects were included. Supporting data that provide mechanisti c i n formation for each selected endpoint were also included. Fo r each included endpoint, all studies reporting data on that endpoint are included regardless of the reported level or statistical significance of the response.
The tables are arr an ged in the order from the health effect with the most data to health effect with
the least data , w it h carcinogenic effects grouped with non

- carcinogenic effects in the same tissue or system . For each endpoint, the studies are presented beginning with sub chr o nic studies followed by chronic exposures. Th e evidence table for all reported genotoxic ity endpoints follows

. The
information in the preliminary evi d ence tables is displayed graphically in preliminary exposure response arrays. In these pr e liminary arrays, the doses are labeled based only on statistical significance as determined by the study’s authors
, wit h out consideration of biological significance.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2-2 DRAFT

— DO NOT CITE OR QUOTE As a consequence, endpoints for which the study’s authors did not report results of statistical 

### 1
2

3 si gnificance tests were not included in these preliminary arrays.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2-3 DRAFT

— DO NOT CITE OR QUOTE 

## 2.2. **Kidney Effects** 1

2 3 **Table 2-1.**  **Evidence pertaining to kidney effects in**  **animals following oral exposure**  **t o**  te rt

- **butanol**

Reference and s tudy d esign Results
Kidney weight (percent change as compared to control)
Lyondell Chemical Company ( 2004 ) Sprague

- Dawley rat; 12/sex/treatment G avage 0, 64, 160, 400, or 1,000 mg/kg -d At least 9 weeks beginning 4 weeks prior to mating through mating (up to 2 weeks), gestation, and lactation. Males

Dose (mg/kg

- d) Left absolute Weight Left relative Weight Right absolute Weight Right relative W eight

0 0 0 0 0

64 +6 +8 +6 +8

160 +9 +14* +6 +11*

400 +12* +14* +14* +17*

1,000 +18* +28* +20* +31*

Females

Dose (mg/kg

- d) Left absolute Weight Left relative Weight Right absolute Weight Right relative Weight

0 0 0 0 0

64

− 1

− 2 +2 0

160 0 0 +1 0

400 +3 +2 +4 +2

1,000 +4 0 +7 +2

NTP ( 1995 ) F344/N rat; 10/sex/treatment Drinking water 0, 2.5, 5, 10, 20, 40 mg/mL M: 0, 230, 490, 840, 1,520, 3,610 a mg/kg -d F: 0, 290, 590, 850, 1,560, 3,620 a mg/kg -d 13 weeks Males Females

Dose (mg/kg

- d) Absolute Weight Relative Weight Dose (mg/kg

- d) Absolute Weight Relative Weight

0 0 0 0 0 0

230 +12* +19* 290 +19* +17*

490 +17* +26* 590 +16* +15*

840 +16* +32* 850 +29* +28*

1,520 +26* +54* 1,560 +39* +40*

3,610 All dead All dead 3,620 +36* +81*


### 4Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2-4 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 1** 

**. Evidence pertaining to k**  **idney effects in**  **animals following oral exposure**  **to** tert

- **butanol (**  **continued )** 

Reference and s tudy d esign Results

NTP ( 1995 ) B6C3F 1 mouse; 10/sex/treatment Drinking water ( 0, 2.5, 5, 10, 20, 40 mg/mL) M: 0, 350, 640, 1,590, 3,940, 8,210 a mg/kg -d F: 0, 500, 820, 1,660, 6,430, 11,620 a mg/kg -d 13 weeks Males Females

Dose (mg/kg

- d) Absolute weight Relative weight Dose (mg/kg

- d) Absolute weight Relative weight

0 0 0 0 0 0

350 +1 +1 500 0

− 3

640 +3 +2 820

− 3

− 1

1,590 +2 +8 1,660 +1 0

3,940 +6 +22* 6,430 +6 +15*

8,210 0 +48* 11,620 +12* +35*

NTP ( 1995 ) F344/N rat; 60/sex/treatment (10/sex/treatment evaluated at 15 months) Drinking water ( 0, 1.25, 2.5, 5, or 10 mg/mL) M: 0, 90, 200, or 420 a mg/kg -d F: 0, 180, 330, or 650 a mg/kg -d 2 years Males Females

Dose (mg/kg

- d) Absolute weight Relative weight Dose (mg/kg

- d) Absolute weight Relative weight

0 0 0 0 0 0

90 +4 +8 180 +8* +14*

200 +11 +15* 330 +18* +21*

420 +7 +20* 650 +22* +42*
Only animals sacrificed at 15 months were evaluated for organ weights.

Histopathology
Acharya et al. ( 1997 ; 1995 )

Wistar rat; 5

– 6 males/treatment Drinking water (0 or 0.5%), 0 or 575 mg/kg -d

10 weeks ↑ tubular degeneration, degeneration of the basement membrane of the Bowman’s
caps ule, diffused glomeruli, and glomerular vacuolation (no incidences reported)

↓ kidney glutathione (~40%)*

Lyondell Chemical Company ( 2004 )

Sprague

- Dawley rat; 12/sex/treatment Gavage 0, 64, 160, 400, or 1,000 mg/kg -d

At leas t 9 weeks beginning 4 weeks prior to mating through mating (up to 2 weeks), gestation, and lactation. There were no changes in kidney histopathology observed.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2-5 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 1** 

**. Evidence pertaining to k**  **idney effects in**  **animals following oral exposure**  **to** tert

- **butanol (**  **continued )** 

Reference and s tudy d esign Results

NTP ( 1995 )

F344/N rat; 10/sex/treatment Drinking water ( 0, 2.5, 5, 10, 20, or 40 mg/mL) M: 0, 230, 490, 840, 1,520, 3,610 a mg/kg -d F: 0, 290, 590, 850, 1,560, 3,620 a mg/kg -d

13 weeks Incidence (severity):

Males Females

Dose (mg/kg

- d) Mineralization Nephropathy Dose (mg/kg

- d) Mineralization Nephropathy

0 0/10 7/10 (1.0) 0 10/10 (1.7) 2/10 (1.0)

230 0/10 10/10 (1.6*) 290 10/10 (2.0) 3/10 (1.0)

490 2/10 (1.5) 10/10 (2.6*) 590 10/10 (2.0) 5/10 (1.0)

840 8/10*(1.4) 10/10 (2.7*) 850 10/10 (2.0) 7/10* (1.0)

1,520 4/10*(1.0) 10/10 (2.6*) 1,560 10/10 (2.0) 8/10* (1.0)

3,610 4/10*(1.0) 7/10 (1.1) 3,620 6/10 (1.2) 7/10* (1.0)

NTP ( 1995 )

B6C3F 1 mouse; 10/sex/treatment Drinking water ( 0, 2.5, 5, 10, 20, or 40 mg/mL ) M: 0, 350, 640, 1,590, 3,940, 8,210 a mg/kg -d F: 0, 500, 820, 1,660, 6,430, 11,620 a mg/kg -d

13 weeks There were no changes in kidney histopathology observed (histopathology data for the 13- week study were not provided)Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2-6 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 1** 

**. Evidence pertaining to k**  **idney effects in**  **animals following oral exposure**  **to** tert

- **butanol (**  **continued )** 

Reference and s tudy d esign Results

NTP ( 1995 )

F344/N rat; 60/sex/treatment (10/sex/treatment evaluated at 15 months) Drinking water ( 0, 1.25, 2.5, 5, 10 mg/mL) M: 0, 90, 200, 420 a mg/kg -d F: 0, 180, 330, 650 a mg/kg -d

2 years Incidence (severity): Males

Dose (mg/kg

- d) Mineralization ( interim ) Mineralization ( terminal ) Linear mineralization ( terminal )

0 1/10 (1.0) 26/50 (1.0) 0/50

90 2/10 (1.0) 28/50 (1.1) 5/50* (1.0)

200 5/10 (1.8) 35/50 (1.3) 24/50* (1.2)

420 9/10* (2.3) 48/50* (2.2) 46/50* (1.7)

Dose (mg/kg

- d) Renal tubule hyperplasia (extended evaluation) Transitional epithelium hyperplasia Nephropathy severity

0 12/50 (2.3) 25/50 (1.7) 3.0

90 16/50 (2.3) 32/50 (1.7) 3.1

200 14/50 (2.2) 36/50* (2.0) 3.1

420 23/50* (2.8) 40/50* (2.1) 3.3*

Females

Dose (mg/kg

- d) Mineralization b
Interim Mineralization b
Terminal Inflammation (suppurative)

incidence

0 10/10 (2.8) 49/50 (2.6) 2/50

180 10/10 (2.9) 50/50 (2.6) 3/50

330 10/10 (2.9) 50/50 (2.7) 13/50*

650 10/10 (2.8) 50/50 (2.9) 17/50*

Dose (mg/kg

- d) Renal tubule hyperplasia Transitional epithelium hyperplasia Nephropathy severity

