SRC TR - 1 3 - 076

HPV 1 /HPV2 TEST RULE DATA ADEQU ACY REVIEW : DECEMBER 2013 UPDATE (DRAFT) Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1 Prepared by: Justine Von Runnen , Julie Klotzbach, Fernando Llados , Ashlee Aldridge, Amy Nguyen,

Jennifer Rayner, William Richards, and Teresa Manyin Defense and Environmental Solutions Division

SRC, Inc. 7502 Round Pond Road North Syracuse, NY 13212 Contract No. EP - W - 09 - 027 Task FG007. 4 .009.004 Submitted to: OCSPP/OPPT/RAD (7403M) U.S . Environmental Protection Agency 1200 Pennsylvania Avenue Washington, D.C . 20460 Bryan Lobar, Project Officer David Brooks, Work Assignment Manager Amy Benson, Alternate Work Ass ignment Manager

December 5 , 201 3Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

i TABLE OF CONTENTS A. SOURCE MATERIAL USED FOR ASSESSMENT ................................ .............................. 1 B. EVALUATING DATA ADEQUACY ................................ ................................ ..................... 5 C. DATA FOR SIDS ENDPOINTS ................................ ................................ .............................. 6 C - 1. Acute Toxicity to Fish ................................ ................................ ................................ ........ 6 C - 2. Acute Toxicity to Aquatic Invertebrates ................................ ................................ .......... 12 C - 3. Chronic Toxicity to Aquatic Invertebrates ................................ ................................ ....... 18 C - 4. Toxicity to Aquatic Plants ................................ ................................ ................................ 25 C - 5. Acute Toxicity ................................ ................................ ................................ .................. 34 C - 6. Repeated - Dose Toxicity ................................ ................................ ................................ ... 46 C - 7. Genetic Toxicity

– Gene Mutation ................................ ................................ ................... 63 C - 8. Genetic Toxicity

– Chromosomal Aberrations ................................ ................................ 77 C - 9. Reproductive/Developmental Toxicity ................................ ................................ ............ 83 D. SUPPLEMENTAL DATA (NON - SIDS ENDPOINTS) ................................ ...................... 104 D - 1. Carcinogenicity ................................ ................................ ................................ .............. 104 D - 2. Skin Irritation ................................ ................................ ................................ ................. 108 D - 3. Eye Irritation ................................ ................................ ................................ .................. 114 E. DATA ADEQUACY SUMMARY TABLE ................................ ................................ ......... 116Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

1 A . SOURCE MATERIAL USED FOR ASSESSMENT In this document, the toxicity of p henol, 4,4' - methylenebis[2,6 - bis(1,1 - dimethylethyl) - (CAS RN 118 - 82 - 1) is assessed based on toxicological data for CASRN 118 - 82 - 1 and the supporting chemical, 2,6 - di - tert - butylphenol ( CASRN 1 2 8 - 39 - 2 ) ; available data are summarized and evaluated for data adequacy . This document represents an update of the data adequacy review written previously for this chemical : EPA . 2007 . Data Adequacy Review and Robust Study Summary Adequacy Requirements : 4,4 ’ - Methylenebis(2,6 - di - tert - butylphenol) ( CAS No. 118 - 82 - 1 ) prepared by Oak Ridge National Laboratory, Oak Ridge, TN . October 18, 2007 . This document includes data submitted in response to HPV TSCA Section 4 Test Rules : (1) Testing of Certain High Production Volume Chemicals; Final Rule, 71 FR 13708 , March 16, 2006; available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ OPPT - 2005 - 0033 - 0197 as of December 4, 2013 ; referred to herein as HPV1 ; and (2) Testing of Certain High Production Volume Chemicals; Second Group of Chemicals; Final Rule, 76 FR 1067, January 7, 2011; available online at http://www.regul ations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2007 - 0531 - 0579 as of December 4, 2013 ; referred to herein as HPV2 . The HPV1 test rule required the following toxicological tests for CASRN 118 - 82 - 1 : D (acute mammalian toxicity), E1 (bacterial reverse mutation test), E2 (chromosomal aberration or micronucleus test), and F2 (reproduction/developmental toxicity screening test) . From July 14, 2006 to May 1 7 , 2007 , Albemarle Corporation, the sponsor for CAS RN 118 - 82 - 1, submitted the following existing information on acute mammalian toxicity, gene mutation , and reproductive/developmental toxicity endpoints in response to the D, E1 , and F2 testing requirements . In addition, the July 14, 2006 letter indicated that Albemarle “will conduct the testing as re quired for E2” (chromosomal aberration or micronucleus test) . Letter to Document Control Office, Office of Pollution Prevention and Toxics, U . S . EPA, Attn : TSCA Section 4, from Kim Boudreaux, Regulatory Advisor, Albemarle Corporation , Baton Rouge, Louisiana , dated July 14, 2006, with three attachments . A vailable online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT 2005 - 0033 - 0181 as of December 4, 2013 . Brooks, TM, and D.E . Wiggins . 1987 . Microbial Mutagenicity Studies with IONOX 220 . Shell Research Limited, Sittingbourne Research Center . Submitted to EPA on May 1 7 , 2007 . Available online at http://www.regulations.gov/#!documentDetail;D=EPA HQ - OPPT - 2005 - 0033 - 0250 as of December 4, 2013 . Following review of this existing information for CASRN 118 - 82 - 1 , EPA determined that the dat a submitted for acute mammalian toxicity and bacterial reverse mutation were adequate to satisfy the D and E1 requirements of the Test Rule . H owever, the existing studies submitted for the reproductive/developmental toxicity endpoint (conducted with CASRN 118 - 82 - 1) w ere not considered to be adequate to satisfy the F2 testing requirement .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

2 In a letter dated May 16, 2007 (available online at http://www.regulations.gov/#!document Detail;D=EPA - HQ - OPPT - 2005 - 0033 - 0247 as of December 4, 2013 ), Albemarle Corporation proposed the use of 2,6 - di - tert - butylphenol (CASRN 128 - 39 - 2) as a supporting chemical for characterizing the effects of phenol, 4,4' - methylenebis[2,6 - bis(1,1 - dimethylethyl) - (CASRN 118 - 82 - 1) on human health . EPA considers CASRN 128 - 39 - 2 to be an appropriate supporting chemical for CASRN 118 - 82 - 1 b ased on structural and functional similarity . Therefore, EPA accepts the use of toxicological data from studies with CASRN 128 - 39 - 2 for the purposes of characterizing the effects of CASRN 118 - 82 - 1 on human health . EPA identified the following gene mutation and reproductive/developmental toxicity studies , conducted with the acceptable supporting chemical (CAS RN 128 - 39 - 2) , which satisfy the F2 testing requirement (reproduction/developmental toxicity screening test) and provide mammalian cell mutagenicity data that supplement the bacterial mutagenicity data for CASRN 118 - 82 - 1, submitted by Albemarle Corporation for the E1 testing requirement. Sandoz Chemical s Ltd . 1991 . Unpublished CCR Report [ on a mammalian cell gene mutation assay] - Sandoz, 243617 . Muttenz, Switzerland . S ummarized in the online ECHA ( http :// echa . europa . eu ) entry for CASRN 128 - 39 - 2 (s tudy indicated as Exp Key Genetic t oxicity in vitro .001 ) . Secondary reference : OECD . 2002 . SIDS/IRPTC Data Profile for 2,6 - Di - tert - butylphenol (CASRN 128 - 39 - 2) . UNEP Publications . 2002 . A vailable online at http://www.chem.unep.ch/irptc/sids/OECDSIDS/128392.pdf as of December 4, 2013 . Sandoz Chemical s Ltd . 1992 . Unpublished CCR Report [on a reproduction/developmental toxicity test] - Sandoz, 321794 . Muttenz, Switzerland . S ummarized in the online ECHA ( http :// echa . europa . eu ) entry for CASRN 128 - 39 - 2 (s tudy indicated as Exp Key Toxicity to reproduction.001 ) . Secondary reference : OECD . 2002 . SIDS/ IRPTC Data Profile for 2,6 - Di - tert - butylphenol ( CAS RN 128 - 39 - 2) . UNEP Publications . 2002 . A vailable online at http://www.chem.unep.ch/irptc/sids /OECDSIDS/128392.pdf as of December 4, 2013 . Due to the identification of adequate studies on acute mammalian toxicity and gene mutation for the test rule chemical (CASRN 118 - 82 - 1) and on reproductive/developmental toxicity for an acceptable supporting ch emical (CASRN 128 - 39 - 2) , EPA revoked the D, E1 , and F2 testing requirements for CASRN 118 - 82 - 1 (Revocation of TSCA Section 4 Testing Requirements for Certain High Production Volume Chemical Substances; Direct Final Rule, 77 FR 15609, March 16, 2012; availa ble online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT 2005 - 0033 - 0402 as of December 4, 2013 ) . To address the E2 testing requirement, Albemarle Corporation contracted TNO Laboratories to conduct a chromosomal aberration study and submitted the following report on June 28, 2007 ; EPA reviewed this chromosomal aberration study and considered it adequate to satisfy the E2 testing requirement promulgated in the HPV1 test rule :Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

3 de Vogel, N . and Usta, B . 2007 . Chromosomal Aberration test with AN - 2 in Cultured Chinese Hamster Ovary (CHO) Cells . TNO Laboratories, Zeist, Netherlands, Project No. 031.11113/01.42, Study No. 6802/24, May 9, 2007 . Submission documented online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0257 as of December 4, 2013 . The HPV2 test rule require d C1 aquatic toxicity testing for CASRN 118 - 82 - 1 . The log K ow value of CASRN 118 - 82 - 1 was experimentally determined to be > 6.50 by SafePharm Laboratories , 1 indicating that testing under Test Group 2, which applies to chemicals with log K ow ≥ 4.2, is required . Test Group 2 includes the following tests : (1) Chronic Toxicity to Daphnia (ASTM E 1193 - 97) and (2) Toxicity to Plants (Algae; ASTM E 1218 - 04) . Albemarle submitted comments (available online at http://www.regulations.gov/#!document Detail;D=EPA - HQ - OPPT - 2007 - 0531 - 0068 as of December 4, 2013 ) in response to the proposed HPV2 test rule on October 21, 2008, suggesting that the following study by Mattock and Girling (1988) would satisfy the toxicity to plants testing requirement; EPA reviewed this study and considered it to be inadequate to satisfy any component of the C1 testing requirement: Mattock, S.D . an d Girling, A.E . 1988 . IONOX 220 : Acute Toxicity ( Salmo gairdneri, Daphnia magna and Selenastrum capricornutum ) and n - octanol/water partition coefficient . Sittingbourne Research Centre, Shell Research Limited . Submitted by Shell Oil Co. to EPA in 1992 as part of a TSCA 8(e) submission (EPA Doc. No. 88 - 920003444 , Fiche No. OTS0537403 ) entitled “Initial submission : acute toxicity of IONOX 220 in Salmo gairdneri, Daphnia magna and Selenastrum capricornutum and n octanol/water partition coef ficient w - letter dated 053092”. The following existing aquatic toxicity data , submitted by Albemarle Corporation on July 14, 2006 , were also reviewed by EPA and considered to be inadequate to satisfy the C1 testing requirement : Stonybrook Laboratories Inc . 1995 . Study No. 6612 5 , conducted for Ethyl Corporation . Source : Ethyl Petroleum Additives International , Bracknell, Berkshire . ( Summarized in IUCLID Dataset for CASRN 118 - 82 - 1 , European Chemical Bureau, 2000) . Robust summary available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ OPPT - 2005 - 0033 - 0181 as of December 4, 2013 . Stonybrook Laboratories Inc . 1995 . Study No. 6612 6, conducted for Ethyl Corporation . Source : Ethyl Petroleum Additives International , Bracknell, Berkshire . ( Summarized in IUCLID Dataset for CASRN 118 - 82 - 1 , European Chemical Bureau, 2000) . Robust summary available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ OPPT - 2005 - 0033 - 0181 as of December 4, 2013 .

1 Study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations. gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013 ; “4,4’ - Methylene bis(2,6 - di tert - butylphenol) : Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/011, study completed on July 18, 2005 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

4 Albemarle Corporation contracted Wildlife International, Ltd . to conduct chronic toxicity to Daphnia and toxicity to algae studies and submitted the following studies o n June 29, 2012 to address the C1 - Test Group 2 testing requirement ; EPA considers these studies adequate to satisfy the C1 testing requirement promulgated in the HPV2 test rule : Minderh out, T., Kendall, T.Z., Gallagher, S. P. and Krueger, H.O . 2012 . AN - 2 : A flow through life - cycle toxicity test with the cladoce ran ( Daphnia magna ) . Final Report . Wildlife International, Ltd. , Easton, MD . Project No. 471A - 120 . Study report a vailable online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2007 - 0531 - 0678 as of December 4, 2013 . Arnie, J.R., Porch, J.R., Kendall, T.Z . and Krueger, H.O . 2012 . AN - 2 : A 96 - hour toxicity test with the freshwater al ga ( Pseudokirchneriella subcapitata ) . Wildlife International, Ltd. , Easton, MD . Project No. 471A - 119 . Study report a vailable online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2007 - 0531 - 0678 as of December 4, 2013 . Although the test rules did not require repeated - dose toxicity testing, the following repeated - dose studies were included herein to provide a complete assessment of toxicological SIDS endpoints : Jolley, W.P., Stemmer, K.L., Grande, F., Richmond, J., and Pfitzer, E.A . 1969 . T he effect s exerted upon beagle dogs during a period of two y ea rs by the introduction of 4,4' - methyle n ebis (2,6 - ditert i ary butylphenol) into their daily diets . Kettering Laboratory, University of Cincinnati, OH . Submitted to EPA in 1992 as part of a TSCA 8 (e) submission ( EPA Doc. No. 88 - 920001353S , Fiche No. OTS0536073) entitled “ Initial sub : the effect exerted upon beagle dogs during a period of 2 - yrs by the introduction of 4,4' - methyle n ebis (2,6 - ditertaryl butylphenol) int o their daily diets (sanitized)” . Jolley, W.P., Stemmer, K.L., Pfitzer, E.A., Grande, F., and Richmond, J . 1975 . The effects upon beagle dogs following the ingestion of 4,4' - methylenebis (2,6 - ditertiary butylphenol) for two years including the changes after an additional seven months on control diet . Kettering Laboratory, University of Cincinnati, OH . Submitted to EPA in 1992 as part of a TSCA 8 (e) submission ( EPA Doc. No. 88 - 920001353S , Fiche No. OTS0536073) entitled “ Initial sub : the effect exerted upon beagle dogs during a period of 2 - yrs by the introduction of 4,4' - methyle n ebis (2,6 - ditertaryl butylphenol) int o their daily diets (sanitized)” . ECHA. Oral repeated - dose toxicity in rats (Exp WoE Repeated dose toxicity: oral.003). Study summary available from the ECHA website at http://apps.echa.europa.eu/registered/data/dossiers/DISS - dcee47d2 - 62ad - 0171 - e044 - 00144f67d031/AGGR - bf547f31 - 7fe6 - 4a5f - a06a - e41a1a9a2ab0_DISS - dcee47d2 - 62ad 0171 - e044 - 00144f67d031.html#AGGR - bf547f31 - 7fe6 - 4a5f - a06a - e41a1a9a2ab0 as of December 4, 2013 . The data source is referred to as a “study report”, with a report date of 1969 - 07 - 08. No other reference information is provided.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

5 Data from all studies submitted by Albemarle Corporation , as well as other available data for SIDS toxi city endpoints (including data evaluated in the previous data adequacy review [ EPA , 2007 1 ]) , are evaluated for adequacy in Section C of this document . Additional data for non SIDS toxicity endpoints (“supplemental” data) , including skin and eye irritation, are reviewed in Section D . Sections C and D incorporate information from the High Production Volume Information System (HPVIS) and the European Chemicals Agency ( ECHA) website ( http://echa.europa.eu/ ) . B . EVALUATING DATA ADEQUACY Each study was summarized according to a standard format and assigned a Klimisch reliability code based on its scientific merit and conduct, using professional judgment . To determine whether data were adequate to satisfy SIDS endpoints, study details were compared to current standard test guidelines to establish whether the study met current testing requirements . Studies for a common endpoint are grouped together in the document and an endpoint adequacy assessment is prov ided near the top of the first page of the summaries for that endpoint . If the data available for the endpoint were determined to be inadequate, a basis for that conclusion is supplied . A table summarizing data availability/adequacy for each SIDS endpoin t is included at the end of the summary document .

1 EPA. 2007. Data Adequacy Review and Robust Study Summary Ad equacy Requirements: 4,4’ - Methylenebis(2,6 - di - tert - butylphenol) (CAS No. 118 - 82 - 1) prepared by Oak Ridge National Laboratory, Oak Ridge, TN. October 18, 2007.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

6 C . DATA FOR SIDS ENDPOINTS C - 1 . Acute Toxicity to Fish Summary of endpoint : Not A dequate . Testing of a cute toxicity to fish is not required for the chemical because the lo g K ow is > 4.2 . 1 Study 1 : Acute toxicity study in rainbow trout ( Mattock and Girling, 1988 ) Title : IONOX 220 : Acute toxicity ( Salmo gairdneri , Daphnia magna , and Selenastrum capricornutum ) and n - octanol/water partition coefficient This study is: Adequate Not Adequate This study is not adequate because a nalytical monitoring was not conducted and the test concentrations (10

– 1000 mg/L) exceeded the water solubility limit of the test substance ( experimentally determined to be < 0.01 mg/L) 2 by many orders of magnitude , although a vehicle was not used . In addition , the study is non - GLP, a test method/guideline was not specified, and the temperature varied by more than 2°C . TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; referred to as IONOX 220 in study report Remarks : 99.04% purity (by GC); pale yellow powder; obtained from San Celoni, Barcelona, Spain METHOD Method/guideline followed : Not specified Test type : Acute T oxicity to F ish ( S emi - S tatic ) GLP compliant? : No Year study was performed : 1988
1 The log K ow value of CASRN 118 - 82 - 1 was experimentally determined to be > 6.50 by SafePharm L aboratories (study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations. gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013; “4,4’ - Methylene bis(2,6 - di tert - butylphenol): Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/011, study completed on July 18, 2005). 2 T he water solubility limit of CASRN 118 - 82 - 1 was experimentally determined to be < 0.01 mg/L by SafePharm Laboratories (study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013; “4,4’ - Methylene bis(2,6 - di - tert - butylphenol): Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/011, study completed on Ju ly 18, 2005).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

7 Test species : Rainbow trout ( Salmo gairdneri ) ; obtained from Itchin Valley Trout Farm, Hampshire ; mean length of 3.6 cm and mean weight of 0.81g (n = 10) Analytical monitoring : No Exposure duration : 96 hours Concentrations tested : 0 (control), 10, 30, 100, 300 , and 1000 mg/L Test method/conditions remarks : Test solutions were renewed daily. Ten fish per concentration were placed in g lass aquaria filled with 10 L of filtered dechlorinated main water . Fish were not fed during exposure . Test Conditions: Temperature : 13

– 17°C pH : 7.7

– 8.3 Hardness : 242

– 260 mg CaCO 3 /L Dissolved Oxygen : 9.6

– 10.4 mg/L Statistics : The 72 - and 96 - hour LC 50 values were calculated using probit analysis after log transformation of the concentrations . RESULTS Endpoint value(s) : Note that analytical monitoring was not conducted and nominal concentrations (10 - 1000 mg/L) were many orders of magnitude greater tha n the water solubility limit of the test substance ( experimentally determined to be < 0.01 mg/L) 1 . Therefore, it is not possible to interpret the reported toxicity values. 72 - hour LC 50 = 870 mg/L; 96 - hour LC 50 = 820 mg/L , based on nominal concentrations . Results remarks : Of the ten fish exposed to the highest concentration (1000 mg/L), three had died within 24 hours, five had died within 48 hours , and six had died within 72

– 96 hours . One death occurred in the 300 mg/L dose group at 96 - hours. Page 2 of the test report, which included studies for fish, Daphnia , and green algae, provided a general comment that the test substance “appeared to be not wholly soluble at concentrations greater than” 10 mg/L .

1 T he water solubility limit of CASRN 118 - 82 - 1 was experimentally determined to be < 0.01 mg/L by SafePharm Laboratories (study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013; “4,4’ - Methylene bis(2,6 - di - tert - butylphenol): Determination of Water Solubility and Partition Coeff icient”, SPL Project Number 1193/011, study completed on July 18, 2005).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

8 CONCLUSIONS The 96 - hour LC 50 for rainbow trout is 820 mg/L (95% CI, 490

– 2600 mg/L) based on nominal concentrations . STUDY RELIABILITY [ 3 ] Not r eliable . Non - GLP study . Test method/guideline was not specified, a nalytical monitoring was not conducted, the temperature varied by more than 2°C, and the test concentrations exceeded the water solubility limit of the test substance ( experimentally determined to be < 0.01 mg/L) 1 by many orders of magnitude , although a vehicle was not used . REFERENCE Mattock, S.D . and Girling, A.E . 1988 . IONOX 220 : Acute Toxicity ( Salmo gairdneri, Daphnia magna and Selenastrum capricornutum ) and n - octanol/water partition coefficient . Sittingbourne Research Centre, Shell Research Limited . Submitted by Shell Oil Co. to EPA in 1992 as part of a TSCA 8(e) submission (EPA Doc . No . 88 - 920003444, Fiche No . OTS0537403) en titled “Initial submission : acute toxicity of IONOX 220 in Salmo gairdneri, Daphnia magna and Selenastrum capricornutum and n - octanol/water partition coefficient w - letter dated 053092”.

1 T he water solubility limit of CASRN 118 - 82 - 1 was experimentally determined to be < 0.01 mg/L by SafePharm Laboratories (study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations.gov/#!documentDe tail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013; “4,4’ - Methylene bis(2,6 - di - tert - butylphenol): Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/011, study completed on July 18, 2005).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

9 Study 2 : Acute toxicity study in sheepshead minnow ( Stonybrook Laboratories Inc. , 1995 ) Title : Acute/Prolonged Toxicity to Fish This study is: Adequate Not Adequate Important study details missing from this report include test substance purity, test method/guidelin e followed, GLP compliance, whether or not a nalytical monitoring was conducted, length, weight and source of test species, control treatment , and detailed numerical results . C oncentrations of the test substance (10

– 1000 mg/L) exceeded the water solubility limit of the test substance ( experimentally determined to be < 0.01 mg/L) 1 by many orders of magnitude and a vehicle was not used . Furthermore, the study report did not provide any information regarding evaluations for physical evidence of insolubility of the test substance in the media (e.g., visible precipitate or residue) . Additionally, the temperature varied by more than 2°C, only three concentrations were tested , and units of salinity measurements were not reported . TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1 Remarks : Purity not provided METHOD Method/guideline followed : Not specified Test type : Acute Toxicity to Fish (Static) GLP compliant? : Not specified Year study was performed : 1995 Test species : Sheepshead Minnow ( Cyprinodon variegatus ) Analytical monitoring : Not specified Exposure duration : 96 hours Concentrations tested : 10, 100 , and 1000 mg/L

1 T he water solubility li mit of CASRN 118 - 82 - 1 was experimentally determined to be < 0.01 mg/L by SafePharm Laboratories (study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013; “4,4’ - Methylene bis(2,6 - di - tert - butylphenol): Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/011, s tudy completed on July 18, 2005).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

10 Test method/conditions remarks : The tests were carried out with a dispersion of the test material in water under static conditions . The test volume was 30 L. Experimental Conditions: Salinity : 21

– 22 (units not specified) Temperature : 19.2

– 22.3°C Dissolved oxygen : 6.7

– 7.6 mg/L pH : 8.55 No . of replicates : 2 No . of fish per replicate : 10 RESULTS Endpoint value(s) : Note that analytical monitoring was not conducted and nominal concentrations (10

– 1000 mg/L) were many orders of magnitude greater tha n the water solubility limit ( experimentally determined to be < 0.01 mg/L) 1 for this compound . Therefore, it is not possible to interpret the reported toxicity values. 96 - hour LC 50 > 1000 mg/L , based on nominal concentrations . Results remarks : No mortalities were observed . No additional information was provided in study summary . CONCLUSIONS The 96 - hour LC 50 for sheepshead minnow is > 1000 mg/L , based on nominal concentrations . STUDY RELIABILITY [3] Not reliable . Important study details missing from this report include test substance purity, t est method/guideline followed, GLP compliance , whether or not a nalytical monitoring was conducted , length, weight and source of test species , control treatment , and detailed numerical results . T est material concentrations exceeded the water solubility limit of the test substance ( experimentally determined to be < 0.01 mg/L) 1 by many orders of magnitude and a vehicle was not used . Furthermore, the study report did not provide any information regarding evaluations for physic al evidence of insolubility of the test substance in the media (e.g., visible precipitate or residue) . Additionally, the temperature varied by more than 2°C, only three concentrations were tested , and units of salinity measurements were not reported.

1 T he water solubility limit of CASRN 118 - 82 - 1 was experimentally determined to be < 0.01 mg/L by SafePharm Laboratories (study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013; “4,4’ - Methylene bis(2,6 - di - tert - butylphenol): Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/011, study completed on July 18, 2005).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

11 REFERENCE Stonybrook Laboratories Inc . 1995 . Study No. 66126, conducted for Ethyl Corporation . Source : Ethyl Petroleum Additives International, Bracknell, Berkshire . ( As s ummarized in IUCLID Dataset for CASRN 118 - 82 - 1, European Chemical Bureau, 2000) . Robust summary available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0181 ( on p. 66 of the document “End_Point_D

– Mammalian_Toxicity_Acute.pdf” ) as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

12 C - 2 . Acute Toxicity to Aquatic Invertebrates 1 Summary of endpoint : Not a dequate . Testing of a cute toxicity to D aphnia is not required for the chemical because the log K ow is > 4.2 . Study 1 : Acute toxicity study in D aphnia (Mattock and Girling, 1988) Title : IONOX 220 : Acute toxicity ( Salmo gairdneri , Daphnia magna , and Selenastrum capricornutum ) and n - octanol/water partition coefficient This study is: Adequate Not Adequate This study is not adequate because c oncentrations of test material (500

– 1000 mg/L) exceeded the water solubility limit of the test substance ( experimentally determined to be < 0.01 mg/L) 2 by many orders of magnitude and a vehicle was not used . Furthermore, the study report did not provide any information regarding evaluations for physical evidence of insolubility of the test substance in the media (e.g., visible precipitate or residue ) . In addition, the study was not conducted under GLP, the t est method/guideline followed and statistical methods used were not specified, only two concentrations were tested, and the temperature varied by more than 2°C . TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; referred to as IONOX 220 in study report Remarks : 99.04% purity (by GC); pale yellow powder; obtained from San Celoni, Barcelona, Spain METHOD Method/guideline followed : Not specified Test type : Acute Toxicity to Daphnia (Static) GLP compliant? : No Year study was performed : 1988 Test species : Water flea ( Daphnia magna ) ; < 24 hours old; obtained from ICI Brixham Laboratory 1 Although acute toxicity testing in Daphnia is not required, test requirement C1 under the Final HPV2 Test Rule dated January 7, 2011 requires chronic toxicity testing in Daphnia according to the ASTM E 1193 guideline. 2 T he water solubility limit of CASRN 118 - 82 - 1 was experimentally determined to be < 0.01 mg/L by SafePharm Laboratories (study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations.gov/#!docume ntDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013; “4,4’ - Methylene bis(2,6 - di - tert - butylphenol): Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/011, study completed on July 18, 2005).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

13 Analytical monitoring : No Exposure duration : 48 hours Concentrations tested : 0 (control), 500 , and 1000 mg/L Test method/conditions remarks : Ten D aphnia were allocated to 150 mL glass crystallizing dishes that contained 100 mL of reconstituted fresh water . T est substance was added to duplicate sets of dishes resulting in two concentrations of 500 and 100 mg/L . The two control dishes did not receiv e test substance . Test Conditions: Temperature : 18

– 22 °C pH : 7. 8

– 8. 0 Hardness : 166 mg CaCO 3 /L Dissolved Oxygen : 9. 0

– 9.2 mg/L Statistics : None specified RESULTS Endpoint value(s) : Note that analytical monitoring was not conducted and nominal concentrations ( 50 0

– 1000 mg/L) were many orders of magnitude greater tha n the wat er solubility limit ( experimentally determined to be < 0.01 mg/L) 1 for th e test compound . Therefore, it is not possible to interpret the reported toxicity value. 48 - hour EC 50 > 1000 mg/L , based on nominal concentrations . Results remarks : Only 1 out of 20 daphni ds was immobilized at 1000 mg/L . Page 2 of t he test report, which included studies for fish, Daphnia , and green algae, provided a general comment that the test substance “ appeared to be not wholly soluble at con centrations greater than ” 10 mg/L . CONCLUSIONS The 48 - hour EC 50 for Daphnia magna is > 1000 mg/L , based on nominal concentrations .

