SRC TR

- 1 3

- 049

HPV2 TEST RULE DATA ADEQUACY REVIEW : JU LY 2013 UPDATE ( FINAL )

Methanone, diphenyl

- , CASRN 119

- 61

- 9

Prepared by:

Teresa Manyin

, Justine Von Runnen, Megan Nelson, Amy Nguyen, and William L. Richards

Defense and Environmental Solutions Division SRC, Inc. 7502 Round Pond Road North Syracuse, NY 13212

Contract No. EP

- W

- 09

- 027 Task FG007. 3 .009.001

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 Assignment Manager

Ju ly 2, 201 3Final HPV2 Chemical Data Adequacy Review (July 2013) CASRN 119

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i TABLE OF CONTENTS

A. SOURCE MATERIAL USED FOR ASSESSMENT

................................ .............................. 1
B. EVALUATING DATA ADEQUACY

................................ ................................ ..................... 1
C. SUMMARY OF DATA ADEQUACY FOR TESTING REQUIRED BY TEST RULE

........ 2

D. DATA FOR SIDS ENDPOINTS

................................ ................................ .............................. 3

D

- 1. Acute Toxicity to Fish

................................ ................................ ................................ ........ 3

D

- 2. Acute Toxicity to Aquatic Invertebrates

................................ ................................ ............ 5

D

- 3. Toxicity to Aquatic Plants

................................ ................................ ................................ 15

D

- 4. Acute Toxicity

................................ ................................ ................................ .................. 23

D

- 5. Repeated

- Dose Toxicity

................................ ................................ ................................ ... 25

D

- 6. Genetic Toxicity

– Gene Mutation

................................ ................................ ................... 27

D

- 7. Genetic Toxicity

– Chromosomal Aberrations

................................ ................................ 30

D

- 8. Reproductive/Developmental Toxicity

................................ ................................ ............ 33
E. SUPPLEMENTAL DATA (NON

- SIDS ENDPOINTS)

................................ ........................ 44

E

- 1. Skin Irritation

................................ ................................ ................................ .................... 44

E

- 2. Eye Irritation

................................ ................................ ................................ ..................... 49

E

- 3. Skin Sensitization

................................ ................................ ................................ ............. 51

E

- 4. Ca rcinogenicity

................................ ................................ ................................ ................. 56

E

- 5. Genetic Toxicity

– Other

................................ ................................ ................................ .. 61
F. DATA ADEQUACY SUMMARY TABLE

................................ ................................ ........... 66Final HPV2 Chemical Data Adequacy Review (July 2013) CASRN 119

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1 A. SOURCE MATERIAL USED FOR ASSESSMENT

This document evaluates a submission from the Benzophenone HPV Consortium of the American Chemistry Council (dated April 10, 2013) that contains data from toxicity testing of methanone, diphenyl

- (CASRN 119

- 61

- 9) and was received by EPA in response to the final test rule, “Testing of Certain High Production Volume Chemicals; Second Group of Chemicals” (76 FR 1067, January 7, 2011; referred to here in as HPV2 ; available online at http://www. regulations.gov/#!documentDetail;D=EPA

- HQ

- OPPT

- 2007

- 0531

- 0579 as of July 1, 2013 ).

T he HPV2 test rule requires the following toxicological testing with CASRN 119

- 61

- 9 : C2 (consisting of acute toxicity to Daphnia and toxicity to algae testing if log K ow < 4.2 or chronic toxicity to Daphnia and toxicity to algae testing if log K ow ≥ 4.2) . Since the log K ow of CASRN 119

- 61

- 9 has been experimentally determined to be 3.18 1

, which is < 4.2, the C 2 aquatic toxicity testing requirement is considered to include acute toxicity to Daphnia and toxicity to algae testing.

The April 10, 2013 submission from the Benzophenone HPV Consortium ( “ Benzophenone : A 96

- hour toxicity test with the freshwater alga ( Pseudokirchneriella subcapitata ) ”, received via email from B. Gulledge [American Chemistry Council] to J. Schaeffer [EPA] ) supplied data fo r the toxicity to algae test. These newly submitted data are eval uated for adequacy herein.

This report also contains data for other ecological and toxicological endpoints that were evaluated previously by EPA and, therefore, provides an update of the data adequacy assessment previously compiled for CASRN 119

- 61

- 9 in the following report: “HPV2 Test Rule Data Adequacy Review: Methanone, diphenyl

- , CASRN 119

- 61

- 9. SRC, Inc., Technical Report #12

- 031. March 16, 2012.” Sources of information for the previous review included submissions from the Benzophenone HPV Consortium of the American Chemistry Council (dated January 17, 2012; available online at http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

- OPPT

2007

- 0531

- 0654 a s of July 1, 2013 ) and Cytec Industries (dated March 8, 2011; available online
at http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

- OPPT

- 2007

- 0531

- 0602 as of Jul y 1, 2013 ), EPA’s High Production Volume Information System (HPVIS)

, the European Chemicals Agency (ECHA) website ( http://echa.europa.eu/ ), publically available literature from scientific journals, and NTP study reports

.
B. EVALUATING DATA ADEQUACY

Data available for SIDS toxicity endpoints are evaluated for adequacy in Section D. 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

1 H ansch. C., Leo , A., and Hoekman , D

. 1995. Exploring QSAR. Hydrophobic, El ectronic, and Steric Constants. ACS Professional Reference Book. Washington , DC: American Chemical Society , as cited in U.S. EPA. (2012) Estimation Programs Interface SuiteTM (EPI Suite) for Microsoft® Windows, v4.11. U.S. Environmental Protection Ag ency, Washington, DC, USA. Available online at http://www.epa.gov/opptintr/exposure/pubs/episuitedl.htm as of June 28, 2013.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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2 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 provided near the top of the first page of the summaries for that endpoint. If the data available for the endpoint we re determined to be inadequate, a basis for that conclusion is supplied. A table summarizing data availability/adequacy for each SIDS endpoint is included on the last page of this document (Section F ).

C. SUMMARY OF DATA ADEQUACY FOR TESTING REQUIRED BY TEST RULE

Data supplied in the new submission identified above from the Benzophenone HPV Consortium of the American Chemistry Council (dated April 1 0 , 2013) are adequate to satisfy the toxicity to aquatic plants (algae) ( component of C2 ) test ing requirement promulgate d in the final HPV2 test rule.

D ata from the ECHA website were previously reviewed (in SRC Technical Report #12

- 031) and found to be adequate to satisfy the acute toxicity to Daphnia (component of C2) testing requirement; these da ta are included in Section D below

. There are no outstanding test rule requirements for toxicity endpoints.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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3 D . DATA FOR SIDS ENDPOINTS

D

- 1. Acute Toxicity to Fish

Summary of endpoint: Adequate

Study 1: Acute toxicity study in fathead minnow (Marchini et al., 1992)

Title: Lethal and sublethal toxicity of benzene derivatives to the fathead minnow, using a short

term test

This study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

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- 9

Remarks: > 99% pure

METHOD

Method/guideline followed: Not available

Test type: Short

- term toxicity to fish (flow

- through)

GLP compliant?: Not specified

Year study was performed: 1992

Test species: Fathead minnow ( Pimephales promelas )

Analytical monitoring: Yes

Exposure duration: 4

- 7 days

Concentrations tested: Not available

Test method/conditions remarks: The dilution water utilized for this test was a filtered, aerated, and heated Lake Superior water (freshwater)

. This 7

- day test followed current EPA short

- term chronic methods. The test lasted 7 days with su rvival and growth as the end points. There was a control and five other test concentrations. The mini

- diluter distributed the test concentrations and the control. Six tanks represented the test concentrations ; they were partitioned to allow for two replicates. Forty fathead minnow larvae (< 24 hours old) were exposed to each test concentration. Fish were fed live < 24 h

- old brine shrimp nauplii ( Artemia salina ), three times a day. However, on day 7 they were only fed once. All of the toxic endpoint values are based on measured concentra tions.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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4 The toxicant concentration in the stock solution and in each exposure tank was analyzed before starting the tests and two or three times during the test.