0 0/50 0/50 1.6

180 0/50 0/50 1.9*

330 0/50 3/50 (1.0) 2.3*

650 1/50 (1.0) 17/50*(1.4) 2.9*Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2-7 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 1** 

**. Evidence pertaining to k**  **idney effects in**  **animals following oral exposure**  **to** tert

- **butanol (**  **continued )** 

Reference and s tudy d esign Results

NTP ( 1995 )

B6C3F 1 mouse; 60/sex/treatment Drinking water ( 0, 5, 10, or 20 mg/mL) M: 0, 540, 1,040, or 2,070 a mg/kg -d F: 0, 510, 1,020, or 2,110 mg/kg -d

2 years No changes in kidney related histopathology observed. c

Tumors

NTP ( 1995 )

F344/N rat; 60/sex/treatment (10/sex/treatment evaluated at 15 months) Drinking water ( 0, 1.25, 2.5, 5, or 10 mg/mL) M: 0, 90, 200, or 420 a mg/kg -d F: 0, 180, 330, or 650 a mg/kg -d

2 years Male Dose (mg/kg

- d) Renal tubule adenoma d Renal tubule adenoma (multiple) d Renal tubule adenoma or carcinoma d

0 7/50 1/50 8/50

90 7/50 4/50 13/50

200 10/50 9/50* 19/50*

420 10/50 3/50 13/50

Results do not include the animals sacrificed at 15 months. Females did not develop kidney tumors.

Hard et al. ( 2011 ) ; reanalysis of the slides in the NTP ( 1995 ) study (see above) Male Dose (mg/kg

- d) Renal tubule adenoma Renal tubular adenoma (multiple) Renal tubule adenoma or carcinoma

0 3/50 1/50 4/50

90 9/50 3/50 13/50*

200 9/50 9/50 18/50*

420 9/50 3/50 12/50*

NTP ( 1995 )

B6C3F 1 mouse; 60/sex/treatment Drinking water ( 0, 5, 10, or 20 mg/mL) M: 0, 540, 1,040, or 2,070 a mg/kg -d F: 0, 510, 1,020, or 2,110 mg/kg -d

2 years No changes in kidney

- related tumorsPreliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2-8 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 1** 

**. Evidence pertaining to k**  **idney effects in**  **animals following oral exposure**  **to** tert

- **butanol (**  **continued )** 

Reference and s tudy d esign Results

Additional studies potentially relevant for informing the mode of action

Williams and Borghoff ( 2001 )

F344 rats; 4/sex Single gavage dose: 500 mg/kg Males: ↑ binding of tert

- butanol to α 2u - globulin compared to females* Females: no change in binding observed

Takahashi et al. ( 1993 )

F344 rats (M); 10/treatment Drinking water (0, 0.25, 0.5, 1, 2 ,4% w/v) 0,230,490, 840, 1,520 or 3,610 a mg/kg

- d

13 weeks ↑ in hyaline droplet accumulation (number and size) with ↑ severity for nephropathy (no incidence data)

Male Dose (mg/kg

- d) Hyaline droplet severity score Hyaline crystals severity score Nephropathy severity score

0 1 0.5 1

230 1.5 2* 2*

490 2* 3* 3*

840 2* 3* 3*

1,520 2* 3* 3*

3,610 >0.25* >0.25* 1

NTP ( 1995 )

F344/N rat; 10/sex/treatment Drinking water ( 0, 2.5, 5, 10, 20, or 40 mg/mL) M: 0, 230, 490, 840, 1,520, 3,610 a mg/kg -d F: 0, 290, 590, 850, 1,560, 3,620 a mg/kg -d

13 weeks Accumulation of hyaline droplets:

Male Dose (mg/kg

- d) Hyaline droplet accumulation

0 0/10

230 + e

490 ++

840 ++
1,520 ++
3,610 0/10
No information provided on females . No results from statistical tests reported.

Hard et al. ( 2011 )

Reanalysis of the slides in the NTP ( 1995 ) study (see above) Males: Confirmed accumulation of hyaline droplets increased with increasing dose

- levels in 13 week study above. No incidence data available.

Females: not evaluated

a The high

- dose group had an increase in mortality. 

### 1 2 3 4 5 6 7 8 9 b Linear mineralization not observed in female rats

. c Organs were not weighed in mice during the 2

- year study. d Standard & extended evaluation combined

. e + or ++ indicated an increased accumulation relative to controls, as reported by the authors ; no additional incidence data and

no results from statistical tests available. * Statistically significant p ≤ 0 .05 as d etermined by the study authors

. Percentage change compared to control = (treated value – c o ntrol value) ÷ control value × 100. Conversions from drinking water concentrations to mg/kg

- d per f ormed by study authors.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2-9 DRAFT

— DO NOT CITE OR QUOTE **Table 2-2** 

**. Evidence pertaining to kidney effects**  **in animals following**  

### 1 2 **inhalation exposure to**  tert

- **butanol**

Reference and s tudy design Results
Kidney weight (percent change compared to control)

NTP ( 1997 )

F344/N rat; 10/sex/treatment Analytical concentration: 0, 134, 272, 542, 1,080, or 2,101 ppm (0, 406, 824, 1,643, 3,273 or 6,368 mg/m 3 ) 6 hr/d, 5 d/wk

13 weeks Males Females

Dose (mg/m 3 ) Absolute weight Relative weight Absolute weight Relative weight

0 0 0 0 0

406 +1 +1

− 4

− 1

824

− 2

− 1 0 +1

1,643 +3 +2 +4 +4

3,273 +11* +8* +2 +2

6,368 +9.8* +9* +4 +9*

NTP ( 1997 )

B6C3F 1 mouse; 10/sex/treatment Analytical concentration: 0, 134, 272, 542, 1,080, or 2,101 ppm (0, 406, 824, 1,643, 3,273 or 6,368 mg/m 3 ) 6 hr/d, 5 d/wk

13 weeks Males Females

Dose (mg/m 3 ) Absolute weight Relative weight Absolute weight Relative weight

0 0 0 0 0

406

− 6

− 4 +1

− 3

824

− 1 +3 +5 +9

1,643 +4 +3 +1

− 2

3,273

− 10

− 3 0 +7

6,368 +3 +6 +3 +15*
Histopathology

NTP ( 1997 )

F344/N rat; 10/sex/treatment Analytical concentration: 0, 134, 272, 542, 1,080, or 2,101 ppm (0, 406, 824, 1,643, 3,273 or 6,368 mg/m 3 ) 6 hr/d, 5 d/wk

13 wee ks Male

Dose (mg/m 3 ) Average severity of chronic nephropathy

0 1.0

406 1.4

824 1.4

1,643 1.6

3,273 1.9

6,36 8 2.0
Severity categories: 1= minimal, 2= mild. No results from statistical tests reported 

### 3Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 10 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 2. Evidence pertaining to kidney effects in animals following**  **inhalation exposure to** tert

- **butanol (continued)**

Reference and s tudy design Results

NTP ( 1997 )

B6C3F 1 mouse; 10/sex/treatment Analytical concentration: 0, 134, 272, 542, 1,080, or 2,101 ppm (0, 406, 824, 1,643, 3,273 or 6,368 mg/m 3 ) 6 hr/d, 5 d/wk

13 weeks There were no kidney effects observed.

Additional studies potentially relevant for informing the mode of action

Borghoff et al. ( 2001 ) F344 rat; 5/sex/treatment Analytical concentration:0, 250, 450, 1
, 750 ppm (0,771, 1,387 or 5,395mg/m 3 ) 6hr/d 10 days Males: positive trend for a ccumulation of protein droplets (p < 0.05), significant increase in accumulation of α 2u - globulin at 5,394 mg/m 3 as compared to controls (no incidence data provided) Females: No positive staining for α 2u - globulin was observed in exposed female rats. * Statistically significant p ≤ 0.05 as determined by study authors

. 