1 T he water solubilit y limit of CASRN 118 - 82 - 1 was experimentally determined to be < 0.01 mg/L by SafePharm Laboratories (study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013; “4,4’ - Methylene bis(2,6 - di - tert - butylphenol): Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/01 1, study completed on July 18, 2005).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

14 STUDY RELIABILITY [3] Not reliable . Non - GLP study . Test method/guideline followed and statistical methods used were not specified, a nalytical monitoring was not conducted, only two concentrations were tested, the temperature varied by more than 2°C, the test concentrations (500

– 1000 mg/L) exceeded the water solubility limit of the test substance (experimentally determined to be < 0.01 mg/L) 1 by many orders of magnitude , and a vehicle was not used . REFERENCE Mattock, S.D . and Girling, A.E . 1988 . IONOX 220 : Acute Toxicity ( Salmo gairdneri, Daphnia magna and Selenastrum capricornutum ) and n - octanol/water partition coefficient . Sittingbourne Research Centre, Shell Research Limited . Submitted by Shell Oil Co . to EPA in 1992 as part of a TSCA 8(e) submission (EPA Doc . No . 88 - 920003444 , Fiche No . OTS0537403 ) entitled “Initial submission : acute toxicity of IONOX 220 in Salmo gairdneri, Daphnia magna and Selenastrum ca

pricornutum and n - octanol/water partition coefficient w - letter dated 053092”.

1 T he water solubility limit of CASRN 118 - 82 - 1 was experimentally determined to be < 0.01 mg/L by SafePharm Laboratories (study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013; “4,4’ - Methylene bis(2,6 - di - tert - butylphenol): Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/011, study completed on July 18, 2005).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

15 Study 2 : Acute toxicity study in mysid shrimp ( Stonybrook Laboratories Inc. , 1995) Title : Acute Toxicity to Aquatic Invertebrates This study is: Adequate Not Adequate Important study details missing from the report include test substance purity, test method/guideline follo wed, GLP compliance, age and source of test species , and detailed numerical results . Additionally, only three concentrations were tested , and units of salinity measurements were not reported . C oncentrations of test material (10

– 1000 mg/L) exceeded the water solubility limit of the test substance ( experimentally determined to be < 0.01 mg/L) 1 by many orders of magnitude and a vehicle was not used . Furthermore, the study report did not provide any information regarding evaluations for physical evidence of insolubility of the test substance in the media (e.g., visible precipitate or residue ) . TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1 Remarks : Purity not provided METHOD Method/guideline followed : Not specified Test type : Mysid acute toxicity test ( s tatic) GLP compliant? : No t specified Year study was performed : 1995 Test species : Mysid shrimp ( Mysidopsis bahia ) Analytical monitoring : No Exposure duration : 96 hours Concentrations tested : 0, 10, 100 , and 1000 mg/L Test method/conditions remarks : The tests were carried out with water - accommodated fractions (WAFs) under static conditions . The test volume was 30 L.

1 T he water solubility limit of CASRN 118 - 82 - 1 was experim entally determined to be < 0.01 mg/L by SafePharm Laboratories (study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.re gulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013; “4,4’ - Methylene bis(2,6 - di - tert - butylphenol): Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/011, study completed on July 18, 2005).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

16 Experimental Conditions: Salinity : 20

– 22 (units not specified) Temperature : 19.0

– 20.9°C Dissolved oxygen : 6.0

– 7.1 mg/L pH : 8.26

– 8.32 No . of replicates : 2 No . of mysids per replicate : 10 RESULTS Endpoint value(s) : Note that analytical monitoring was not conducted and nominal concentrations (10

– 1000 mg/L) were many orders of magnitude greater tha n the reported water solubility limit ( experimentally determined to be < 0.01 mg/L) 1 for this compound . Therefore, it is not possible to interpret reported toxicity values. 96 - hour L C 50 > 1000 mg/L , based on nominal concentrations . Results remarks : No mortalities were observed . No additional information was provided in study summary . CONCLUSIONS The 96 - hour L C 50 for Mysidopsis bahia is > 1000 mg/L , based on nominal concentrations . STUDY RELIABILITY [3] Not reliable . Important study details missing from the report include test substance purity, test method/guideline followed, GLP compliance, and detailed numerical results . C oncentrations of the test material exceeded the water solubility limit of the test substance ( experimentally determined to be < 0.01 mg/L) 1 by many orders of magnitude and a vehicle was not used . Furthermore, the study report did not provide any information regarding evaluations for physical evidence of insolubility of the test substance in the media (e.g., visible precipitate or residue) . Additionally, only three concentrations were tested , rather than five or more concentrations, and units of salinity measurements were not reported .

1 T he water solubility limit of CASRN 118 - 82 - 1 was experimentally determined to be < 0.01 mg/L by SafePharm Laboratories (study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013; “4,4’ - Methylene bis(2,6 - di - tert - butylphenol): Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/011, study completed on July 18, 2005).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

17 REFERENCE Stonybrook Laboratories Inc . 1995 . Study No . 66125, conducted for Ethyl Corporation . Source : Ethyl Petroleum Additives International, Bracknell, Berkshire . (Summarized in IUCLID Dataset for CASRN 118 - 82 - 1, European Chemical Bureau, 2000) . Robust summary available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0181 ( on p. 66 of the document “End_Point_D

– Mammalian_Toxicity_Acute.pdf” ) as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

18 C - 3 . Chronic Toxicity to Aquatic Invertebrates Summary of endpoint : Adequate Study 1 : Chronic toxicity study in D aphnia (M inderhout et al.

, 2012 ) Title : AN - 2 : A flow - through life - cycle toxicity test with the cladoceran ( Daphnia magna ) This stud y is: Adequate Not Adequate TEST SUBSTANCE Identity : 4,4’ - methylenebis ( 2,6 - di - tert - butyl[U 14 C]phenol) , Batch number CFQ41115 , from Quotient Bioresearch . Test substance was a 14 C - labeled form of CASRN 118 - 82 - 1; referred to as AN - 2 in study report . Remarks : Properties of Quotient Research Batch number CFQ41115 were : L iquid , 99.8% radiochemical purity by high performance liquid chromatography ( HPLC ) , specific activity of 60 mCi/mmol by mass spectrometry and 55 mCi/mmol by gravimetric analysis, MW 426.6 at this specific activity . “The material co - chromatographs with client supplied material in the above chromatographic [HPLC] system . The mass spectrum is consistent with the proposed structur e and a non labelled reference . The 1 H - NMR [nuclear magnetic resonance] spectrum is consistent with the proposed structure and with a non label l ed reference.” [Test substance properties and quoted material are from the Certificate of Analysis for Batch number CFQ41115 presented in Appendix 2, p. 44 of : Arnie, J.R., Porch, J.R., Kendall, T.Z . and Krueger, H.O . 2012 . AN - 2 : A 96 - Hour Toxicity Test with the Freshwater Alga ( Pseudokirchneriella subcapitata ) . Wildlife International, Ltd . Projec t No. 471A - 119 . Submitted to Albemarle Corporation . Available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ OPPT - 2007 - 0531 - 0678 as of December 4, 2013 . ] METHOD Method/guideline followed : OECD TG 211, EPA OPPTS 850.1300 , and ASTM E 1193 - 97 Test type : Chronic Toxicity to Daphnia (Flow - Through ) GLP compliant? : Yes Year study was performed : 2012 Test species : Water flea ( Daphnia magna ); neonates ( < 24 hours old at initiation); obtained from in - house cultures at Wildlife International, Ltd . , Easton, MD. Analytical monitoring : Yes Exposure duration : 21 daysDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

19 Concentrations tested : Nominal : 0 (negative control), 0 (solvent control), 1.6, 3.1, 6.3, 13, 25 , and 50 ng/L ; m ean m easured concentrations based on 14 C : < LOQ 1 , < LOQ, 1.4, 2.4, 5.5, 12, 21 , and 43 ng/L . Note : M easurement of 14 C does not distinguish between test substance and degradation products; mean measured concentrations of the intact test substance were not reported. Vehicle : HPLC - grade dimethylformamide (DMF) Test method/conditions remarks : “Daphnids were exposed to a geometric series of six test concentrations, a negative control (dilution water) and a solvent control (0.025 mL/L HPLC grade dimethylformamide) . Two replicate test chambers were tested for each treatment and control group . Each replicate contained two c ompartments with five daphnids, resulting in a total of 20 daphnids in each treatment and control group . Nominal test concentrations were selected in consultation with the Sponsor based on exploratory range finding toxicity data and the solubility limit i n the test system . The nominal test concentrations were 1.6, 3.1, 6.3, 13, 25 and 50 ng AN - 2/L . Mean measured test concentrations [ based on measurement of 14 C] were determined from samples of test water collected from each treatment and control group at test initiation, at approximately weekly intervals during the test and at test termination.” [ Minderhout et al . (2012), p . 10] “Delivery of the test substance to the test chambers was initiated seven days prior to the introduction of the daphnids to the test water in order to achieve equilibrium of the test substance in the test chambers . At test initiation, neonate daphnids were impartially assigned to exposure chambers . First - generation daphnids were observed daily during the test for mortality, the o nset of reproduction, and clinical signs of toxicity . Following the onset of reproduction, the numbers of second - generation daphnids were counted three times per week (i.e., Monday, Wednesday and Friday), and at test termination (Day 21) . Body lengths an d dry weights of the surviving first generation daphnids were measured at the end of the exposure period . Observations of the effects of AN - 2 on survival, reproduction and growth were used to determine the no - observed effect concentration (NOEC), the lowe st - observed - effect concentration (LOEC), and the maximum acceptable toxicant concentration (MATC) . EC50 values were determined based on reproduction and on first - generation immobility at test termination.” [ Minderhout et al . (2012), p p . 10 - 11] Preparati on of Test Concentrations: “ Individual stock solutions were prepared for each of the six concentrations tested, and were prepared once during the study . Each test stock solution was prepared by mixing a calculated amount of radiolabelled AN - 2 into HPLC - grade dimethylformamide (DMF) at a nominal concentration of 0.064, 0.124, 0.252, 0.52, 1.0 and 2.0 μg/mL . The stock solutions were mixed by inversion . The stock solutions at 0.064 to 1.0 μg/mL appeared clear and colorless, while the stock solution at 2.0 μg/mL appeared clear with a slight purple tint . Stock solutions were stored refrigerated in glass amber bottles, and aliquots of each stock were placed in the syringe pump every 4 to 7 days during the study.” [ Minderhout et al . (2012), p. 14]

1 LOQ = 0. 873 ng/LDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

20 “The five test substance stock solutions were injected into the diluter mixing chambers at a rate of 2.5 μL/minute where they were mixed with dilution water delivered at a rate of 100 mL/minute to achieve the desired test concentrations . The negative control received dilution water only . The solvent control was prepared by delivering HPLC - grade DMF to the mixing cha mber for the solvent control . The concentration of DMF in the solvent control and all AN - 2 treatment groups was 0.025 mL/L.” [ Minderhout et al . (2012), p. 14] Analytical Sampling: “Samples were collected from each treatment and control group five days prior to the start of the test after conditioning the diluter for two days . Water samples also were collected from alternating replicate test chambers in each treatment and control group at the beginning of the test, at approximately weekly intervals duri ng the test and at the end of the test to measure concentrations of the test substance . Samples of the stock solutions being delivered to the diluter were collected for analysis on Day

– 11 to confirm the stock concentrations, after the stocks were prepare d . Additional samples of test solutions were collected on Day 22 to confirm concentration of the 1.6 ng/L treatment level when the analytical result of this treatment level on Day 21 was suspect . All test solution samples were collected from mid - depth, p laced in separatory funnels, and processed immediately for analysis by LSC [liquid scintillation counting] .” [Mindehout et al . (2012 ) , p. 14] Analytical Method: “ The concentrations of the test substance in freshwater samples were verified by liquid scintillation counting (LSC) . Samples were extracted two times with dichloromethane, evaporated to dryness and reconstituted using acetonitrile . Extracts were transferred into scintillation vials containing 13 mL of Ultima Gold XR scintillation cocktail and submitted for analysis by LSC . A Packard 2500TR Liquid Scintillation Analyzer was used to determine disintegrations per minute (dpm) . In addition to the test samples, a 13 mL sample of Ultima Gold XR scintillation cocktail was analyzed at each interv al to determine background radioactivity. ” [Mindehout et al . (2012 ) , p. 15 ] Biological Observations and Measurements : “ Observations of each first - generation daphnid were made daily during the test . At these times, the numbers of immobile daphnids were recorded along with any clinical signs of toxicity (e.g., inability to maintain position in the water column, uncoordinated swimming or cessation of feeding) . Immobility was defined as a lack of movement, except for minor spontaneous random movement of th e appendages . The presence of eggs in the brood pouch, aborted eggs, males or ephippia also were recorded daily . With the onset of reproduction, neonates produced by the first - generation daphnids were counted and then discarded every Monday, Wednesday an d Friday during the test . The body length and the dry weight of each surviving first - generation daphnid were measured at the end of the test. ” [ Minderhout et al . (2012), p. 16] Statistical Analyses: “Test endpoints analyzed statistically for first - genera tion daphnids were survival, reproduction, and growth (length and dry weight) . Reproduction was based on the number of live young produced per reproductive day . Reproductive days were defined as the number of days that the adult daphnid was alive from the day the first brood was released from any adult daphnid in the test until test termination . If an adult daphnid died, the number of reproductive days, for that adult, ended on the last day it was alive.” [ Minderhout et al . (2012), p. 16 ]Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

21 “ Negative control and solvent control data for each parameter were compared using an appropriate statistical test . No significant differences between the control groups were fo und for any parameter tested (p > 0.05) . Therefore, the control data for all parameters were pooled for comparison with the treatment groups.” [ Minderhout et al . (2012), p. 17] “Survival data was considered to be discrete - variable data, while reproductio n and growth data were considered continuous - variable data . Discrete - variable data were analyzed using Chi square and Fisher’s Exact test to identify treatment groups that showed a statistically significant difference (p ≤ 0.05) from the control . All con tinuous - variable data were evaluated for normality using Shapiro - Wilk’s and for homogeneity of variance using Levene’s (α = 0.01) . When the data passed the assumptions of normality and homogeneity, those treatments that were significantly different from t he control means were identified using Dunnett’s test (α = 0.05) . All statistical tests were performed using a personal computer with SAS software.” [ Minderhout et al . (2012), p. 17] “The results of the statistical analyses were used to aid in the determination of the NOEC, LOEC and MATC . However, scientific judgment was used to determine if statistical differences were biologically meaningful, and if the data followed a concentration - dependent response . The NOEC was defined as the highest test co ncentration that produced no significant treatment related effects on survival, reproduction or growth . The LOEC was defined as the lowest test concentration that produced a significant treatment - related effect on survival, reproduction or growth . The MA TC was calculated as the geometric mean of the NOEC and LOEC . EC50 values were determined based on reproduction and on the immobility observed in the first generation daphnids at the end of the test.” [ Minderhout et al . (2012), p. 17] Test Conditions: Temperature : 19.6

– 20.4 °C pH : 8.0

– 8.2 Hardness : 134

– 141 mg CaCO 3 /L D issolved Oxygen : 5.6

– 8.6 mg/L Dilution water TOC : < 1mg C/L Light Intensity : 171 lux RESULTS Endpoint value(s) : B ased on mean measured concentrations of 14 C , the 21 - day EC 50 for immobility , reproduction , and growth is greater than 43 n g/L; the 21 - day NOEC for immobility , reproduction , and growth is 43 n g/L based on no significant effects at the highest concentration tested . Note : Measurement of 14 C does not distinguish between test substance and degradation products; mean measured concentrations of the intact test substance were not reported.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

22 Results remarks : Test solutions appeared clear and colorless with no evidence of precipitation . Measurement of Test Concentrations: “Samples of the stock solutions being delivered to the diluter system had measured [ 14 C] concentrations that ranged from 100 to 105% of nominal concentrations, confirming the concentrations being delivered to the diluter system ... . The meas ured [ 14 C] concentrations of samples collected to verify the diluter system prior to the test ranged from 76.9 to 96.7% of nominal concentrations ... . Samples of the test solutions collected during the test had measured concentrations [of 14 C] that ranged from 66.0 to 177% of nominal concentrations ... . Since the analytical result of the sample [ for the ] 1.6 ng/L treatment group on Day 21 was suspect (the recovery was 177%), [an] additional sample was collected from this treatment group to confir m the test concentration on Day 22 . The results of the sample collected on Day 22 confirmed the test concentration; the result of the sample for the 1.6 ng/L treatment group collected on Day 21 was excluded from the calculation of mean measured test concentration . When the measured concentrations of test solution samples collected on Days 0, 7, 14, 21 and 22 of the test were averaged for each treatment group, the mean measured test concentrations [based on concentrations of 14 C] were 1.4, 2.4, 5.5, 1 2, 21 and 43 ng/L, which represented 88, 77, 87, 92, 84 and 86% of nominal concentrations, respectively . The results of the study were based on the mean measured concentrations [of 14 C] .” [ Minderhout et al . (2012), p. 18] Survival and Clinical Observations: “After 21 days of exposure, survival in the negative control and solvent control groups were 75 and 95%, respectively . There was statistically no significant difference in survival between the negative and solvent control groups . Therefore, the control data were pooled for comparison with the treatment groups . Survival in the 1.4, 2.4, 5.5, 12, 21 and 43 ng/L treatment groups at test termination was 90, 90, 100, 95, 90 and 90%, respectively . Fisher’s Exact test indicated that no statistica lly significant decrease in survival was detected in any of the AN - 2 treatment groups in comparison to the pooled control (p > 0.05) . Consequently, the NOEC for survival was 43 ng/L and the LOEC was greater than 43 ng/L . Based on the immobilities observe d in the treatment groups, the 21 - day EC50 value was greater than 43 ng/L, the highest concentration tested.” [ Minderhout et al . (2012), p p. 18 - 19] “Daphnids in the 1.4, 2.4, 5.5, 12, 21 and 43 ng/L treatment groups that survived to test termination generally appeared normal . During the test pale, small and/or lethargic daphnids were observed in the 1.4, 2.4, 12, 21 and 43 ng/L treatment groups . However, the numbers of sublethal effects noted in the 1.4, 12 and 21 ng/L treatment groups were infreque nt . At test termination, approximately 33% of surviving first - generation daphnids in the 43 ng/L treatment group appeared pale in comparison to the control organisms . While the discoloration of the daphnids in the 43 ng/L treatment group were not analyze d statistically, it is important to note that the condition appeared to follow the dose - response pattern . All surviving daphnids in the 2.4 and 5.5 ng/L treatment groups were normal in appearance at test termination.” [ Minderhout et al . (2012), p. 19]Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

23 Re production: “ The first day of brood production in the negative and solvent control replicates and in all AN - 2 treatment replicates was Day 7, 8 or 9 of the test, indicating that there was no apparent delay in the onset of production at any AN - 2 concentrati on tested . With the exception of two immobile neonates in the negative control and solvent control groups and three immobile neonates in the 43 ng/L treatment group on Day 9 or 11 of the test, no immobile neonates were noted in the 1.4, 2.4, 5.5, 12 and 21 ng/L treatment groups during the test (Appendix 9 [of the study report] ) . No aborted or shed eggs were present in any of the control or treatment groups . No males or ephippia were produced during the test.” [ Minderhout et al . (2012), p. 19]

“ Adult da phnids in the negative and solvent control groups produced an average of 8.1 and 8.7 live young per reproductive day, respectively . There was no significant difference in reproduction between the negative and solvent control groups . Therefore, the contro l data were pooled for comparison with the treatment groups . Adult daphnids in the 1.4, 2.4, 5.5, 12, 21 and 43 ng/L treatment groups produced an average of 8.9, 6.7, 8.1, 6.9, 7.7 and 8.3 live young per reproductive day, respectively . Dunnett’s test ind icated there were no statistically significant decreases in mean neonate production in any of the AN - 2 treatment groups in comparison to the pooled control (p > 0.05) . Consequently, the NOEC for reproduction was 43 ng/L and the LOEC was greater than 43 ng /L . Based on the reproduction observed in the treatment groups, the 21 - day EC50 value was greater than 43 ng/L, the highest concentration tested.” [ Minderhout et al . (2012), p. 20] Growth: “ Daphnids in the negative and solvent control groups averaged 4.6 mm in length and 0.63 mg in dry weight, while daphnids in the solvent control group averaged 4.5 mm in length, and 0.69 mg in dry weight, respectively . There were no significant differences in the growth parameters between the negative and solvent con trol groups . Therefore, the control data were pooled for comparison with the treatment groups . Daphnids in the 1.4, 2.4, 5.5, 12, 21 and 43 ng/L treatment groups had mean lengths of 4.6, 4.3, 4.4, 4.4, 4.4 and 4.5 mm, respectively, and mean dry weights o f 0.76, 0.60, 0.74, 0.76, 0.63 and 0.60 mg, respectively . Dunnett’s test indicated there were no statistically significant decreases in mean dry weight in any of the AN - 2 treatment groups in comparison to the pooled control (p > 0.05) but statistically si gnificant reductions were noted in mean total length in the 2.4, 5.5, 12 and 21 ng/L treatment groups when compared to the pooled controls (p ≤ 0.05) . Since the reductions in mean total length of daphnids in the 2.4, 5.5, 12 and 21 ng/L treatment groups d id not follow a dose response pattern, the slight differences noted were not considered to be biologically meaningful . In addition, the mean length of the daphnids in the 2.4, 5.5, 12 and 21 ng/L treatment groups were within the ranges of the mean length of daphnids in the historical control data (3.6 to 6.3 mm) . Consequently, the NOEC for growth (length and dry weight) was 43 ng/L, and the LOEC was greater than 43 ng/L.” [ Minderhout et al . (2012), p. 20] CONCLUSIONS “The cladoceran, Daphnia magna , was exposed to AN - 2 at mean measured concentrations [based on concentrations of 14 C] of 1.4, 2.4, 5.5, 12, 21 and 43 ng/L under flow - through conditions for 21 days . There were no statistically significant treatment - related effects on survival, reproduct ion or growth at concentrations ≤ 43 ng/L . Consequently, the NOEC, based on survival, reproduction and growth, was 43 ng/L . The LOEC was greater than 43 ng/L and theDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

24 MATC was calculated to be greater than 43 ng/L . The 21 - day EC50 values for adult immobi lity and reproduction [and growth] were both greater than 43 ng/L, the highest concentration tested.” [ Minderhout et al . (2012), p. 21] STUDY RELIABILITY [ 2 ] Reliable with restrictions . Following the preparation of test solutions containing radiolabelled test substance [ 4,4’ - methylenebis (2,6 - di - tert - butyl[U 14 C]phenol)], t he method used for monitoring test solutions over the course of the study for the presence of radiolabel (extraction of organic components from the aqueous samples into dichloromethane, evaporation of the dichloromethane, reconstitution of dried material in acetonitrile, and use of liquid scintillation counting [ LSC ] to analyze total 14 C radioactivity) did not distinguish between the intact test substance and degradation products . However, this deficiency does not appear to affect the validity of the EC 50 , NOEC, LOEC, and MATC values that were derived from the data of t his stud y . REFERENCE Minderhout, T., Kendall, T.Z., Gallagher, S. P. and Krueger, H.O . 2012 . AN - 2 : A flow - through life - cycle toxicity test with the cladoceran ( Daphnia magna ) . Final Report . Wildlife International, Ltd., Easton, MD . Project No. 471A - 120 . Study report available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2007 - 0531 - 0678 as of December 4, 2013 . Text in quotes is taken directly from Minderhout et al ., 2012.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

25 C - 4 . Toxicity to Aquatic Plants Summary of endpoint : Adequate Study 1 : Toxicity study in green algae (Arnie et al. , 2012) Title : AN - 2 : A 96 - hour toxicity test with the freshwater alga ( Pseudokirchneriella subcapitata ) This study is: Adequate Not Adequate TEST SUBSTANCE Identity : 4,4’ - methylenebis ( 2,6 - di - tert - butyl[U 14 C]phenol ) , Batch number CFQ41115 , from Quotient Bioresearch . Test substance was a 14 C - labeled form of CASRN 118 - 82 - 1; referred to as AN - 2 in study report . Remarks : Properties of Quotient Research Batch number CFQ41115 : L iquid , 99.8% radiochemical purity by high performance liquid chromatography ( HPLC ) , specific activity of 60 mCi/mmol by mass spectrometry and 55 m Ci/mmol by gravimetric analysis, MW 426.6 at this specific activity . “The material co - chromatographs with client supplied material in the above chromatographic [HPLC] system . The mass spectrum is consistent with the proposed structure and a non labelled reference . The 1 H - NMR [nuclear magnetic resonance] spectrum is consistent with the proposed structure and with a non labe l led reference.” [Arnie et al. , 2012, Certificate of Analysis, Appendix 2 , p. 44 ] . METHOD Method/guideline followed : EPA OPPTS 850.5400; OECD TG 201; ASTM E1218 - 04 Test type : Toxicity to Green Algae (Static) GLP compliant? : Yes Year study was performed : 2012 Test species : Freshwater green algae ( Pseudokirchneriella subcapitata ); original algal cultures were obtai ned from the University of Toronto Culture Collection, and had been maintained in culture medium in - house at Wildlife International, Ltd., Easton, Maryland. Analytical monitoring : Yes Exposure duration : 96 hours Concentrations tested : Nominal : 0 (negative control), 0 (solvent control), 2, 4, 8, 16 , and 32 ng/L; Initial (day 0) m easured concentrations based on 14 C : < LOQ 1 (negative control), 1 LOQ = 0.582 ng/LDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

26 < LOQ (solvent control), 1.50, 3.35, 6.84, 12.9 , and 26.5 ng/L . Note : Measurement of 14 C does not distinguish between test substance and degradation products; mean measured concentrations of the intact test substance were not reported. Vehicle : D imethylformamide (DMF) Test method/conditions remarks : “ The green alga, Pseudokirchneriella subcapitata , was exposed to five test concentrations, a solvent control (0.1 mL N,N - dimethylformamide per L) and a negative control (culture medium) for 96 hours . Nominal test concentrations were selected in consultation with the Sponsor and were based upon the resu lts of an exploratory range - finding toxicity test . Nominal test concentrations selected were 2.0, 4.0, 8.0, 16.0 and 32.0 nanograms per liter (ng/L) . Measured concentrations [based on measurement of 14 C] were determined from samples of test medium collec ted from each treatment and control group at 0 and 96 hours of the test. ” [Arnie et al . (2012), p. 11] “Three replicate test chambers were maintained in each treatment group and six replicates were maintained in the negative control group . At test initiation, an inoculum of the algal cells was added to each test chamber to achieve a nominal concentration of approximately 10,000 cells/mL . Samples were collected from each replicate test chamber at approximately 24 - hour intervals during the te st to determine cell densities, yield and growth rates . Cell densities, yields and growth rates were used to calculate percent inhibition values relative to the control over the 96 - hour exposure period . EC 50 and E r C 50 values (i.e., the theoretical toxica nt concentrations that would produce a 50% reduction in cell density and growth rate, respectively) were calculated, when possible, for each 24 - hour interval of the exposure period . E y C 50 values (i.e., the theoretical toxicant concentrations that would pr oduce a 50% reduction in yield) were calculated, when possible, at 72 and 96 hours of exposure . No - observed - effect concentrations (NOEC) were determined at 72 and 96 hours through statistical evaluation of cell densities, yields and growth rates, as well as examination of the concentration - response pattern.” [Arnie et al . (2012), p p. 11 - 12] Test Apparatus: “Test chambers were sterile, 250 - mL Erlenmeyer flasks plugged with foam stoppers, and contained 100 mL of test or control medium . The test chambers were labeled with the project number, test concentration and replicate, and were indiscriminately positioned daily on a mechanical shaker in an environmental chamber designed to maintain the desired test temperature throughout the test . The test flasks we re shaken continuously at 100 rpm.” [Arnie et al . (2012), p. 13] Culture Medium: “The algal cells were cultured and tested in freshwater AAP algal medium . Stock nutrient solutions were prepared by adding reagent - grade chemicals to purified well water (N ANOpure ® water) . The test medium was then prepared by adding appropriate volumes of the stock nutrient solutions to NANOpure ® water ... The pH of the medium was adjusted to 7.5 ± 0.1 using 10% hydrochloric acid and was sterilized by filtration (0.22 μm) prior to use. ” [Arnie et al . (2012), p. 13] A nalyses performed to measure selected contaminants (pesticides, organics, and metals) in the well water used to prepare the algal medium showed acceptable concentrations.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