Alkalinity was 39 mg CaCO 3 /L, water hardness was 44.5 to 46.5 mg CaCO 3 /L, dissolved oxygen was 7.2 to 7.4 mg/L, pH was 7.15 to 8.25, and temperature was 24 to 26 °C. The photoperiod was 16 hours light and 8 hours dark.

The trimmed Spearman

- Karber method was used to calculate the LC 50 at 96 hours.

RESULTS

Endpoint value(s): Based on measured concentrations: 96

- hour LC 50 = 10.89 mg/L; 7

- day LOEC (survival) = 9.24 mg/L; 7

- day NOEC (survival) = 5.86 mg/L

Results remarks: The confidence interval for the 96

- hour LC 50 was 9.64 to 12.31 mg/L.

CONCLUSIONS

The 96

- hour LC 50 for fathead minnow was 1 0.89 mg/L.

STUDY RELIABILITY

[1] Reliable without restrictions. Data appeared in a peer

- reviewed journal article. The test method is well described, and the test solutions were measured.

REFERENCE

Marchini, S., Tosato, M.L., Norberg

- King, T.J., Hammermeister, D.E., and Hoglund, M.D. 1992. Lethal and sublethal toxicity of benzene derivatives to the fathead minnow, using a short

term test. Environmental Toxicology and Chemistry 11: 187

- 195. As summarized in HPVIS (summary available at http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

- OPPT

- 2007

- 0531

- 0057 as of July 2, 2013)

.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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5 D

- 2. Acute Toxicity to Aquatic Invertebrates

Sum mary of endpoint: A dequate

NOTE: Acute toxicity to Daphnia data supplied by the Benzophenone HPV Consortium ( Studies 2

- 4 , below) were inadequate to fulfill this endpoint. However, EPA located an Acute Toxicity to Daphnia study on the ECHA website ( Study 1 , below) that is adequate to fulfill the test requirement; the Benzophenone HPV Consortium does not need to conduct an additional study for the Acute Toxicity to Daphnia endpoint.

Study 1 : Acute toxicity study in Daphnia magna (From ECHA website, Exp Key Short

- term toxicity to aquatic invertebrates.001)

Title: Not provided

This study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

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- 9

Remarks: Purity not provided

METHOD

Method/guideline followed: OECD TG 202

Test type: Daphnia sp., Acute Immobilization Test ( Semi

- s tatic)

GLP compliant?: Yes

Year study was performed: 2011

Test species: Water flea ( Daphnia magna ), 4

- 24 hours old, bred at test facility

Analytical monitoring: Yes

Exposure duration: 48 hours

Concentrations tested: Nominal concentrations: 2.1, 4.7, 10.3, 22.7, and 50.1 mg/L ; Me a sured concentrations: 2.1, 4.47, 8.78,17.9, and 37.4 mg/L.

Test method/conditions remarks:

“ The test item was distributed to the replicate beakers using an aqueous stock solution. The stock solution (50.1 mg test item/L) was prepared in dilution water and stirred for 24 h. TheFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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6 individual test solutions were prepared by dilution of the stock sol ution with dilution water and distributed to the test beakers. ” [ECHA ]

“

Four replicates per treatment with 5 specimens per replicate were conducted. ” [ECHA]

“ The total hardness of the dilution water was 1.3 mmol/L ( equivalent to 107 mg CaCO 3 /L )...” [ECH A ]

“ The water temperature ranged between 20.3°C and 20.7°C. ” [ECHA]

“ The pH values throughout the test were within a range of pH 7.6

– 8.6 at all treatment levels. ” [ECHA ]

“ The oxygen saturation was between 7.8 mg/L and 8.7 mg/L. ” [ECHA ]

“

Conductivity was 314.0 μS/cm

. ” [ECHA]

“ The concentration of the test item in the aqueous phase of the test was assessed by chemical analysis. Test samples were analysed from fresh media at test start and at 24h. Samples of aged test media were analy z ed after 24h and at the end of the test. All test concentrations and the control were analy z ed. ” [ECHA ]

“... Due to recovery rates outside the range of 80

– 120 % of nominal for the highest treatment

, mean measured concentrations were applied for effect assessment. ” [ECHA ]

K 2 Cr 2 O 7 was used as a reference substance (positive control).

RESULTS

Endpoint value(s): 48

- hour EC 50 = 6.784 mg/L ( based on mean measured concentrations) ; 48

- hour NOEC = 4.47 mg/L

Results remarks:

“ The means of initial and aged measured concentrations were 2.1, 4.47, 8.78, 17.9, and 37.4 mg/L [ at nominal concentrations of 2.1, 4.7, 10.3, 22.7, and 50.1 mg/L , respectively ]

, corresponding with 100, 95, 85, 79, and 75 % of the nominal loading. ” [ECHA ]

Confidence limits of the 48

- hour EC 50 were 5.514

- 8.327 mg/L. Percent incidence of i mmobilization after 24 hours was 0, 0, 5, 45, 100, and 100% at 0, 2.1, 4.47, 8.78, 17.9, and 37.4 mg/L, respectively. Percent incidence of i mmobilization after 48 hours was 0, 5, 10, 70, 100, and 100% at 0, 2.1, 4.47, 8.78, 17.9, and 37.4 mg/L, respectively.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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7 CONCLUSIONS

The 48

- hour EC 50 for Daphnia magna was 6.784 mg/L.

STUDY RELIABILITY

[2] Reliable with restrictions. The summary does not provide a full citation f or the study report. Data, including immobilization incidence and water quality values, for each replicate are not provided in the summary.

REFERENCE

Study available from ECHA website, http://echa.europa.eu

, fourth row of the “Search results” table resulting from a “Chemical Substance Search” for CASRN 119

- 61

- 9. Study indicated as Exp Key Short

- term toxicity to aquatic invertebrates.001 . The data source is referred to as a “study report”, with a report date of 2011

- 11

- 10. No other reference information is provided.

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).

Text in quotes is taken directly from ECHA ( no date provided ) study summary (available online
at http://apps.echa.europa.eu/registered/ data/dossiers/DISS

- b6b4ef07

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- 7379

- e044

- 00144f67d031_DISS

- b6b4ef07

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- e044

- 00144f67d031.html as of June 28, 2013 )

.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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8 Study 2 : Acute toxicity study in Daphnia magna ( Tosato et al., 1991 )

Title: A new strategy for ranking chemical hazards. Framework and a pplication.

This study is: Adequate Not Adequate

This study is not adequate because t he duration of exposure is 24 hours (per OECD TG 202 [1984] ), which is shorter than the 48

- hour duration specified by the test rule guideline, ASTM E 729

- 96. In addition, study details are limited; missing information i ncludes concentrations tested, measurements of dissolved oxygen and pH during the test, and immobilization counts at each test concentration.