### 1 2 3 4 5 Conversion from ppm to mg/m 3 is 1 ppm = 3.031 mg/m 3

. Percentage change compared to control = (treated v a lue – control value) ÷ control value × 100.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 11 DRAFT

— DO NOT CITE OR QUOTE 

### 1 2 3

4 5 6 10 100 1,000 10,000 100,000Dose (mg/kg-day)Absolute weight; M Rat (D)Nephropathy severity; M Rat (D)Relative weight; M Rat (D)Decreased glutathione; M Rat (A)Mineralization; M Rat (D)Absolute weight; F Rat (D)Relative weight; F Rat (D)Nephropathy incidence; F Rat (D)Mineralization; F Rat (D)Absolute weight; M Mouse (D)Relative weight; M Mouse (D)Relative weight; F Mouse (D)Absolute weight; F Mouse (D)Linear mineralization; M Rat (D)Absolute weight; M Rat (D)Relative weight; M Rat (D)Transitional epithelium hyperplasia; M Rat (D)Interim/terminal mineralization; M Rat (D)Nephropathy severity; M Rat (D)Renal tubular hyperplasia; M Rat (D)Nephropathy severity; F Rat (D)Relative weight; F Rat (D)Absolute weight; F Rat (D)Inflammation; F Rat (D)Transitional epithelium hyperplasia; F Rat (D)Interim/terminal mineralization; F Rat (D)Renal tubule hyperplasia; F Rat (D)Relative weight; M Rat (C)Absolute weight; M Rat (C)Absolute weight; F Rat (C)Relative weight; F Rat (C)Renal tubular adenoma or carcinoma; M Rat (B)Renal tubular adenoma or carcinoma; M Rat (D)Renal tubular adenoma or carcinoma; F Rat (D)Renal tubular adenoma or carcinoma; M Mouse (D)Renal tubular adenoma or carcinoma; F Mouse (D)  = exposures at which the endpoint was reported statistically significant by study authors  = exposures at which the endpoint was reported not statistically significant by study authors

× = exposures at which all animals were dead and unable to be examined for the endpoint

SUBCHRONIC CHRONIC CANCER REPRO

DUCTIVE Sources: (A) Acharya et al. ( 1997 ; 1995 ) ; (B) Hard et al. ( 2011 ) *; (C) Lyondell ( 2004 ); (D) NTP ( 1995 ) *: reanalysis of NTP ( 1995 )

**Figure 2-1**  **. Exposure response array of**  **kidney effects in rats and mice**  **following oral exposure to** tert

- **buta nol.** Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 12 DRAFT

— DO NOT CITE OR QUOTE 

### 1 2

3 4 5

6 100 1,000 10,000Absolute weight; M RatRelative weight; M RatAbsolute weight; F RatRelative weight; F RatAbsolute/relative weight; M MouseAbsolute weight; F MouseRelative weight; F MouseConcentration (mg/m3)

 = exposures at which the endpoint was reported statistically significant by study authors  = exposures at which the endpoint was reported not statistically significant by study authors

Source: NTP ( 1997 )

**Figure 2-2**  **. Exposure**

**- response array of kidney effects in rats**  **and mice following subchr**  **onic inhalation exposure to** tert

- **butanol.**Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 13 DRAFT

— DO NOT CITE OR QUOTE 

## 2.3. **Thyroid Effects** 1

2 3

4 **Table 2-3**  **. Evidence pertaining to thyroid effects in animals following oral exposure to**  tert

- **bu t**  **anol**

Reference and s tudy design Results Follicular cell hyperplasia

NTP ( 1995 ) F344/N rat; 60/sex/treatment (10/sex/treatment evaluated at 15 months) Drinking water ( 0, 1.25, 2.5, 5, or 10 mg/mL) M: 0, 90, 200, or 420 a mg/kg -d F: 0, 180, 330, or 650 a mg/kg -d 2 years Incidence b

Males Females

Dose (mg/kg

- d) Follicular cell hyperplasia Dose (mg/kg

- d) Follicular cell hyperplasia

0 3/50 0 0/50

90 0/49 180 0/50

200 0/50 330 0/50

420 0/50 650 0/50

NTP ( 1995 ) B6C3F 1 mouse; 60/sex/treatment Drinking water ( 0, 5, 10, or 20 mg/mL ) M: 0, 540, 1,040, or 2,070 a mg/kg -d F: 0, 510, 1,020, or 2,110 mg/kg -d 2 years Incidence (severity)

Males Females

Dose (mg/kg

- d) Follicular cell hyperplasia Dose (mg/kg

- d) Follicular cell hyperplasia

0 5/60 (1.2) 0 19/58 (1.8)

540 18/59* (1.6) 510 28/60 (1.9)
1,040 15/59* (1.4) 1,020 33/59* (1.7)
2,070 18/57* (2.1) 2,110 47/59* (2.2)

Tumors

NTP ( 1995 ) F344/N rat; 60/sex/treatment (10/sex/treatment evaluated at 15 months) Drinking water ( 0, 1.25, 2.5, 5, or 10 mg/mL) M: 0, 90, 200, or 420 a mg/kg -d F: 0, 180, 330, or 650 a mg/kg -d 2 years Incidence b

Dose (mg/kg

- d) Follicular cell adenoma Follicular cell carcinoma

Male

0 2/50 2/50

90 0/49 0/49

200 0/50 0/50

420 0/50 0/50

Female

0 1/50 1/50

180 0/50 0/50

330 1/50 1/50
6,500 0/50 0/50Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 14 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 3**  **. Evidence pertaining to thyroid effects in animals following oral exposure to**  tert

- **butanol (**  **continued )** 

Reference and s tudy design Results

NTP ( 1995 ) B6C3F 1 mouse; 60/sex/treatment Drinking water ( 0, 5, 10, or 20 mg/mL ) M: 0, 540, 1,040, or 2,070 a mg/kg -d F: 0, 510, 1,020, or 2,110 mg/kg -d 2 years Incidence

Dose (mg/kg

- d) Follicular cell adenoma Mortality

adjusted rate (%) Follicular cell carcinoma or

adenoma Mortality

adjusted rate (%)

Male

0 1/60 3.6 1/60 3.6

540 0/59 0.0 0/59 0.0
1,040 4/59 10.1 4/59 10.1
2,070 1/57 5.9 2/57 8.7

Female

0 2/58 5.6 2/58 5.6

510 3/60 8.6 3/60 8.6
1,020 2/59 4.9 2/59 4.9
2,110 9/59* 19.6 9/59* 19.6

a There was a significant decrease in survival in the high

- dose group. 

### 1 2 3 4 5 6
7
8 b Results do not include the animals sacrificed at 15 months. * Statistically significant p ≤ 0 .05 as determined by the study authors

. Conversions f ro m drinking water concentrations to mg/kg

- d performed by study authors.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 15 DRAFT

— DO NOT CITE OR QUOTE 

### 1 2

3 4 5

6 10 100 1,000 10,000Hyperplasia; M mouseHyperplasia; F mouseHyperplasia; M ratHyperplasia; F ratAdenoma; M mouseAdenoma; F mouseAdenoma; M ratAdenoma; F ratDose (mg/kg-day)

NONCANCER CANCER  = exposures at which the endpoint was reported statistically significant by study authors  = exposures at which the endpoint was reported not statistically significant by study authors

Source: NTP ( 1995 )

**Figure 2-3** 

**. Exposure** 

**- response array of**  **thyroid follicular cell effects in rats**  **and mice following chronic oral exposure to**  tert

- **but anol** 

**.**Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 16 DRAFT

— DO NOT CITE OR QUOTE 

## 2.4. **Reproductive, Developmental, and Neurodevelopmental Effects** 1

2 3 **Table 2-4**  **. Evidence pertaining to r**  **eproductive effects in**  **animals following**  **exposure to** tert

- **but a**  **nol**

Reference and s tudy design Results


### Oral

Billitti et al. ( 2005 )

CD- 1 male mice; number unclear Gavage, 0, 400, 1,000, 2,000 mg/kg

Single dose no effect on testosterone levels in feces or serum (data not provided) no effect on body weight (statistical tests not reported) ↑ testes weights (averaged weights in the 1,000 and 2,000 mg/kg groups +14% higher relative to control s; p ≤ 0.05) higher percentage (+7%; p ≤ 0.05) of sloughing in the seminiferous epithelium in the control animals
Lyondell Chemical Company ( 2004 )

Sprague

- Dawley rat; 12/sex/treatment Gavage 0, 64, 160, 400, or 1,000 mg/kg -d

P0 Males and Females: At least 9 weeks beginning 4 weeks prior to mating through mating (up to 2 weeks), gestation, and lactation. P0 reproductive effects M: No effects weights of male reproductive organs or sperm observed

F:

Dose (mg/kg

- d) 0 64 160 400 1000
Pregnancy index

91.7% 91.7% 100% 100% 91.7%

Live pups/litter response relative to control

0

− 9

− 11

− 7

− 33*

NTP ( 1995 )

F344/N rat; 10/sex/treatment Drinking water ( 0, 2.5, 5, 10, 20, or 40 mg/mL ) M: 0, 230, 490, 840, 1,520, 3,610 a mg/kg -d F: 0, 290, 590, 850, 1,560, 3,620 a mg/kg -d

13 weeks No significant effect on weights of male reproductive organs or sperm obse rved

No significant effect on female estrous cycle (0, −2, −4, 0, +8 % change relative to control)

NTP ( 1995 )

B6C3F 1 mouse; 10/sex/treatment Drinking water ( 0, 2.5, 5, 10, 20, or 40 mg/mL ) M: 0, 350, 640, 1,590, 3,940, 8,210 a mg/kg -d F: 0, 500, 820, 1,660, 6,430, 11,620 a mg/kg -d

13 weeks No significant effect on weights of male reproductive organs or sperm observed

↑ length of estrous cycle Response relative to control : 0, +5, +5, +5, +6, +28*%



### 4
5Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 17 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 4** 

**. Evidence pertaining to r**  **eproductive effects in**  **animals following**  **exposure to** tert

- **butanol (continued)** 

Reference and s tudy design Results


### Inhalation

NTP ( 1997 )

F344/N rat; 10/sex/treatment Analytical concentration: 0, 134, 272, 542, 1,080, or 2,101 ppm (0, 406, 824, 1
, 643, 3,273 or 6,368 mg/m 3 ) 6 hr/d, 5 d/wk

13 weeks No significant effect on weights of male reproductive organs or sperm observed No significant effect on female estrous cycle (0, −4, +2, +4 % change relative to control)

Evaluations were only performed for concentrations ≥54 2 ppm

NTP ( 1997 )

B6C3F 1 mouse; 10/sex/treatment Analytical concentration: 0, 134, 272, 542, 1,080, or 2,101 ppm (0, 406, 824, 1,643, 3,273 or 6,368 mg/m 3 ) 6 hr/d, 5 d/wk

13 weeks No significant effect on weights of male reproductive organs or sperm observed No significant effect on female estrous cycle (0, −3, −9, −5 % change relative to control)

Evaluations were only performed for concentrations ≥54 2 ppm

* Statistically significant p ≤ 0.05 as determined by the study authors

. 