27 Environmental Conditions: “Test chambe rs were held in an environmental chamber at a temperature of 24 ± 2 ° C . The temperature of a container of water adjacent to the test chambers in the environmental chamber was recorded twice daily during the test using a liquid - in - glass thermometer . In addition, temperature in the environmental chamber was measured continuously using a minimum - maximum thermometer . The algae were held under continuous cool - white fluorescent lighting throughout the test . The target light intensity was 6,000 lux ± 10% . L ight intensity was measured at test solution level at five locations surrounding the test flasks at test initiation using a SPER Scientific 840006C light meter . The pH of the medium in each treatment and control group was measured at test initiation and e xposure termination using a Thermo Orion Model 525 APlus . At test initiation, pH was measured in the individual batches of test solution prepared for each treatment and control group . At exposure termination, pH was measured in pooled samples of test sol ution collected from each of the replicates of each treatment and control group.” [Arnie et al . (2012), p. 13] Preparation of Test Concentrations: “A primary stock solution was prepared by dissolving 0.213 mL of radiolabelled AN - 2 in 50 mL of N,N - diem e th ylformamide to achieve a nominal concentration of 32.0 μg/mL . The primary stock was mixed by inversion and appeared clear and purple in color upon preparation . A secondary stock was prepared by diluting 1.0 mL of 32.0 μg/mL primary stock in 100 mL of N,N - dimethylformamide to achieve a nominal concentration of 0.319 μg/mL . Additional stocks were prepared at nominal concentrations of 0.16, 0.08, 0.04, 0.02 μg/mL in N,N - dimethylformamide by serial dilution . The 2.0, 4.0, 8.0, 16.0 and 32.0 ng/L test soluti ons were prepared by diluting 100 μL of each respective stock solution in 1000 mL of freshwater AAP medium . The solvent control solution was prepared by diluting 100 μL of N,N dimethylformamide in 1000 mL of freshwater AAP medium to achieve a nominal conc entration of 0.1 mL N,N - dimethylformamide per liter, equivalent to the solvent concentration in all of the test solutions except the negative control . The negative control solution consisted of freshwater AAP algal medium without test substance or solvent added . All test solutions were mixed by inversion, and appeared clear and colorless and were otherwise unremarkable after mixing.” [Arnie et al . (2012), p. 14] Analytical Sampling : “ Samples of the test solutions were collected at approximately 0 and 96 hours to measure concentrations of the test substance . Samples at test initiation were collected from the individual batches of test solution prepared for each treatment and control group after distribution into the test chambers . At exposure termination, samples were collected from the pooled replicates from each treatment and control group . Samples were processed immediately for LSC [liquid scintillation counting] analysis. ” [Arnie et al . (2012), p. 14] Analytical Method : “ The concentratio ns of the test substance in freshwater algal medium samples were verified by liquid scintillation counting (LSC) . Samples were extracted two times with dichloromethane, rotary evaporated to dryness and reconstituted using acetonitrile . Extracts were quantitatively transferred into scintillation vials containing 10 mL of Ultima Gold XR scintillation cocktail and submitted for analysis by LSC . A Packard 2500TR Liquid Scintillation Analyzer was used to determine disintegrations per minute (dpm) . In add ition to the test samples, a 10 mL sample of Ultima Gold XR scintillation cocktail was analyzed at each interval to determine background radioactivity. ” [Arnie et al . (2012), p. 14]Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

28 Algal Growth Measurements and Observations: “Test medium samples were co llected from each replicate of the treatment and control groups for the determination of algal cell densities . Samples were collected at approximately 24 - hour intervals during the 96 - hour exposure and were held for a maximum of three days under refrigerat ed conditions sufficient to inhibit growth until cell counts could be performed.” [Arnie et al . (2012), p. 15] “Samples of test solution were collected from each replicate at the end of the test . These samples were pooled within their respective treatments, and sub - samples were removed and examined microscopically for atypical cell morphology (e.g., changes in cell shape, size or color) . Cells in the replicate test chambers also were assessed for aggregation or flocculation of cells, and adherenc e of the cells to the test chamber.” [Arnie et al . (2012), p. 15] Statistical Analyses: “ The calculation of cell densities, yield, growth rates and percent inhibition values, as well as all statistical analyses, were conducted using ‘ The SAS System for W indows, Version 8.2 ’ . Growth rate was calculated for each replicate of the control and treatment groups...” [Arnie et al . (2012), p. 15] “Yield was calculated for each replicate of the control and treatment groups as the final biomass (cell density) in the exposure period minus the initial biomass (cell density) . Inhibition values were calculated for each treatment group as the percent reduction in cell density and growth rate relative to the negative control replicates for each 24 - hour interval, and yield at 72 and 96 hours of exposure.” [Arnie et al . (2012), p. 16] “EC 50 values in this study represent the theoretical test concentrations that would produce a 50% reduction in a variable of interest relative to the negative control . The E y C 50 and E r C 50 values and their 95% confidence intervals were calculated, when possible, using non - linear regression with replicate data (cell density, yield or growth rate) and day 0 measured test concentrations.” [Arnie et al . (2012), p. 16] “The negative and solvent control data was compared using a student’s t - test (α = 0.05) . No statistically significant differences were detected between the negative and solvent control data . The treatment group responses were compared to the negative control response for statist ical comparisons and EC 50 calculations, when possible . The 72 and 96 - hour cell density, yield and growth rate data were evaluated for normality and homogeneity of variance (α = 0.01) using Shapiro - Wilk’s and Levene’s tests, respectively . All data met ass umptions or normal distribution and equal variance; therefore, the treatment groups were compared to the negative control using Dunnett’s one - tailed t - test (α = 0.05) . The results of the statistical analyses, as well as an evaluation of the concentration - response pattern, were used to determine the NOEC relative to each parameter at 72 and 96 hours.” [Arnie et al . (2012), p. 16] Test Conditions: Temperature : 22.4

– 25.5°C pH : 7.1

– 8.5 Light Intensity : 5870

– 6430 luxDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

29 RESULTS Endpoint value(s) : Based on initial [day 0] measured concentrations of 14 C : 96 - hr EC 50 ( cell density, growth rate , and yield) > 26.5 ng/L ; NOEC (cell density, growth rate, and yield) = 26.5 ng/L Results remarks : Measurement of Test Concentrations: “ The test solutions appeared clear and colorless at the time of preparation, and there was no evidence of surface slicks or precipitation at exposure termination . Measured concentrations [of 14 C] on Day 0 ranged from 75 to 86% of nominal . Measured concentrations [of 14 C] de clined by Day 4, and ranged from 16 to 24% of nominal . The results of the study [i.e., the EC 50 values and NOEC] are based on day 0 measured concentrations [of 14 C] of 1.50, 3.35, 6.84, 12.9 and 26.5 ng/L, representing 75, 84, 86, 81 and 83% of the target nominal test concentrations [of 14 C] . Measured concentrations of 14 C AN - 2 by LSC analysis in the negative and solvent controls were less than the limit of quantitation (LOQ) at both sampling intervals.” [Arnie et al . (2012), p p. 16 - 17] Observations and Measurements: “After 72 hours of exposure, inhibition of cell density in the 1.50, 3.35, 6.84, 12.9 and 26.5 ng/L treatment groups was 15, 8, 49, - 9 and - 3%, respectively, relative to the negative control . Inhibition of yield in the 1.50, 3.35, 6.84, 12. 9 and 26.5 ng/L treatment groups was 15, 8, 49, - 9 and - 3 [%] , relative to the negative control . Inhibition of growth rate in the 1.50, 3.35, 6.84, 12.9 and 26.5 ng/L treatment groups was 4, 1, 13, - 2 and - 1% , respectively, relative to the negative control . Dunnett’s test indicated that there were significant reductions ( p ≤ 0.05) in cell density, yield and growth rate in the 6.8 ng/L treatment group . Since this reduction was not dose - responsive and appeared analogous to the data collected from the other treatment groups, it was not considered to be treatment related . Consequently, the 72 - hour NOEC for cell density, yield and growth rate was determined to be 26.5 ng/L based on evaluation of the dose - response.” [Arnie et al . (2012), p p. 17 - 18] “After 96 hours of exposure, inhibition of cell density in 1.50, 3.35, 6.84, 12.9 and 26.5 ng/L treatment groups was - 23, - 22, 12, - 10 and 5%, respectively, relative to the negative control . Inhibition of yield in 1.50, 3.35, 6.84, 12.9 and 26.5 ng/L treatment groups was - 24, - 22, 12, - 10 and - 5%, respectively, relative to the negative control . Inhibition of growth rate in the 1.50, 3.35, 6.84, 12.9 and 26.5 ng/L treatment groups was - 3, - 4, 2, - 2 and - 1%, respectively, relative to the negative control . No significant reductions in cell density, yield or growth rate were detected (Dunnett’s test, p > 0.05) when the responses of the treatment groups were compared to the negative control respon se . Consequently, the 96 - hour NOEC for cell density, yield and growth rate was 26.5 ng/L.” [Arnie et al . (2012), p. 18] “After 96 hours of exposure, cells present in the 1.50, 3.35, 6.84, 12.9 and 26.5 ng/L treatment groups appeared normal when compared to cells present in the negative and solvent control groups during microscopic observations of test solution samples . Adherence to the test chambers and flocculation or aggregation of cells were not observed in any of the experimental groups.” [Arnie et a l . (2012), p. 18]Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

30 Validity of the Test: The mean cell density in the negative control replicates increased by a factor of 189 within 72 hours and by a factor of 382 within 96 hours . The coefficient of variation of average specific growth rates in the control replicates was 5.45% within 72 hours , and the mean coefficient of variation for section - by - section specific growth rates (days 0

– 1, 1

– 2, and 2

– 3) was 26.3% . These values conformed to the OECD and ASTM criteria for validity of an algae test. CONCL USIONS The 96 - hr EC 50 for cell density, growth rate , and yield is > 26.5 ng/L ( based on initial [day 0] measured concentrations of 14 C ); the NOEC is 26.5 ng/L . STUDY RELIABILITY [ 2 ] R eliable with restrictions . Following the preparation of test solutions containing radiolabelled test substance [ 4,4’ - methylenebis (2,6 - di - tert - butyl[U 14 C]phenol) ], t he method used for monitoring test solutions at 0 and 96 hours for the presence of radiolabel ( extraction of organic components from the aqueous samples into dichloro m ethane , evaporation of the dichloromethane, reconstitution of dried material in acetonitrile, and use of liquid scintillation counting [ LSC ] to analyze total 14 C radioactivity ) did not distinguish between test substance and degradatio n products . Furthermore , the test report did not provide an explanation of why the c oncentrations of 14 C were not stable over the 4 - day exposure period . Measured concentrations of 14 C on Day 0 ranged from 75 to 86% of nominal , declining to 16 to 24% of nominal on Day 4 . There is unce rtainty about whether this decline represents an actual loss of test substance (or its degradates) from the test vessels or only an apparent loss , that is possibly due to : (1) the interference of increasing algae biomass with the extraction of the test substance or its degradates from aqueous solution into dichlorom ethane (e.g., does test substance radioactivity remain in the water phase with the algae cells rather than being e xtracted into the dichloromethane phase?) , or (2) interference with the quantitation of radioactivity by LSC (e.g., does color from the algae cause a reduction of counting efficiency [ i.e., quenching ] by LSC?) . However, these deficiencies do not appear to affect the validity of the EC 50 values and NOEC values , which were calculated using day 0 “measured” values that are based on radioactivity measurements . REFERENCE Arnie, J.R., Porch, J.R., Kendall, T.Z . and Krueger, H.O . 2012 . AN - 2 : A 96 - hour toxicity test with the freshwater alga ( Pseudokirchneriella subcapitata ) . Wildlife International, Ltd., Easton, MD . Project No. 471A - 119 . Study report available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2007 - 0531 - 0678 as of December 4, 2013 . Text in quotes is taken directly from Arnie et al ., 2012.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

31 Study 2 : Toxicity study in green algae (Mattock and Girling, 1988) Title : IONOX 220 : Acute toxicity ( Salmo gairdneri , Daphnia magna , and Selenastrum capricornutum ) and n - octanol/water partition coefficient This study is: Adequate Not Adequate This study is not adequate because the test concentrations exceeded the water solubility limit of the test substance ( experimentally determined to be < 0.01 mg/L) 1 by many orders of magnitude . Furthermore, this is a n on - GLP study , the t est method/guideline followed and statistical methods used were not specified, a nalytical monitoring was not conducted, water chemistry parameters were not provided (hardness), and control treatment and control responses were not specified . TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; referred to as IONOX 220 in study report Remarks : 99.04% purity (by GC); pale yellow powder; obtained from San Celoni, Barcelona, Spain METHOD Method/guideline followed : Not specified Test type : Toxicity test to freshwater algae GLP compliant? : No Year study was performed : 1988 Test species : Freshwater green algae ( Selenastrum capricornutum ); from the axenic laboratory culture obtained from the American Type Culture Collection, Maryland, U.S.A. Analytical monitoring : No Exposu re duration : 96 hours Concentrations tested : 0 (control), 1, 2.2, 4.6, 10, 22, 46, 100, 220, 460 , and 1000 mg/L

1 T he water solubility limit of CASRN 118 - 82 - 1 was experimentally determined to be < 0.01 mg/L by SafePharm Laboratories (study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013; “4,4’ - Methylene bis(2,6 - di - tert - butylphenol): Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/011, study completed on July 18, 2005).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

32 Test method/conditions remarks : IONOX 220 in Analar acetone was added to Erlenmeyer flasks containing 50 mL of culture medium to give an approximately logarithmically spaced series of concentrations ranging from 1 to 1000 mg/L . The concentration of Analar acetone in all test flasks, including the controls, was adjusted to 0.1 mL/L . Each flask was inoculated with S . capricornutum to give a n initial concentration of 500 cells/mL . Flasks were then incubated under constant illumination at 22

– 27.6°C for 4 days . Following the 4 - day incubation period, the algal biomass in each flask was estimated based on the concentration of Chlorophyll a . Th e pH of the test solutions during the test was 7.6

– 7.9 . Statistics : None specified RESULTS Endpoint value(s) : Note that analytical monitoring was not conducted and nominal concentrations (1

– 1000 mg/L) were many orders of magnitude greater than the water solubility limit ( experimentally determined to be < 0.01 mg/L) 1 for this compound . Therefore, it is not possible to interpret reported toxicity values. 96 - hour EC 50 (Chlorophyll a concentrations) > 1000 mg/L , based on nominal concentrations . Results remarks : The Chlorophyll a concentrations in all of the test solutions, except for 4.6 mg/L, were higher than the mean concentration in the controls . Page 2 of the test report, which included studies for fish, Daphnia , and green algae, provided a general comment that the test substance “appeared to be not wholly soluble at concentrations greater than” 10 mg/L. CONCLUSIONS The 96 - hr EC 50 is > 1000 mg/L (nominal concentration) based on Chlorophyll a concentrations. STUDY RELIABILITY [3] Not reliable . Non - GLP study . Test method/guideline followed and statistical methods used were not specified, a nalytical monitoring was not conducted, water chemistry parameters were not provided (hardness), control treatment and control responses were not sp ecified , and the test concentrations exceeded the water solubility limit of the test substance ( experimentally determined to be < 0.01 mg/L) 1 by many orders of magnitude .

1 T he water solubility limit of CASRN 118 - 82 - 1 was experimentally determined to be < 0.01 mg/L by S afePharm Laboratories (study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 4, 2013; “4,4’ - Methylene bis(2,6 - di - tert - butylphenol): Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/011, study completed on Ju ly 18, 2005).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

33 REFERENCE Mattock, S.D . and Girling, A.E . 1988 . IONOX 220 : Acute Toxicity ( Salmo gairdneri, Daphnia magna and Selenastrum capricornutum ) and n - octanol/water partition coefficient . Sittingbourne Research Centre, Shell Research Limited . Submitted by Shell Oil Co . to EPA in 1992 as part of a TSCA 8(e) submission (EPA Doc . No. 88 - 9 20003444 , Fiche No. OTS0537403 ) entitled “Initial submission : acute toxicity of IONOX 220 in Salmo gairdneri, Daphnia magna and Selenastrum capricornutum and n - octanol/water partition coefficient w - letter dated 053092”.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

34 C - 5 . Acute Toxicity Summary of endpoint : Adequate Study 1 : Acute oral toxicity study in rats ( Ethyl Petroleum Additives International, 1994 ) Title : Acute Oral Toxicity Study This study is: Adequate Not Adequate TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1 Remarks : Purity not provided METHOD Method/guideline followed : OECD 401 Test type : Acute Oral - Rat GLP compliant? : Yes Year study performed : 1994 Test species : Rats (strain not specified) Number of animals/sex/dose : 5 Route of administration : Oral gavage Vehicle : Not specified Doses/concentrations tested : 5000 mg/kg - bw Statistical methods used : None specified Test method/conditions remarks : Ten rats (5/sex) were administered a single limit dose of 5000 mg/kg - bw by gavage, according to OECD TG 401 . Animals were observed for 14 days post - dosing . RESULTS Endpoint value(s) : LD 50 > 5000 mg/kg - bwDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

35 Results remarks : No animals died . Signs of toxicity included d iarrhea and yellow urine . No visible lesions were noted . No additional details were available from this study summary. CONCLUSIONS The oral LD 50 is > 5000 mg/kg - bw . STUDY RELIABILITY [ 2 ] Reliable with restrictions . Although this is an OECD guideline study , i mportant study details were missing from the study summary , includ ing test substance purity and use of a vehicle . Data are considered adequate based on weight of evidence. REFERENCE Ethyl Petroleum Additives International . 1994 . Study No. PH - 402 - ET - 005 - 94 for Ethyl Corporation, Pharmakon Research International, Waverly, PA . (Cited in IUCLID Dossier, European Chemical Bureau, 2000) . Submitted by A l bemarle Corporation . Robust summary located at h ttp://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0181 ( on p. 5 of the document “End_Point_D

– Mammalian_Toxicity_Acute.pdf” ) as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

36 Study 2 : Acut e oral toxicity study in rats (Carter and Dharmarajan, 1976 ) Title : A Report on the Acute Toxicity of AN - 702 This study is: Adequate Not Adequate TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1 Remarks : Purity not provided METHOD Method/guideline followed : Not indicated Test type : Acute Oral - Rat GLP compliant? : Not indicated Year study performed : 1976 Test species : Sprague - Dawley rats Number of animals/sex/dose : 10 males/dose Route of administration : Oral gavage Vehicle : Peanut Oil Doses/concentrations tested : 3000 mg/kg - bw Statistical methods used : None specified Test method/conditions remarks : Ten male Sprague - Dawley rats (weight of 200

– 300 g) were administered by gavage a single dose of 3000 mg/kg - bw of the test substance as a 10% w/v solution in peanut oil and were observed for 14 days following dosing . At termination, the survivors were sacrific ed and necropsied. RESULTS Endpoint value(s) : LD 50 > 3000 mg/kg - bw Results remarks : No animals died or exhibited any signs of toxicity . Extensive d ark areas were observed on the liver . Treatment with the test material did not affect body weight gain during the observation period . No additional details were available from this study summary.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

37 CONCLUSIONS The oral LD 50 is > 3000 mg/kg - bw. STUDY RELIABILITY [2] Reliable with restrictions . Important study details were missing from the study summary , includ ing test substance purity, method/guideline followed, and GLP compliance . Data are considered adequate based on weight of evidence. REFERENCE Carter, G . and V . Dh ar marajan, “ A Report on the Acute Toxicity of AN - 702, ” Tul ane University School of Medicine, for Ethyl Corporation, 1 976 . Submitted by Albemarle Corporation . Study report available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT 2005 - 0033 - 0181 ( on p p. 3, 7 - 13 of the document “End_Point_D

– Mammalian_Toxicity_Acute.pdf” ) as of Decembe r 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

38 Study 3 : Acute oral toxicity study in guinea pigs ( Haring and Robbins, 1965 ) Title : Acute Oral Toxicity of SCH 12150 in Guinea Pigs This study is: Adequate Not Adequate TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; referred to as SCH 12150 in study report ; Batch No . 3004 - 95 Remarks : Purity not provided METHOD Method/guideline followed : Not indicated Test type : Acute Oral - Guinea Pig GLP compliant? : N/A (Pre - GLP) Year study performed : 1965 Test species : Albino guinea pigs; from Gordon’s Biological Supply Company; average weight of 282 g Number of animals/sex/dose : 6 males/group Route of administration : Oral gavage Vehicle : Methyl Cellulose Doses/concentrations tested : 0 (control), 1000, 1780, 3170, 5630, or 10,000 mg/kg - bw Statistical methods used : None specified Test method/conditions remarks : Single doses of t he test substance were administered by gavage as a 20% suspension in 0.5% methyl cellulose to groups of 6 male guinea pigs . Animals were observed for 14 days following dos ing . Following the observation period, all surviving ani mals were autopsied and gross observations were recorded . Liver sections were retained for microscop ic examination . Other tissues which showed gross pathology were preserved. RESULTS Endpoint value(s) : LD 50 > 10,000 m g/kg - bwDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

39 Results remarks : Two deaths occurred in treated animals : one in the 1000 mg/kg - bw group (day 10) and one in the high est - dose group (day 12) . Cause of death in both cases was reported as “secondary infection” . One control animal died on day 11 , and pneumonia was present on autopsy . Pneumonia was found randomly in all groups . N o treatment - related clinical signs were observed . R educed weight gain (approximately 16% compared to control; statistical significance not reported ) occurred in the high est - dose group . Absolute l iver weights were not significantly different in treated versus control animals . Portions of the liver fro m five animals in each dose group , as well as any gross lesions, were processed for microscopic exam ination . No significant treatment - related changes were seen in microscopic examination of the liver; cellular fat and liver glycogen content appeared normal . Results of a follow - up study conducted in 1966 were included in this study report . The follow - up study was a repeat of the original experiment using two groups of animals at the high - dose level . One group was given chlortetracycline in the drink ing water to en sure that endemic infection was not present . There were no differences in the two groups with respect to infection or body weight gain . CONCLUSIONS The oral LD 50 is > 10,000 mg/kg - bw. STUDY RELIABILITY [2] Reliable with restrictions . Important study details missing from the study summary included test substance purity, method/guideline followed, statistical methods used and detailed numerical findings . Data are considered adequate based on weight of evidence. REFERENCE Haring, G.D . and G.R . Robbins, “ Acute Oral Toxicity of SCH 12150 In Guinea Pigs, ” Schering Laboratories, 19 65 . Submitted by Albemarle Corporation . Study report available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0181 ( on p p. 4 - 5, 41 - 55 of the document “End_Point_D

– Mammalian_Toxicity_Acute.pdf” ) as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

40 Study 4 : Acute dermal toxicity study in rabbits (Carter and Dharmarajan , 1976) Title : A Report on the Acute Toxicity of AN - 702 This study is: Adequate Not Adequate TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1 ; referred to as AN - 702 in study report Remarks : Purity not provided METHOD Method/guideline followed : Not specified Test type : Acute Dermal - Rabbit GLP compliant? : Not specified Year study performed : 1976 Test species : New Z ealand White rabbits; 2.3

– 3.0 kg Number of animals/sex/dose : 4 (sex not specified) Route of administration : Dermal , occluded Vehicle : Peanut Oil Doses/concentrations tested : 1500 m g/kg - bw Statistical methods used : No information available Test method/conditions remarks : The test substance (10% w/v solution in peanut oil) was injected under a plastic sleeve wrapped around the clipped trunks of four rabbits ( two with intact skin and two with abraded skin) for an exposure period of 24 hours . Eval uation s of erythema and edema were made 24 and 72 hours after application of the test material . After removal of the sleeves, the animals were observed for 14 days. RESULTS Endpoint value(s) : LD 50 > 1500 mg/kg - bwDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

41 Results remarks : No mortalities occurred . Well defined erythema was observed on the first day of observation. CONCLUSIONS The dermal LD 50 is > 1500 mg/kg - bw. STUDY RELIABILITY [ 2 ] R eliable with restrictions . Important study details that w e re missing from the study summary included test substance purity, methods/guidelines followed, GLP compliance, sex of test animals , statistical methods, detailed findings , and numerical results . Data are considered adequate based on weight of evidence. REFERENCE Carter, G . and V . Dh ar marajan, “ A Report on the Acute Toxicity of AN - 702, ” T ul ane University School of Medicine, for Ethyl Corporation, 1 976 . Submitted by Albemarle Corporation . Study summary available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT 2005 - 0033 - 0181 ( on p p. 7 - 13 of the document “End_Point_D

– Mammalian_Toxicity_Acute.pdf” ) as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

42 Study 5 : Acute oral toxicity study in rats (Cleveland and Witherup, 1962) Title : The Immediate Oral Toxicity of 4,4' - Methylenebis(2,6 - Di - tert - Butylphenol) This study is: Adequate Not Adequate The post - dosing observation period was less than 14 days . Also, i mportant study details were missing from the study summary , includ ing test substance purity, method/guideline followed, and statistical methods used. TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1 Remarks : Purity not provided METHOD Method/guideline followed : Not indicated Test type : Acute Oral - Rat GLP compliant? : N/A (Pre - GLP) Year study performed : 1962 Test species : Rats (strain not specified) Number of animals/sex/dose : 10 males/dose (definitive study); 1

– 2 males/dose (preliminary study) Route of administration : Oral gavage Vehicle : Peanut Oil Doses/concentrations tested : 180, 1000, 1600, 2400, 3200, 4700, or 10,000 mg/kg - bw (preliminary study); 3200, 4700, 7000, 10,000, 16,000 , or 24,000 mg/kg - bw (definitive study) Statistical methods used : None specified Test method/conditions remarks : In a preliminary study, m ale rats (10

– 14 weeks old) were administered the test substance as a 10% suspension in peanut oil by gavage at dose s ranging from 180 to 10,000 m g/kg - bw . In the definitive study, the rats received various amounts of a suspension containing 30 g of the test substance diluted to 100 mL with peanut oil ; this provided doses of the test substance ranging from 3200 to 24,000 m g/kg - bw . At the end of theDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

43 observation period, animals were necropsied and microscopic examination of the viscera was conducted. RESULTS Endpoint value(s) : LD 50 > 24,000 mg/kg - bw Results remarks : The duration of the observation period was not specified, but it appears to have been 4 weeks in the preliminary study and 5 days in the definitive study (from Tables in the report) . No animals died in either study . No signs of intoxication were observed, and only mild transient diarrhea was observed in animals administered the highe r doses (not specified) . Postmortem examination revealed no gross pathology findings ; no histopathological changes to the viscera were observed . No additional details were available from this study summary. CONCLUSIONS The oral LD 50 is > 24,000 mg/kg - bw . STUDY RELIABILITY [ 3 ] Not r eliable . The post - dosing observation period was less than 14 days . Also, i mportant study details were missing from the study summary , includ ing test substance purity, method/guideline followed, and statistical methods used. REFERENCE Cleveland, F. P. and S . Witherup, “ The Immediate Oral Toxicity of 4,4' - Methylenebis(2,6 - Di tert - Butylphenol), ” The Kettering Laboratory in the Department of Preventive Medicine and Industrial Health, College of Medicine, University of Cincinnati, Cincinnati, Ohio, for Ethyl Corporation, 1962 . (Submitted as part of Food Additive Petition No. 4B 1469 and Food Additive Petition 1397, Food and Drug Administration) . Study report available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0181 (on p p. 3, 14 - 17 of the document “End_Point_D

– Mammalian_Toxicity_Acute.pdf”) as December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

44 Study 6 : Acute oral toxicity study in dogs (Vymetal and Robbins, 1965) Title : Acute Oral Toxicity of SCH 12150 in Dogs This study is: Adequate Not Adequate The number of animals per group (one male and one female) is not sufficient to evaluate acute toxicity . Furthermore, statistical methods could not be used because of the small number of animals per group . Important study details missing from the study summary included test substance purity and method/guideline followed . TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; referred to as SCH 12150 in study report; Batch No. 3004 - 96; supplied as a white powder which was not soluble in water Remarks : Purity not provided METHOD Method/guideline followed : Not indicated Test type : Acute Oral - Dogs GLP compliant? : N/A (Pre - GLP) Year study performed : 1965 Test species : Beagle Dogs; 8

– 16 months old; obtained from Ubrian Farm Kennels Number of animals/sex/dose : 1 Route of administration : Oral - capsule Vehicle : No Doses/concentrations tested : 0 (control), 100, 251, 631, 1585 , and 2512 mg/kg - bw Statistical methods used : Not applicable given the use of one dog/sex in each dose group. Test method/conditions remarks : Groups of o ne male and one female beagle were administered a single dose of the test substance in gelatin capsules via the oral route and were observed for 14 days post - administration . Endpoints monitored included clinical signs, food consumption, body weight, as we ll as hematology, clinical chemistry, and urinalysis . AtDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

45 termination, the dogs were necropsied and the liver, kidneys, heart, gall bladder, pancreas, adrenals, and pituitary were prepared for microscopic examination. RESULTS Endpoint value(s) : LD 50 > 2512 mg/kg - bw Results remarks : There w ere no adverse clinical signs during the study . There were no treatment - related changes in any biochemical parameters measured (blood glucose, BUN, SGPT [ALT] , SGOT [AST] , alkaline phosphatase, icteric index) with the exception of an insignificant temporary rise in BUN in the male dog at the 631 mg/kg dose level . No treatment - related hematological effects were noted (hematocrit, hemoglobin, total WBC and differential coun ts, prothrombin time and platelet count) . Urinalysis revealed no treatment - related findings . No significant pathology was found at necropsy and microscopic examination of the selected organs . CONCLUSIONS The oral LD 50 is > 2512 mg/kg - bw. STUDY RELIABILITY [ 3 ] Not reliable . The number of animals per group (one male and one female) is not sufficient to evaluate acute toxicity . Furthermore, statistical methods could not be used because of the small number of animals per group . Important study d etails missing from the study summary included test substance purity and method/guideline followed . REFERENCE Vymetal, R . and G.R . Robbins, “ Acute Oral Toxicity of SCH 12150 in Dogs, ” Schering Laboratories, 1965 . Submitted by Albemarle Corporation . Study report available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0181 (on p p. 4, 18 - 40 of the document “End_Point_D

– Mammalia n_Toxicity_Acute.pdf”) as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

46 C - 6 . Repeated - Dose Toxicity Summary of endpoint : Adequate Study 1 : 2 - Year repeated - dose oral toxicity study in dogs ( Jolley et al.