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9 (referred to as benzophenone in study description )

Remarks : 99% pur e

METHOD

Method/guideline followed: OECD TG 202 (1984)

Test type: Daphnia sp., Acute Immobilization Test and Reproduction Test: Part I

– 24h EC 50 Acute Immobilization Test (Static)

GLP compliant?: Not specified

Year study was performed: 1991

Test species: Water flea ( Daphnia magna )

, < 24 hours old

Analytical monitoring: Yes

Exposure duration: 24 hours

Concentrations tested: Not specified

Test method/conditions remarks: Dil ution water was prepared by mixing Milli

- Q and tap water in approximately a 1:1 ratio, adjusted to attain a hardness of 130

- 150 mg CaCO 3 /L , with a pH of 8.2 ± 0.5 and conductivity of 400 ± 25 μS/cm . Stock solutions were prepared by adding the chemicals to the dilution water with mechanical stirring.

“ Test solutions were obtained by diluting the stock solutions. The concentrations of the test solutions were determined at the beginning and end of the experiment. Quantitative analyses were carried out on Varian 5000 HPLC equipment under the following conditions: C

- 18 reverse

-Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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- 9

9 phase column (Micro Pack MCH

- 5, length 15 cm); eluting mixture methanol

- water (3:1); flow rate 1 mL/min; volume injected, as adequate (mainly 10 μL). Chromatographic peaks were recorded on a Varian 4270 integrator and quantified by the external standard method. ” [Tosato et al. 1991, p. 700]

Daphnids ( Daphnia magna ) < 24 hours old were obtained from an in

- house laboratory cul ture maintained in the same water as the dilution water and fed yeast and Selenastrum capricornutum

.

“ Ten animals, subdivided in two replicates, were used for each of the concentrations tested (a minimum of five) and the control. ” [Tosato et al. 1991, p. 700]

“ The tests were performed in closed bottles (150 mL) filled to the top in order to avoid loss of chemical due to volatilization. The temperature was 20 ± 1 °C and the photoperiod was 16 h light

, 8 h dark. ” [Tosato et al. 1991, p. 700]

“ The 24

- h our EC 50 was estimated by the Litchfield and Wilcoxon method. ” [Tosato et al. 1991, p. 700]

RESULTS

Endpoint value(s): 24

- hour EC 50 = 0.28 mg/L (use of nominal or measured concentrations not specified)

Results remarks: Confidence limits of the 24

- hour EC 50 were 0.21

- 0.37 mg/L.

CONCLUSIONS

The 24

- hour EC 50 for Daphnia magna was 0.28 mg/L

.

STUDY RELIABILITY

[ 3 ] Not r eliable. The duration of exposure is 24 hours (per OECD TG 202 [1984] ) , which is shorter than the 48

- hour duration specified by the test rule guideline, ASTM E 729

- 96. In addition, study details are limited; missing information includes concentrations tested, measurements of dissolved oxygen and pH during the test , and immobilization counts at each test concentration

.

REFERENCE

Tosato, M.L., Vigano, L., Skagerberg, B., and Clementi, S. 1991. A new strategy for ranking chemical hazards. Framework and a pplication. Environmental Science & Technology 25: 695

- 702. ( In a submission dated March 8, 2011, Cytec Industries [a member of the Benzophenone HPV Consortium] presented the results of this study. In a submission dated January 17, 2012,
t he Benzophenone HPV Consortium provided a link to a summary of this study at http://apps.echa.europa.eu/registered/data/ dossiers/DISS

- 9c7935fb

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- e044

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- 04cb11d3

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- 84b1

- 084d82237941_DISS

- 9c7935fb

- 2f34

- 5de5

-Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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10 e044

- 00144f67d249.html#GEN_RESULTS_HD . Additional details were obtained from the primary source [Tosato et al., 1991].)

Text in quotes is taken directly from Tosato et al. (1991).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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11 Study 3 : Acute toxicity study in Daphnia magna ( Marchini et al., 1993 )

Title: Comparison of the susceptibility of daphnids and fish to benzene derivatives

This study is: Adequate Not Adequate

This study is not adequate because t he duration of exposure is 24 hours, which is shorter than the 48

- hour duration specified by the test rule guideline, ASTM E 729

- 96. In addition, study details are limited; missing information includes concentrations tested, measurements of dissolved oxygen, pH, and temperature during the test, and mortality at each test co ncentration.

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9 (referred to as benzophenone in study description)

Remarks: > 99% pure

METHOD

Method/guideline followed: Not specified

Test type: Acute static daphnid test

GLP compliant?: Not specified

Year study was performed: 199 3

Test species: Water flea ( Ceriodaphnia dubia ); < 24 hours old ; supplier: EPA Environmental Resea r ch Laboratory

– Duluth

Analytical monitoring: Yes

Exposure duration: 24 hours

Concentrations tested: Not specified

Test method/conditions remarks: Dilution water consisted of sand

- filtered, aerated water from Lake Superior. Water was characterized as follows: temperature = 25 ± 1 °C, pH = 7.6 ± 0.6, conductivity = 111 ± 9 μΩ - 1 , dissolved oxygen = 7.3 ± 1 mg/L, alkalinity = 41 ± 5 mg CaCO 3 /L, and hardness = 45.5 ± 1 mg CaCO 3 /L.

Daphnids were exposed in a static system to five concentrations of the test substance, with a dilution factor of 0.65 and two replicates per concentration . Five daphnids per replicate were exposed in 25 mL of test solution in a tightly closed chamber to avoid chemical volatilization during exposure. Measurements at the beginning and end of the test were conducted in on eFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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12 vessel at each concentration and included: toxicant concentration, temperature, pH, and dissolved
oxygen.

Water samples for analytical measurements of test substance concentrations were collected into reagent bottles containing a known volume of hexane, followed by stirring for 45 minutes to facilitate extraction and subsequent concentration. Chemical analysis was conducted using high

performance liquid chromatography using 70

- 80% acetonitrile in water as mobile phase, a C 18 reverse phase column, and a UV

- visible variable wavelength detector. Exposure concentrations were determined using linear regression analysis of the peak area responses versus four external standards.

The trimmed Spearman

- Karber method was used to estim ate LC 50 values and confidence limits.

RESULTS

Endpoint value(s): 24

- hour L C 50 = 7.6 mg/L (use of nominal or measured concentrations not specified)

Results remarks: Based on the comparison of 24

- hour EC 50 /LC 50 values, Daphnia magna were significantly more sensitive (by a factor of 1.4 log units) than Ceriodaphnia to b enzophenone (see Tosato et al., 1991 for EC 50 in Daphnia magna )

.

CONCLUSIONS

The 24

- hour L C 50 for Ceriodaphnia dubia was 7.6 mg/L.

STUDY RELIABILITY

[ 3 ] Not r eliable. The duration of exposure is 24 hours, which is shorter than the 48

- hour duration specified by the test rule guideline, ASTM E 729

- 96. In addition, study details are limited; missing information includes concentrations tested, measurements of dis solved oxygen

, pH , and temperature during the test, and mortality at each test concentration.

REFERENCE

Marchini, S., Hoglund, M.D., Broderius, S.J., and Tosato, M.L. 1993. Comparison of the susceptibility of daphnids and fish to benzene derivatives. The Science of the Total Environment, Supplement 1993, pp. 799

- 808. (In a submission dated March 8, 2011

, Cytec Industries [a member of the Benzophenone HPV Consortium ] presented the results of this study. Additional details were obtained from the primary source [ Marchini et al., 199 3 ].)