### 1 2 3 4 5 6 Conversions from drinking water concentrations to mg/kg

- d performed by study authors. Conversion from ppm to mg/m 3 is 1 ppm = 3.031 mg/m 3

. Percentage change compared to control = (treated value – c o ntrol value) ÷ control value × 100.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 18 DRAFT

— DO NOT CITE OR QUOTE **Table 2-5**  **. Evidence pertaining to developmental**  **effects in**  **animals following**  **exposure to** tert

- **butanol** 

### 1 2

R eference and study design Results

Oral

Daniel and Evans ( 1982 )

Swiss Webster (Cox) mouse; 15 pregnant dams/treatment Liquid diet (0, 0.5, 0.75, 1.0%, w/v) 0 (isocaloric amounts of maltose/dextrin), 3,324, 4,879, 6,677 mg/kg -d

GD 6 –20 No statistical analysis was conducted on any of these data

Maternal
Percent change compared to control:

Dose (mg/kg

- d) Body weight on GD 20 Body weight gain Number of litters (% pregnant dams)

0 0 0 11 (77%)

3,324 +2

− 3 12 (80%)

4,879

− 5

− 19 8 (53%)

6,677

− 10

− 20 7 (47%)

Fetal
Percent change compared to control:

Dose (mg/kg

- d) Number of neonates/litter Fetal body weight on PND 2

0 0 0

3
, 324

− 1

− 7

4,879

− 29

− 19

6,677

− 49

− 38
Number of stillborn also increased with dose (3, 6, 14, and 20, respectively), but appropriate denominators not reported so statistical significance of this change could not be calculated. The high dose also caused a delay in eye opening and a lag in weight gain during PND 2

– 10 (information was only provided in text or figures)

Faulkner et al. ( 1989 )

CBA/J mouse; 7 pregnant females in control, 12 pregnant females in treated Gavage ( 10.5 mmoles/kg twice a day) ; 0 (tap water), 1,556 mg/kg -d

GD 6 –18 Maternal results not reported.

Fetal
Percent change compared to control: Incidence:

Dose (mg/kg

- d) L ive fetuses/litter Fetal weight S ternal variations S kull variations

0 0 0 4/28 1/28

1,556

− 41 *

− 4 7/30 3/30

S ternal variations: misaligned or unossified sternebrae S kull variations: moderate reduction in ossification of supraoccipital bone
Number of total resorptions (10 resorptions/66 implants in controls, 37/94 implants in treated) and resorptions per litter resorptions per litter ( +118%) increased ( p < 0.05)Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 19 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 5** 

**. Evidence pertaining to d**  **evelopmental effects in**  **animals following**  **exposure to** tert

- **butanol (continued)** 

R eference and study design Results

Faulkner et al. ( 1989 )

C57BL/6J mouse; 5 pregnant female s in controls, 9 pregnant females treated Gavage ( 10.5 mmoles/kg twice a day) 0 (tap water), 1,556 mg/kg -d

GD 6 –18 Maternal results not reported.

Fetal

Percent change compared to control: Incidence:

Dose (mg/kg

- d) L ive fetuses/litter Fetal weight S ternal variations S kull variations

0 0 0 5/21 1/21

1,556

− 58%*

− 4 9/16 7/16

S ternal variations: misaligned or unossified sternebrae S kull variations: moderate reduction in ossification of supraoccipital bone
Number of total resorptions (4 resorptions/44 implants in controls, 38/68 implants in treated) and resorptions per litter resorptions per litter ( +428%) increased ( p < 0.05)
Lyondell Chemical Company ( 2004 )

Sprague

- Dawley rat; 12/sex/treatment Gava ge 0, 64, 160, 400, or 1,000 mg/kg -d

P0 Males and Females: At least 9 weeks beginning 4 weeks prior to mating through mating (up to 2 weeks), gestation, and lactation.

F1 Males and Females: 7 weeks (throughout gestation and lactation; 1 male and 1 female from each litter was treated themselves from PND 21 -28) Response relative to control

Dose (mg/kg

- d) 0 64 160 400 1000

Maternal effects
Body weight gain GD 0

- 20

0

− 3

− 4 0

− 16*
Body weight gain PND 1

- 21

0 +3

− 10 +3 +100*
F1 Reproductive effects

0 64 160 400 1000
Viability index

96.4% 98.7% 98.2% 99.4% 74.1%*

Lactation index

100% 100% 100% 99.2% 98.8%
Sex ratio ( % males)

54.4 52.3 50.9 53.4 52.1
Pup weight/ litter PND 1 response relative to control

0 +6 +4 +7

− 10
Pup weight PND 28 response relative to control

M: 0 +2 0 0

− 12*

F: 0 0

− 4

− 2

− 8 

### 1Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 20 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 5** 

**. Evidence pertaining to d**  **evelopmental effects in**  **animals following**  **exposure to** tert

- **butanol (continued)** 

R eference and study design Results

I nhalation

Nelson et al. ( 1989 )

Sprague

- Dawley rat; 15 pregnant dams/treatment Whole body inhalation Analytical concentration: 0, 2,200, 3,510, 5,030 ppm (0, 6,669, 10,640, 15,248 mg/m 3 ), 7 hr/d

GD 1 –19 Maternal: Unsteady gait (no statistical tests reported) , dose

- dependent ↓ in body weight gain (results presented in figure only), dose

- dependent ↓ in food consumption ranging from 7

– 36% depending on dose and time

Fetal
Percent change compared to control:

Dose (mg/m 3 ) Number of live fetuses/litter Resorptions per litter

0 0 0

6,669 0 +9

10,640 +15

− 18

15,248 +8 0
Percent change compared to control: Incidence:

Dose (mg/m 3 ) Fetal weight (males) Fetal weight (females) Skeletal variation by litter Skeletal variation by fetus

0 0 0 10/15 18/96

6,669

− 9*

− 9* 14/17 35/104

10,640

− 12*

− 13* 14/14 53/103*

15,248

− 32*

− 31* 12/12 76/83*
Skeletal variation by litter refers to the number of variations observed in the number of litters examined. Skeletal variation by fetus refers to the number of variations observed in the total number of fetuses examined. Fetuses are not categorized by litter. * Statistically significant p ≤ 0.05 as determined by study authors

. 

### 1 2 3 4 5 6 Conversions from diet concentrations to mg/kg

- d performed by study authors. Conversion from ppm to mg/m 3 is 1 ppm = 3.031 mg/m 3

. Percentage change compared to control = (treated value – c o ntrol value) ÷ control value × 100.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 21 DRAFT

— DO NOT CITE OR QUOTE **Table 2-6**  **. Evidence pertaining to neurodevelopmental**  **effects in**  **animals** 

### 1 2 **following exposure to** tert

- **butanol**

Reference and s tudy design Results

Oral

Daniel and Evans ( 1982 )

Liquid diet (0, 0.5, 0.75, or 1.0%, w/v); GD6 –20; Swiss Webster (Cox) mouse; 15 pregnant dams/treatment; after birth half the pups were nursed with their treated dams and the other half were fostered by untreated dams who recently gave birth

0 (isocaloric amounts of maltose/dextrin), 3,324, 4,879, or 6,677 mg/kg -d Results were presented in figures only with no comparison to control or in tables with only comparison between pups fostered with their maternal dams and those fostered with untreated dams with no indication of dose.