, 1969) Title : The effects exerted upon beagle dogs during a period of two years by the introduction of 4,4' - methylenebis (2,6 - ditertiary butylphenol) into their daily diets This study is: Adequate Not Adequate TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1 Remarks : 99% pure; Sample No. 718 - 65; Appearance : fine, pale yellow colored, crystalline solid. METHOD Method/guideline followed : Not specified Test type : 2 - Year Oral Repeated - Dose Toxicity GLP compliant? : N/A (pre - GLP) Year study performed : 1969 Test species : Beagle dogs; obtained from Atlantic Kennels of Zionville, Indiana; 6.5

– 8.5 months old at study initiation Number of animals/sex/dose : 3/sex in each of the treated groups, 6/sex in the control group Route of administration : Oral - diet Vehicle : Corn oil (prior to incorporation into diet at a level of 5% corn oil by weight ) Exposure duration : 2 years Doses/concentrations tested : 0, 15, 60, 100, 500, and 3000 ppm (0, 0.395, 1.40, 2.52, 14.0, and 96.9 mg/kg - bw/day, respectively, for males and 0, 0.372, 1.74, 2.57, 16.3, and 99.7 mg/kg bw/day, respectively, for females; estimated by the investigators based on mean body weight and food consumption throughout the study) Statistical methods used : None specifiedDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

47 Test method/conditions remarks : Beagle dogs were fed diets containing the test substance at various concentrations for a period of 2 years . Dogs were observed daily for clinical signs of toxicity . Individual body weights and food consumption were recorded weekly . Analyses of hematological (packed cell volume, hemoglobin concentration, total leukocyte count, percentage of differentiated leukocytes, and prothrombin time) and clinical chemistry param eters (total serum cholesterol, total lipids, alkaline phosphatase, and serum glutamic - pyruvic transaminase [SGPT]) as well as urinalysis (color, appearance, specific gravity, pH, albumin, sugar, acetone, and microscopic assessment) were conducted prior to treatment (baseline; two measurements per dog), after 2 weeks , and after 1, 6, 12, 18, and 24 months of feeding on the test diets . Measurements of alkaline phosphatase, SGPT, and prothrombin time were not performed at the 2 - week feeding period but were p erformed at all other time points . Additional tests (serum bilirubin, blood urea nitrogen, bromsulfalein retention, calcium, inorganic phosphate, total protein, albumin, uric acid, glucose, and glutaminic - oxalacetic transaminase [SGOT]) were conducted to assess the possible effects on liver function at 12, 18, and/or 24 months . At termination, all dogs were subjected to gross examination and the major organs (liver, heart, lungs, kidneys, spleen, brain, prostate, testes, ovaries, pituitary, adrenals, and thyroids) were weighed . These organs, as well as the aorta, gall bladder, stomach, duodenum - pylorus, small intestine (ileum), large intestine (colon), mesenteric lymph nodes, urinary bladder, uterus, pancreas, thymus, spinal cord, eye, bone marrow, bone, skeletal muscle, and skin were processed for microscopic examination . Examination of feces at 6 months revealed hookworms, roundworms, and whip - worms present . Although not all of the dogs had these parasites, all were treated with the same worming trea tment . RESULTS Endpoint value(s) : LOAEL = 500 ppm (14.0 and 16.3 mg/kg - bw/day for males and females, respectively) based on reduced body weight gains (in the absence of a treatment - related effect on food consumption), increased relative liver weights, and histopathological changes in the liver (increased fat in hepatic cells [males] and increased prominence of Glisson’s capsule [females]); NOAEL = 100 ppm (2.52 and 2.57 mg/kg - bw/day for males and females, respectively). Results remarks : Note that the study authors did not report group means or percent change from baseline (either for individual dogs or for treatment groups) . Therefore, quantitative results reported as percent change from baseline or percent change from control were calculated by the EPA reviewers for this review . If baseline data were reported, the percent change relative to the baseline (mean of the two baseline values for each dog) for individual dogs was determined, and then group means for the percent change were calculated for e ach treatment group . If baseline data were not reported, group means for treatment groups were calculated and compared to control means for each time point. At 3000 ppm, one male died during week 40 and another was sacrificed moribund during week 57 . Bo th deaths were considered to be due to liver cirrhosis resulting from dietary administration of the test substance . No deaths occurred at dietary concentrations of 500 ppm and lower in males, and no deaths occurred in females.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

48 Dogs from all groups, inclu ding the control group, exhibited one or more clinical signs including patches of alopecia, enlarged nipples and/or mammary glands, excess of tartar, undershot jaw, and tonsillitis . The study authors did not consider any of these signs to be treatment - rel ated; however, no incidence data were provided to support this claim . Body weight loss was observed in two dogs . After 2 years of treatment, body weight of the surviving male in the 3000 ppm group was 22% lower and body weight of one female in the 500 ppm was 5.3% lower, compared to body weights at the start of treatment . Body weight gain in males was decreased by 46% in the 500 ppm group, compared to the control group . Body weight gain in females in the 500 and 3000 ppm groups was decreased by 54 and 66%, respectively, relative to female controls . No adverse treatment - related effects on body weight gain were apparent in males or females at dietary concentrations of 15, 60, or 100 ppm . Food consumption did not appear to be affected by treatment . E valuation of hematological parameters showed decreased packed cell volume and hemoglobin concentration in males in the 3000 ppm group, compared to baseline, but not at lower dietary concentrations or in any group for females . Packed cell volume in males w as decreased by approximately 26 (n=2), 34 (n=1), 33 (n=1), and 33% (n=1) after 12, 14 (early death), 18, and 24 months of treatment, respectively . Hemoglobin concentration in males was decreased by approximately 35 (n=2), 41 (n=1), 38 (n=1), and 31% (n=1 ) after 12, 14 (early death), 18, and 24 months, respectively . Due to early deaths of 2 out of 3 males, all measurements after 12 months were for only 1 dog; therefore, it is difficult to draw conclusions regarding the potential for treatment - related effe cts on packed cell volume and hemoglobin concentration . Study authors stated that these hematological changes were not clinically significant based on the absence of reticulocytosis . No treatment - related effects were observed on other hematologic parameters . Urinalysis revealed no treatment - related findings. Serum cholesterol was decreased, compared to baseline, throughout the treatment period in males and females in the 3000 ppm group . Decreases were 47, 53, 72, 71, 86, and 66% in males and 38, 54, 61, 60, 76, and 66% in females after 2 weeks and after 1, 6, 12, 18, and 24 months of treatment, respectively . Other treatment groups displayed more modest decreases in serum cholesterol; these changes were not consistently observed throughout the st udy period and their relationship to treatment was unclear . Total serum lipids were decreased (relative to baseline) in males and females in the 3000 ppm group, although changes were not consistent in males over the 2 - year treatment period . In males, lip ids were decreased by 33, 41, 21, 43, and 9% after 1, 6, 12, 18, and 24 months of treatment, respectively . In females, lipids were decreased by 32, 32, 36, and 39% after 1, 6, 18, and 24 months of treatment, respectively, with no decrease at 12 months . T otal serum lipids were not affected in any other treatment group . Study authors stated that a “questionable lowering” of serum total protein was observed in dogs in the 3000 ppm group after 24 months of treatment (not measured at earlier times), compare d to controls (no baseline measurement reported) . In the one male dog surviving until 24 months, protein was decreased by 13% . In female dogs, there was no apparent treatment - related effect on serum total protein, with a decrease of only 8% at 24 months . There were variations in totalDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

49 protein values among animals from other groups but they were very slight and did not show a dose - response relationship . Clinically significant increases ( ≥ 3 - fold) in serum alkaline phosphatase levels were observed in ma les and females in the 3000 ppm group, relative to baseline, at several time points . In males, alkaline phosphatase levels were increased by 4.1 - , 6.9 - , and 5.2 - fold at 6, 12, and 24 months, respectively, with a smaller increase of 2.5 - fold after 1 and 18 months of exposure . In females, levels were increased by 3.0 - , 4.3 - , and 3.7 - fold after 6, 12, and 18 months of exposure, respectively, with smaller increases of 1.7 - and 2.2 - fold after 1 and 24 months of exposure . In the 500 ppm group, no clinically si gnificant increases in alkaline phosphatase were observed; the highest increases were 1.6 - fold in males at 24 months and 2.3 - fold in females at 12 months . Alkaline phosphatase levels were not increased at dietary concentrations ≤ 100 ppm . Increases in me an SGPT levels were observed at all concentrations at varying time points throughout the 2 - year study period; however, these increases showed no clear, consistent dose - response relationship and were less than 3 - fold compared to baseline . No treatment - re lated effects were observed for serum bilirubin, blood urea nitrogen, bromsulfalein retention, calcium, inorganic phosphate, albumin, uric acid, glucose, or SGOT. At the end of treatment (upon early death or after 24 months on treatment), liver weights were increased in dogs fed diets containing 500 and 3000 ppm compared to controls . In the 500 ppm group, relative liver weights were increased by 32 and 28% in males and females, respectively, while in the 3000 ppm group, relative liver weights were incre ased by 92 and 61% in males and females, respectively . In the 500 ppm group, absolute liver weights were increased by 20 and 7% in males and females, respectively; similar increases were observed at lower doses and absolute liver weights did not show a do se - response relationship at dietary concentrations ≤ 500 ppm . In the 3000 ppm group, absolute liver weights were increased by 53 and 31% in males and females, respectively . The study authors considered the increase in relative liver weight in the 500 ppm group to be of questionable toxicological significance . However, since histopathological findings in the liver also were observed in the 500 ppm group, the EPA reviewers consider the increase in relative liver weight to be toxicologically significant . Increased absolute and relative kidney weights were observed in male dogs in the 3000 ppm group (37% increase in absolute weight and 72% increase in relative weight, compared to controls), with a smaller increase in females in the 3000 ppm group (10% inc rease in absolute kidney weight and 31% increase in relative kidney weight, compared to controls) . The toxicological significance of this finding is questionable, as no treatment - related changes in renal function or histopathological effects in the kidney were observed . No consistent dose - related changes were observed for weights of other organs . Post - mortem examination of all dogs receiving the 3000 ppm diet revealed marked cirrhosis of the liver and hyperplasia of the gall bladder mucosa . In the 500 ppm group, histopathological changes in the liver were observed in one male dog (increased fat in the hepatic cells) and one female dog (increased prominence of the Glisson’s capsule) . Study authors described severity of these findings as “slight” . Thes e changes in the liver were not observed in control animals . In the kidney, vacuolation of the epithelial cells of the Loop of Henle was noted in one female dog at 500 ppm and two female dogs at 3000 ppm, as well as one female dog in the control group;Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

50 th e toxicological significance of this finding is uncertain . There were no treatment - related histopathological findings at concentrations less than 500 ppm. CONCLUSIONS The LOAEL is 500 ppm (14.0 and 16.3 mg/kg - bw/day for males and females, respectively) based on reduced body weight gains (in the absence of a treatment - related effect on food consumption), increased relative liver weights, and histopathological changes in the liver (increased fat in hepatic cells [males] and increased prominence of Glisson’ s capsule [females]) . T he NOAEL is 100 ppm (2.52 and 2.57 mg/kg - bw/day for males and females, respectively) . STUDY RELIABILITY [2] Reliable with restrictions . No information on statistical analyses was reported (although some effects were reported as “significant”) and a small number of animals (3 dogs/sex/group) was used . The full study, although not a guideline study, was well documented and adequate based on the information provided. REFERENCES Jolley, W.P., Stemmer, K.L., Grande, F., Richmond, J., and Pfitzer, E.A . 1969 . The effects exerted upon beagle dogs during a period of two years by the introduction of 4,4' - methylenebis (2,6 - ditertiary butylphenol) into their daily diets . Kettering Laboratory, University of Cincinnati, OH . Submitted to EPA in 1992 as part of a TSCA 8(e) submission (EPA Doc. No. 88 - 920001353S, Fiche No. OTS0536073) entitled “Initial sub : the effect exerted upon beagle dogs during a period of 2 - yrs by the introduction of 4,4' - methylenebis (2,6 - ditertaryl butylphenol) into their daily diets (sanitized)”.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

51 Study 2 : 2 - Year repeated - dose oral toxicity study in dogs ( Jolley et al.

, 1975) Title : The effects upon beagle dogs following the ingestion of 4,4' - methylenebis (2,6 - ditertiary butylphenol) for two years including the changes after an additional seven months on control diet This study is: Adequate Not Adequate TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1 Remarks : 99% pure; Sample No. 1519 - 66; Appearance : fine, pale yellow - colored, crystalline solid. METHOD Method/guideline followed : Not specified Test type : 2 - Year Oral Repeated - Dose Toxicity GLP compliant? : Not specified Year study performed : 1975 Test species : Beagle dogs; obtained from American Animal Industries Inc., Indiana; 10

– 11 months old at study initiation . Number of animals/sex/dose : 6 Route of administration : Oral - diet Vehicle : Corn oil (prior to incorporation into diet at a level of 5% corn oil by weight ) Exposure duration : Up to 27 months; at one dose level (35 mg/kg - bw/day), a 27 - month dosing period was followed by 7 months on control diet Doses/concentrations tested : 0, 7, 35, and 75 mg/kg - bw/day; estimated by the investigators based on mean food consumption and body weight throughout the study. Statistical methods used : None specified Test method/conditions remarks : Beagle dogs (6/sex/dose) were fed diets containing the test substance in corn oil at 0, 7, 3 5, or 75 mg/kg - bw/day for periods up to 27 months . After 6 months on the test diet, 3 animals/sex/dose were sacrificed . The group of animals (3/sex) that was dosed at 35 mg/kg - bw/day for 27 months was then shifted to the control diet for 7 months and sacrificed at 34 months . The remaining animals (3/sex/dose) were sacrificed after 27 monthsDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

52 at 0 and 7 mg/kg - bw/day and after 25 months at 75 mg/kg - bw/day . Dogs were observed daily for clinical signs of toxicity . Individual body weights and food consump tion were recorded weekly . Analyses of hematological (packed cell volume, hemoglobin concentration, total leukocyte count, percentage of differentiated leukocytes, and prothrombin time) and clinical chemistry parameters (total serum cholesterol, alkalin e phosphatase, SGPT, blood urea nitrogen, fasting blood sugar, and total protein) as well as urinalysis (color, appearance, specific gravity, pH, albumin, sugar and microscopic appearance) were conducted prior to treatment (baseline), after 2 weeks, and af

ter 1, 3, 6, 12, 18, 24, and/or 27 months of feeding on the test diets; the hematological parameters were also evaluated during recovery at 33 months and the above clinical chemistry parameters were evaluated during recovery at 27.5, 28, 28.75, 33, and/or 34 months . Prior to the start of treatment and at 24 months, the following clinical chemistry parameters were assessed : total free serum lipids, serum sodium, serum potassium, thymol turbidity, and protein bound iodine . Additional clinical chemistry ana lyses (albumin, calcium, inorganic phosphate, SGOT, serum bilirubin, uric acid, and bromosulfalein retention) were conducted at 12, 18, 24, 27, 27.5, 28, 28.75, 33, and/or 34 months . At termination, all dogs were subjected to autopsy, the viscera were exa mined for gross pathological changes , and the major organs were weighed and processed for microscopic examination . The following organs were weighed individually : liver, heart, lungs, kidneys, spleen, brain, gonads, pituitary, adrenals, and thyroids . Th e following organs and tissues were examined microscopically : heart, lung, spleen, gall bladder, liver, kidney, stomach, two levels of small intestine, large intestine, pancreas, brain (cortex, brain stem, cerebellum, spinal cord), pituitary, thyroid glan d, adrenal gland, gonads, uterus, mammary glands, aorta, bone, lens of the eyes, urinary bladder, lymph nodes, skin, muscle, and fat. RESULTS Endpoint value(s) : LOAEL = 35 mg/kg - bw/day based on clinically significant increases in serum alkaline phosphatase activity and decreases in serum cholesterol levels, as well as histopathological changes in the liver (cirrhosis [with abundance of fibrous tissue, duplication of bile ducts, and cellular degeneration], marked lymphocytic cellular infiltration, dark brown pigmentation in Glisson’s c apsule, and vacuolation of hepatocytes); NOAEL = 7 mg/kg - bw/day . Results remarks : Note that the study authors did not report grou p means or percent change from baseline (either for individual dogs or for treatment groups) . Therefore, quantitative results reported as percent change from baseline or percent change from control were calculated by the EPA reviewers for this review . If baseline data were reported, the percent change relative to the baseline (mean of the two or three baseline values for each dog) for individual dogs was determined, and then group means for the percent change were calculated for each treatment group . If baseline data were not reported, group means for treatment groups were calculated and compared to control means for each time - point. One female dog from the 75 mg/kg - bw/day group developed marked ascites, was moribund , and was sacrificed during the 14 th m onth . Necropsy of this female revealed marked cirrhosis of the liver and congestion of the spleen . A male dog from the same group died during the 21 st monthDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

53 of treatment . The death of the male was considered to be due to intra - intestinal hemorrhages and marked cirrhosis of the liver resulting from treatment with the test substance . No clinical signs of toxicity were observed in surviving dogs. Weight loss was observed in the one male dog of the 75 mg/kg - bw/day group that died . No other effects on body weight were observed . No clear treatment - related effects were observed on growth rate (based on body weight gain) or food consumption . No treatment - related effects on hematology or urinalysis parameters were apparent. Clinically significant increases ( ≥ 3 - fold) in serum alkaline phosphatase levels, compared to baseline levels, were observed in male and female dogs in the 35 and 75 mg/kg - bw/day groups, with greater increases observed in females compared to males . In males in the 35 mg/kg - bw/day group, alkaline phosphatase levels were increased by approximately 3.3 - and 3.4 - fold after 6 and 12 months, respectively, with subclinical increases of 2.5 - , 2.6 - , and 2.8 - fold at 3, 18, and 24 months, respectively . In males in the 75 mg/kg - bw/day group, alkalin e phosphatase was increased by 3.6 - , 4.6 - , 3.8 - , and 3.7 - fold after 3, 6, 12, and 24 months, respectively, with subclinical increases of 2.7 - , 2.6 - , and 2.9 - fold after 0.5, 1, and 18 months of treatment, respectively . In females in the 35 mg/kg - bw/day gro up, alkaline phosphatase was increased by 4.8 - , 4.3 - , 7.3 - , 9.0 - , 5.9 - , and 7.6 - fold after 0.5, 3, 6, 12, 18, and 24 months of treatment, respectively; increases after 1 month of treatment were subclinical (2.6 - fold increase) . In females in the 75 mg/kg - b w/day group, alkaline phosphatase was increased by 3.0 - , 4.4 - , 5.9 - , 8.9 - , 4.6 - , and 4.6 - fold after 0.5, 3, 6, 12, 18, and 24 months of treatment, respectively; increases after 1 month of treatment were subclinical (2.4 - fold increase) . Serum alkaline phos phatase in males and females in the 35 mg/kg - bw/day group returned to approximately baseline levels within 1.75 months into the recovery period . No consistent treatment - related, clinically significant increases ( ≥ 3 - fold) in SGPT levels, relative to bas eline, were observed . In the 75 mg/kg - bw/day group, SGPT levels were increased by 3.8 - fold in both males and females at the 12 - month time - point only . Measurements of SGOT at 12, 18, and 24 months showed no consistent treatment - related pattern of change, relative to controls (baseline data not reported) . The two dogs that died at 75 mg/kg - bw/day showed increases in SGOT at 12 months of 3.6 - fold (male) and 3.0 - fold (female) relative to the control mean . Any increases in SGOT observed in other animals did not exceed 2 - fold (relative to the control mean) at any time point. Decreases (relative to baseline) in serum total cholesterol levels were observed in dogs in the 35 mg/kg - bw/day group from 6 through 24 months and in the 75 mg/kg - bw/day group from 3 thro ugh 24 months . During months 6

– 24 of treatment with 35 mg/kg - bw/day, decreases in serum total cholesterol ranged from 43 to 60% in male dogs and from 35 to 42% in female dogs; decreases did not exhibit duration - dependence in the 35 mg/kg - bw/day group . In male dogs in the 75 mg/kg - bw/day group, cholesterol was decreased by 52% after 3 months, with duration dependent decreases up to 79% of pre - treatment levels at 24 months . In female dogs in the 75 mg/kg - bw/day group, cholesterol was decreased by 37% after 3 months, with decreases ranging from 61 to 75% during months 6

– 24, with no duration - dependence . Serum total cholesterol in dogs in the 35 mg/kg - bw/day group returned to pre - treatment levels within 1.75 months into the recovery period . Serum total free lipids were decreased relative to baselineDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

54 in the 75 mg/kg - bw/day group at 6 and 24 months (not measured at other time points) by 33 and 49%, respectively, in males and by 39 and 43%, respectively, in females . No consistent changes from baseline levels we re observed for dogs in the 7 and 35 mg/kg - bw/day groups . Elevated bromsulfalein retention values (an indication of liver toxicity) were noted in all treatment groups; however, there was no dose response - relationship, the increases were less than 2 - fold, and there was “considerable variation” in measurements within groups . No treatment related effects were observed on serum bilirubin, serum uric acid, or thymol turbidity . For serum total protein levels, study authors stated that levels were sli ghtly decreased in dogs in the 75 mg/kg - day group, although they did not indicate if protein levels in treatment groups were compared to baseline levels or controls . Changes in serum total protein levels, relative to controls, were calculated for this rev iew; decreases were found to be ≤ 13% in males and ≤ 19% in females . Comparison of treatment groups relative to baseline levels, conducted by the EPA reviewers, revealed a decrease in protein early in the study, followed by an increase; all groups display ed a similar pattern and, therefore, a treatment - related effect on protein was not apparent . Serum albumin levels at 12, 18 , and 24 months were decreased in a dose - related manner, compared to controls (baseline levels not reported); no pattern of durati on - dependence was observed . In male dogs during months 12 to 24, decreases ranged from 7 to 11%, 16 to 36%, and 22 to 49% in the 7, 35, and 75 mg/kg - bw/day groups, respectively . In females, decreases ranged from 21 to 27%, 24 to 35%, and 43 to 50% in the 7, 35, and 75 mg/kg - bw/day groups, respectively . Changes at 75 mg/kg - bw/day were of likely toxicological significance, while the toxicological significance of changes at 7 and 35 mg/kg - bw/day is unclear . For serum urea nitrogen, study authors stated t hat levels in the 35 and 75 mg/kg - bw/day groups “tended to be slightly higher than controls” at 1, 3, and 6 months . However, review of the data for this report showed that levels in treatment groups were not consistently higher or lower than controls thro ughout the study . The largest increases in serum urea nitrogen at 35 and 75 mg/kg bw/day, relative to controls, were observed at 3 months and were 14 and 43%, respectively, in males and 33 and 57%, respectively, in females . Decreased serum urea nitrogen was observed later in the study period; at 24 months, decreases at 35 and 75 mg/kg - bw/day, relative to controls, were 24 and 35% in males, respectively, and 19 and 32%, respectively, in females . Changes relative to baseline values showed a similar pattern . In the absence of consistent effects on serum urea nitrogen, the toxicological significance of these changes is not clear. Serum sugar levels were slightly and sporadically increased in the 35 and 75 mg/kg - bw/day groups, compared to controls as well as baseline values, at 1, 3, and/or 6 months . Study authors stated that values were within the normal range, although the normal range was not reported . Increases in serum sugar levels were not observed after 12 months, leading study authors to conclude th at changes in serum sugar levels were not clinically significant . Serum protein - bound iodine levels after 24 months of treatment (only time point tested ) were decreased , based on comparison with baseline values, by 19, 27, 39, and 47% in males and by 0, 21, 34, and 53% in females at 0, 7, 35, and 75 mg/kg - bw/day , respectively . The clinical significance of this finding is uncertain .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

55 No treatment - related effects were observed for serum sodium, potassium, calcium, or inorganic phosphorus levels at 12, 18, or 24 months, relative to baseline levels . Study authors stated that slight decreases in calcium were observed in dogs in the 75 mg/kg - bw/day group; however, study authors do not indicate how comparisons between treatment groups were made . Comparison of the 75 mg/kg - bw/day group relative to control values, conducted for this review, showed decreases that were ≤ 13% and were not considered clinically significant by the EPA reviewers . Comparison of treatment groups relative to baseline levels, conducte d by the EPA reviewers, revealed that calcium increased in all groups during the first 12 months , and all groups showed a similar pattern of change compared to baseline . Absolute and relative liver weights were increased , relative to controls , in male a nd female dogs in the 75 mg/kg - bw/day group after 25 months . In high - dose males, absolute and relative liver weights were increased by 86 and 91%, respectively, and in females by 42 and 49%, respectively . At 35 mg/kg - bw/day, small increases in absolute ( 7 and 14% in males and females, respectively) and relative (13 and 20% in males and females, respectively) liver weights were observed after 6 months; the toxicological significance of these changes was unclear . The remaining dogs in the 35 mg/kg - bw/day g roup were sacrificed after the 7 - month recovery period and had liver weights similar to control values . Liver weights at 7 mg/kg - bw/day did not appear to be affected by treatment . No consistent dose - related changes were observed for weights of other orga ns . Histopathological examination revealed morphological changes in the liver s of dogs from the 35 and 75 mg/kg - bw/day groups . After 25 months at 35 mg/kg - bw/day, three dogs (one male and two females) showed moderately increased prominence of Glisson’s c apsule (also observed in some control animals); two other dogs (males) showed liver cirrhosis (with abundance of fibrous tissue, duplication of bile ducts, and cellular de generation) accompanied by marked lymphocytic cellular infiltration and dark brown pigment in Glisson’s c apsule . The sixth animal in this dose group (a female) showed no remarkable histological findings in the liver at 25 months . Vacuolation of hepatocyt es was observed in one male and one female at 35 mg/kg bw/day . The majority of dogs fed 75 mg/kg - bw/day suffered from moderate to severe liver cirrhosis by 2 years, accompanied by mild - to - moderate lymphocytic cellular infiltration, dark brown pigmentation in Glisson’s c apsule, and vacuolation of hepatocytes . Mild to moderate prominence of Glisson’s c apsule and mild lymphocytic cellular infiltration were observed in the controls . Changes similar to those in the controls were observed in livers of dogs in the 7 mg/kg bw/day group . After being fed the control diet for 7 months, the severity of the liver lesions was markedly decreased in all affected animals of the 35 - mg/kg - bw/day group . Additionally, the characteristics of the lesions changed; the fibrou s tissue became denser and was in most instances a fine band . Therefore, the hepatic lesions in dogs receiving 35 mg/kg - bw/day appear to be partially reversible . The residual lesions in the liver were not considered by study authors to be cirrhotic . Com plete recovery of the liver did not occur following a 7 - month period on the control diet . Adverse, treatment - related histopathological changes were not seen in the other organs.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