Supporting Information: Tosato, M.L., Vigano, L., Skagerberg, B., and Clementi, S. 1991. A new strategy for ranking chemical hazards. Framework and a pplication. Environmental Science & Technology 25: 695

- 702.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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13 Study 4 : Acute toxicity study in Daphnia magna ( Ministry of the Environment, Japan, 1998 )

Title: Not available

This study is: Adequate Not Adequate

This study is not adequate because study details were not provided. Missing details included water qualit y of test solutions (e.g., pH, temperature, water hardness, and dissolved oxygen), test substance purity, details regarding analytical monitoring, experimental conditions (e.g., number of replicates, number of daphnids per replicate, chamber volume

, whethe r chambers were sealed to prevent evaporation of test substance, and use of static, static renewal, or flow

- through conditions), and results for individual replicates. According to Japan’s Ministry of the Environment, “because most of these data have not been evaluated by experts, confirmation of test results is needed if these data are used for assessment” and “tests conducted before FY 2002 need confirmation of test results, because some of these tests were conducted using a dispersant”. Therefore, eval uation of the full study report would be necessary before a reliability rating other than 4 (not assignable) could be designated

.

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9 (referred to as benzophenone in study description)

Remarks: Purity not provided

METHOD

Method/guideline followed: OECD TG 202

Test type: Daphnia sp., Acute Immobilisation Test

GLP compliant?: Yes

Year study was performed: 199 8

Test species: Water flea ( Daphnia magna )

Analytical monitoring: Not specified

Exposure duration: 48 hours

Concentrations tested: Not specified

Test method/conditions remarks: None specifiedFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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14 RESULTS

Endpoint value(s): 48

- hour E C 50 > 10 mg/L (use of nominal or measured concentrations not specified)

Results remarks: None specified

CONCLUSIONS

The 48

- hour L C 50 for daphnids was > 10 mg/L.

STUDY RELIABILITY

[4] Not assignable. Study details were not provided in the summaries. Missing details included water quality of test solutions (e.g., pH, temperature, water hardness, and dissolved oxygen), test substance purity, details regarding analytical monitoring, experimental conditions (e.g., number of replicates, number of daphnids per replicate, chamber volume, whether chambers were sealed to prevent evaporati on of test substance , and use of static, static renewal, or flow

- through conditions ), and results for individual replicates. According to Japan’s Ministry of the Environment, “because most of these data have not been evaluated by experts, confirmation of test results is needed if these data are used for assessment” and “tests conducted before FY 2002 need confirmation of test results, because some of these tests were conducted using a dispersant”. Therefore, evaluation of the full study report would be necessary before a reliability rating other than 4 (not assignable) could be designat ed

.

REFERENCE

In a submission dated M arch 8, 2011, Cytec Industries ( a member of the Benzophenone HPV Consortium ) presented the results of this study. In a submission dated January 17, 2012, t he Benzophenone HPV Consortium provided a link to a summary of this study at the website for Japan’s Ministry of the Environment ( http://www.env.go.jp/chemi/sesaku/02e.pdf ) ; summaries of this study are also available at websites for Japan’s National Institute of Technology and Evaluation ( http://www.safe.nite.go.jp/english/sougou/view/ComprehensiveInfoDispl ay _en.faces ) and the European Chemicals Agency ( http://echa.europa.eu/ ; Exp WoE Short

- term toxicity to aquatic invertebrates.00 3

, fourth row of the “Search results” table resulting from a “Chemical Substanc e Search” for CASRN 119

- 61

- 9 ; http://apps.echa.europa.eu/re gistered/ data/dossiers/DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031/DISS

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- 00144f67d031_DISS

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. None of the summaries contain sufficient data to evaluate study adequacy.

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 pr ovided written notification is given to the ECHA Communication Unit (info (at) echa.europa.eu).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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15 D

- 3

. Toxicity to Aquatic Plants

Summary of endpoint: Adequate

Study 1: Growth inhibition toxicity test to Pseudokirchneriella subcapitata (Jacobs et al., 2013)

Title: Benzophenone: A 96

- hour toxicity test with the freshwater alg a ( Pseudokirchneriella subcapitata )

This study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9 (referred to as benzophenone in study description )

Remarks: Purity: 99.6%; white powder ; supplied by Sigma Aldrich; Lot No. MKBG2920V; stored under room temperature (ambient) and dark conditions

METHOD

Method/guideline followed: OECD TG 201 (2006; “Freshwater Alga and Cyanobacteria, Growth Inhibition Test”); EC Directive 92/69/EEC, Method C.3 ( “Algal Inhibition Test” ) and OPPTS 850.4500 (“Algal Toxicity, Tiers I and II”)

Test type: Toxicity Test to Freshwater Algae (Static Cond itions)

GLP compliant?: Yes (definitive test; range

- finding test was non

- GLP compliant) ; the following exceptions were noted: “Periodic analyses of well water for potential contaminants were not performed according to Good Laboratory Practice Standards, but were performed using a certified laboratory and standard US EPA analytical methods. The characterization and stability of the test substance under conditions of storage at the test site were not determined in accordance with Good Laboratory Practice standards.” [Jacobs et al. 2013, p. 3]

Year study was performed: 2012

Test species: Green algae ( Pseudokirchneriella subcapitata ); obtained from the University of Toronto (maintained at Wildlife International in Easton, MD since March 2000).

Analyti cal monitoring: Yes, only in the definitive test (at 0 and 96 hours)

Exposure duration: 96 hoursFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

16 Concentrations tested: Nominal concentrations: 0, 0.26, 0.64, 1.6, 4.0, and 10 mg/L; Mean measured concentrations : < LOQ 1 , 0.23, 0.55, 1.4, 3.8, and 9.1 mg/L

Test method/conditions remarks: In the range

- finding test, Pseudokirchneriella subcapitata cells were exposed to a series of nominal test concentrations of 0.10, 1.0, and 10 mg/L. Negative control cells were not exposed to the test item.

Based o n the results of the range

- finding test, nominal test concentrations selected for the 96

- hour definitive test were 0.26, 0.64, 1.6, 4.0, and 10 mg/L (mean measured concentrations: 0.23, 0.55, 1.4, 3.8, and 9.1 mg/L).

“Test chambers were sterile, 250

- mL g lass Erlenmeyer flasks plugged with sterile foam stoppers, and contained 100 mL of test or control medium. The test chambers were... indiscriminately positioned daily in an environmental chamber designed to maintain the desired test temperature throughout t he test. The test flasks were continuously shaken on a mechanical shaker at 100 rpm throughout the duration of the test.” [Jacobs et al. 2013, p. 13]

“Test chambers were held in an environmental chamber at a temperature of 24 ± 2oC. 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

ma ximum thermometer. The algae were held under continuous cool

- white fluorescent lighting throughout the test. The target light intensity was 6,000 lux ± 10%. Light intensity was measured at test solution level at five locations surrounding the test flask s 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 exposure termination using a Thermo Orion Model 4Star plus pH meter. 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 solution collected from each of the replicates of each treatment and control group.” [J acobs et al. 2013, p. 13]

“The algal cells were cultured and tested in freshwater AAP medium [ according to ASTM E1218

- 04 ]... [Stock nutrient solutions and test medium were prepared with purified well water (NANOpure ® water) ]... The pH of the medium was adjusted to 7.5 ± 0.1 using 10% hydrochloric acid and [the medium] was sterilized by filtration (0.22 μm) prior to use.” [Jacobs et al. 2013, p. 13]

“Algal cells used in this test... had been actively growing in culture medium unde r similar environmental conditions as used in this test for at least two weeks prior to test initiation. Algal cells for this study were taken from a culture that had been transferred to fresh medium three days prior to test initiation.” [Jacobs et al. 2 013, p. 12]

1 LOQ = limit of quantification; d efined as 0.100 mg/L for CASRN 119

- 61

- 9 in the study report.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

17 “Prior to test initiation, the algal cells in the stock culture... were verified in freshwater algal medium at a concentration of approximately 1.36 x 10 6 cells/mL... using a hemacytometer and microscope

... ” [Jacobs et al. 2013, p. 15]