Results indicate

• a dose

- dependent increase in the time it took for the righting reflex with more time needed in animals maintained with mat ernal dams

• a dose

- dependent decrease in open field behavior with less activity with pups maintained with maternal dams

• a dose

- dependent decrease in rotorod performance with the pups from maternal dams having lower performances

• a dose

- dependent decrease in the amount of time the pups were able to avoid a cliff with animals maintained with their maternal dams having less avoidance time

Grant and Samson ( 1982 )

Surgical cannulation; male and female Long

Evans rat fetus; 8 control and 12 treated

PND 4

– 7 (a period of brain growth) with consecutive daily doses of 1,440, 2,160, 600, and 2,690 mg/kg

- d tert

- butanol on PND 4, 5, 6, and 7, respectively (average daily dose of 1,723 mg/kg

- d) ↓ Absolute and relative brain weights Response relativ e to control (only relative data provided, but no changes were noted in body weight) : 0, −16%*

Brain biochemical parameters included : ↓ DNA levels in the hindbrain (−16%*) and ↓ protein levels in the forebrain (−15%*).

Inhalation

Nelson et al. ( 1991 )

Whole body inhalation; Sprague

- Dawley rat; 15 pregnant dams/treatment

Analytical concentration: 0, 6,000, or 12,000 mg/m 3 ; 7 hr/d GD 1

– 19 Data were not presented specifically by dose nor were any tables or figures of the data provided

Maternal toxicity was noted by decreased food consumption and body weight gains

Results in offspring

• increase in rotorod performance in high

- dose group (16 versus 26 revolutions/min for controls and 3,500 mg/m 3 animals, respectively)

• decreased time held on wire in the performance ascent test in the low

- dose group (16 sec versus 10 sec for controls and 1,750 mg/m 3 animals, respectively)

There were some differences in neurochemical measurements in the brain betwee n control and treated offspring, but they were not related to dose



### 3Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 22 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 6** 

**. Evidence pertaining to**  **neurodevelopmental effects**  **in animals**  **following exposure to** tert

- **butanol (continued)** 

Reference and s tudy design Results

Nelson et al. ( 1991 )

Whole body inhalation; adult male Sprague

Dawley rats (18/treatment) mated to untreated females

Analytical concentration: 0, 6
, 000, or 12,000 mg/m 3 ; 7 hr/d for 6 wk Data were not presented specifically by dose nor were any tables or figures of the data provided

Results (generally only specified as paternally treated versus controls) in offspring indicate

• increase in rotorod performance (16 versus 20 revolutions/min for controls and 3,500 mg/m 3 animals, respectively)

• decreased time in open field (less time to reach the outer circle of the field, 210 sec versus 115 seconds for controls and 3,500 mg/m 3 animals, respectively) Th ere were some differences in neurochemical measurements in the brain between control and treated offspring, but they were not related to dose * Statistically significant p ≤ 0.05 as determined by study authors

. 

### 1 2 3 4
5
6 Conversions from diet concentrations to mg/kg

- d performed by study authors. Percentage change compared to control = (treated value – c o ntrol value) ÷ control value × 100.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 23 DRAFT

— DO NOT CITE OR QUOTE 

### 1

2 3 4

5 6 7 10 100 1,000 10,000 100,000Dose (mg/kg-day)Testes weight; M mouse (A)Reproductive organs or sperm; M rat (E)Reproductive organs or sperm; M rat (D)Number of live pups per litter; F rat (D)Estrous cycle length; F rat (E)Pregnancy index; F rat (D)Reproductive organs or sperm; M mouse (E)Estrous cycle length; F mouse (E)Body weight gain (gestation days 0-20); F rat (D)Body weight gain (lactation days 1-21); F rat (D)Pup weight per litter (postnatal day 28); M rat (D)Pup weight per litter (postnatal day 28); F rat (D)Viability index; M+F rat (D)Pup weight per litter (postnatal day 1); M+F rat (D)Sex ratio; M+F rat (D)Neurodevelopment; M+F rat (C)Lactation index; M+F rat (D)Number ofresorptions; M+F mouse (B)Number of live fetuses per litter; M+F mouse (B)Pup weight; M+F mouse (B)Skeletal variations; M+F mouse (B)

REPRODUCTIVE DEVELOPMENTAL Acute Sub

- chronic Fetal Maternal  = exposures at which the endpoint was reported statistically significant by study authors  = exposures at which the endpoint was reported not statistically significant by study authors

Sources: (A) Billitti et al. ( 2005 ) ; (B) Faulkner et al. ( 1989 ) ; ( C ) Grant and Samson ( 1982 ) ; (D) Lyondell Chemical Company ( 2004 ) ; (E) NTP ( 1995 )

**Figure 2-4** 

**. Exposure** 

**- response array of reproductive, developmental**  **and neurod**  **evelopmental effects following oral exposure to** tert

- **but anol.** Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 24 DRAFT

— DO NOT CITE OR QUOTE 

### 1 2 3

4 5 6 100 1,000 10,000 100,000Reproductive organs or sperm; M rat (A)Estrous cycle; F rat (A)Reproductive organs or sperm; M mouse (A)Estrous cycle; F mouse (A)Body weight; M mouse (B)Body weight; F mouse (B)Skeletal variation; M+F mouse (B)Number of resorptions per litter; M+F mouse (B)Number of live pups per litter; M+F mouse (B)Concentration (mg/m3)

REPRODUCTIVE (Parental) DEVELOPMENTAL (Fetal)  = exposures at which the endpoint was reported statistically significant by study authors  = exposures at which the endpoint was reported not statistically significant by study authors

Sources: (A) NTP ( 1997 ) ; (B) Nelson et al. ( 1989 ) ; (C) Nelson et al. ( 1991 )

**Figure 2-**  **5**

**. Exposure** 

**- response array of reproductive**  **and developmental**  **effects following inhalation exposure to** tert

- **but anol** 

**.**Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 25 DRAFT

— DO NOT CITE OR QUOTE 

## 2.5. **C entral Nervous System (C**  **NS )**  **Effects** 1

2 3 **Table 2-7** 

**. Evidence pertaining to CNS**  **effects in**  **animals following exposure to**  tert

- **bu tanol** 

Reference and s tudy design Results
Oral

Grant and Samson ( 1981 )

Male Long

- Evans rat ; 4

– 6/treatment; daily intakes were estimated to be 700 to 2, 800 mg/kg depending on the treatment and time

Drinking water Scheduled

- induced condition (maintained at 80% body weight): Treatment 1: 0.5% for 5 days, then 1% for 60 days Tre atment 2:Drinking water stepwise increase with 0.25%, 0.50%, 1.0%, 2.0%, and 2.5% for 5 days each and 3.0%, and 3.5% for 10 days each, then 3.0% for 40 days Treatment 3: 1.0% for 5 days, 2.0% for 5 days, then 1.0% for 5 days and 3.0% for 90 days

Home ca ge condition: Treatment 1: 80% body weight administered 0.5% 5 days, then 1.0% for 90 days Treatment 2: free access to food, 1.0% for 5 days, 2.0% for 10 days, and 3.0% for 90 days Treatment 3: 80% body weight on water

After treatment, all animals were gi ven water to replace t

- butanol • 5 of 15 animals treated with 3% t

- butanol had to be removed from the experiment based on self

- withdrawal and/or self

- mutilation (2 died), trend of increased t

- butanol intake and decreased food intake prior to removal. Sympto ms were not observed with 1% t

- butanol or with control.

• 1 of 4 animals receiving 3.5% t

- butanol self

- mutilated, ceased drinking, and was removed from the experiment.

• 60 days of 3% t

- butanol was related to moderate seizure activity (withdrawal scores of 9, 5, and 6).

• 90 days of 3% t

- butanol had withdrawal scores of 12, 6, and 7

• At least one animal from every group had a withdrawal score of 0, and only animals exposed to 3% t

- butanol had withdrawal scores of 6 or higher. Significantly higher withdrawal scores were observed in animals receiving greater amounts of t

- butanol (Fischer’s exact p<0.002).

• Schedule

- induced conditioning resulted in greater withdrawal symptoms over the home cage conditions.

• Withdrawal symptoms were considered less than those observed with ethanol.



### 4Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2

- 26 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 7** 

**. Evidence pertaining to CNS**  **effects in**  **animals following exposure to**  tert

- **butanol (continued)** 

Reference and s tudy design Results
Wood and Laverty ( 1979 )

Male Wistar rat; 12 rats/ 20

- day treatment and 24 rats on time

- course withdrawal sequence (4 rats withdrawn on days 4, 6, 8, 10, 12, and 14)

Liquid diet; 44

– 54 mmol/kg (mean 47 mmol/kg); daily consumption was variable with spontaneous periods of abstinence in a few rats; blood alcohol levels 2

– 25 mmol/L

4

– 20 days • Signs of intoxication in the 20

- day study included docility and slight ataxia while on the diet.

• One rat out of 12 in the 20

- day study spontaneously withdrew from the diet on Day 10, and had signs of irritability, hyperactivity, and muscular rigidity.

• Following rem oval of t

- butanol after 20 days, animals initially appeared intoxicated. Head bobbing and paw shaking were observed 3

– 4 hours after withdrawal. M uscular rigidity, tail signs, abnormal gait, tremor, and irritability were apparent 5

– 6 hours after withdrawal.