56 CONCLUSIONS The LOAEL is 35 mg/kg - bw/day based on clinically significant increases in serum alkaline phosphatase activity and decreases in serum cholesterol levels, as well as histopathological changes of the liver ( cirrhosis [ with abundance of fibrous tissue, duplica tion of bile ducts, and cellular degeneration ] , marked lymphocytic cellular infiltration, dark brown pigmentation in Glisson’s c apsule, and vacuolation of hepatocytes ) ; the NOAEL is 7 mg/kg - bw/day . STUDY RELIABILITY [2] Reliable with restrictions . No information on statistical analyses was reported . The full study, although not a guideline study, was well documented and adequate based on the information provided. REFERENCES Jolley, W.P., Stemmer, K.L., Pfitzer, E.A., Grande, F., and Richmond, J . 19 75 . The effects upon beagle dogs following the ingestion of 4,4' - methylenebis (2,6 - ditertiary butylphenol) for two years including the changes after an additional seven months on control diet . Kettering Laboratory, University of Cincinnati, OH . Submitte d to EPA in 1992 as part of a TSCA 8(e) submission (EPA Doc. No. 88 - 920001353S, Fiche No. OTS0536073) entitled “Initial sub : the effect exerted upon beagle dogs during a period of 2 - yrs by the introduction of 4,4' - methylenebis (2,6 - ditertaryl butylphenol) into their daily diets (sanitized)” .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

57 Study 3 : Combined repeated - dose and carcinogenicity toxicity study (ECHA website, Exp WoE Repeated dose toxicity : oral.003 ) Title : Not provided This study is: Adequate Not Adequate TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1 ; referred to as 4,4' - Methylenebis(2,6 - di - tert - butylphenol) in the study report (other synonyms : BHD, AN2, Ethyl Antioxidant 702). Remarks : Purity not specified; crystalline solid METHOD Method/guideline followed : Similar to OECD TG 453 “Combined Chronic Toxicity/Carcinogenicity Studies” Test type : Combined Repeated - Dose Toxicity/Carcinogenicity Study GLP compliant? : N/A (Pre - GLP) Year study performed : 1969 Test species : Crj : CD(SD) rat; s ource : Charles River Breeding Laboratories Inc., Brookline, MA; age : weanlings at initiation; weight (average during week 1) : 394

– 436 g (males) and 248

– 269 g (females) . Number of animals/sex/dose : 25 in the test groups; 50 in the control group Route of administration : Oral - Diet Vehicle : Mazola corn oil (reported to be free of any antioxidants) Exposure duration : 2 years Doses/concentrations tested : 0, 15, 60, 100, 500, and 3000 ppm (corresponding approximate dose levels were 0, 0.55, 2.14, 3.56, 18.1, and 107 mg/kg - bw/day, respectively, for males and 0, 0.67, 2.63, 4.35, 22.2, and 138 mg/kg - bw/day, respectively, for females) Statistical methods used : Not specifiedDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

58 Test method/conditions remarks : Dosing and Environmental Conditions: Rats (25/sex/dose) were fed diets containing the test substance at various concentrations for up to 2 years . A control group of 50 rats was fed a diet containing only the corn oil vehicle . Four rats per sex were assigned to subgroups of each dose group for interim kills at 13, 26, 52, and 78 weeks (3, 6, 12 , and 18 months) . Terminal sacrifice was performed at 104 weeks (24 months) . The animals were acclimated for 2 weeks and were not fasted prior to the start of the study . The test animals were housed three males and four females per cage prior to the s tart of the study . Diet and water were provided ad libitum throughout the study . During the study, the test temperature was maintained at 76 ± 2 °F and the humidity was not controlled. Preparation of Test Diets: For preparation of the test diets, “1 kg of corn oil was added to 19 kg of the base diet with or without the test substance . The mixture was stirred for 20 min in a Hobart mixer (stainless steel) . [Five] diets containing the test item were prepared... The diet was freshly prepared every week.” [ ECHA ] The test diets were prepared by “mixing appropriate amounts with [a] special preparation of Rockland Mouse and Rat diet containing no more than 1% of fat.” [ECHA ] The rodent diet food was obtained fresh once each month. Clinical Observations, Body Weights, and Food Consumption: Cage - side observations were made on a daily basis for general appearance and behavior . Animals were observed weekly for a detailed assessment of clinical signs . Body weight and food consumption were recorded weekly . Mean test substance intake for males and females in each group was calculated by an unspecified method . Ophthalmological examinations were performed weekly , and all groups were examined for the presence of cataracts . Hematology and Clinical Chemistry Measurements: Hematological examinations were performed on unfasted treated and control animals (10 rats/sex/dose) at 1 week before the start of dosing, and again at 13, 26, 52, 78, and 104 weeks . Hematological parameters examined included the following : prothrombin time, hemoglobin, packed cell volume of erythrocytes, and total and differential leukocytes . Clinical chemistry investigations were performed on blood samples collected at scheduled sacrifice of all test animals . Clinical chemistry paramete rs examined included the following : total lipids, cholesterol, and alkaline phosphatase . No urinalysis assessments were performed. Gross Necropsy and Histopathology: It appears that organ weights were recorded, but, with the exception of the liver, the organs weighed were not specified in the ECHA study summary (the full study report was not available for review) . Pathological examinations were performed on the following organs at gross necropsy : liver, heart, lung, kidneys, spleen, brain, gonads, pituitary, adrenals, and thyroid . Histological examinations were performed on selected tissues, including the heart, aorta, lung, liver, spleen, kidney, stomach, duodenum, pylorus, smallDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

59 intestine (ileum), large intestine (colon), mesenteric lymph nodes, urinary bladder, prostate, testes, ovaries, uterus, pancreas, thyroid, thymus, adrenals, pituitary, brain, spinal cord, eye, bone marrow, bone, skeletal muscle, and skin. RESULTS Endpoint value(s) : LOAEL (males) = 500 ppm (18.1 mg/kg - bw/ day) based on increased incidence and severity of fatty degeneration of the liver and increased incidence of bile duct multiplication; NOAEL (males) = 100 ppm (3.56 mg/kg - bw/day) . LOAEL (females) = 100 ppm (4.35 mg/kg - bw/day) based on increased incidence and severity of bile duct multiplication; NOAEL (females) = 60 ppm (2.63 mg/kg - bw/day). Results remarks : Mortality: Mortalities occurred at all dose levels at incidences similar to controls . The mortality percentages at dietary concentration s of 0, 15, 60, 100, 500, and 3000 ppm were 54.9, 46.2, 57.1, 44.0, 61.5, and 46.2 % , respectively, for males and 52.9, 40.0, 44.0, 68.0, 38.5, and 52.0%, respectively, for females . Several rats in all groups had various infectious diseases (not specified on the ECHA website), which resulted in death . These illnesses were not considered to be related to treatment . Clinical Observations, Body Weight, and Food Consumption: Greenish - yellow discoloration of the fur was observed at 500 and 3000 ppm beginning at 1 year and 6 months of treatment, respectively . Occasional discoloration was also observed in some rats at 100 ppm after 15 months of treatment . No treatment - related effects were found on mean body weight, mean body weight gain, or food cons umption in any group . Although variations were observed in individual body weight, body weight gain, and food consumption for some test animals, these effects were considered to be related to illness and were not considered to be treatment - related . Food efficiencies in all treated groups were comparable to controls . No treatment - related findings were observed during ophthalmological examinations . Hematology and Clinical Chemistry Measurements: Mild to moderate leukopenia was observed in some females a f

t er up to 78 weeks of feeding, but this finding was comparable to controls . Anaemia or polychromasia, which was not considered to be treatment - related, was noted in individual males and females in all groups, including controls, at weeks 78 and 104 . On e week prior to treatment, a statistically significant difference (p - value was not provided) in prothrombin time was noted in males at 15 and 500 ppm, and in females at 60, 100, 500, and 3000 ppm, relative to controls (the direction and magnitude of these changes were not provided on the ECHA website) . Additionally, this same effect was noted in males at 15, 100, and 3000 ppm at 13 weeks and in females at 500 ppm at 26 weeks . No other differences in prothrombin time relative to controls were noted at any other observation times (weeks 52, 78, and 104) . “As this finding was not dose related and did not persist throughout the course of the study, it was not considered treatment - related.” [ECHA ] No other effects were observed for any other hematological par ameters that were analyzed.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

60 A statistically significant increase (p - value was not provided) in alkaline phosphatase levels was observed in males at 3000 ppm at week 104 . “Due to major fluctuations in the measurements this finding was not considered treat ment related by the [study] authors.” [ ECHA ] However, due to the histopathological effects observed in the liver of rats in this group, the EPA reviewers consider the increase in alkaline phosphatase to be toxicologically significant . “No consistent treatment related effect was observed on the serum cholesterol and total lipid levels.” [ECHA ] Gross Necropsy and Histopathology: “All relative organ weights and most absolute organ weights in the treated groups were comparable to the res pective control values . Single rats receiving 3000 ppm of the test substance were [reported] to have enlarged livers, but the group averages did not reach statistical significance, with the exception of females killed after week 13 and males killed after week 78.” [ECHA ] A further description of the incidence and statistical significance of enlarged livers after these time points was not provided on the ECHA website . Mean relative liver weight was statistically significantly reduced (p - value was not prov ided) compared to controls in females fed 15 ppm of the test diet for 104 weeks . Since a dose - related trend was not observed and the finding appeared to be an isolated observation, the effect was no t considered to be treatment - related . Necropsy showed no treatment - related gross organ changes in male or females. Histopathological examination revealed that all of the animals (treatment and control) killed at 24 months had pneumonia or interstitial nephritis or both . Throughout the study, 183 rats died . “[For] 52 [rats] no postmortem pathology could be done due to autolysis . For 131 [rats] postmortem pathology revealed in most instances massive pulmonary infection as the cause of death . Liver lesions were the most prominent findings and consisted of fa tty degeneration of the hepatic [cells] and a proliferation of the bile ducts . Fatty degeneration [ranged] from small fat vacuoles to a fatty liver . There was no special localization of the [degenerated] cells in the liver lobe... Two types of proliferati on of the bile ducts could be distinguished morphologically : 1 . multiplication of the bile ducts with retainment of their normal structure occurred in small clusters . 2 . A dilatation and multiplication of the bile ducts with fibrotic walls without epithe lium . In some of the dilated bile ducts mucous plaques were observed . No inflammatory reaction was present . [With] [u]p to 3 months of feeding no alterations in the livers were observed . Both lesions [increased] in control and treated rats from 18 mont hs of treatment . After 24 [months] more severe lesions were observed in the 500 and 3000 ppm dose groups.” [ECHA ] At 12 months in males, the incidences of fatty degeneration of the liver at dietary concentrations of 0, 15, 60, 100, 500, and 3000 ppm wer e 0/4, 1/4, 0/4, 0/4, 4/4, and 4/4 (0, 25, 0, 0, 100, and 100%) , respectively . At 18 months in males, the incidences at 0, 15, 60, 100, 500, and 3000 ppm were 0/4, 0/4, 0/1 1 , 0/4, 3/3, and 2/3 (0, 0, 0, 0, 100, and 67%) , respectively . At 24 months in males, the incidences at 0, 15, 60, 100, 500, and 3000 ppm were 8/22, 8/14, 5/12, 8/14, 6/11, and
1 Only one male was examined histologically in the 60 ppm group at 18 months , according to the data tables for fatty degeneration and bile duct multiplication provided on the ECHA website . The reason (s) for the sm all number of males in this sub group was not explained .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

61 14/14 ( 36, 57, 42, 57, 55, and 100%), respectively . Statistical analyses 1 by Fisher’s exact test revealed significant increases in incidence of hepatic fatty degeneration in males at 500 ppm at 12 (p = 0.03) and 18 months (p = 0.03) and at 3000 ppm at 12 (p = 0.03) and 24 months (p < 0.0001) . In addition, there appeared to be a dose - related increase in severity of hepatic fatty degeneration in males at ≥ 500 ppm at 24 months . At 12 months in males, the incidences of bile duct multiplication at 0, 15, 60, 100, 500, and 3000 ppm were 0/4, 1/4, 1/4, 2/4, 3/4, and 4/4 ( 0, 25, 25, 50, 75, and 100%), respectively . At 18 months in males, the incidences at 0, 15, 60, 100, 500, and 3000 ppm were 0/4, 0/4, 0/1, 0/4, 3/3, and 3/3 ( 0, 0, 0, 0, 100, and 100%) , respectively . At 24 months in males, the incidences at 0, 15, 60, 100, 500, and 3000 ppm were 5/23, 3/14, 2/12, 5/14, 5/11, and 10/14 ( 22, 21, 17, 36, 45, and 71%), respectively . Statistical analyses by Fisher’s exact test revealed significant increases in incidence of bile duct multiplication in males at 500 ppm at 18 months (p = 0.03) and at 3000 ppm at 12 (p = 0.03), 18 (p = 0.03), and 24 months (p = 0.005) . No dose - related increase in severity of bile duct multiplication in males was apparent. At 12 months in females, the incidences of fatty degeneration of the liver at 0, 15, 60, 100, 500, and 3000 ppm were 0/4, 0/4, 0/4, 0/4, 1/4, and 3/4 ( 0, 0, 0, 0, 2 5, and 75%), respectively . At 18 months in females, the incidences at 0, 15, 60, 100, 500, and 3000 ppm were 0/4, 0/4, 1/4, 0/4, 3/4, and 3/3 ( 0, 0, 25, 0, 75, and 100%), respectively . At 24 months in females, the incidences at 0, 15, 60, 100, 500, and 3 000 ppm were 4/24, 3/14, 4/14, 2/8, 6/16, and 8/12 ( 17, 21, 29, 25, 38, and 67%), respectively . Statistical analyses by Fisher’s exact test revealed significant increases in incidence of hepatic fatty degeneration in females at 3000 ppm at 18 (p = 0.03) a nd 24 months (p = 0.007) . No dose - related increase in severity of hepatic fatty degeneration in females was apparent. At 12 months in females, the incidences of bile duct multiplication at 0, 15, 60, 100, 500, and 3000 ppm were 0/4, 0/4, 1/4, 1/4, 1/4, and 3/4 ( 0, 0, 25, 25, 25, and 75%), respectively . At 18 months in females, the incidences at 0, 15, 60, 100, 500, and 3000 ppm were 1/4, 0/4, 3/4, 0/4, 3/4, and 4/4 ( 25, 0, 75, 0, 75, and 100%), respectively . The incidences in females at 24 months at 0, 15, 60, 100, 500, and 3000 ppm were 2/24, 3/15, 4/14, 4/8, 11/16, and 8/12 ( 8, 20, 29, 50, 69, and 67%), respectively . Statistical analyses by Fisher’s exact test revealed significant increases in incidence of bile duct multiplication in females at 100 p pm at 24 months (p = 0.02), at 500 ppm at 24 months (p = 0.0001), and at 3000 ppm at 18 (p = 0.05) and 24 months (p = 0.0006) . In addition, there appeared to be a dose - related increase in severity of bile duct multiplication in females at ≥ 100 ppm at 24 months. CONCLUSIONS The LOAEL was 500 ppm (18.1 mg/kg - bw/day) in males based on increased incidence and severity of fatty degeneration of the liver and increased incidence of bile duct multiplication . The NOAEL was 100 ppm (3.56 mg/kg - bw/day) in males .

1 All statistical analyses of histopathological results were conducted by EPA reviewers using GraphPad Software (Scientific Software) , a vailable at http://graphpad.com/quickcalcs/index.cfm .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

62 The LOAEL was 100 ppm (4.35 mg/kg - bw/day) in females based on increased incidence and severity of bile duct multiplication . The NOAEL was 60 ppm (2.63 mg/kg - bw/day) in females. STUDY RELIABILITY [2] Reliable with restrictions . The study is considered to be reliable with restrictions based on weight of evidence . When compared to the TSCA 799.9430 guideline, the following deviations were noted : ( 1) the total number of animals at test initiation was not specified ; ( 2) the lot or batch number of the test substance was not provided; ( 3) the environmental test temperature (76 ± 2 ° F) exceeded the recommended temperature of 22 ± 3 °C (71.6 ° F); ( 4) humidity was not controlled ; ( 5) limited hematology and clinical chemistry parameters were examined ; ( 6 ) urinalysis was not conducted ; ( 7) organs weighed at necropsy were not specified ; and ( 8) group mean data for endpoints other than histopathological lesions were not reported . Inconsistencies in some of the data tables on the ECHA website for severity grades of fatty degeneration of the liver and bile duct multiplication (incidences and percentages) render some of this information of questionable value for fully describing the potential toxicity of CASRN 118 - 82 - 1 . However, these inconsistencies did not impact the assignment of the endpoint values . REFERENCE Study summary available from the ECHA website at http://apps. echa.europa.eu/registered/data/dossiers/DISS - dcee47d2 - 62ad - 0171 - e044 - 00144f67d031/AGGR - bf547f31 - 7fe6 - 4a5f - a06a - e41a1a9a2ab0_DISS - dcee47d2 - 62ad - 0171 - e044 - 00144f67d031.html#AGGR - bf547f31 - 7fe6 - 4a5f - a06a - e41a1a9a2ab0 (study indicated as Exp WoE Repeated dose t oxicity: oral.003 in online entry for CASRN 118 - 82 - 1) as of December 2, 2013. The data source is referred to as a “study report”, with a report date of 1969 - 07 - 08 . No other reference information is provided . Text in quotes is taken directly from the ECHA website. Copyright Notice : © European Chemicals Agency, 2007 - 2011 . Reproduction is authorised provided the source is fully acknowledged in the form “ Source : European Chemicals Agency, http://echa. europa.eu/ ” , and provided written notification is given to the ECHA Communication Unit (info (at) echa.europa.eu).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

63 C - 7 . Genetic Toxicity – Gene Mutation Summary of endpoint : Adequate Study 1 : Mutagenicity studies in Salmonella typhimurium and E scherichia coli ( Brooks and Wiggins, 1987

) Title : Microbial mutagenicity studies with IONOX 220 This study is : Adequate Not Adequate TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; referred to as IONOX 220 in study report. Remarks : 99.04% purity; Appearance : pale yellow powder; obtained from San Celoni, Barcelona, Spain. METHOD Method/guideline followed : Other (based on Ames et al ., 1975) ; similar to guideline study OECD 471 Test type : Ames reverse gene mutation assay GLP compliant? : Yes Year study performed : 1987 Test species : Salmonella typhimurium strains TA1535, TA153 7 , TA1538, TA98, and TA100 and Escherichia coli strain W P2uvrA pKM 101 Metabolic activation : Without and with (S - 9 metabolic activation from rat livers) Concentrations tested : 0 (solvent control), 31.25, 62.5, 125, 250, 500, 1000, 2000, or 4000 μg/plate (nominal) ; precipitate was observed at concentrations ≥ 500 μg/plate . Statistical methods used : Not specified Test method/conditions remarks : S . typhimurium and E . coli strains were exposed to the test substance in acetone both in the presence and absence of rat liver S - 9 mix and incubated at 37 ° C for 48 to 72 hours in a plate - incorporat ion assay . The test was conducted twice . Three plates per test concentration were u sed for each test . The initial number of cells per plate was not reported . The number of revertants per plate and ratio of the number of revertants for the test substance to the number of revertants for control (including means and standard deviations) were reported .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

64 Increases in the ratio of revertants per treated plate to revertants per control plate of ≥ 2.5 - fold were considered a positive mutagenic response . Positive Controls: TA98, TA100 , and TA1538 : Benzo(a)pyrene ( 20 μg ) TA1537 : Neutral red ( 20 μg ) TA1535 : Sodium azide ( 5 μg ) WP2 uvrA pKM 101 : Potassium dichromate ( 20 μg ) RESULTS Genotoxicity result : Negative in the presence and absence of metabolic activation Cytotoxic concentration (if applicable) : > 4 000 μg/plate (nominal) , in the absence and presence of metabolic activation Results remarks : No effects of the test substance on mutation frequency were observed with or without activation . Precipitation of the test substance was observed at nominal concentrations ≥ 500 μg/plate . The addition of 2000 μg/mL test article (equivalent to 4000 μg/plate) changed media pH from 7.49 to 7.45 . No information was reported on the pH at other concentrations. Solvent controls responded appropriately. Positive control substances responded appropriately for the following strains and activation combinations : WP2 uvrA pKM 101 without activation; TA1535 with and without activation; TA98, TA100, TA1537, and TA1538 with activation . Positive control substances did not produce positive responses for WP2 uvrA pKM 101 with activation and TA98, TA100, TA1537, and TA1538 without activation. CONCLUSIONS The test article did not induce gene mutations in any strain in the presence or absence of metabolic activation. STUDY RELIABILITY [ 2 ] Reliable with restrictions . The study meets the current guidelines for the assay with the exception that the investigators did not achieve appropriate positive control responses for all strains with and without activation , as indicated by the lack of a positive response for some combinations of positive control substance, strain, and presence/absence of activation . Therefore, the results are only valid for the combinations of strain and activation for which positive controls responded appropriately.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

65 REFERENCE Brooks, T . M . and D.E . Wiggins . 1987 . Microbial Mutagenicity Studies with IONOX 220 . S hell Research Limited , Sittingbourne Research Center . Submitted to EPA on May 1 7 , 2007 . Available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0250 as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

66 Study 2 : Mutagenicity stud y in Saccharomyces cerevisiae (Brooks and Wiggins, 1987 ) Title : Microbial mutagenicity studies with IONOX 220 This study is: Adequate Not Adequate TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; referred to as IONOX 220 in study report. Remarks : 99.04% purity; Appearance : pale yellow powder; obtained from San Celoni, Barcelona, Spain. METHOD Method/guideline followed : Other (based on Zimmermann, 1977 and Davies, P. J . et al. , 1975) Test type : Saccharomyce s mitotic gene conversion assay GLP compliant? : Yes Year study performed : 1987 Test species : Saccharomyces cerevisiae strain JD1 Metabolic activation : W ithout and w ith (S - 9 metabolic activation from rat livers) Concentrations tested : 0 (solvent control), 0.01, 0.1, 0.5, 1.0 , and 2.5 mg/mL . Th e study report does not provide any information regarding precipitation of the test substance . Statistical methods used : Not specified Test method/conditions remarks : Liquid suspension cultures of log - phase cells of Saccharomyces cerevisiae JD1 were exposed to the test substance in acetone in the presence and absence of rat liver S9 fraction . Cells were incubated for 18 hours at 30 ° C in the absence of S 9 or for 2 hours at 37°C followed by 16 hours at 30°C in the presence of S9 . Cultures were then seeded onto the appropriate culture media and incubated for 3 days at 30°C prior to counting prototrophic colonies . The test was conducted twice . Three plates per concentrat ion were used for each test . The number of p r ototrophs per plate at the histidine locus and at the tryptophan locus and the ratios of number of p r ototrophs for the test substance to the number of p r ototrophs for control for each locus were reported . A 2 - fold increase above control was considered a positive mutagenic response .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

67 Positive controls: 4 - nitroquinoline - N - oxide (0.00025 mg/mL) without metabolic activation Cyclophosphamide (1.25 mg/mL) with metabolic activation RESULTS Genotoxicity result : Negative in the presence and absence of metabolic activation Cytotoxic concentration (if applicable) : > 2.5 mg/mL Results remarks : There was no increase in the rate of mitotic gene conversion either in the presence or absence of S9 . No effect on cell viability was observed . Positive and solvent controls responded appropriately. CONCLUSIONS The test substance did not induce mitotic gene conversions in Saccharomyces either in the presence or absence of metabolic activation. STUDY RELIABILITY [ 2 ] Reliable with restrictions . Similar to OECD guideline study 481 . This study is rated as “reliable with restriction s ” because t he established limit concentration for this assay wa s not identified and no justification was provided for choice of the highest concentration tested . (OECD TG 481 does not specify a limit concentration.) REFERENCE Brooks, T . M . and D.E . Wiggins . 1987 . Microbial Muta genicity Studies with IONOX 220 . S hell Research Limited, Sittingbourne Research Center . Submitted to EPA on May 1 7 , 2007 . Available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0250 as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

68 Study 3 : Mammalian gene mutation assay ( Sandoz Chemical Ltd, 1991 , as summarized in the online ECHA entry for 2,6 - di - tert - butylphenol, Exp Key Genetic toxicity in vitro .001 ) Title : Mammalian gene mutation assay This study is: Adequate Not Adequate TEST SUBSTANCE Identity : Analog : 2,6 - D i - tert - butylphenol , CASRN 128 - 39 - 2 Remarks : 98.7% purity ; Lot/batch No. : Charge 10/6; solid at 20°C; stability in solvent : at least 48 h ours in water and DMSO METHOD Method/guideline followed : OECD 476; 40 CFR, Ch . I part 798 ; EEC Directive 87/302, L 133 Test type : Mammalian cell gene mutation assay GLP compliant? : Yes Year study performed : 1991 Test species : Chinese hamster lung fibroblasts ( V79 ) Metabolic activation : With and without S9 mix ( liver microsome preparations ) Concentrations tested : Pre - experiment for toxicity (colony forming ability ) : 0 ( negative [ untreated ] control), 0 (solvent control), 0.1, 1.0, 3.0, 6.0, 10.0, 30.0, 50.0 , and 100 μg/mL without and with S9 mix Main experiment: 0 ( negative [untreated] control ), 0 (solvent control), 0.3, 1.0, 2.0, 4.0, 6.0 , and 8.0 μg/mL without S9 mix; 0 ( negative [untreated] control ), 0 (solvent control), 3 .0, 10.0, 20.0, 30.0, 40.0 , and 50.0 μg/mL with S9 mix Statistical methods used : “ Since the distribution of mutant cells does not follow known statistical models, an adequate statistical method is not available. ” [ECHA] Test method/conditions remarks : A pre - experiment study of cytotoxicity (plating efficiency assay) was conducted to determine the toxicity of the test substance both with and without metabolic activati on . The main study consisted of two independent experiments, using identical procedures, each conducted with and without the presence of metabolic activation . In the main study, Chinese hamster V79 cells were exposed to the test substance for 4 hours at concentrations of 0.3, 1.0, 2.0, 4.0, 6.0 , and 8.0 μg/m L without S9 mix and at 3.0, 10.0, 20.0, 30.0, 40.0 , and 50.0 μg/m L with S9 mix.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