“A primary stock solution was prepared by dissolving 0.0100 g of benzophenone in 1000 mL of freshwater AAP medium to achieve a nominal concentration of 10 mg/L . The primary stock [solution] was mixed by inversion and was sonicated for ~forty three minutes, as well as stirred with a Teflon ®

- coated stirbar and magnetic stirplate for ~fourteen minutes prior to use. The 10 mg/L primary stock [solution] appeared clear and colorless and was otherwise unremarkable. The 10 mg/L stock was proportionately diluted with freshwater AAP medium to prepare [the] test solutions

... All test solutions were mixed by inversion, and appeared clear and colorless and were otherwise unremarkable after mixing. The negative control solution consisted of fre shwater AAP medium.” [Jacobs et al. 2013, p. 14]

“Four replicate test chambers were maintained in each treatment group and six replicates were maintained in the control group. At test initiation, an inoculum of the algal cells was added to each test cha mber to achieve a nominal concentration of approximately 10,000 cells/mL.” [Jacobs et al. 2013, p. 11]

“ Test medium samples were collected from each replicate of the treatment and control groups for the determination of algal cell densities. Samples wer e collected at approximately 24

- hour intervals during the 96

- hour exposure and were held for a maximum of four days under refrigerated conditions sufficient to inhibit growth until cell counts could be performed. Cell counts were performed using an electr onic particle counter (Coulter Electronics, Inc.)... A primary counting standard containing Pseudokirchneriella subcapitata cells was prepared at a target nominal concentration of 100,000 cells/mL.” [Jacobs et al. 2013, p. 16]

“Samples of the test solutio ns were collected at approximately 0 and 96 hours to measure concentrations of the test substance [in the definitive test]. Samples at test initiation were collected from the individual batches of test solution prepared for each treatment and control
grou p prior to distribution into the test chambers. At exposure termination, samples were collected from the pooled replicates from each treatment and control group. Samples collected at test initiation and test termination were collected into glass vessels and were processed immediately for analysis.” [Jacobs et al. 2013, p. 14]

“Concentrations of benzophenone in the samples were determined by HPLC [with UV detection at 220 nm] using an Agilent Series 1260 High Performance Liquid Chromatograph (HPLC) equipped with an Agilent Series 1260 Variable Wavelength Detector. Chromatograph ic separations were achieved using a YMC

- PACK ODS AM column (150 mm x 4.6 mm, 3

- μm particle size).” [Jacobs et al. 2013, p. 14]

“The method Limit of Quantitation (LOQ) was defined as 0.100 mg/L

... ” [Jacobs et al. 2013, p. 15]Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

18 Statistical Analysis: “The c alculation of cell densities, area under the growth curve [biomass], yield, growth rates and percent inhibition values, as well as all statistical analyses, were conducted using ‘The SAS System for Windows, Version 8.2’.” [Jacobs et al. 2013, p. 17]

“ EC 50 values, (i.e., the theoretical test concentrations that would produce a 50% reduction in area under the growth curve (E b C 50 ), growth rate (E r C 50 ) and yield (E y C 50 ), in comparison to the negative control) and the corresponding 95% confidence limits for t he 72 and 96 hour intervals were calculated by non

- linear regression with replicate data (area under the growth curve, growth rate and yield, respectively) and mean measured test concentrations.” [Jacobs et al. 2013, p. 18]

“The 72 and 96

- hour area under the growth curve, 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 assumptions of normal distribution and homogeneity of variance. The
trea tment 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 para meter at 72 and 96 hours.” [Jacobs et al. 2013, p. 18]

Validation Criteria: The results of the test were considered valid if the following criteria were met:

• “The mean cell density in the control replicates increased by a factor greater than 16 within th ree days and at least 100 within 4 days

. ” [Jacobs et al

. 2013, p. 20]

• “The coefficient of variation of average specific growth rate in the negative control replicates during the first 72 hours did not exceed 7%

. ” [Jacobs et al. 2013, p. 20]

• “The mean percent coefficient of variation for section

- by

- section specific growth rates (days 0

- 1, 1

- 2 and 2

- 3) in the negative control did not exceed 35%

. ” [Jacobs et al. 2013, p. 20]

RESULTS

Endpoint value(s): The following results were based on mean measured c oncentrations:

Growth Rate: 72

- hour E r C 50 = 3.0 mg/L (95% confidence limits: 2.3

- 4.0 mg/L) 72

- hour NOEC = 0.23 mg/L; 72

- hour LOEC = 0.55 mg/L 96

- hour E r C 50 = 2.9 mg/L (95% confidence limits: 2.2

- 3.9 mg/L) 96

- hour NOEC = 0.23 mg/L; 96

- hour LOEC = 0.55 m g/L

Yield: 72

- hour E y C 50 = 1.7 mg/L (95% confidence limits: 1.2

- 2.4 mg/L) 72

- hour NOEC = 0.23 mg/L; 72

- hour LOEC = 0.55 mg/L 96

- hour E y C 50 = 1.8 mg/L (95% confidence limits: 0.96

- 3.4 mg/L) 96

- hour NOEC = 0.23 mg/L; 96

- hour LOEC = 0.55 mg/LFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

19 Biomass: 72

- hour E b C 50 = 1.7 mg/L (95% confidence limits: 1.4

- 2.2 mg/L) 72

- hour NOEC = 0.23 mg/L; 72

- hour LOEC = 0.55 mg/L 96

- hour E b C 50 = 1.8 mg/L (95% confidence limits: 1.3

- 2.4 mg/L) 96

- hour NOEC = 0.23 mg/L; 96

- hour LOEC = 0.55 mg/L

Results remarks:

Range

- F inding Test Results: Mean percent inhibition of cell density was

- 8, 26, and 99% at nominal concentrations of 0.10, 1.0, and 10 mg/L, respectively. All test solutions were clear and colorless, with no precipitation noted.

Definitive Test Results:

“

Measured concentrations of benzophenone ranged from 96 to 105% of nominal in samples collected on Day 0 and from 74 to 83% of nominal at test termination at 96 hours... Test solution[s] appeared clear and colorless after mixing, and no surface films or part iculate matter were observed in the test chambers at study termination

... The results of the study were based on mean, measured concentrations. ” [Jacobs et al. 2013, p. 18]

“After 72 hours of exposure, inhibition of area under the growth curve [biomass] in the 0.23, 0.55, 1.4, 3.8 and 9.1 mg/L treatment groups was 5, 12, 28, 97 and 98%, respectively, relative to the negative control. Inhibition of growth rate in the 0.23, 0.55, 1.4, 3.8 and 9.1 mg/L treatment groups was 1, 2, 6, 74 and 78%, respectively, relative to the negative control. Inhibition of yield in the 0.23, 0.55, 1.4, 3.8 and 9.1 mg/L treatment groups was 6, 11, 29, 99 and 99%, respectively, relative to the negative control. The 72

- hour EC 50 values and 95% confidence intervals for area under the growth curve [biomass], growth rate and yield were 1.7 (1.4 to 2.2) mg/L, 3.0 (2.3 to 4.0) mg/L and 1.7 (1.2 to 2.4) mg/L, respectively. Dunnett’s test indicated that there were significant reduction s ( p ≤ 0.05) in mean area under the growth curve [biomass], growth rate and yield in the 0.55, 1.4, 3.8 and 9.1 mg/L treatment groups [for the 72

- hour exposure]. Therefore, the 72

- hour NOEC for area under the growth curve [biomass], growth rate and yield was 0.23 mg/L.” [Jacobs et al. 2013, p. 19]