• 4/12 treated rats had spontaneous forelimb convulsions

• 5/12 treated rats had audiogenic convulsions; resulted in 3 deaths.

• In the surviving 8/12 rats, irritability, hyperreactivity, and muscular rigidity were considerable at 24 hours post

- withdrawaland i n some animals lasted for up to 72 hours.

• In the time

- course group, withdrawal symptoms were seen with as little as 4 days of t

- butanol intake. S everity rating increased with increasing days on the diet.
Snell and Harris ( 1980 )

Male DBA/2J mouse; 20 mice/treatment

1.25% t

- butanol via liquid diet 3,390 mg/kg

- day (average)

7 days • Shock avoidance behavior was impaired after cessation of treatment.

• Latencies for experimental group was significantly ( p < 0.05) longer than the controls.

• One day after withdrawal, the number of trials where animals avoided shock was significantly ( p < 0.05) greater in the control group (control: 35/150; t

- butanol: 6/150).

• Even 2 days after withdrawal, treated animals showed a significant ( p < 0.01) deficit in avoidance response measured by both latency and number of avoidances (control: 51/100; t

- butanol: 35/100).

• At time of withdrawal, there was no difference in body temperature and withdrawal scores were low (0.20 ± 0.13, no t significantly different from the control). Body temperature

Control t

- butanol

3 hours after withdrawal 37.6 ± 0.1 34.9 ± 0.4*

8 hours after withdrawal 37.4 ± 0.1 35.7 ± 0.3*

Withdrawal scores

Control t

- butanol

3 hours after withdrawal 0 0.30 ± 0.21

8 hours after withdrawal 0 0.30 ± 0.15Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 27 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 7** 

**. Evidence pertaining to CNS**  **effects in**  **animals following exposure to**  tert

- **butanol (continued)** 

Reference and s tudy design Results

Thurman et al. ( 1980 )

Female Sprague

- Dawley rat; 12

– 14/treatment

Gavage; 5.7% in saline every 8 hours for up to 6 days 14 symptoms of withdrawal were scored according to severity on a 1

– 3 point scale (every 2 hours beginning 6

- 8 hours after last treatment.

• Treatment caused a steady increase in withdrawal score (results presented in a figure)

Inhalation

McComb and Goldstein ( 1979a )

Male Swiss

- Webster mouse ; 14 mice/treatment

Two 3

- day cycles separated by 24 hours of no treatment (withdrawal reaction observed for 24 hours following each exposure period); mice were maintained on either a low (range 3.5

– 6.4 mM) or high (range 6.5

– 10.0 mM) b lood alcohol level Withdrawal reaction consisted of signs of CNS hyperexcitability, including tremors, spontaneous convulsions, and c onvulsions elicited by handling Withdrawal peak height

Low blood alcohol High blood alcohol 3 days of exposure (n=11) 0.79 ± 0.12 1.43 ±0.07
6 days of exposure (n=13) 1.49 ± 0.08 1.63 ±0.10

McComb and Goldstein ( 1979b )

Male Swiss

- Webster mouse; 24 mice/treatment

Mice receive d an initial i.p. priming dose of 6.8 or 10.1 mmol/kg t- butanol; t

butanol vapor concentration was maintained between 50 and 80 μmol/m 3 for 24 hours then were steadily increased to maintain a constant blood alcohol level

Withdrawal was measured after 1, 3, 6 or 9 days • Withdrawal reaction consisted of signs of CNS hyperexcitability, including tremors, spontaneous convulsions, and convulsions elicited by handling.

• Intensity of withdrawal symptoms was related to blood levels and duration of exposure (results provided in figure only)

. o At mean blood levels of 5mM t

- butanol (range 2.4

- 6.6 mM), severity of withdrawal reaction rose steadily through day 9.

o At mean blood levels of 8.5 mM (range 6.8

- 10.7 mM) and above, a ceiling of withdrawal levels occurred at a height of approximately 1.5.

• Peak withdrawal reactions occurred 3

– 5 hours after removal of t

butanol and was 4

- 5 times greater than with ethanol.

* Statistically significant p ≤ 0.05 as determined by study authors

. 

### 1 2Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 28 DRAFT

— DO NOT CITE OR QUOTE 

## 2.6. **Other Systemic Effects ( Body Weight, Liver, and Urinary**  **Bladder)** 1

2 3 **Table 2-8** 

**. Evidence pertaining to effects on body weight**  **in animals following**  **oral exposure t**  **o** te rt

- **butanol**

Reference and s tudy design Results Acharya et al. ( 1995 )

Wistar rat; 5

– 6 males/treatment Drinking water (0 or 0.5 %), 0 or 575 mg/kg -d

10 weeks Body weight in treated animals lower than controls by ~7% (p< 0.05); (results only provided in a figure)

Lyondell Chemical Company ( 2004 )

Sprague

- Dawley rat; 12/sex/treatment Gavage 0, 64, 160, 400, or 1,000 mg/kg -d

At least 9 weeks beginning 4 weeks prior to mating through mating (up to 2 weeks), gestation, and lactation. Percent change compared to control:

Males Females

Dose (mg/kg

- d) Body weight Dose (mg/kg

- d) Body weight

0 0 0 0

64

− 2 64 0

16 0

− 4 16 0

− 2

40 0 +2 40 0 +1
1,00 0

− 7 1,00 0 +4

NTP ( 1995 )

F344/N rat; 10/sex/treatme nt Drinking water ( 0, 2.5, 5, 10, 20, or 40 mg/mL) M: 0, 230, 490, 840, 1,520, 3,610 a mg/kg -d F: 0, 290, 590, 850, 1,560, 3,620 a mg/kg -d

13 weeks Percent change compared to control:

Males Females

Dose (mg/kg

- d) Body weight Dose (mg/kg

- d) Body weight

0 0 0 0

230

− 4 290 +2

490

− 5* 590 +1

840

− 12* 850 +1
1,520

− 17* 1,560

− 2
3,610 All dead 3,620

− 21*

NTP ( 1995 )

B6C3F 1 mouse; 10/sex/treatment Drinking water ( 0, 2.5, 5, 10, 20, or 40 mg/mL) M: 0, 350, 640, 1,590, 3,940, 8,210 a mg/kg -d F: 0, 500, 820, 1,660, 6,430, 11,620 a mg/kg -d

13 weeks Percent change compared to control:

Males Females

Dose (mg/kg

- d) Body weight Dose (mg/kg

- d) Body weight

0 0 0 0

350

− 1 500 +3

640 +1 820

− 1
1,590

− 4 1,660 +4
3,940

− 14* 6,430

− 6
8,210

− 24* 11,620

− 15* High

- dose females had a significantly lower initial weight, but also had a significantly lower body weight gain indicating that there was some effect of
treatmentPreliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 29 DRAFT

— DO NOT CITE OR QUOTE 

### 1
**Table 2** 

**- 8** 

**. Evidence pertaining to e**  **ffects on body weight in**  **animals following oral exposure**  **to** tert

- **butanol (continued)** 

Reference and s tudy design Results

NTP ( 1995 )

F344/N rat; 60/sex/treatment (10/sex/treatment evaluated at 15 months) Drinking water ( 0, 1.25, 2.5, 5, 10 mg/mL) M: 0, 90, 200, 420 a mg/kg -d F: 0, 180, 330, 650 a mg/kg -d

2 years Percent change compared to control:

Males Females

Dose (mg/kg

- d) Body weight Dose (mg/kg

- d) Body weight

0 0 0 0

90

− 15 180

− 2

200

− 18 330

− 5

420

− 24 650

− 21
Only animals that survived at the end of 2 years were evaluated for body weight. Note: statistical significance not determined by study authors.

NTP ( 1995 )

B6C3F 1 mouse; 60/ sex/treatment Drinking water ( 0, 5, 10, 20 mg/mL) ; M: 0, 540, 1,040, 2,070 a mg/kg

- d F: 0, 510, 1,020, 2,110 mg/kg -d

2 years Percent change compared to control:

Males Females

Dose (mg/kg

- d) Body weight Dose (mg/kg

- d) Body weight

0 0 0 0

540 +1 510

− 2
1,040

− 2 1,020

− 3
2,070

− 1 2,110

− 12 Only animals that survived at the end of 2 years were evaluated for body weight. Note: statistical significance not determined by study authors.

a There was a significant decrease in survival in the high

- dose group. 