69 Cells: V79 cells were stored as frozen stock cultures, and subcultures were made f rom these stocks . Cultures were periodically checked for mycoplasma contamination and karyotype stability . The medium for culturing the V79 cells was MEM (minimal essential medium), supplemented with 10% fetal calf serum (FCS) . The cells were subculture d twice per week and were incubated at 37°C with 4.5% carbon dioxide atmosphere . For the selection of mutants at the HGPRT locus , the medium was supplemented with 11 μg thioguanine/mL. Preparation of Dose Levels : Dimethyl sulfoxide (DMSO) was chosen for the vehicle based on its solubility properties and its non - toxicity to cells. Solvent (Vehicle) Control Substance: DMSO Positive Control Substances : Without metabolic activation : EMS ( indicated to be used at a concentration of 0.6 μg/mL in Tables I through V but 0.6 mg/mL in the “Controls” section, which also named this positive control “1890” rather than EMS ); With metabolic activation : DMBA (3.85 μg/mL ; the “Controls” section named this positive control “26” rather than DMBA ) . Method s of Application: “2 or 3 days old logarithmically growing stock cultures more than 50% confluent were trypsinized at 37°C for 5 min . Then digestion [was] stopped by adding complete medium and a single cell suspension prepared . Prior to trypsin treatmen t cells were rinsed with Ca - Mg - free salt solution containing 200 mg/l EDTA . The cell suspension was seeded into plastic culture flasks (Greines, D - 7440 Nürtingen, Germany) . Approx . 8 × 100000 [in 30 mL medium for the mutagenicity tests] and 5 × 100 cells [i n 5 mL medium for the determination of cytotoxicity] , respectively, were seeded in each flask . The medium was MEM with 10% FCS (complete medium).” [ECHA] After a 24 - hour preincubation period, the medium was replaced with serum - free medium containing the test substance, either with or without 50 μL/mL S9 mix . After a 4 - hour exposure period, the treatment medium was replaced with normal medium after the cells had been rinsed twice with “saline G” . Evaluation Criteria: “A test article is classified posi tive if it induces either a significant concentration - related increase in the mutant frequency or a reproducible and significant positive response for at least one of the test points.” [ECHA] “A test article producing neither a significant concentration r elated increase in the mutant frequency nor a significant and reproducible positive response at any one of the test points is considered non - mutagenic in this system.” [ECHA] “A significant response is described as follows: The test article is classified as mutagenic if it induces reproducibly with at least one of the concentrations a mutation frequency that is three times higher than the spontaneous mutation frequency in the experiment.” [ECHA]Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

70 “The test article is classifi ed as mutagenic if there is a reproducible concentration - related increase in the mutation frequency . Such evaluation may be considered independently of an enhancement factor for induced mutants.” [ECHA] “However, in a case by case evaluation this decision depends on the level of the corresponding negative control data . If there is by chance a low spontaneous mutation rate as compared to the range normally found (5

– 45 mutants per 1000000 cells) a seemingly concentration - related increase of the muta tions within this range will be regarded as to be irrelevant.” [ECHA] RESULTS Genotoxicity result : Negative in the presence and absence of metabolic activation . Cytotoxic concentration (if applicable) : 6.0 μg/mL without metabolic activation and 50.0 μg/mL with metabolic activation based on relative plating efficiency in the pre - experiment for toxicity. Results remarks : “Taking into account the mutation rates found in the groups treated with the test article compared to the negative and solvent con trols it can be concluded that no biologically relevant increase of point mutations was observed . The test article did not induce reproducible concentration - related increase in the mutant colony numbers . The mutant values of the groups treated with the t est article were in the range of the negative controls.” [ECHA] “In this study in both experiment [s] (with and without S9 mix) the range of the negative controls was from 0.7 up to 49.1 mutants per 1000000 cells, the range of the groups treated with the test article was from 0.5 up to 36.8 mutants per 1000000 cells.” [ECHA] The positive control substanc es showed a distinct increase in induced mutant colonies. Negative and solvent controls responded appropriately. CONCLUSIONS Under the experimental conditions reported, the test substance did not induce point mutations at the HGPRT locus in V79 cells ei ther in the presence or absence of metabolic activation. STUDY RELIABILITY [ 2 ] Reliable with restrictions . The test concentrations used were below the maximum recommended in the test guidelines; however, the c ytotoxic ity findings from the pre - experiment justify the concentrations tested . The ECHA summary was inconsistent about the concentration of the substance (EMS) that was used as a positive control in absence of metabolic activation . The study summary did not indica te whether or not precipitation of test substance was observed at the concentrations tested . In the ECHA summary, the final table, which should have presented mutation rates found in experiment II, was missing . ECHA did not provide a full citation for th is study (the ECHA Reference section indicates only “study report” and “1991”), but it appears to be the same study that was attributed to Sandoz Chemical Ltd . (1991) by the OECD/SIDSDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

71 documents for CASRN 128 - 39 - 2 [ 2,6 - di - tert - butylphenol ] . These factors/d eviations do not appear to affect the validity of the conclusion of this study . REFERENCE Sandoz Chemical Ltd . 1991 . Unpublished CCR Report [on a mammalian cell gene mutation assay] - Sandoz, 243617 . Muttenz, Switzerland . As summarized in the online ECHA ( http://apps.echa.europa.eu/registered/data/dossiers/DISS - 97d6a15b - c218 - 5de0 - e044 - 00144f67d031/AGGR - 0d490876 - 6ff9 - 4b92 - be19 - 3c34473107ba_DISS - 97d6a15b - c218 - 5de0 - e044 - 00144f67d031.html#AGGR - 0d490876 - 6ff9 - 4b92 - be19 - 3c34473107ba ) entry for CASRN 128 - 39 - 2 (s tudy indicated as Exp Key Genetic t oxicity in vitro.001 , available online as of December 2, 2013 ) . Secondary reference : OECD . 2002 . SIDS/IRPTC Data Profile for 2,6 - Di tert - butylphenol (CASRN 128 - 39 - 2) . UNEP Publications . 2002 . A vailable online at http://www.chem.unep.ch/irptc/sids/OECDSIDS/128392.pdf as of December 4, 2013 . Text in quotes is taken directly from ECHA. Copyright Notice : © European Chemicals Agency, 2007 - 2011 . Reproduction is authorized provided the source is fully acknowledged in the form “ Source : European Chemicals Agency, http :// echa . europa . eu / ” , and provided written notification is given to the ECHA Communication Unit (info (at) echa.europa.eu).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

72 Study 4 : Mutagenicity study in Salmonella typhimurium (Johnson, 1984 ) Title : Genetic Toxicology : Salmonella /Microsomal Assay This study is: Adequate Not Adequate N umerous experimental details were not included in the study report, including test substance purity, test method/guideline followed, statistical methods , criteria to define a positive response, identi t y of positive control (s) , quantitative results for untreated control, detailed results for triplicate plating, and quantitative data on cytotoxicity at 25 μg/plate in strains TA1535, TA1537 , and TA100 . TEST SUBSTANCE Identity : Analog : 2,6 - Di - tert - butylphenol , CASRN 128 - 39 - 2 ; referred to as AN 701 in the study report. Remarks : Purity not provided; received as a yellow - brown liquid METHOD Method/guideline followed : Other ( plate incorporation Ames Assay was carrie d out according to S.O. P. ETTOX - 027 ) Test type : Salmonella Gene Mutation Assay GLP compliant? : Yes Year study performed : 1984 Test species : S almonella typhimurium strains TA1535, TA1537, TA98 , and TA100 Metabolic activation : W ithout and w ith ( Aroclor 1254 induced rat liver S - 9 fraction ) Concentrations tested : 0.25, 0.5, 2.5, 5 , and 25 μg/plate ; test substance was soluble at all concentrations tested. Statistical methods used : Not specified Test method/conditions remarks : The compound was tested using the Ames plate incorporation assay . Salmonella strains were exposed to the test substance in acetone in the presence and absence of metabolic activation . Each concentration was tested in triplicate . An untreated control, solvent control , and positive control were tested concurrently . The positive control was not identified .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

73 RESULTS Genotoxicity result : Negative in the presence and absence of metabolic activation Cytotoxic concentration (if applicable) : 25 μg/plate in strains TA1535, TA1537 , and TA100 with and without metabolic activation ; cytotoxicity not reported for TA98 Results remarks : The test substance did not induce a dose - related increase in mutant colonies in any strains tested with or without metabolic activation. Numerous experimental details were not included in the study report, including test substance purity, test method/guideline followed, statistical methods , a clear statement of cr iteria used to define a positive response, identi t y of positive control (s) , quantitative results for untreated control, detailed results for triplicate plating, and quantitative data on cytotoxicity at 25 μg/plate in strains TA1535, TA1537 , and TA100 . CONCLUSIONS The test substance did not induce gene mutations in Salmonella typhimurium strains TA1535, TA1537, TA98 , or TA100 in the absence or presence of metabolic activation. STUDY RELIABILITY [4] Not assignable. N umerous experimental details were not included in the study report, including test substance purity, test method/guideline followed, statistical methods , a clear statement of criteria t o define a positive response, identi t y of positive control (s) , quantitative results for untreated control, detail ed results for triplicate plating , and quantitative data on cytotoxicity at 25 μg/plate in strains TA1535, TA1537 , and TA100 . REFERENCE Johnson, D.E . 1984 . Genetic Toxicology Salmonella /Microsomal Assay . Performing Laboratory : Genetic Toxicology Laboratory , Baton Rouge, LA . Study No. : Ames 086 - 068 . Submitted by Ethyl Corporation to EPA in 19 86 as part of a TSCA 8(e) submission (EPA Doc. No. 40 - 8627016 , Fiche No. OTS0526562 ) entitled “ Letter from Ethyl Corp to US EPA regarding submission of 2,6 - di - tert - butylphenol toxicological studies with attachments ”.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

74 Study 5 : Mutagenicity study in S . typhimurium and E . coli ( Hachiya and Takizawa

, 1994 ) Title : Mutagenicity of plastic additives This study is: Adequate Not Adequate Information for this study was reviewed in HPVIS (4/8/2005) based on a partial English translation of the original study; a complete translation of this study was not available . The HPVIS (4/8/2005) summary indicated that positive controls were not included in this assay . Positive controls, with and without metabolic activation, ar e required to validate assay performance . Since none of the 20 chemicals tested yielded a positive response, performance of this assay cannot be validated . Additional reporting inadequacies include lack of information on replicate plating, purity of the test substance, cytotoxic concentrations, quantitative results, and justification for testing up to a concentration that was only 40% of the recommended limit concentration of 5 000 μg/plate . TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - di methylethyl) - , CASRN 118 - 82 - 1 Remarks : Purity not provided METHOD Method/guideline followed : Not specified Test type : S . typhimurium and E . coli gene conversion assay GLP compliant? : Not specified Year study performed : 1994 Test species : S . typhimurium strains TA97, TA98, TA100, and TA102 and E . coli strain WP2 uvrA (pKM101) Metabolic activation : With and without Concentrations tested : 0, 50, 100, 200, 500, 1000 , and 2000 μg/plate Statistical methods used : Not specified Test method/conditions remarks : No details on test method/ conditions were reported . Solvent was acetone and Tween 80.

“ There was no indication of either cytotoxicity or solubility problems beyond the statement that the test substance was insoluble but homogeneously dispersed in phosphate buffer in the assayDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

75 with metabolic activation . It thus appears that the chemical should have been tested up to the limit concentration of 5000 μg/plate. ” (HPVIS, 4/8/2005 )

“ There was no repeat experiment to confirm the negative result . Especially since the original experiment only tested up to 40% of the limit concentration, a justification for having no repeat experiment should have been provided . It will be important to know if they u sed more than one plate per concentration; they should have used triplicate plating with the reported values being the mean and the standard error of the mean for those three values . They provided only one number per plate, with no measure of the variabil ity . It will be important to know how they prepared their S - 9 mix to provide endogenous metabolic activation . The purity of the test chemical is unknown. ” (HPVIS, 4/8/2005 ) Note that 20 d ifferent chemicals were evaluated for mutagenicity in this study . None of the chemicals had a positive response. RESULTS Genotoxicity result : Negative Cytotoxic concentration (if applicable) : Not specified Results remarks : The authors reported negative results for the subject chemical in all strains of S . typhimurium and E . coli. CONCLUSIONS T his test was not adequate to rule out induction of gene mutations in bacteria with or without metabolic activation due to the lack of positive controls . However, results that were reported for the test substance were negative for induction of gene mutations in bacteria up to a concentration that was 40% of the guideline - recommended limit concentration of 5 000 μg/plate . STUDY RELIABILITY [ 4 ] Not assignable . Information for this study was reviewed in HPVIS ( 4/8/2005 ) based on a partial English translation of the original study; a complete translation of this study was not available . Th e HPVIS ( 4/8/2005 ) summary indicated that positive controls were not included in this assay . Positive controls, with and without metabolic activation, are required to validate assay performance . Since none of the 20 chemicals tested yielded a positive response, performance of this assay cannot be validated . Additional reporting inadequacies include lack of information on r eplicate plating, purity of the test substance, cytotoxic concentrations, quantitative results, and justification for testing up to a concentration that was only 40% of the recommended limit concentration of 5 000 μg/plate .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

76 REFERENCES Hachiya, N . and Y. Takizawa . 1994 . Mutagenicity of Plastic Additives (translation) . Hen'lgensei Shiken 3(3):147 - 154 . Study summary a vailable online at http://www.regulations.gov/# !documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0181 (summarized in “End_Point_E1

_ - _ Mammalian_Toxicity_ - _Genotoxicity.pdf”) as of December 4, 2013 . US EPA . OPPT . High Production Volume Information System (HPVIS) . Submission ID : 24929673 . Submission Name : Phenol, 4,4’ - methylenebis[2.6 - bis(1,1 - dimethylethyl) - . Sponsor : Orphan Chemical . Submission created on 04/08/2005. Text in quotes is taken directly from HPVIS (4/8/2005).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

77 C - 8 . Genetic Toxicity – Chromosomal Aberrations Summary of endpoint : Adequate Study 1 : Chromosomal aberrations study in CHO cells ( de Vogel and Usta, 2007) Title : Chromosomal Aberration test with AN - 2 in Cultured Chinese Hamster Ovary (CHO) Cells Th is study is: Adequate Not Adequate TEST SUBSTANCE Identity : 4,4' - methylenebis(2,6 - di - tert - butylphenol) (AN - 2; Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - ), CAS RN 118 - 82 - 1 Remarks : 99.45% purity; white powder; production lot : 5127; MW = 424.7; supplier : Albemarle Corporation; TNO Dispense No. 060123; vehicle : DMSO METHOD Method/guideline followed : OECD TG 473, 40 CFR 799.9537 Test type : Chromosomal Aberration Test in Cultured Chinese Hamster Ovary Cells GLP compliant? : Yes Year study performed : 200 6 Test species : Chinese hamster ovary (CHO) cells Metabolic activation : With and without Concentrations tested : First test (pulse exposure; with and without activation) : 0, 0.1, 0.2, 0.39, 0.78, 1.56, 3.13, 6.25, 12.5, 25, 50, 100, and 200 μg/mL; second test (pulse exposure; with activation) : 0, 5, 10, 30, 50, 75, 100, 150, 200, and 300 μg/mL; second test (continuous exposure; without activation) : 0, 1.25, 2.5, 5, 10, 30, 50, 75, 100, 150, 200, and 300 μg/mL . Statistical methods used : Fisher’s exact probability test (two - sided) was conducted to determine significant differences between treated and control cultures.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

78 Test method/conditions remarks : Cells: Chinese Hamster Ovary cells (CHO K - 1 line) were stored as frozen stock cultures, and subcultures were made from these stocks . Each passage was tested for mycoplasma contamination and karyotype stability . The medium for culturing the CHO cells was Ham's F - 12 medium (with Glutamax - I) suppleme nted with heat - inactivated fetal calf serum (10%), penicillin , and streptomycin . Metabolic Activation System : The S - 9 mix consisted of a liver homogenate fraction and cofactors, derived from Aroclor 1254 - induced Wistar rats . The final concentration of S - 9 mix in culture medium used in the tests requiring activation was 4% . Preparation of Dose Levels : Dimethyl sulfoxide (DMSO) was chosen for the vehicle . Based on the solubility pre - test, 200 μg/m L final concentration in the culture medium was chose n as the highest dose level for the first test. Methods for 1 st Chromosomal Aberration Test : Exponentially growing cells were seeded in sterile tissue culture flasks containing culture medium, and incubated at 37 °C in humidified air containing 5% CO 2 . Approximately 24 hours after seeding, the cells were exposed to the test substance or control substance, with or without S - 9 mix . In the presence of S - 9, the culture medium with fetal calf serum was replaced by culture medium with penicillin and streptomy cin, but without calf serum . In the first test (pulse exposure), the cells were exposed (in duplicate cultures) to the test substance for 4 hours at the following concentrations, either with or without activation : 0 (vehicle control), 0.1, 0.2, 0.39, 0.7 8, 1.56, 3.13, 6.25, 12.5, 25, 50, 100, and 200 μg/mL . At the end of the exposure, the cultures were checked, the medium with test substance was removed, and the cells were washed twice and supplied with fresh culture medium . The cells were incubated for another 14 hours at 37 °C in humidified air with 5% CO 2 . Two hours before the end of the culture period, the cells and culture media were checked visually and selected cultures were arrested in metaphase by the addition of colcemid . The cells were harvested at 18 hours by trypsinization . The following concentrations were selected for cell sampling : 0, 6.25, 12.5, 25, 50, 100, and 200 μg/mL. Methods for 2 nd Chromosomal Aberration Test: In the second test, the 4 - hour pulse exposure with activation was repeated at the following concentrations : 0, 5, 10, 30, 50, 75, 100, 150, 200, and 300 μg/mL . The following concentrations were selected for cell sampling : 0, 50, 75, 100, 150, 200, and 300 μg/mL . In the absence of activation, the cells were expos ed continuously for 18 hours and harvested immediately after the exposure period . The concentrations used for continuous exposure were : 0, 1.25, 2.5, 5, 10, 30, 50, 75, 100, 150, 200, and 300 μg/mL . The concentrations selected for cell sampling were : 0, 1.25, 2.5, 5, 10, and 30 μg/mL. Positive Control Substance: The positive control substances were : cyclophosphamide (5 μg/mL) with activation and mitomycin C (0.1 μg/mL) without activation. Methods for Determining Mitotic Index: Two slides were prepared from each culture that had been selected for cell sampling . For each culture, at least 1000 nuclei were examined (500 per slide) to determine the mitotic index (percentage of cells in mitosis) . On the basis of the mitotic index observed, three c oncentrations of the test substance were selected for the scoring ofDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

79 chromosomal aberrations . If possible, the highest concentration chosen reduced the mitotic index by at least 50% (but not more than 70%) when compared to the negative control, or exhibit ed some other clear indication of cytotoxicity . For each chosen concentration, 200 well spread metaphases (100 metaphases/culture, each containing 20

– 22 centromeres) were examined for chromatid - type aberrations (gaps, breaks, fragments, and interchanges) and chromosome - type aberrations (gaps, breaks, minutes, rings, and dicentrics) . Heavily damaged cells were not included in the 200 analyzed cells. Evaluation of Test Results: T he criteria for a clastogenic response are : a concentration - related increase in the percentage of cells with chromosomal aberrations over the concurrent control frequencies or a single positive test point in both tests at approximately the same dose level . Cells with only gaps (achromatic lesions) were not included in the final a ssessment of clastogenic activity . If the increase in cells with chromosomal aberrations is marginally significant (0.05 < p < 0.1), then the response is considered equivocal. RESULTS Genotoxicity result : Negative Cytotoxic concentration (if applicable): 1 ≥ 100 μg/mL in pulse exposures with or without activation; ≥ 10 μg/mL in continuous exposure without activation Results remarks : 1 st Chromosomal Aberration Test: In the first test, in the pulse treatment group with metabolic activation, the three concentrations of test material chosen for chromosomal aberration analysis were 50, 100, and 200 μg/mL . The mitotic index of the highest concentration analyzed (200 μg/mL ) was reduced to 71% of the concurrent vehicle control (DMSO) . The test substance did not induce a statistically significant increase in the number of aberrant cells at any of the dose levels analyzed when compared to the number of aberrant cells obser ved in the concurrent control (DMSO) . In the first test, in the pulse treatment group without activation, the three concentrations of test material chosen for chromosomal aberration analysis were 50, 100, and 200 μg/mL . The mitotic index of the lowest concentration analyzed (50 μg/mL) was not reduced; the mitotic indices of the middle concentration (100 μg/m L ) and the highest concentration (200 μg/mL) were reduced to 59 and 54% , respectively , of that of the concurrent vehicle control . The test substanc e did not induce a statistically significant increase in the number of aberrant cells at any of the dose levels analyzed when compared to the number of aberrant cells in the vehicle control group. 2 nd Chromosomal Aberration Test : In the second test, in the pulse treatment group with metabolic activation, dose - related toxicity could be demonstrated . The test concentrations chosen for chromosomal aberration analysis were 100, 200 , and 300 μ g/mL . The mitotic index of the lowest concentration analyzed ( 100 μ g/ mL ) was reduced to 80%; the mitotic index of the mid - concentration analyzed (200 μ g/mL) was reduced to 49% ; and the mitotic index of the highest concentration analyzed was reduced to 40% of that of the concurrent vehicle control . 1 Mitotic index was ≤ 80% of control v alue at cytotoxic concentrations.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

80 The test substance did not induce a statistically significant increase in the number of aberrant cells at any of the dose levels analyzed, when compared to the aberrant cells found in the concurrent (DMSO) control cultures. In the second test, in the continuous treatment gr oup of 18 hours without metabolic activation, dose - related toxicity could be demonstrated . Dose levels chosen for chromosomal aberration analysis were 2.5, 5 , and 10 μg/mL . The mitotic index of the lowest concentration analyzed ( 2.5 μ g/ mL ) was 91%, the mitotic index of the mid - concentration analyzed (5 μ g/mL) was reduced to 87%, and the mitotic index of the highest concentration analyzed ( 10 μ g/mL) was reduced to 47% of that of the concurrent vehicle control . At the next higher concentration se lected for cell sampling (30 μ g/m L ), the mitotic index was reduced to 11% . This strong reduction was above the acceptable toxicity level and thus this dose was not selected for chromosomal aberration analysis . The test substance did not induce a statisti cally significant increase in the number of aberrant cells at any of the dose levels analyzed, when compared to the number of aberrant cells found in the concurrent control cultures. Precipitation of the test substance was observed at 100 and 200 μg/mL in the first test and at 300 μg/mL in the second test. Criteria for validity of the study were met by the positive controls (mitomycin C without S - 9, and cyclophosphamide with S - 9) giving the expected statistically increased numbers of aberrant cells and the negative (vehicle) control results fallin g within the historical range. CONCLUSIONS Under the conditions of the study, the test subs tance was not clastogenic to CHO cells. STUDY RELIABILITY [1 ] Reliable with out restrictions . REFERENCE de Vogel, N . and B. Usta . 2007 . Chromosomal Aberration test with AN - 2 in Cultured Chinese Hamster Ovary (CHO) Cells . TNO Laboratories, Zeist, Netherlands, Project No. 031.11113/01.42, Study No. 6802/24, May 9, 2007 . Submission documented online at http://www.regulations.gov/#!d ocumentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0257 as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

81 Study 2 : Chromosomal aberrations study in Chinese hamster V79 cells (Sandoz Chemical Ltd, 1992) Title : In vitro mammalian chromosome aberration test Th is study is: Adequate Not Adequate Important study details missing from the study summary included description of the metabolic activation sy stem, solvent and positive control information (named by strain and metabolic activation), statistical methods used, and additional quantitative data on the results and information on cytotoxicity . Other deficiencies include : (1) T he shortest exposure pe riod was 18 hours rather than the OECD - recommended 3

– 6 hour exposure period . (2) The SIDS/IRPTC Data Profile stated that the test substance negatively affected the plating efficiency of the cells at concentrations greater than 3 μ g /mL in the absence of S - 9 mix and at concentrations greater than 30 μ g /mL in the presence of S - 9 mix . It is uncertain whether or not this is indicative of cytotoxicity . (3) A limit concentration of 5 mg/mL is recommended by OECD TG 473; however, no rational e was provided for testing only up to 10.0 μ g/mL without activation and only up to 50.0 μg /mL with activation . TEST SUBSTANCE Identity : Analog : 2,6 - Di - tert - butylphenol, CASRN 128 - 39 - 2 Remarks : 98.7% purity METHOD Method/guideline followed : OECD TG 473 (EEC Directive 84/449, B 10) Test type : In vitro mammalian chromosomal aberration test GLP compliant? : Yes Year study performed : 1992 Test species : Chinese hamster V79 cells Metabolic activation : With and without S - 9 mix Concentrations tested : 0.3, 3.0, 6.0, 10.0 μ g/mL for 18 hr s and 6.0 μg /mL for 28 hr s without S9 mix; 3.0, 30.0, 50.0 μg /mL for 18 hr s and 50.0 μg /mL for 28 hr s with S9 mix . Statistical methods used : Not specified Test method/conditions remarks : The exposure period was 18 to 28 hours.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

82 RESULTS Genotoxicity result : Negative with and without metabolic activation Cytotoxic concentration (if applicable) : > 3.0 μg /mL without S - 9 mix and > 30.0 μg /mL with S - 9 mix. Results remarks : No mutagenic effects were obtained with or without metabolic activation . CONCLUSIONS The test substance did not induce chromosomal aberrations in Chinese hamster V79 cells either in the absence or presence of metabolic activation . STUDY RELIABILITY [3] Not reliable . Important study details missing from the study summary included description of the metabolic activation system, solvent and positive control information (named by strain and metabolic activation), statistical methods used, and additional quantitative data on the results and information on cytotoxicity. Other deficiencies include : (1) T he shortest exposure period was 18 hours rather than the OECD - recommended 3

– 6 hour exposure period . (2) The SIDS/IRPTC Data Profile stated that the test substance negatively affected the plating efficiency of the cells at concentrations greater than 3 μ g /mL in the absence of S - 9 mix and at concentrations greater than 30 μ g /mL in the presence of S - 9 mix . It is uncertain whether or not this is indicative of cytotoxicity . (3) A limit concentration of 5 m g/mL is recommended by OECD TG 473 ; however, no rationale was provided for testing only up to 10.0 μ g/mL without activation and only up to 50.0 μg /mL with activation . REFERENCE Sandoz Chemical Ltd . 1992 . Unpublished CCR Report [on a chromosomal aberration assay] - Sandoz, 243628 (full study report not available) . Muttenz, Switzerland . As summarized in : OECD . 2002 . SIDS/IRPTC Data Profile for 2,6 - Di - tert - butylphenol (CASRN 128 - 39 - 2) . UNEP Publications . 2002 . A vailable online at http://www.chem.unep.ch/irptc/sids/OECDSIDS/128392.pdf as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

83 C - 9 . Reproductive /Developmental Toxicity Summary of endpoint : A dequate Study 1 : Preliminary reproduction/developmental toxicity screening test in rats (Sandoz Chemical Ltd., 1992

, as summarized in the online ECHA entry for 2,6 - di - tert - butylphenol, Exp Key Toxicity to reproduction.001 ) Title : Not provided Th is study is: Adequate Not Adequate TEST SUBSTANCE Identity : Analog : 2,6 - Di - tert - butylphenol , CASRN 128 - 39 - 2 Remarks : 98.7% purity; Lot/batch No. 10/6; solid at 20°C (colorless solid) METHOD Method/guideline followed : OECD Preliminary Reproduction /Developmental Toxicity Screening Test (Precursor Protocol of OECD TG 421 [1995] ) Test type : One - Generation Reproduction /Developmental Toxicity GLP compliant? : Yes Year study performed : 1992 Test species : Wistar rats ; s ource : Biolog ical Research Laboratories Ltd., Switzerland ; a ge: at least 11 weeks old ; w eight: 304

– 339 g (males) or 1 82

– 229 g (females) at study initiation. Number of animals/sex/dose : 10 Route of administration : Oral gavage Vehicle : Bi - distilled water with 4% carboxymethyl cellulose ( CMC ) Exposure duration : P arental females were exposed for 2 weeks prior to mating, throughout the mating period (up to 13 days) , and through gestation and lactation up to postpartum day 4 . P arental males were exposed for 2 weeks prior to mating, throughout the mating period , and following mating for a total exposure period of 43 days. Doses/concentrations tested : 0 (vehicle con trol) , 30, 150 , and 750 mg/kg - bw /dayDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