“After 96 hours of exposure, inhibition of area under the growth curve [biomass] in the 0.23, 0.55, 1.4, 3.8 and 9.1 mg/L treatment groups was 5, 11, 26, 98 and 99%, respectively, relative to the negative contr ol. Inhibition of growth rate in the 0.23, 0.55, 1.4, 3.8 and 9.1 mg/L treatment groups was 1, 2, 4, 76, and 80%, respectively, relative to the negative control. Inhibition of yield in the 0.23, 0.55, 1.4, 3.8 and 9.1 mg/L treatment groups was 5, 11, 23, 99 and 100%, respectively, relative to the negative control. The 96

- hour EC 50 values and 95% confidence intervals for area under the growth curve [biomass] , growth rate and yield were 1.8 (1.3 to 2.4) mg/L, 2.9 (2.2 to 3.9) mg/L and 1.8 (0.96 to 3.4) mg/ L, respectively. Dunnett’s test indicated that there were significant reductions ( p ≤ 0.05) in mean area under the growth curve [biomass]

, growth rate and yield in the 0.55, 1.4, 3.8 and 9.1 mg/L treatment groups [for the 96

- hour exposure]

. Therefore, th e 96

- hour NOEC for area under the growth curve [biomass] , growth rate and yield was 0.23 mg/L. ” [Jacobs et al. 2013, p. 19

- 20]Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

20 “At test initiation algal cells appeared normal. After 96

- hours of exposure there were no noticeable changes in cell morphology in any of the tested concentrations when compared to the control and there were no noticeable aggregations and flocculations of cells, or adherence to the test chambers, in any treatment or the control group. The test solution appeared normal with no surface slicks or particulate matter present at the end of exposure.” [Jacobs et al. 2013, p. 20]

The test was considered valid because:

• The cell density increased by a factor of 336 in three days and a factor of 557 in 4 days.

• The coefficient of variation for average specific growth rate for controls after 72 hours was 1.1%.

• The mean coefficient of variation for section

- by

- section growth rates for controls over 0

- 72 hours was 24 %.

“Temperatures remained within the 24 ± 2°C range established for the test [range: 23.3 to 23.9°C during 0

- 96 hours]. The pH of the test solutions ranged from 7.3 to 7.4 at test initiation and ranged from 8.0 to 9.8 in the test solutions at test termin ation. The observed increase in pH is typical for tests conducted with P. subcapitata and is attributed to the photosynthetic activity of the algae. The light intensity [at initiation] ranged from 5,590 to 6,520 lux, which was within the desired range of 6000 lux ± 10%.” [Jacobs et al. 2013, p. 18]

CONCLUSIONS

The 72

- and 96

- hour NOEC s were both determined to be 0.23 mg/L. The 72

- hour EC 50 values for growth rate, yield, and biomass were 3.0 mg/L, 1.7 mg/L, and 1.7 mg/L, respectively. The 96

- hour EC 50 values for growth rate, yield, and biomass were 2.9 mg/L, 1.8 mg/L, and 1.8 mg/L, respectively.

STUDY RELIABILITY

[1] Reliable without restrictions.

REFERENCE

Jacobs, A.M., Kendall, T.Z., and Porch

, J.R.

. 2013. Benzophenone : A 96

- hour toxicity te st with the freshwater alga ( Pseudokirchneriella subcapitata ). Performing Laboratory: Wildlife International, Easton, MD. Project No. 264P

- 101. Sponsor: American Chemistry Council, Washington, DC.

Text in quotes is taken directly from Jacobs et al. (2013).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

21 Study 2 : Toxicity to Pseudokirchneriella subcapitata ( Ministry of the Environment, Japan, 1998 )

Title: Not available

This study is: Adequate Not Adequate

This study is not adequate because s tudy details were not provided. Missing details included water quality of test solutions (e.g., pH, temperature, water hardness, and dissolved oxygen), test substance purity, details regarding analytical monitoring, experimental conditions (e.g., light intensity, number o f replicates, chamber volume, and whether chambers were sealed to prevent evaporation of test substance ), and results for individual replicates. According to Japan’s Ministry of the Environment, “because most of these data have not been evaluated by exper ts, confirmation of test results is needed if these data are used for assessment” and “tests conducted before FY 2002 need confirmation of test results, because some of these tests were conducted using a dispersant”. Therefore, evaluation of the full stud y report would be necessary before a reliability rating other than 4 (not assignable) could be designated

.

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9 (referred to as benzophenone in study description)

Remarks: Purity not provid ed

METHOD

Method/guideline followed: OECD TG 201

Test type: Freshwater Alga and Cyanobacteria, Growth Inhibition Test

GLP compliant?: Yes

Year study was performed: 1998

Test species: Freshwater green algae ( Pseudokirchneriella subcapitata )

Analytical monitoring: No t specified

Exposure duration: 72 hours

Concentrations tested: Not specified

Test method/conditions remarks: No ne specifiedFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

22 RESULTS

Endpoint value(s): 72

- hour EC 50 (growth rate) = 3.5 mg/L; 72

- hour NOEC (growth rate) = 1 mg/L; 72

- hour EC 50 (biomass) = 1.8 mg/L; 72

- hour NOEC (biomass) = 0.46 mg/L (use of nominal or measured concentrations not specified)

Results remarks: None specified

CONCLUSIONS

The 72

- hour EC 50 s for growth rate and biomass were 3.5 and 1.8 mg/L, respectively. The 72

- hour NOEC s for growth rate and biomass were 1 and 0.46 mg/L , respectively

.

STUDY RELIABILITY

[ 4] Not assignable

. Study details were not provided in the summaries

. Missing details included water quality of test solutions (e.g., pH, temperature, water hardness, and dissolved oxygen), test substance purity, details regarding analytical monitoring, experimental conditions (e.g., light intensity, number of replicates, chamber volume, and whether chambers were sealed to preven t evaporation of test substance ), and results for individual replicates. According to Japan’s Ministry of the Environment , “because most of these data have not been evaluated by experts, confirmation of test results is needed if these data are used for as sessment” and “tests conducted before FY 2002 need confirmation of test results, because some of these tests were conducted using a dispersant”. Therefore, evaluation of the full study report would be necessary before a reliability rating other than 4 (no t assignable) could be designated

.

REFERENCE

In a submission dated M arch 8, 2011, Cytec Industries ( a member of the Benzophenone HPV Consortium ) presented the results of this study. In a submission dated January 17, 2012, the Benzoph enone HPV Consortium provided link s to summar ies of this study at the website s for Japan’s Ministry of the Environment ( h ttp://www.env.go.jp/chemi/sesaku/02e.pdf ) and the European Chemicals Agency ( http://apps.echa. europa.eu/registered/data/dossiers/DISS

9875cc3a

- 73ed

- 5dcf

- e044

- 00144f67d031/AGGR

- 83a4ad65

- b0f5

- 4029

- 8d5e

c262ccb1985d_DISS

- 9875cc3a

- 73ed

- 5dcf

- e044

- 00144f67d031.html#AGGR

- 83a4ad65

- b0f5

- 4029

- 8d5e

- c262ccb1985d ; Exp Key Toxicity to aquat ic algae and cyanobacteria.001); a

summar y of this study is also available at the website for Japan’s National Institute of Technology and Evaluation ( http://www.safe.nite.go.jp/engli sh/sougou/view/ ComprehensiveInfoDisplay_en.faces ). None of the summaries contain sufficient data to evaluate study adequacy.

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).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

23 D

- 4

. Acute Toxicity

Summary of endpoint: Adequate

Study 1: Acute oral toxicity study in mice ( Caprino et al., 197 6

)

Title: Toxicological studies of photosensitizer agents and photodegradable polyolefins

Th is study is: Adequate Not Adequate

Although t he post

- dosing observation period was 7 days, rather than the 14

- day period required by OECD TG 401 , this study is considered acceptable on a weight

- of

- evidence basis

.