### 2 3 4 5 6 7 * Statistically significant p ≤ 0.05 as determined by study authors

. Conversion s from drinking water concentrations to mg/kg -d performed by study authors. Percentage change compared to control = ( treated value – control value) ÷ control value × 100.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 30 DRAFT

— DO NOT CITE OR QUOTE **Table 2-9** 

**. Evidence pertaining to liver effects**  **in animals following oral**  **exposure to**  tert

- **butanol** 

### 1 2

Reference and s tudy design Results
Liver weight
Acharya et al. ( 1995 )

Wistar rat; 5

– 6 males/treatment Drinking water (0 or 0.5%), 0 or 575 mg/kg -d

10 weeks No significant treatment

- related effects (results were only provided in a figure)

Lyondell Chemical Company ( 2004 )

Sprague

- Dawley rat; 12/sex/treatment Gavage 0, 64, 160, 400, or 1,000 mg/kg -d

At least 9 weeks beginning 4 weeks prior to mating through mating (up to 2 weeks), gestation, and lactation. Percent change compared to control:

Males Females

Dose (mg/kg

- d) Absolute weight Relative weight Dose (mg/kg

- d) Absolute weight Relative weight

0 0 0 0 0 0

64

− 1 0 64

− 4

− 4

16 0

− 3 +1 16 0

− 7

− 5

40 0

− 2

− 1 40 0 +2 +1

1,00 0 +8 +16* 1,00 0 +8 +3

NTP ( 1995 )

F344/N rat; 10/sex/treatment Drinking water ( 0, 2.5, 5, 10, 20, or 40 mg/mL) M: 0, 230, 490, 840, 1,520, 3,610 a mg/kg -d F: 0, 290, 590, 850, 1,560, 3,620 a mg/k g-d

13 weeks Percent change compared to control:

Males Females

Dose (mg/kg

- d) Absolute weight Relative weight Dose (mg/kg

- d) Absolute weight Relative weight

0 0 0 0 0 0

230

− 2 +4 290 +11* +9*

490 +1 +8* 590 +10* +9*

840 +5 +20* 850 +12* +11*

1,520 +8 +31* 1,560 +15* +16*

3,610 All dead All dead 3,620 +9* +41*

NTP ( 1995 )

B6C3F 1 mouse; 10/sex/treatment Drinking water ( 0, 2.5, 5, 10, 20, or 40 mg/mL) M: 0, 350, 640, 1,590, 3,940, 8,210 a mg/kg -d F: 0, 500, 820, 1,660, 6,430, 11,620 a mg/kg -d

13 weeks Percent change compared to control:

Males Females

Dose (mg/kg

- d) Absolute weight Relative weight Dose (mg/kg

- d) Absolute weight Relative weight

0 0 0 0 0 0

350 +2 +3 500

− 1

− 4

640

− 1

− 2 820

− 5

− 3

1,590

− 1 +5 1,660

− 8

− 9*

3,940 0 +14* 6,430

− 2 +6

8,210

− 16 +22* 11,620

− 6 +13*Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 31 DRAFT

— DO NOT CITE OR QUOTE 

### 1
**Table 2-9** 

**. Evidence pertaining to liver effects**  **in animals following oral**  **exposure to**  tert

- **butanol (continued)** 

Reference and s tudy design Results

NTP ( 1995 )

F344/N rat; 60/sex/treatment (10/sex/treatment evaluated at 15 months) Drinking water ( 0, 1.25, 2.5, 5 or 10 mg/mL) M: 0, 90, 200, or 420 a mg/kg -d F: 0, 180, 330, or 650 a mg/kg -d

2 years Percent change compared to control:

Males Females

Dose (mg/kg

- d) Absolute weight Relative weight Dose (mg/kg

- d) Absolute weight Relative weight

0 0 0 0 0 0

90 +2 +7 180

− 14*

− 8

200 +8 +11 330

− 3

− 1

420 +1 +14* 650

− 6 +9*

Only animals sacrificed at 15 months were evaluated for organ weights.
Histopathology
Acharya et al. ( 1997 ; 1995 )

Wistar rat; 5

– 6 males/treatment Drinking water (0, 0.5%), 0, 575 mg/kg -d

10 weeks ↑ liver glycogen (~ 7 fold) *

↑incidence of centrilobular necrosis, vacuolation of hepatocytes, loss of hepatocyte architecture, peripheral proliferation, and lymphocyte infiltration (incidences and results of statistical tests not reported)

Lyondell Chemical Company ( 2004 )

Sprague

- Dawley rat; 12/sex/treatment Gavage 0, 64, 160, 400, or 1
, 000 mg/kg -d

At least 9 weeks beginning 4 weeks prior to mating through mating (up to 2 weeks),
ge station, and lactation. No treatment

- related effects observed.

NTP ( 1995 )

F344/N rat; 10/sex/treatment Drinking water ( 0, 2.5, 5, 10, 20, or 40 mg/mL) M: 0, 230, 490, 840, 1,520, 3,610 a mg/kg -d F: 0, 290, 590, 850, 1,560, 3,620 a mg/kg -d

13 weeks No treatment

- related effects observed (histopathology data for the 13

- week study were not provided)

NTP ( 1995 )

B6C3F 1 mouse; 10/sex/treatment Drinking water ( 0, 2.5, 5, 10, 20, 40 mg/mL) M: 0, 350, 640, 1,590, 3,940, 8,210 a mg/kg -d F: 0, 500, 820, 1,660, 6,430, 11,620 a mg/kg -d 13 weeks No treatment

- related effects observed (histopathology data for the 13

- week study were not provided)Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 32 DRAFT

— DO NOT CITE OR QUOTE **Table 2-9** 

**. Evidence pertaining to liver effects**  **in animals following oral**  **exposure to**  tert

- **butanol (continued)** 

Reference and s tudy design Results

NTP ( 1995 )

F344/N rat; 60/sex/treatment (10/sex/treatment evaluated at 15 months) Drinking water ( 0, 1.25, 2.5, 5, 10 mg/mL) M: 0, 90, 200, or 420 a mg/kg -d F: 0, 180, 330, or 650 a mg/kg -d

2 years No treatment

- related effects observed.

NTP ( 1995 ) B6C3F 1 mouse; 60/sex/treatment Drinking water ( 0, 5, 10, 20 mg/mL) M: 0, 540, 1,040, or 2,070 a mg/kg

- d

F: 0, 510, 1,020, or 2,110 mg/kg -d

2 years Males Females

Dose (mg/kg

- d) Incidence of fatty change Dose (mg/kg

- d) Incidence of fatty change

0 12/59 0 11/60

540 5/60 510 8/60
1,040 8/59 1,020 8/60
2,070 29/59* 2,110 6/60

a The high

- dose group had an increase in mortality. 

### 1 2 3 4 5 * Statistically significant p ≤ 0.05 as determined by study authors

. Conversions from drinking water concentrations to mg/kg

- d per f ormed by study authors. Percentage change compared to control = (treated value – c o ntrol value) ÷ control value × 100.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 33 DRAFT

— DO NOT CITE OR QUOTE **Table 2-**  **10**

**. Evidence pertaining to l**  **iver effects in**  **animals following** 

### 1 2 **i nhalation exposure**  **to** tert

- **butanol**

Reference and s tudy design Results
Liver Weight

NTP ( 1997 )

F344/N rat; 10/sex/treatment Analytical concentration: 0, 13 5 , 27 0 , 54 0
, 1,080, or 2,101 ppm (0, 406, 824, 1,643, 3,273 or 6,368 mg/m 3 ) 6 hr/d, 5 d/wk

13 weeks Percent change compared to control:

Males Females

Dose (mg/ m 3 ) Absolute weight Relative weight Absolute weight Relative weight

0 0 0 0 0

406

− 8

− 8 0 +3

824

− 2

− 1 0 0

1,643 +1

− 1 +3 +2

3,273 +10 +7 +9 +9*

6,368 +5 +5 +4 +8*

NTP ( 1997 )

B6C3F1 mouse; 10/sex/treatment Analytical concentration: 0, 134, 272, 542, 1,080, or 2,101 ppm (0, 406, 824, 1,643, 3,273 or 6,368 mg/m 3) 6 hr/d, 5 d/wk

13 weeks Percent change compared to control:

Males Females

Dose (mg/ m 3 ) Absolute weight Relative weight Absolute weight Relative weight

0 0 0 0 0

406

− 1 0 +1

− 4

824 +4 +9 +1 +5

1,643 +7 +5 +5 +1

3,273

− 8

- 2 +2 +9*

6,368 +5 +7 +8 +21* * Statistically significant p ≤ 0.05 as determined by the study authors

. 