84 Statistical methods used : Statistical analyses were performed on data for b ody weight, food consumption, organ weights, reproduction , and breeding . If the variables followed a normal distribution, the Dunnett ’s many - to - one t - test, based on a pooled variance estimate, was used for intergroup comparisons (i.e . , single treatment groups against the control group) . The Steel - test (many - to - one rank test) was applied when the data did not follow a normal distribution . For the spontaneous mortality of pups data, Fisher's Exa ct test ( 2 × 2 tables ) was applied. Test method/conditions remarks : Dose selection was based on the results of a suba cute 28 - day toxicity study (RCC NOTOX Project 057454) . M ixtures of test substance and vehicle were prepared daily before administration . Male and female Wistar rats (10 / sex /dose ) were administered the test substance via gavage at doses of 0, 30, 150 , and 750 mg/kg - bw /day daily . Females received the test substance for 2 weeks prior to mating , throughout the mating period (up to 13 days , except for one female which was paired for 17 days ) , and through gestation and lactation up to postpartum day 4 . Males received the test substance for 2 weeks prior to mating, throughout the mating p eriod , and following mating for a total exposure period of 43 days . T he day on which spermatozoa were found in the vaginal smear or a vaginal plug was obs erved was designated as day 0 post coitum . After successful mating, each pregnant female was separated and housed individually . Animals were observed at least twice daily for mortality and clinical signs; body weights were recorded daily . F emales were observed for signs of difficult or prolonged parturition . Food consumption in males was reco rded weekly during the pre - mating and postmating periods . Food consumption in females w as recorded for days 1

– 8 and 8

– 14 of the pre - mating period , days 0

– 7, 7

– 14 , and 14

– 21 post coitum , and days 0

– 4 postpartum . No food consumption was recorded during the mating period . Male s were sacrificed when they were no longer necessary for the assessment of reproductive effects , and dams were sacrificed on day 4 postpartum . All parental animals were necropsied and examine d macroscopically for any structural abnormalities or pathological changes . U ter i of non - pregnant females were placed in a solution of ammonium sulfide to visualize possible h emorrhag ic areas of implantation sites . The testes and epididymides of all parent al males were weighed . Histo patho logical examination of the testes and ovaries was performed on animals from the control and high - dose group. The following reproductive and developmental parameters were recorded : number of corpora lutea, number of implantations, l itter size, number of live birth s , number of stillbirth s, sex ratio , number of pups living on day 4 postpartum , and pup body weights on days 0, 1, and 4 postpartum . Dead pups (except if excessively cannibalized) a nd pups killed at day 4 of lactation were examined macroscopically for gross abnormalities . RESULTS Endpoint value(s) : The LOAEL for parental toxicity wa s 750 mg/kg - bw / day based on severe clinical signs of toxicity ( sedation, ataxia, lateral or ventral recumbency, tremor, clonic spasms of the forepaws, hunched posture , and salivation ) in both sexes , reduced food consumption in females during the pre - mating period , and reduced body weight gain in both sexes (statisticalDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

85 significance not reported) . The NOAEL for parental toxicity wa s 150 mg/kg - bw/day . The LOAEL for reproductive and developmental toxicity wa s 750 mg/kg - bw/day based on reduced viability index and a reduction in pup body weight gain . The NOAEL for reproductive and developmental tox icity wa s 150 mg/kg - bw/day. Results remarks : No deaths of parental animals occurred during the course of the study . Severe clinical signs of toxicity were observed in high - dose males and females . In males, the signs were observed from day 2 of the pre - mating period until day 5 of the mating period . Signs included ruffled fur, sedation, ataxia, lateral or ventral recumbency, tremor, clonic spasms of the forepaws, hunched posture , and salivation . After day 5 of the mating period, until termination of th e study, the number and intensity of the signs diminished continuously and /or disappeared . In parental females, the identical signs were noted during the pre - mating, mating , and gestation period s . During the lactation period, only ruffled fur was observed . There was a marginal reduction in body weight gain for males of the high - dose group during the pre - mating and mating periods . During the g estation period (days 0

– 21 post coitum), the mean body weight gain in females from the vehicle control, an d low - , mid - , and high - dose groups was 5 1.1, 52.8, 55.2 , and 47.3% , respectively . During days 1

– 4 of lactation, mean body weight gains of 8.3 , 4.5 , 6.9 , and 2.8% were observed in control, and low - , mid - , and high - dose females, respectively . Neither secondary source (OECD/SIDS or ECHA) reported statistical significance for these findings . T he mean food consumption of high - dose males was statistically significantly increased from days 8 to 14 of the pre - mating period and from days 1 to 10 af ter the mating period ; secondary sources did not report the magnitude of changes . No differences were observed among mean food consumption rates during both periods in males from the vehicle control , low - dose , and mid - dose groups . T he mean food consumpti on of high - dose females was statistically significantly reduced between days 1 and 8 of the pre - mating period (magnitude of change not reported in secondary sources) . Du ring days 8 to 14 of the pre - mating period , the mean food consumption in high - dose females was only slightly increa sed (compensatory reaction; magnitude not reported) . No test article - related differences in the mean food consumption of the females were noted during the gestation and lactation periods . S tatistically significantly increa sed mean food consumption was observed in females from the mid - dose group between days 14 and 21 post coitum ; however, this increase “is considered to be incidental because from the beginning of the study in this group, the highest food consumption was rec orded .” [ECHA] The mean organ weights and organ/body weight ratios for testes and epididymides of males were similar in all groups . Macroscopic examination of parent al animals revealed no test substance related effects . The microscopic examination of te stes and ovaries of parent al animals from the control and high - dose groups showed no indication of test substance - related effects . A significant (p ≤ 0.01) decrease in viability index (survival from postnatal day 0 to 4) was observed in pups from the hi gh - dose group (90.3%) compared to controls (99.1%) . No test substance - related effects were observed for pre - coital time, percentage mating, fertility index , and conception rate in any dose group .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

86 Pup body weight at birth was similar among all groups . However, b ody weight gain of pups from days 1 to 4 postpartum was reduced in the high - dose group (23. 3 % weight gain) compared to controls (38.3% weight gain ) . No effect s w ere observed up on external examination or necropsy of the pups in any group. CONCLU SIONS The LOAEL for parental toxicity wa s 750 mg/kg - bw/day based on severe clinical signs of toxicity ( sedation, ataxia, lateral or ventral recumbency, tremor, clonic spasms of the forepaws, hunched posture , and salivation ) in both sexes , reduced food consumption in females during the pre - mating period , and reduced body weight gain s in both sexes (statistical significance not reported) . The NOAEL for parental toxicity wa s 150 mg/kg - bw/day . The LOAEL for reproductive and developmental toxicity wa s 750 mg/kg - bw/day based on reduced viability index and a reduction in pup body weight gain . The NOAEL for reproductive and developmental toxicity wa s 150 mg/kg - bw/day. STUDY RELIABILITY [2] Reliable with restrictions . Histological examination of the epididymides, which is recommended by OECD TG 421, was not conducted . Also, r eviews of this study reported by secondary sources did not include results of statistical analyses for all variables examined, including maternal weight gain, postnatal loss duri ng lactation, and fetal body weight . REFERENCE Sandoz Chemical Ltd . 1992 . Unpublished CCR Report [on a reproduction/developmental toxicity test] - Sandoz, 321794 . Muttenz, Switzerland . As summarized in the online ECHA ( http://apps. echa.europa.eu/registered/data/dossiers/DISS - 97d6a15b - c218 - 5de0 - e044 - 00144f67d031/AGGR - e91a95e9 - 69e4 - 4f56 - a8ac - 562a89c8c06f_DISS - 97d6a15b - c218 - 5de0 - e044 - 00144f67d031.html#AGGR - e91a95e9 - 69e4 - 4f56 - a8ac - 562a89c8c06f ) entry for CASRN 128 - 39 - 2 (s tudy indicated as Exp Key Toxicity to reproduction. 001 , available online as of December 2, 2013 ) . Secondary Reference : OECD . 2002 . SID S/IRPTC Data Profile for 2,6 - Di tert - butylphenol (CASRN 128 - 39 - 2) . UNEP Publications . 2002 . A vailable online at http://www.chem.unep.ch/irptc/sids/OECDSIDS/128392.pdf as of December 4, 2013. Text in quotes is taken directly from ECHA. Copyright Notice : © European Chemicals Agency, 2007 - 2011 . Re production is authorized provided the source is fully acknowledged in the form “ Source : European Chemicals Agency, http :// echa . europa . eu / ” , and provided written notification is given to the ECHA Communication Unit (info (at) echa.europa.eu).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

87 Study 2 : Fertility screening study in rats ( Davis and Jolley, 1961) Title : A comparison of the effects of two antioxidants on gestation and parturition in the rat when fed several concentrations in the diet Th is study is: Adequate Not Adequate The study was not conducted according to standard guidelines . The highest dose tested did not induce toxi c effects as recommended in guideline 40 CFR 799.9355 and was below the recommended limit dose of 1000 mg/kg - bw/day . Male and female parental animals were not dosed for a minimum of 2 weeks prior to mating as recommended in the guideline (females were exposed for only 1 week before mating and males were exposed only during mating) . The number of pregnant females per group was less than the minimum number of eight that is recommended in guideline 40 CFR 799.9355 . Only two males were used per group, whereas the guideline recommends ten . Guideline - required parameters that were not evaluated included body weight, food consumption, clinical signs of parental rats , and weights and histology of reproductive organs . Pups were not examined for g ross abnormalities . I mportant study details that were missing from the study summary included test substance purity and statistical methods used. TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; Referred to as Antioxidant 702 and 4,4’ - methylenebis(2,6 - di - tert - butylphenol) in study report Remarks : Purity not provided ; Light yellow crystalline solid METHOD Method/guideline followed : Not specified Test type : Reproductive /Developmental Toxicity Study - Rats GLP compliant? : N/A (pre - GLP) Year study performed : 1961 Test species : CFN rats Number of animals/sex/dose : 2 males and 10 females /group Route of administration : Oral - Diet Vehicle : Alphacel (non - nutritive cellulose flour)Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

88 Exposure duration : Females were exposed during a pre - mating period for 7 days and throughout mating (12 days), gestation, and parturition to day 4 of lactation . Males were exposed only during mating . Doses/concentrations tested : 0 (control), 0.125, 0.5 , and 1.25% in feed ( approximately 62.5, 250, and 625 mg/kg - bw/day, calculated for this review, using a food factor of 0.05) 1 Statistical methods used : None specified Test method/conditions remarks : Females were exposed during a pre - mating period for 7 days and throughout mating (12 days), gestation , and parturition to day 4 of lactation . Males were exposed only during the mating period . Concentrations in feed were 0.125, 0.5 , and 1.25% . A negative control group was maintained on a basal diet and a positive control group was maintained on the basal diet plus 1.25% N,N' - diphenyl - p - phenylenediamine (DPPD) . Females were housed in groups of five per cage for the duration of the experiment . During the mating period, one male was housed with each group of five females. RESULTS Endpoint value(s) : N OAEL (reproductive/developmental toxicity) = 1.25% (equivalent to 625 mg/kg - bw /day; based on no effects at the highest dose tested) . Results remarks : The test substance had no effect on fertility, length of gestation, number of live or dead pups per litter at birth , pup weight, or the viability of offspring during the initial 4 days after delivery. Results of reproductive performance are provided in th e table below.
% compound in diet No. of females Avg . length of gestation (days) No. of

litters Avg .

weight of pups (g) Avg . no s . per litter Avg . no . of surviv ing pups at postpartum day 4 Bred Pregnant Live births Still births 0 10 7 22 7 5.8 10.9 0.7 9.3 1.25 10 6 22 6 5.9 11.8 0.2 11.3 0.5 10 10 22 10 6.2 9.9 0.3 9.1 0.125 10 9 22 9 6.2 9.6 1.3 8.7 Rats fed DPPD (positive control) had an increased incidence of stillborn pups ( ~ 50%) , but there was no effect on the number of young or the weight of offspring . The period of gestation of 3 / 10 rats fed on the diet containing DPPD was prolonged (24

– 25 days) . Two of these rats died during the 25 th day of pregnancy and prior to parturition . At autopsy, their unborn litters were found to be normal in number and size .

1 1 mg/kg in f

ee d = 0.05 mg/kg - bw/day .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

89 CONCLUSIONS The N OAEL for reproductive and developmental toxicity was 1.25% (equivalent to 625 mg/kg bw /day; based on no effects at the highest dose tested). STUDY RELIABILITY [3] Not reliable . The study was not conducted according to standard guidelines . The highest dose tested did not induce toxic effects as recommended in guideline 40 CFR 799.9355 and was below the recommended limit dose of 1000 mg/kg - bw/day . Male and female parental animals were not dosed for a minim um of 2 weeks prior to mating as recommended in the guideline (females were exposed for only 1 week before mating and males were exposed only during mating) . The number of pregnant females per group was less than the minimum number of eight that is recomm ended in guideline 40 CFR 799.9355 . Only two males were used per group, whereas the guideline recommends ten . Guideline - required parameters that were not evaluated included body weight, food consumption, clinical signs of parental rats , and weights and histology of reproductive organs . Pups were not examined for gross abnormalities . I mportant study details that were missing from the study summary included test substance purity and statistical methods used. REFERENCE Davis, R.K . and W . Jolley . 1961 . “ A Comparison of the Effects of Two Antioxidants on Gestation and Parturition in the Rat When Fed Several Concentrations in the Diet, ” The Kettering Laboratory in the Department of Preventive Medicine and Industrial Health, College of Medicine, University of Cincinnati, Cincinnati, Ohio, for Ethyl Corporation . (Submitted as part of Food Additive Petition No. 4B1469 and Food Additive Pet ition 1397, Food and Drug Administration) . Available online at http://www.regulations.gov/#!documentDetail;D=EPA HQ - OPPT - 2005 - 0033 - 0181 ( on p p. 1 0 - 21 of “End_Point_F2

– Mammalian_Toxicity_ - _Repeated_Dose_Reproduction_Developmental.pdf”) as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

90 Study 3 : Three - generation reproductive toxicity study in rats ( Witherup et al. , 1969 ) Title : The effects exerted upon the fertility of rats and upon the viability of their offspring by the introduction of 4,4' - methylenebis (2,6 - ditertiary butylphenol) into their diets Th is study is: Adequate Not Adequate The highest dose tested did not induce toxic effects as recommended in guideline 40 CFR 799.9355 and was below the recommended limit dose of 1000 mg/kg - bw/day . In addition, r esults of this study are considered equivocal due to a potential treatment - related decrease in the lactation index in F2b pups; therefore, NOAEL and LOAEL values for developmental toxicity cannot be determined . Important study details that were missing from the study summary incl uded test substance purity, test method/guideline followed , and statistical methods used . Measurements of body weights and food consumption were not conducted for the parental animals . Additionally, extraneous disease and underlying environmental factors complicate the interpretation of the results . TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; Referred to as BHD in study report Remarks : Purity not provided ; obtained from the Ethyl Corporation; finely divided crystalline cream - colored solid METHOD Method/guideline followed : Not specified Test type : Three - Generation Reproduction Toxicity Study - Rats GLP compliant? : N/A (pre - GLP) Year study performed : 1969 Test species : CD Rats ; obtained from the Charles River Breeding Laboratories, Inc. , Brookline, Massachusetts. Number of animals/sex/dose : 20 females and 10 males/group Route of administration : Oral - Diet Vehicle : Corn oil (test substance was dissolved in corn oil prior to incorporation into the diet)Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

91 Exposure duration : P males were exposed for 8 weeks during a pre - mating period and throughout mating; P females were exposed for 8 weeks during a pre - mating period and during mating, gestation, lactation, and weaning of the F 1 offspring (until postnatal day [PND] 21); F 1 and F 2 animals were exposed for the same duration as the P animals , starting at the time of weaning ( i.e., at 3 weeks old). Doses/concentrations tested : 0, 15, 60, 100, 500, and 3000 ppm in feed (approximately equivalent to 0, 1. 3 , 5.2 , 8.6 , 43 , and 258 mg/kg - bw/day for males; 1. 5 , 5.9 , 9.8 , 4 9 , and 294 mg/kg - bw/day for females 1 ) Statistical methods used : Not specified Test method/conditions remarks : “ The feeding of the experimental diets was started when the initial animals were approximately 7 weeks of age; thereafter, the initial progenitors and 3 successive generations of offspring were maintained exclusively on the respectively assigned diets . After the initial animals had fed on the respective diets for 8 weeks and were approximately 105 days of age, the 2 sexes were housed together, 2 females and 1 male being fed on th e same diet per cage . The male rats were kept with the femal es until the pregnancy of each female was ascertained by a s harp increase in body weight . Th e pregnant females were placed in individual nesting cages and were observed daily th ereafter . If neither female was evidently pregnant after 4 weeks, another male t hat had already sired offspring was pl aced with the females and the delinquent male was housed with females of established fertility. ” [Witherup et al . (1969), p. 4 - 5]

“ Shortly after birth, each pup was examined for anomalies in the form, size, and color of the trunk; the number, size, formation, and location of the various appendages (head, feet, limbs, toes, and tail), and the number and location of the rudimentary ears, eyes, nose, and mouth were examined also . The pups were counted and weighed as a gr oup. ” [Witherup et al . (1969), p. 5]

“ The initial progenitors and 2 generations of offspring were given 2 opportunities to mate [resulting in two litters, “a” and “b”] ; the first litter of each initial female [from the P generation] was permitted to suckle for 7 days [or 4 days for pups from F1 females] ; the pups then were killed and their viscera were examined for gross anatomical anomalies and pathological abnormalities . After an interval of 1 week, the dams again were housed with the males . The o ffspring resulting from the second period of mating were permitted to suckle for 3 weeks, the normal age of weaning . The pups then were grouped according to their sex and were counted and weighed as a group . The offspring of the [P and F1] animals fed on each diet were reduced in number by random selection to 20 females and 10 males [per dietary concentration] ; these [F1 and F2 animals] were continued on the same diets as had been fed to their progenitors and in turn were mated when about 3 months of age. ” [Witherup et al . (1969), p. 5]

“ Pups in the first litters of the F1 and F2 females were counted and weighed when 4 days of age; those of the Fl females were killed and the dams were remated . The pups in both litters of the F2 1 Doses were converted from ppm to mg/kg - bw/day using the SRC Dose Calculator v 1.0 , with exposure duration set to “subchronic” and using default body weights and food consumption values for Sprague - Dawley rats because CD rats are Sprague - Dawley - derived.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

92 females and those in the second litters of the F1 females were reduced to no more than 10 pups per litter and continued to suckle on the dams until they were weaned at 21 days of age.” [Witherup et al . (1969), p p . 5 - 6] “Representative animals of the F3 ge nerations when 21 days of age were killed by intraperitoneal injections of Nembutal . Their viscera were examined for gross pathological changes . Tissues of the brain, heart, lungs, liver, kidneys, spleen, intestinal tract and the adrenal glands were prepared for microscopic examination; these were fixed in neutral formalin, embedded in paraffin and cut in sections of 6 to 8 microns in thickness . They were stained with hematoxylin and eosin.” [Witherup et al . (1969), p. 6] RESULTS Endpoint value(s) : The NOAEL for systemic and reproductive toxicity was 3000 ppm ( 258 and 294 mg/kg - bw/day for males and females, respectively ) based on no observed treatment - related effects up to the highest dietary concentration tested . D ue to a potential treatment - related decrease in the lactation index observed only in F2b pups, results of this study are considered equivocal and NOAEL and LOAEL values for developmental toxicity cannot be determined. Results remarks : There were no treatment - related effects on survival of parental rats . Occasional fatalities in the procreating animals were attributed to dystocia or pneumonia. Fertility indices of the P generation (both test substance - exposed and control) were poor (53.8

– 87.2%) compared to subsequent generations (80

– 100%) . Variations in gestation index were noted in test substance - exposed and control animals from successive generations (75

– 100%) . Variations in the viability index were noted in the first generation progeny from all groups, including controls ( 20.8

– 83.7 % ) . The differences in fertility, gestation , and viability indices showed no dose - relationship and were attributed to the influence of extraneous diseases unrelated to the test substance; therefore, these effects a re not considered to be treatment - related. A dose - related decrease in survival from postpartum days 4 to 21 [i.e., the lactation index , calculated as : (number of pups alive on postpartum day 21/number of pups entered into the lactation phase on postpartum day 4) × 100 % ] was observed in the F2b generation at dietary concentrations of 100, 500 , and 3000 ppm ; lactation indices were 88.9, 91.9, 83.2, 67.1, 62.8, and 47.3% at dietary concentrations of 0, 15, 60, 100, 500, and 3000 ppm , respectively . The s tudy authors did not regard the observed decrease as treatment - related because no dose - related changes were observed in the lactation index of F3 offspring . However, the study authors d id not provide additional information or data to evaluate other possib le cause s of th e dose - related effect in the F2b generation . A relationship between the decrease in lactation index and treatment with the test diets cannot be ruled out; therefore, results of this study are considered equivocal. There were no effects on body weights of offspring . Approximately 5400 rats that were progeny of animals fed on diets containing the test article were examined for anatomical anomalies; none were found . All of the offspring were normal inDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

93 appearance of the trunk and appendages , whether examined at birth, 4 days, 7 days, or 3 weeks of age . In the control offspring (1100) , there were 5 pups in the first litter of one of the second generation females that were dismembered by the dam . No gross pathological changes were observed i n the viscera of pups killed at 4, 7, or 21 days of age . H istological examination of the brain, heart, lungs, kidneys, liver, spleen, intestinal tract, and adrenal glands of the third generation of offspring (F3a, b) revealed no pathology . CONCLUSIONS Throughout the study, the animals were affected by extraneous diseases complicated by environmental factors that were unrelated to the content of test substance in the diets . The test substance had no effect on fertility and did not produce anatomical an omalies in offspring . The NOAEL for systemic and reproductive toxicity was 3000 ppm ( 258 and 294 mg/kg - bw/day for males and females, respectively ) based on no observed treatment - related effects up to the highest dietary concentration tested . However, due to a potential treatment - related decrease in the lactation index observed only in F2b pups, results of this study are considered equivocal and NOAEL and LOAEL values for developmental toxicity cannot be determined. STUDY RELIABILITY [3] Not reliable . The highest dose tested did not induce toxic effects as recommended in guideline 40 CFR 799.9355 and was below the recommended limit dose of 1000 mg/kg - bw/day . In addition, r esults of this study are considered equivocal due to a potential treatment - related decrease in the lactation index in F2b pups; therefore, NOAEL and LOAEL values for developmental toxicity cannot be determined . Important study details that were missing from the study summary included test substance purity, test method/ guideline followed , and statistical methods used . Measurements of body weights and food consumption were not conducted for the parental animals . Additionally, extraneous disease and underlying environmental factors complicate the interpretation of the re sults . REFERENCE Witherup, S., K.L . Stemmer, P. Taylor, and E.A . Pfitzer . 1969 . “ The Effects Exerted Upon the Fertility of Rats and Upon the Viability of Their Offspring by the Introduction of 4,4' - Methylenebis (2,6 - Ditertiary Butylphenol) Into Their Diets, ” The Kettering Laboratory in the Department of Preventive Medicine and Industrial Health, College of Medicine, University of Cincinnati, Cincinnati, Ohio, for Ethyl Corporation . Available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0181 ( on p p. 22 - 61 of “End_Point_F2

– Mammalian_Toxicity_ - _Repeated_Dose_Reproduction _Developmental.pdf” ) as of December 4, 2013 . Text in quotes was taken directly from Witherup et al ., 1969 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

94 Study 4 : Reproductive toxicity study in rats and dogs (Schering Corporation, 1966) Title : Not provided Th is study is: Adequate Not Adequate The original study report was not available for review . This wa s n ot a standard study of reproductive toxicity endpoints ; the endpoints assessed and numbers of animals used were insufficient to satisfy guideline requirements . Important study details that were missing from the summary included test substance purity, test methods/guidelines followed, type of oral route used in the rat study , dog strain and sex , statistical methods used, test methods/conditions , and detailed results/findings . Additionally, LOAEL/NOAEL values were not reported and could not be determined based on the limited study details provided. TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; Synonym : SN# 1002, AN 702; Sch 12150 Remarks : Purity not provided METHOD Method/guideline followed : Not specified Test type : Reproductive Toxicity GLP compliant? : N/A (pre - GLP) Year study performed : 1966 Test species : Rats (ovariectomized females) and Dogs (strain and sex not specified) Number of animals/sex/dose : 5/group (rats); Not specified (dogs) Route of administration : Oral or subcutaneous injection (rat studies) ; In capsules or as pure compound dissolved in edible oil and fed in the diet (dog studies) Vehicle : Edible Oil ( for dietary study in dogs) Exposure duration : 4 weeks (rats); 20 days (dogs) Doses/concentrations tested : Rats : 100 or 300 mg/kg - bw /day (subcutaneous); 100, 300 , or 900 mg/kg - bw /day (oral route) Dogs : 15, 30 , or 45 mg/kg - bw /dayDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

95 Statistical methods used : Not specified Test method/conditions remarks : “Numerous studies were conducted on AN 702 to determine an appropriate species for inve stigation of the pharmacologic activity and mechanism of action of AN 702 . Most of the studies were performed for Ethyl Corporation by Schering Corporation (thus the Sch 12150 test article identification) . Some endpoints relative to reproduction were ass essed: a . Five rats per group were ovariectomized and given AN702 orally [at 1 00, 300, or 900 mg/kg bw/day] or subcutaneously [at 100 or 300 mg/kg - bw/day] for four weeks to assess hypocholesterol effects . Effects on uterus, vagina, pituitary , and adrenal were also measured ... b . 24 dogs were administered AN 702 for 20 days as micronized, dispersible formulation in capsules or as pure compound dissolved in edible oil and fed in the diet . Doses were 15 mg/kg, 30 mg/kg, and 45 mg/kg . Six weeks after discontinuing the drug, eyes, liver, kidneys , and testes were examined histologically.” [EPA - HQ - OPPT - 2005 - 0033 - 0181, p. 8 ] RESULTS Endpoint value(s) : N/A Results remarks : a . Ovariectomized r ats injected subcutaneously with AN 702 at 100 or 300 mg/kg - bw /day for 4 weeks had slightly higher uterine weights than control rats (0.136 and 0.145 g at 100 and 300 mg/kg - bw/day, respectively, versus 0.108 g in controls ) . Ovariectomized r ats given AN 702 orally for 4 weeks at doses of 100, 300, or 900 mg/ kg - bw /day did not show a trend in uterine weight differences (0.117, 0.138, and 0.101 g at 100, 300, and 900 mg/kg - bw /day , respectively, versus 0.115 g in controls ) . N o differences from control were seen in either group in vaginal cytology, pituitary, or adrenal weights . b . No abnormalities associated with the test article were found upon histological examination of the eyes, liver, kidneys , and testes of dogs s ix weeks after discontinuing oral administration of AN 702 . CONCLUSIONS Increased uterine weights (statistical significance not indicated) were observed in ovariectomized rats that were injected subcutaneously with 100 or 300 mg/kg - bw /day AN 702 for a period of 4 weeks . No effects were observed in ovariectomized rats that wer e dosed orally with up to 900 mg/kg - bw /day AN 702 for 4 weeks or in dogs dosed orally with up to 45 mg/kg - bw/day AN 702 for 20 days followed by six weeks of observation .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

96 STUDY RELIABILITY [3] Not reliable . This was n ot a standard study of reproductive toxicity endpoints ; the endpoints assessed and numbers of animals used were insufficient to satisfy guideline requirements . Important study details that were missing from the summary (the original study report was not available for review) in cluded test substance purity, test methods/guidelines followed, type of oral route used in the rat study , dog strain and sex, statistical methods used, test methods/conditions , and detailed results/findings . Additionally, LOAEL/NOAEL values were not reported and could not be determined based on the limited study details provided. REFERENCE Schering Corporation . 1966 . “ AN 702 (Sch 12150) Schering Corporation for Ethyl Corporation Taken from ‘ Preclinical Information for EY 2333 (702) ’ Vol . II, Book 1 of 3, 1966. ” Study summary available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT 2005 - 0033 - 0181 ( on p p. 7 - 8 of “End_Point _F2

– Mammalian_Toxicity_ - _Repeated_Dose_Reproduction_Developmental.pdf” ) as of December 4, 2013 . Text in quotes is taken directly from EPA - HQ - OPPT - 2005 - 0033 - 0181 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