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks: Purity not provided

METHOD

Method/guideline followed: S imilar to OECD TG 401

Test type: Acute Oral Toxicity

GLP compliant?: Not applicable (pre

- GLP)

Year study performed: 197 5

Test species: Swiss mice

Number of animals/sex/dose: 8/dose (sex not specified)

Route of administration: Oral gavage

Vehicle: 5% Gum arabic

Doses/concentrations tested: Not specified

Statistical methods used: The LD 50 was calculated using probit analysis.

Test method/conditions remarks: Swiss mice, fasted for 16 hours, were administered by gavage with the test material suspended in 5% gum Arabic. The animals were observed for 7 days. Necropsies were performed on all animals at death or at the end of the study. The LD50 was calculated by probit analysis. A so lvent control was also tested. Groups of eight animals were tested, but it is not clear h ow many groups were tested. The dose levels were not provided.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

24 RESULTS

Endpoint value(s): LD 50 = 2895 mg/kg

- bw

Results remarks: Sedation, progressive depression of motor activity, unstable gait, tremors, and respiratory impairment were noted at lethal doses. No lesions or abnormalities were noted grossly in the visceral organs at necropsy.

CONCLUSIONS

The oral LD 50 for mice was 2895 mg/kg

- bw (95% C.L. 2441

- 3433 mg/kg

- bw ). Although the post

- dosing observation period was only 7 days, this study is considered acceptable based on a weight of evidence. Opdyke (1979) reported an acute oral LD 50 of >10,000 mg/kg

- bw in the rat and an acute dermal LD 50 of 3535 mg/kg

- bw in the rabbit ( no additional details ; Opdyke, D.L.J. 1979. Benzophenone. Monographs on Fragrance Raw Materials. Pergamon Press. p. 124). Eastman Kodak Company (1991; unpublished) reported an acute oral LD 50 of 1900 mg/kg

- bw in the rat (summarized in NTP Report No. TR

- 533

, 2004

, “ Toxicological and carcinogenesis studies on benzophenone (CAS 119

- 61

- 9) in F344/N rats and B6C3F1 mice (feed studies) ”)

.

STUDY RELIABILITY

[2] Reliable with restrictions. Although t he post

- dosing observation period was 7 days, rather than the 14

- day period required by OECD TG 401 , this study is considered accept able on a weight

- of

- evidence basis

. Missing details included test substance purity , doses tested, sex of test animals , and effects observed at each dose level.

REFERENCE

Caprino, L., Togna, G., and Mazzei, M. 1976. Toxicological studies of photosensitizer agents and photodegradable polyolefins. European Journal of Toxicology 9: 99

- 103. As summarized in HPVIS (summary available at http://www.regulati ons.gov/#!documentDetail;D=EPA

- HQ

OPPT

- 2007

- 0531

- 0057 as of July 2, 2013) and at http://echa.europa.eu/ (Exp Key Acute toxicity: oral.001

, fourth row of the “Search results” table resulting from a “Chemical Substance Search” for CASRN 119

- 61

- 9 ; http://apps.echa.europa.eu/regi stered/data/dossiers/DISS

b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031/DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031_DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031.html )

. 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 Communicat ion Unit (info (at) echa.europa.eu).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

25 D

- 5

. Repeated

- Dose Toxicity

Summary of endpoint: Adequate

Study 1: Repeated

- dose oral toxicity study in rats ( NTP, 2000 )

Title: NTP Technical Report on the toxicity studies of benzophenone (CAS No. 119

- 61

- 9) administered in feed to F344/N rats and B6C3F1 mice

Th is study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks: > 99% pure

METHOD

Method/guideline followed: S imilar to EPA 870.3100

Test type: 90

- Day Oral Toxicity in Rodents (dietary)

GLP compliant?: Yes

Year study performed: 2000

Test species: Fischer 344 rats (male and female)

Number of animals/sex/dose: 10

Route of administration: Oral

– dietary

Vehicle: None

Exposure duration: 14 weeks

Doses/concentrations tested: 0, 1250, 2500, 5000, 10,000, 20,000 ppm (equivalent to 75, 150, 300, 700, or 850 mg/kg

- bw /day for males and 80, 160, 300, 700, or 1000 mg/kg

- bw /day for females)

Statistical methods used: Not available

Test method/conditions remarks: Groups of 10 rats/sex/group were given test material in the diet for 14 weeks. The rats were observed daily for clin i cal signs of toxicity and were weighedFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

26 weekly. At sacrifice, blood, tissues, and organs were removed for gros s and histopathological examination.

Endpoint value(s): LOAEL = 1250 ppm (75 and 80 mg/kg

- bw/day in males and females, respectively) based on hypertrophy and cytoplasmic vacuolization in the liver , increased bile salts, and increased liver CYPIIB activity, as well as decreased body weights in females ; NOAEL = not established

Results remarks: All 20,000 ppm rats were terminated for humanitarian reasons before the end of the study. Significantly decreased body weights were found for all treated fe male rats and for male rats receiving >1250 ppm. The liver and kidney were identified as the target organs and the weight of both organs were increased in all treated animals. In the liver of all treated rats, hypertrophy and/or cytoplasmic vacuolization were found. In the kidney of 2500 ppm males and females of the 10,000 and 20,000 groups, unique lesions were observed that were well

demarcated, wedge

- shaped areas of prominent tubule dilitation. Bile salts were increased in all treated groups, consiste nt with hepatic congestion. Liver CYPIIB activity was increased in all
groups.

CONCLUSIONS

Under the conditions of the study, the target organs of toxicity were the liver and kidney. The test material induced the formation of renal tubule adenomas in m ale rats, and there was equivocal evidence for increased incidences of mononuclear cell leukemia and histiocytic sarcoma in females.

STUDY RELIABILITY

[ 1 ] Reliable with out restrictions.

REFERENCE S

National Toxicology Program (NTP Report) Apr. 2000. NTP Technical Report on the toxicity studies of benzophenone (CAS No. 119

- 61

- 9) administered in feed to F344/N rats and B6C3F1 mice. NIH Publication No. 00

- 3943, NTP Report No. 61. As summarized in HPVIS (summary available at http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

- OPPT

- 2007

- 0531

- 0057 as of July 2, 2013)

. Supporting Information: NTP Report No. TR

- 533. 2004. Toxicological a nd carcinogenesis studies on benzophenone (CAS 119

- 61

- 9) in F344/N rats and B6C3F1 mice (feed studies).

Burdock, G.A., D.H. Pence, and R.A. Ford. 1991. Safety evaluation of ben zophenone. Fd Chem. Toxic. 29 : 741

- 750.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

27 D

- 6

. Genetic Toxicity

– Gene Muta tion

Summary of endpoint: Adequate

Study 1: Mutagenicity study in Salmonella typhimurium ( Mortelmans et al., 1986 )

Title: Salmonella mutagenicity tests: II. Results from the testing of 270 chemicals

Th is study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks : Supplied by Aldrich; according to the vendor it was 99% pure.

METHOD

Method/guideline followed: Similar to OECD TG 471

Test type: Bacterial Reverse Mutation Test

G LP compliant?: No data

Year study performed: 1986

Test species: Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537

Metabolic activation: With and without

Concentrations tested: 0, 3, 10, 33, 100, 166, 333, and 1000 μg/plate (Not all concentrations were used in every trial.)