### 3 4 5 6 7
8 Conversion from ppm to mg/m 3 is 1 ppm = 3.031 mg/m 3

. Percentage change compared to control = (treated value – c o ntrol value) ÷ control value × 100.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 34 DRAFT

— DO NOT CITE OR QUOTE **T able**  **2- 11** 

**. Evidence**  **pertaining to u rinary bladder effects**  **in animals**  

### 1 2 **following oral exposure to**  tert

- **butanol**

Reference and s tudy design Results
Histopathology

NTP ( 1995 )

F344/N rat; 10/sex/treatment Drinking water ( 0, 2.5, 5, 10, 20, 40 mg/mL) M: 0, 230, 490, 840, 1,520, 3,610 a mg/kg -d F: 0, 290, 590, 850, 1,560, 3,620 a mg/kg -d

13 weeks Incidence (severity) :
Males Females

Dose (mg/kg

- d) T ransitional epithelial hyperplasia Dose (mg/kg

- d) T ransitional epithelial hyperplasia

0 0/10 0 0/10

230 n ot evaluated 290 not evaluated

490 n ot evaluated 590 not evaluated

840 0/10 850 not evaluated

1,520 1/10 (3.0) 1,560 0/10

3,610 7/10* (2.9) 3,620 3/10 (2.0)
Severity: 1 = minimal, 2 = mild, 3 = moderate, 4 = marked

NTP ( 1995 )

B6C3F 1 mouse; 10/sex/treatment Drinking water ( 0, 2.5, 5, 10, 20, 40 mg/mL) M: 0, 350, 640, 1,590, 3,940, 8,210 a mg/kg -d F: 0, 500, 820, 1,660, 6,430, 11,620 a mg/kg -d

13 weeks Incidence (severity):

Males Females

Dose (mg/kg

- d) Transitional epithelial hyperplasia Inflam

mation Dose (mg/kg

- d) Transitional epithelial hyperplasia Inflam

mation

0 0/10 0/10 0 0/10 0/10

350 not evaluated 500 0/1 0/1

640 not evaluated 820 not evaluated

1,590 0/10 0/10 1,660 not evaluated

3,940 6/10* (1.3) 6/10* (1.3) 6,430 0/10 0/10

8,210 10/10* (2.0) 10/10* (2.3) 11,620 3/9 (2.0) 6/9* (1.2)
Severity: 1 = minimal, 2 = mild, 3 = moderate, 4 = marked

NTP ( 1995 )

F344/N rat; 60/sex/treatment (10/sex/treatment evaluated at 15 months) Drinking water ( 0, 1.25, 2.5, 5, or 10 mg/mL) M: 0, 90, 200, 420 a mg/kg -d F: 0, 180, 330, 650 a mg/kg -d

2 years No treatment

- related effects observed



### 3Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 35 DRAFT

— DO NOT CITE OR QUOTE **Table 2** 

**- 11** 

**. Evidence pertaining to u**  **rinary bladder effects in**  **animals following oral**  **exposure to**  tert

- **butanol (continued)** 

Reference and s tudy design Results

NTP ( 1995 )

B6C3F 1 mouse; 60/sex/treatment Drinking water ( 0, 5, 10, or 20 mg/mL) M: 0, 540, 1,040, 2,070 a mg/kg

- d F: 0, 510, 1,020, 2,110 mg/kg -d

2 years Incidence (s everity) :
Males Females

Dose (mg/kg

- d) Transitional epithelial hyperplasia Inflam

mation Dose (mg/kg

- d) Transitional epithelial hyperplasia Inflam

mation

0 1/59 (2.0) 0/59 0 0/59 0/59

540 3/59 (1.7) 3/59 (1.7) 510 0/60 0/60

1,040 1/58 (1.0) 1/58 (1.0) 1,020 0/59 0/59

2,070 17/59* (1.8) 37/59* (2.0) 2,110 3/57 (1.0) 4/57* (2.0)

S everity: 1 = minimal, 2 = mild, 3 = moderate, 4 = marked

a The high

- dose group had an increase in mortality. 

### 1 2 3 4 * Statistically significant p ≤ 0.05 as determined by study authors

. Conversions from drinking water concentrations to mg/kg

- d per formed by study authors.Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 36 DRAFT

— DO NOT CITE OR QUOTE 

### 1 2

3 4 5

6 10 100 1,000 10,000 100,000Dose (mg/kg-day)M rat (D)M rat (A)F rat (D)M mouse (D)F mouse (D)M rat (C)F rat (C)Transitional epithelium hyperplasia; M rat (D)Transitional epithelium hyperplasia; F rat (D)Transitional epithelium hyperplasia; M mouse (D)Inflammation; M mouse (D)Inflammation; F mouse (D)Transitional epithelium hyperplasia; F mouse (D)Transitional epithelium hyperplasia; M mouse (D)Inflammation; M mouse (D)Inflammation; F mouse (D)Transitional epithelium hyperplasia; F mouse (D)Increased glycogen; M rat (B)Relative weight; M rat (B)Relative weight; M rat (D)Absolute weight; F rat (D)Relative weight; F rat (D)Absolute weight; M rat (D)Relative weight; M mouse (D)Relative weight; F mouse (D)Absolute weight; M mouse (D)Absolute weight; F mouse (D)Relative weight; M rat (D)Relative weight; F rat (D)Absolute weight; M rat (D)Absolute weight; F rat (D)Fatty tissue; M mouse (D)Fatty tissue; F mouse (D)Relative weight; M rat (C)Absolute weight; M rat (C)Absolute weight; F rat (C)Relative weight; F rat (C)

BODY WEIGHT Subchronic URINARY BLADDER LIVER Subchronic Chronic Subchronic ChronicReproductive

Reproductive  = exposures at which the endpoint was reported statistically significant by study authors  = exposures at which the endpoint was reported not statistically significant by study authors

× = exposures at which all animals were dead and unable to be examined for the endpoint

Sources: (A) Acharya et al. ( 1995 ) ; (B) Acharya et al. ( 1997 ; 1995 ) ; ( C) Lyondell ( 2004 ) ; (D) NTP ( 1995 )

**Figure 2-6** 

**. Exposure** 

**- response**  **array of body weight, liver effects, and urinary**  **bladder effects following oral exposure to**  tert

- **bu tanol.** Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 37 DRAFT

— DO NOT CITE OR QUOTE 

## 2.7. **Genotoxic Effects**

**Table 2-**  **12**

**. Evidence pertaining to genotoxic effects of**  tert

- **butanol in vitro**  **and in vivo studies**  

### 1

2 3

Species/ cell line Test system (strain/species) Exposure (dose/ concentration) Results Metabolic activation
Reference

– S9 +S9
In vitro Bacterial systems
Salmonella typhimurium Reverse mutation (TA98, TA100, TA1535, TA1537, TA1538) 10 mg/plate

– – Zeiger et al. ( 1987 )

2.9

– 10,000 μ g/plate

– – ARCO ( 1994e )

Reverse mutation (TA102) 1

– 4 mg/plate ND + Williams

- Hill et al. ( 1999 )

Reverse mutation (TA98, TA100, TA102, TA1535, TA1537) 5

– 5,000 μg/plate

– – McGregor et al. ( 2005 )

E.coli Reverse mutation (WP2 uvrA/PKM101) 5

– 5,000 μg/plate

– – McGregor et al. ( 2005 )

Neurospora crassa Reverse mutation, ad

3A locus (allele 38701) 1.75mol/L

– – Dickey et al. ( 1949 )

Saccharomyces cerevisiae Mitochondrial mutation (K5 -5A, MMY1, D517

- 4B and DS8) 4.0% (vol/vol) + a ND Jimenez et al. ( 1988 )

Mammalian cells

– rodent Mouse lymphoma cells L5178Y TK +/

– Gene mutation 625

– 5,000 mg/m L

– – McGregor et al. ( 1988 )

2.4

– 32 μ L /m L

– – ARCO ( 1994a )
Chi nese hamster
ovary Sister

- chromatid exchange 1.25

– 5 mg/m L

– – NTP ( 1995 )

0.31

– 20 μ L /m L (- S9); 0. 625

– 20 μL /m L (+S9) – b

– ARCO ( 1994b )

Chromosomal aberrations 1.25

– 5 mg/m L

– – NTP ( 1995 )

Rat fibroblasts DNA damage (comet assay) 0.44 mmol/L (IC 50 ) + c ND Sgambato et al. ( 2009 )Preliminary Materials for the IRIS Toxicological Review of **tert**

- Butanol

This document is a draft for review p urposes only and does not constitute Agency policy. 2- 38 DRAFT

— DO NOT CITE OR QUOTE Species/ cell line Test system (strain/species) Exposure (dose/ concentration) Results Metabolic activation
Reference

– S9 +S9
Mammalian cells

– human

Human HL

- 60 leukemia cells DNA damage (comet assay) 1

– 30 mmol/L + ND Tang et al. ( 1997 )

In vivo
Mammalian

– rodent Male Kunming mouse liver, kidney and lung cells DNA adducts 0.1

– 1,000 μg/kg b.w. + NA Yuan et al. ( 2007 )

B6C3F 1 mouse peripheral blood cells Micronucleus formation 2.5

– 40 mg/m L drinking water – NA NTP ( 1995 )

ND = not determined; NA = not applicable 

### 1 2 3 4 5 6
7 a Effect is predicted to be due to mitochondrial membrane composition

. b Results were stated to be statistically increased in the 20 μg/m L with and without activation and the 10 μg/m L with activation,

but results did not meet positive criteria for the assay.

c DNA damage was completely reversed with increase in time of exposure

.