97 Study 5 : Teratology study in rats (Powers, 1970a ) Title : Teratology Study - Rats, EY 2333 Final Report Th is study is: Adequate Not Adequate The highest dose tested did not induce toxic effects as recommended in guideline 40 CFR 799.9355 and was below the recommended limit dose of 1000 mg/kg - bw/day . Additionally, important det ails that were missing from the study includ ed test method/guidelines followed and statistical methods used. TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; r eferred to as EY 2333 in study report Remarks : 100% purity was assumed by study authors; Sample No . 158 - 70; Fine white powder with noticeable odor METHOD Method/guideline followed : Not specified Test type : Developmental Study - Rats GLP compliant? : N/A (pre - GLP) Year study performed : 1970 Test species : Charles River Rats ; obtained from Charles River Breeding Laboratories Number of animals/sex/dose : 18 (control and low - dose) or 19 (high - dose); females only Route of administration : Oral gavage Exposure duration : 10 days ( gestational days 6

– 15 ) Doses/concentrations tested : 0 (vehicle control), 15 , and 150 mg/kg - bw /day Vehicle : Corn Oil S tatistical methods used : None specified Test method/conditions remarks : Rats were treated by oral gavage with 0, 15 , or 150 mg/kg bw /day in corn oil on gestational days 6 through 15 . The control group received corn oil . Two females were placed with one male in a breeding cage until the day sperm were noted in aDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

98 vaginal smear . Females were then housed singly . From a total of 55 fe males in which sperm were noted, animals were randomly assigned to test group s , with 18 females each in the control and 15 mg/kg - bw/day groups and 19 females in the 150 mg/kg - bw/day group. “Mortality, appearance, and behavior of the females were observed daily during the mating and gestation periods . Body weights were recorded after observation of sperm, at initiation of compound administration, at a five - day interval during treatment, at termination of treatment, and prior to sacrifice for Caesarean deli very . Food consumption was recorded for the period prior to treatment, at two intervals during treatment, and for the period following treatment to time of sacrifice” [Powers (1970a), p. 4]

“ Females were sacrificed on Day 19 of gestation . Caesarean sect ions were performed, and the following observations were made : number and placement of implantation sites, resorption sites, and live and dead fetuses ; individual fetal weight and length (crown - rump); and external fetal anatomical structure . ” [Powers (19 70a), p. 4] After external observations, about one - third of the fetuses from each litter were fixed with Bouin ’ s solution and evaluated for visceral changes by Wilson's technique . The remaining two - thirds of the fetuses were prepared for skeletal examination. Examination of the visceral structures by Wilson's sectioning technique was performed on 58, 61 , and 68 fetuses from the control, low - dose, and high - dose groups, respectively. RESULTS Endpoint value(s) : NOAEL (maternal and developmental toxicity) = 150 mg/kg - bw/day based on no treatment - related effects observed at the highest dose level tested. Results remarks : Maternal Toxicological Data: There were n o maternal deaths during the study . Pregnancy was observed in a total of 53 out of 55 rats (18 control, 17 low - dose , and 18 high - dose animals) . One low - dose female and one high - dose female were not pregnant . There were no treatment - related changes in appearance, behavior, body weight gain , and food consumption . Incidental findings consisted of wheezing observed in one low - dose female on day 11 and soft feces in one high dose female on day 19 . The low - dose animal which was not pregnant exhibited distended uterine horns with clear fluid . No other findings were no ted at necropsy. Caesarean Data: There were no effects on the number and placement of uterine implantation and resorption sites or the number, weight, and length of live fetuses in the test groups . One dead fetus, which was normally developed, was found in the control group . Three late resorption sites were noted in one control female , and one late resorption site was noted in a second control female . Two low - dose females exhibited one late resorption site each .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

99 External Visceral Data: A few fetuse s from all groups, including the control, appeared small in size . One of the small fetus es from the high - dose level and another fetus from the same dose group were pale in color . One fetus from the low - dose group was missing due to a technical error . Findings included clotted blood in the thoracic region in 15 control, 28 low - dose , and 18 high dose fetuses , clotted blood in the abdominal or intestinal region in 6 control fetuses and 1 high dose fetus, clotted blood in the cranial region in 1 control fetus , dilation of one or both renal pelves in 2 control, 3 low - dose , and 8 high - dose fetuses, dilation of the lateral ventricles of the brain in 1 control and 3 high - dose fetuses , and light colored areas in the liver of one control fetus. Skelet al Data: “ Single ribs [seen in 1 fetus at the high dose] and ribs described as ‘ floating ’ in appearance or unattached from the vertebral column [seen in 1 fetus in the low dose] were located at the 20th vertebra in all animals in which they were observed . The 14th pair of ribs noted in one control fetus was small in size . Rudimentary bone structures were observed at the 20th vertebra on both sides of the vertebral column in one high level fetus and on the left side in another . The interparietal skull b one was missing or underdeveloped in two control fetuses and one high level fetus . Three control and two high level fetuses had the occipital skull bone missing or underdeveloped . The centrum of the first lumbar vertebra was split in two fetuses and smal l in one, while t he centra of the 11th, 12th, and 13th thoracic vertebrae were split in one fetus from one control litter . A fetus from another control litter had the centrum of the 12th thoracic vertebra small and split. ” [Powers (1970a), p. 8]

“ There was no evidence of a compound - related effect among the test fetuses . There were no meaningful differences in the developmental indices of the test groups as compared to the controls . No trends toward lesser or greater ske letal development were observed . ” [Powers (1970a), p. 9] CONCLUSIONS Based on the results of this study, the test substance did not produce maternal toxicity or developmental effects in rats . The NOAEL for maternal and developmental toxicity in rats was 150 mg/kg - bw/day (highest dose tested). STUDY RELIABILITY [3] Not reliable . The highest dose tested did not induce toxic effects as recommended in guideline 40 CFR 799.9355 and was below the recommended limit dose of 1000 mg/kg - bw/day . Additionally, important details that were missing from the study includ ed test method/guidelines followed and statistical methods used.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

100 REFERENCE Powers, M.B . 1970a . “ Teratology Study

– Rats : EY 2333 Final Report, ” Hazleton Laboratories, Inc., for Ethyl Corporation . Available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0181 ( on p p. 84 - 100 of “End_Point_F2

– Mammalian_Toxicity_ - _Repeated_Dose_Reproduction_Developmental. pdf”) as of December 4, 2013 . Text in quotes is taken directly from Powers, 1970a.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

101 Study 6 : Teratology study in rabbits (Powers, 1970b ) Title : Teratology Study - Rabbits, EY 2333 Final Report Th is study is: Adequate Not Adequate A method/guideline was not identified and no ratio nale was provided for selection of the doses that were tested . In addition, t his study had the following deficiencies compared with guidelines for the conduct of a prenatal developmental toxicity test (OECD TG 414, OPPTS 870.3700, and 40 CFR 799.9370) : (1) Each test and control group should contain a sufficient number of females to result in approximately 20 animals with implantation sites at necropsy , whereas initial group size in the current study was 15 females and two maternal animals died (one at 0 mg/kg bw/day and one at 15 mg/kg - bw/day) and seven were non - pregnant (one at 15 mg/kg - bw/day and six at 150 mg/kg - bw/day) ; (2) Two doses were tested whereas at least three doses should be used and the highest dose should produce some evidence of developmen tal and/or maternal toxicity; (3) Statistical analyses of the results were not provided; (4) Maternal body weight and food consumption data were recorded but were not presented in the study report that was available; (5) Pre - and post - implantation loss cal culations were not provided; and (6) Fetal sex ratio data were not presented in the study report that was available . Also , historical control data were not presented to allow interpretation of the following findings of this study : (1) abortion occurred i n ~20% of the control animals but not in dosed groups; and (2) 40% of the high - dose animals were non - pregnant compared with 0% in controls and ~7% at the low dose . TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; Referred to as EY 2333 in study report Remarks : 100% purity was assumed by study authors METHOD Method/guideline followed : Not specified Test type : Prenatal Developmental Study - Rabbits GLP compliant? : N/A (pre - GLP) Year study performed : 1970 Test species : New Zealand White rabbits ; obtained from Rowmar Rabbitry Number of animals/sex/dose : 15 females /dose Route of administration : Oral ( gelatin capsules)Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

102 Exposure duration : G estational day s 6

– 18 Doses/concentrations tested : 0 (control), 15 , and 150 mg/kg - bw/day Vehicle : None S tatistical methods used : None specified Test method/conditions remarks : Following the injection of 45 female New Zealand W hite rabbits with p ituitary l uteinizing h ormone to stimulate ovulation , each animal was impregnated by artificial insemination . “The day of insemination was designated as Day 0 of gestation.” [Powers (1970b), p. 3 ] . From the group of 45 impregnated females , 15 each were r andomly assigned to the control, 15 - , and 150 - mg/kg - bw/day treatment groups . Daily, from days 6 through 18 of gestation, the impregnated females were administered gelatin capsules “containing the appropriate dosage levels” [Powers (1970b), p. 4] of test material . Animals of the control group received empty capsules . “Mortality, appearance, and behavior of the animals were observed daily . Body weights were recorded weekly and terminally . Food consumption was recorded daily during the treatme nt period.” [Powers (1970b), p. 4] The females were sacrificed on day 28, 29 , or 30 of gestation . Pups were removed and the following observations were made : number of corpora lutea ; number and placement of implantation sites, resorption sites, and live and dead fetuses ; number of live and dead fetuses after a 24 - hour incubation period ; individual fetal weights and lengths ; and external fetal anatomy . Pups were examined for visceral and skeletal abnormalities . RESULTS Endpoint value(s) : NOAEL (maternal and developmental toxicity) = 150 mg/kg - bw/day based on no treatment - related effects observed at the highest dose level tested. Results remarks : “One control doe and one low level doe died during the study; in both cases, it was too early in ge station to determine pregnancy . Among the surviving animals, evidence of abortion was noted in three control animals . One control doe delivered her litter naturally . The remaining control and all test litters were delivered by Caesarean section . Pregna ncy was observed in a total of 36 of the 45 does used in this study (14 control, 13 low level, and nine high level animals).” [Powers (1970b), p. 1] “Appearance, behavior, body weight gain, and food consumption were comparable among control and test anim als . All uterine, litter, and Caesarean data were also comparable among the test and control groups.” [Powers (1970b), p. 1] “There was no indication of a compound - related effect in the external appearance, visceral anatomy, development, and skeletal st ructure of the test fetuses as compared to those of the control fetuses.” [Powers (1970b), p. 1]Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

103 CONCLUSIONS The NOAEL for maternal and developmental toxicity in rabbits was 150 mg/kg - bw/day (highest dose tested). STUDY RELIABILITY [3] Not reliable . A method/guideline was not identified and no rationale was provided for selection of the doses that were tested . In addition, t his study had the following deficiencies compared with guidelines for the conduct of a prenatal developmental toxicity test (OEC D TG 414, OPPTS 870.3700, and 40 CFR 799.9370) : (1) Each test and control group should contain a sufficient number of females to result in approximately 20 animals with implantation sites at necropsy , whereas initial group size in the current study was 15 females and two maternal animals died (one at 0 mg/kg - bw/day and one at 15 mg/kg - bw/day) and seven were non - pregnant (one at 15 mg/kg - bw/day and six at 150 mg/kg - bw/day) ; (2) Two doses were tested whe reas at least three doses should be used and the highest dose should produce some evidence of developmental and/or maternal toxicity; (3) Statistical analyses of the results were not provided; (4) Maternal body weight and food consumption data were recorde d but were not presented in the study report that was available; (5) Pre - and post - implantation loss calculations were not provided; and (6) Fetal sex ratio data were not presented in the study report that was available . Also , historical control data were not presented to allow interpretation of the following findings of this study : (1) abortion occurred in ~20% of the control animals but not in dosed groups; and (2) 40% of the high - dose animals were non - pregnant compared with 0% in controls and ~7% at th e low dose . REFERENCE Powers, M.B . 1970b . “ Teratology Study - Rabbits, EY 2333 Final Report, ” Hazleton Laboratories, Inc., for Ethyl Corporation . Available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0181 ( on p p. 62 - 83 of “End_Point_F2

– Mammalian_Toxicity_ - _Repeated_Dose_Reproduction_ Developmen tal.pdf” ) as of December 4, 2013 . Text in quotes is taken directly from Powers, 1970b.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

104 D . SUPPLEMENTAL DATA (NON - SIDS ENDPOINTS) D - 1 . Carcinogenicity Study 1 : Combined r epeated - d ose and c arcinogenicity t oxicity s tudy (

ECHA website, Exp Key Carcinogenicity .00 1 ) Title : Not provided TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1 ; referred to as 4,4' - Methylenebis(2,6 - di - tert - butylphenol) in the study report (other s ynonym s : BHD, AN2, Ethyl Antioxidant 702 ). Remarks : Purity not specified ; crystalline s olid METHOD Method/guideline followed : Similar to OECD TG 453 “ Combined Chronic Toxicity/Carcinogenicity Studies ” Test type : Combined Repeated - Dose Toxicity/Carcinogenicity Study GLP compliant? : N/A ( Pre - GLP) Year study performed : 1969 Test species : Crj : CD(SD) rat ; s ource : Charles River Breeding Laboratories Inc. , Brookline, M A ; age : weanlings at initiation; weight (average during week 1) : 394

– 436 g (males) and 248

– 269 g (females) . Number of animals/sex/dose : 25 in the test groups; 50 in the control group Route of administration : Oral - Diet Vehicle : Mazola corn oil (reported to be free of any antioxidants) Exposure duration : 2 years Doses/concentrations tested : 0, 15, 60, 100, 500, and 3000 ppm ( corresponding approximate dose levels were 0, 0.55 , 2.14, 3.56, 18.1, and 107 mg/kg - bw/day, respectively, for males and 0, 0.67 , 2.63 , 4.35, 22.2 , and 138 mg/kg - bw/day, respectively , for females ) Statistical methods used : Not specifiedDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

105 Test method/conditions remarks : Dosing and Environmental Conditions: Rats (25/sex/dose) were fed diets containing the test substance at various concentrations for up to 2 years . A control group of 50 rats was fed a diet containing only the corn oil vehicle . Four rats per sex were assigned to subgroups of each dose group for interim kills at 13, 26, 52, and 78 weeks (3, 6, 12, and 18 months) . Terminal sacrifice was performed at 104 weeks (24 months) . The animals were acclimated for 2 weeks and were not fasted prior to the start of the study . The test animals were housed three males and four females per cage prior to the start of the study . Diet and water were provided ad libitum throughout the study . During the study, the test tem perature was maintained at 76 ± 2 °F and the humidity was not controlled. Preparation of Test Diets: For preparation of the test diets, “1 kg of corn oil was added to 19 kg of the base diet with or without the test substance . The mixture was stirred for 20 min in a Hobart mixer (stainless steel) . [Five] diets containing the test item were prepared... The diet was freshly prepared every week.” [ ECHA ] The test diets were prepared by “mixing appropriate amounts with [a] special preparation of Rockland Mouse and Rat diet containing no more than 1% of fat.” [ ECHA ] The rodent diet food was obtained fresh once each month. Clinical Observations, Body Weights, and Food Consumption: Cage - side observations were made on a daily basis for general appearance and beha vior . Animals were observed weekly for a detailed assessment of clinical signs . Body weight and food consumption were recorded weekly . Mean test substance intake for males and females in each group was calculated by an unspecified method . Ophthalmological examinations were performed weekly and all groups were examined for the presence of cataracts . Hematology and Clinical Chemistry Measurements: Hematological examinations were performed on unfasted treated and control animals (10 rats/s ex/dose) at 1 week before the start of dosing, and again at 13, 26, 52, 78, and 104 weeks . Hematological parameters examined included the following : prothrombin time, hemoglobin, packed cell volume of erythrocytes, and total and differential leukocytes . Clinical chemistry investigations were performed on blood samples collected at scheduled sacrifice of all test animals . Clinical chemistry parameters examined included the following : total lipids, cholesterol, and alkaline phosphatase . No urinalysis assessments were performed. Gross Necropsy and Histopathology: It appears that organ weights were recorded, but, with the exception of the liver, the organs weighed were not specified in the ECHA study summary (the full study report was not available for review) . Pathological examinations were performed on the following organs at gross necropsy : liver, heart, lung, kidneys, spleen, brain, gonads, pituitary, adrenals, and thyroid . Histological examinations were performed on selected tissues, including t he heart, aorta, lung, liver, spleen, kidney, stomach, duodenum, pylorus, smallDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

106 intestine (ileum), large intestine (colon), mesenteric lymph nodes, urinary bladder, prostate, testes, ovaries, uterus, pancreas, thyroid, thymus, adrenals, pituitary, brain, s pinal cord, eye, bone marrow, bone, skeletal muscle, and skin. RESULTS Endpoint value(s) : No evidence of carcinogenicity Results remarks : “ Neoplastic lesions observed in the study were subcutaneous or in females mammary gland adeno [ fibroma ] , pituitary adenoma, one adenoma in the parathyroid, thyroid adenoma, adrenal adenoma, [and] liver angioma that wer e all benign tumors . Leukemia, [s] eminoma and ovary carcinoma were single findings in control animals only . No test substance related incre ased incidence of any tumo r was observed and it can be concluded that the test substance did not induce neoplastic lesions under the conditions of this study. ” [ ECHA ]

“ The histopathology findings were confirmed in a second reading of the data with independent pathologists and reported in a supplementary report of the same laboratory ...” [ ECHA ] CONCLUSIONS The study summary concluded that the test substance did not induce an increase in tumor incidence under the conditions of the study . STUDY RELIABILITY [2] Reliable with restrictions . When compared to the TSCA 799.9430 guideline, the following deviations were noted : ( 1) the total number of animals at test initiation was not specified ; ( 2) the lot or batch number of the test substance was no t provided ; ( 3) the environmental test temperature (76 ± 2 ° F) exceeded the recommended temperature of 22 ± 3 °C (71.6 ° F); ( 4) humidity was not controlled ; ( 5) limited hematology and clinical chemistry parameters were examined ; ( 6) urinalysis was not conducted ; ( 7) organs weighed at necropsy were not specified ; and ( 8) group mean data for endpoints other than histopathological lesions were not reported . REFERENCE Study summary available from the ECHA website, http://apps.echa.europa.eu/registered/data/dossi ers/DISS - dcee47d2 - 62ad - 0171 - e044 - 00144f67d031/AGGR - b809391a - f592 - 4fd4 - b07c - 046e2cb18268_DISS - dcee47d2 - 62ad - 0171 - e044 - 00144f67d031.html#AGGR - b809391a - f592 - 4fd4 - b07c - 046e2cb18268 . The data source is referred to as a “study rep ort”, with a report date of 1969 - 07 - 08 . No other reference information is provided . Text in quotes is taken directly from the ECHA website.Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

107 Copyright Notice : © European Chemicals Agency, 2007 - 2011 . Reproduction is authorised provided the source is fully acknowledged in the for m “ Source : European Chemicals Agency, http://echa.europa.eu/ ” , and provided written notification is given to the ECHA Communication Unit (info (at) echa.europa.eu).Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

10 8 D - 2 . Skin Irritation Study 1 : Acute dermal toxicity study in rabbits (Carter and Dharmarajan , 1976) Title : A Report on the Acute Toxicity of AN - 702 TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; referred to as AN - 702 in study report Remarks : Purity not provided METHOD Method/guideline followed : Not specified Test type : Acute Dermal - Rabbit GLP compliant? : Not specified Year study performed : 1976 Test species : New Zealand White rabbits; 2.3

– 3.0 kg Number of animals/sex/dose : 4 (sex not specified) Route of administration : Dermal Vehicle : Peanut Oil Doses/concentrations tested : 1500 m g/kg - bw Statistical methods used : No information available Test method/conditions remarks : In an acute dermal toxicity test, t he test substance (10% w/v solution in peanut oil) was injected under a plastic sleeve wrapped around the clipped trunks of two intact and two abraded rabbits for an exposure period of 24 hours . Evaluations of erythema and edema were made 24 and 72 hours after application of the test material . After removal of the sleeves, the animals were observed for 14 days. RESULTS Endpoint value(s) : IrritatingDraft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

109 Results remarks : No mortalities occurred . Well defined erythema was observed on the first day of observation. CONCLUSIONS The test substance was irritating to rabbit skin . STUDY RELIABILITY [ 2 ] R eliable with restrictions . Important study details that were missing from the study summary included test substance purity, methods/guidelines followed, GLP compliance, sex of test animals , statistical methods, detailed findings , and numerical results . REFERENCE Carter, G. and V . Dh ar marajan . 1976 . “ A Report on the Acute Toxicity of AN - 702, ” Tulane University School of Medicine, for Ethyl Corporation . Submitted by Albemarle Corporation . Study summary available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ OPPT - 2005 - 0033 - 0181 (on p p. 7 - 13 of the document “End_Point_D

– Mammalian_Toxicity_Acute.pdf”) as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

110 Study 2 : Skin irritation study in rabbits ( Ethyl Petroleum Additives International, 1991) Title : Acute Dermal Irritation/Corrosion Test in the Rabbit TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1 Remarks : Purity not provided METHOD Method/guideline followed : OECD 404 “Acute Dermal Irritation/Corrosion” Test type : Primary Dermal Irritation - Rabbit GLP compliant? : Yes Year study performed : 1 991 Test species : New Zealand White rabbits Number of animals/sex/dose : 3 males Route of administration : Dermal Vehicle : None specified Doses/concentrations tested : 0. 5 mL Statistical methods used : None specified Test method/conditions remarks : Three male rabbits were exposed to the test substance for 4 hours and observed for 16 days thereafter . RESULTS Endpoint value(s) : I rritating Results remarks : No deaths were observed . Well - defined to moderate erythema was observed in all animals during the first 9 day s following treatment . Slight erythema was noted in all animals 12 days after treatment and in one animal at study termination . Very slight or slight edema was noted in two animals during the first 4 days following treatment . Very slight edema was noted at all test s ites 2 days later, and persisted in one animal up to 12 days . Tw o animals exhibited e xfoliation on days 6 , 9 , and/or 12 post - exposure .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

111 CONCLUSIONS The test substance was irritating to rabbit skin. STUDY RELIABILITY [ 2 ] Reliable with restrictions . Only the study summary was available . Important study details that were missing from the study summary included test substance purity, concentration of test solution, interpretation of irritation scores , and detailed numerical results . REFERENCE Ethyl Petroleum Additives International . 1991 . Acute D ermal I rritation/ C orrosion T est in the R abbit . Study No. 90/ETP012/1368 . Study conducted by Life Science Research Limited . (Cited in IUCLID Dossier, European Chemical Bureau, 2000) . Study summary available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0181 ( on p. 71 of the document “End_Point_D

– Mammalian_Toxicity_Acute.pdf ) as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

112 Study 3 : Dermal irritation study in rabbits ( Carter and Dharmarajan , 1976) Title : A Report on the Acute Toxicity of AN - 702 TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; referred to as AN - 702 in study report Remarks : Purity not provided METHOD Method/guideline followed : Not specified Test type : Primary Dermal Irritation - Rabbit GLP compliant? : Not specified Year study performed : 1976 Test species : New Zealand White rabbits Number of animals/sex/dose : 6 (sex not specified) Route of administration : Dermal Vehicle : None Doses/concentrations tested : 0.5 g Statistical methods used : None specified Test method/conditions remarks : The test substance was applied as a neat powder to four clipped skin areas of each animal, two of which were abraded . The areas were covered with gauze pads for 24 hours . Observations for erythema and edema were made at 24 and 72 hours following exposure . RESULTS Endpoint value(s) : Not irritating Results remarks : No erythema or edema was observed (Draize score of 0) .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

113 CONCLUSIONS The test substance was not irritating to rabbit skin when applied as a nea t powder . However, the investigators noted that when the test substance was applied as a 10% w/v solution in peanut oil there was defined erythema. STUDY RELIABILITY [ 4 ] Not assignable . Important study details that were missing from the study summary included test substance purity, methods/guidelines followed, GLP compliance, and sex of test species . In addition, the results conflict with a different test conducted with the same chemical. REFERENCE Carter, G . and V . Dh ar marajan . 1976 . “ A Report on the Acute Toxicity of AN - 702, ” Tulane University School of Medicine, for Ethyl Corporation . Submitted by Albemarle Corporation . Study summary available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ OPPT - 2005 - 0033 - 0181 ( on p p. 7 - 13 of the document “End_Point_D

– Mammalian_Toxicity_ Acute.pdf ) as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

114 D - 3 . Eye Irritation Study 1 : Eye irritation study in rabbits ( Carter and Dharmarajan , 1976 ) Title : A Report on the Acute Toxicity of AN - 702 TEST SUBSTANCE Identity : Phenol, 4,4’ - methylenebis[2,6 - bis(1,1 - dimethylethyl) - , CASRN 118 - 82 - 1; referred to as AN - 702 in study report Remarks : Purity not provided METHOD Method/guideline followed : Not specified Test type : Primary Eye Irritation - Rabbit GLP compliant? : Not specified Year study performed : 197 6 Test species : New Zealand White rabbits Number of animals/sex/dose : 6 (sex not specified) Route of administration : Ocular Vehicle : None Doses/concentrations tested : 0.1 g Statistical methods used : None specified Test method/conditions remarks : One eye of each of six New Zealand White rabbits was treated with 0.1 g of test substance as a neat powder . The other untreated eye served as the control . Observations for damage and irritation were made at 1, 2, 3 , and 7 days following administration .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

115 RESULTS Endpoint value(s) : Not Irritating Results remarks : Slight corneal opacity was observed in one rabbit and s light conjunctival irritation was observed in five rabbits on the first day of observation . Mean scores for cornea, iris , and conjunctivae at 1, 2, 3 , and 7 days post - exposure , respectively, were 3.5, 1.5, 0.8 , and 0 out of a total of 110 . CONCLUSIONS The test substance caused slight corneal opacity and slight conjunctival irritation. STUDY RELIABILITY [ 4 ] Not assignable . Important study details that were missing from the study summary included test substance purity, methods/guidelines followed, GLP compliance, sex of test species , and definition of the scoring system . REFERENCE Carter, G . and V . Dh ar marajan . 1976 . “ A Report on the Acute Toxicity of AN - 702, ” T ul ane University School of Medicine, for Ethyl Corporation . Study summary available online at http://www.regulations.gov/#!documentDetail;D=EPA - HQ - OPPT - 2005 - 0033 - 0181 ( on p p. 7 - 13 of the document “End_Point_D

– Mammalian_Toxicity_Acute.pdf ) as of December 4, 2013 .Draft HPV1/HPV2 Test Rule Data Adequacy Review (December 2013) CASRN 118 - 82 - 1

116 E . DATA ADEQUACY SUMMARY TABLE

Endpoint Study Information Available Acceptable Data Gap Aquatic Effects Acute Toxicity to Fish 1 Yes No No Acute Toxicity to Aquatic Invertebrates 1 Yes No No Chronic Toxicity to Aquatic Invertebrates 2 Yes Yes No Toxicity to Aquatic Plants 2 Yes Yes No Health Effects

Acute Toxicity Yes Yes No Repeated - Dose Toxicity Yes Yes No Gene Mutation Yes Yes No Chromosomal Aberrations Yes Yes No Reproductive Toxicity 3 Yes Yes No Developmental Toxicity 3 Yes Yes No 1 A cute toxicity testing in fish and aquatic invertebrates is not required under the HPV2 test rule because the log K ow of CASRN 118 - 82 - 1 is ≥ 4.2. (The log K ow value of CASRN 118 - 82 - 1 was experimentally determined to be > 6.50 by SafePharm Laboratories [study submitted to EPA by Albemarle Corporation on May 12, 2006, available online at http://www.regulations.gov/ #!documentDetail ;D=EPA - HQ - OPPT - 2005 - 0033 - 0163 as of December 5 , 2013; “4,4’ - Methylene bis(2,6 - di - tert - butylphenol): Determination of Water Solubility and Partition Coefficient”, SPL Project Number 1193/011, study completed on July 18, 2005].) 2 The HPV2 test rule, “Testing of Certain High Production Volume Chemicals; Second Group of Chemicals” (76 FR 1067, January 17, 2011), requires C1 aquatic toxicity testing for Phenol, 4,4' - methylenebis[2,6 - bis (1,1 - dimethylethyl) - (CASRN 118 - 82 - 1). The log K ow for CASRN 118 - 82 - 1 is > 6.50, indicating that testing under Test Group 2, which applies to chemicals with log K ow ≥ 4.2, is required. Test Group 2 includes the following tests: (1) Chronic Toxicity to Daphnia (ASTM E 1193 - 97) and (2) Toxicity to Plan ts (Algae; ASTM E 1218 - 04). 3 Adequate data were available for an acceptable supporting compound (2,6 - di - tert - butylphenol; CASRN 128 - 39 - 2).