Statistical methods used: None specified

Test method/conditions remarks: The method was similar to OECD TG 471. A preincubation
assay was used, as described earlier in Haworth et al . (1983), but with some differences as noted below. Test chemical dilution or solvent (0.05 m L ), overnight Salmon ella culture (0.05 m L

), and S

- 9 mix or buffer (0.50 m L ) were combined, mixed, and incubated for 20 minutes at 37°C, without shaking. Then 2.0 m L of molten (45°C) top agar supplemented with 0.5 mM L

- histidine and 0.5 mM D

- biotin was added. The contents of the tubes were mixed and poured onto the surface of 15 x 100 mm Petri dishes that contained Vogel

- Bonner minimal glucose bottom agar. After the top agar solidified, the plates were inverted. The histidine

- revertant (his+) colonies that arose on these pl ates were counted after two days of incubation at 37°C.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

28 The test chemical was initially tested in a preliminary cytotoxicity test using TA100 to determine the appropriate concentration range for the mutagenicity assay. Toxic concentrations were those
t hat produced his+ pinpoint colonies, a decrease in the number of his+ colonies, or thinning or absence of the background lawn. Cytotoxic chemicals like this one were tested up to a concentration that elicited some degree of toxicity. All tests used triplicate plating, and results were presented as the mean number of his+ revertant colonies for three plates and the standard

error o f the mean. Chemicals that were poorly soluble were tested up to concentrations defined by their solubili ties. Chemicals with good solubility and no cytotoxicity were tested up to a limit concentration of 10 mg/plate. Concurrent solvent and positive controls were run with every trial.

Experiments were repeated no less than one week following the initial experiment. The S

- 9 (9,000g supernatant) fractions were derived from livers of Aroclor 1254

- induced male Sprague

Dawley rats and male Syrian hamsters. The S

- 9 fractions were mixed with appropriate co

factors immediately prior to each assay, and they all contained 10% S

- 9. All chemicals were tested without exogenous metabolic activation and with 10% rat and hamster S

- 9. Because the chemical was provided as a coded sample, the laboratory did not know what chemical it was testing. At least 5 concentration s of the test chemical were tested in every trial.

Strain text description: TA98, TA100, TA1535, and TA1537 Vehicle used: Yes Vehicle Name: DMSO

Criteria for Postive Results: Requires reproducible concentration

- related response in replicate trials in at least one strain

. It was not necessary for a response to reach two

- fold over background for a chemical to be considered mutagenic. Initial classification of a response as mutagenic, nonmutagenic, or equivocal was made at the testing laboratory, but Errol Zeiger, who was the project officer, made the final classification. The chemical was then decoded. When, as was the case here, onl y the results of the second experiment were displayed in the paper, it is implied that those results confirmed the results of the first experiment at each laboratory.

Positive Control Substance s : Without metabolic activation: 9

- aminoacridine with TA97, 4

- nitro

o

- phenylenediamine with TA98, and sodium azide with TA100 and TA1535. With metabolic activation: 2

- aminoanthracene with all 4 strains.

Negative Control Substance Remarks: The solvent control was DMSO, with a maximum of 0.05 mL added to each plate.

RESULTS

Genotoxicity result: Negative with and without metabolic activation

Cytotoxic concentration (if applicable): 166 μg/plate without S

- 9; 1000 μg/plate with S

- 9

Results remarks: The test chemical showed no indication of induction of mutations with or without metabolic activation in any of the four strains tested. Solvent and positive controls gaveFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

29 appropriate responses. Thus

, none of the 24 trials gave any indication of inductio n of mutations; half of those results were not displayed in the paper. Precipitate was not seen on any plates. In the absence of metabolic activation, slight cytotoxicity, as evidenced by slight clearing of the background lawn and a reduction in the numb er of his+ revertants, was seen in all strains at 166 μg/plate, which was the highest concentration tested under that test condition. With rat S

- 9 mix, slight cytotoxicty was seen in all strains at 1000 μg/plate. The highest concentration tested with ham ster S

- 9 mix was 333 μg/plate, and it caused no cytotoxicity.

CONCLUSIONS

The test chemical does not induce gene mutations in bacteria either with or without metabolic activation.

STUDY RELIABILITY

[ 2 ] Reliable with restrictions. This was not a guideline study ; only four bacterial strains were tested, whereas OECD TG 471 indicates that five strains should be tested

. However, it was an excellent study with a clear result. The details of the study were clearly presented.

REFERENCES

Mortelmans , K., Hayworth, S., Lawlor, T., Speck, W., Tainer, B., and Zeiger, E. 1986. Salmonella mutagenicity tests: II. Results from the testing of 270 chemicals. Environ. Mutagen. 8 (Suppl. 7): 1

- 119. As summarized in HPVIS (summary available at http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

- OPPT

- 2007

- 0531

- 0057 as of July 2, 2013)

. Supporting Reference (displays some additional data from the same exper iment): NTP. 2000. NTP technical report on the toxicity studies on benzophenone (CAS No. 119

- 61

- 9) administered in feed to F344/N rats and B6C3F1 mice. Toxicology Report Series No. 61.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

30 D

- 7

. Genetic Toxicity

– Chromosomal Aberrations

Summary of endpo int: A dequate

Study 1: Micronucleus assay in mice ( NTP, 2000 )

Title: NTP technical report on the toxicity studies on benzophenone (CAS No. 119

- 61

- 9) administered in feed to F344/N rats and B6C3F1 mice

Th is study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks: The sample was obtained from Aldrich Chemical Company. The supplier indicated that the sample had purity greater than 99% based on gas chromatographic analysis.

METHOD

Method/guideline followed: Similar to OECD TG 474

Test type: Mammalian Erythrocyte Micronucleus Test ( in vivo )

GLP compliant?: No data

Year study performed: 200 0

Test species: B6C3F1 male mice

Number of animals/sex/dose: 5 males /dose

Route of administration: Intraperitoneal

Vehicle: Corn oil

Exposure duration: 72 hours

Doses tested: 0, 200, 300, 400, and 500 mg/ kg

- bw (The NTP report did not specify whether these values represent dose levels for each injection or the total dose received by the test animals.)

Statistical methods used: The frequencies of micronucleated PCEs were compared statistically using a software package (ILS, 1990) that tested for increasing trend over the dose groups by using a one

- tailed Cochran

- Armitage trend test, followed by pairwise comparisons between each d ose group and the solvent control. A binomial dispersion test was included that adjusted forFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

31 excess binomial variation. Statistical significance was initially declared for an individual trial if the trend test P

- value was less than or equal to 0.025 or i f the P

- value for any single dose group was less than or equal to 0.025 divided by the number of dose groups. The NTP scientific staff makes the final determination if a result is positive based on statistical analysis, the reproducibility of any effects observed, and the magnitude of those effects.

Test method/conditions remarks: The method was similar to OECD TG 474. It followed NTP’s standard protocol, which was described in detail by Shelby et al. (1993). There was a preliminary range

- finding stu dy, and for the test chemical the limiting factor was toxicity. The test chemical was dissolved in corn oil and injected intraperitoneally into male mice three times at 24

- hour intervals. The total dosing volume each time was 0.4 mL. Mice were killed 24 hours after the final injection, and bone marrow was extracted from their femurs to make blood smears. Those smears were air

- dried, fixed, and stained. Two thousand polychromatic erythrocytes (PCEs) from each mouse were examined for the presence of micr onuclei. Results were tabulated as the mean and the standard error of the mean for the pooled results of all 5 animals within each group. Nothing was specified about the ratio of polychromatic erythrocytes to normochromatic erythrocytes (PCE:NCE), which is used as a measure of bone marrow toxicity.

Positive Control Substance: Cyclophosphamide at 25 mg/kg

- bw Negative Control Substance: Corn oil at 0.4 mL per injection

One limitation of this experiment was that there was no testing of females. The test ing in females becomes much less important when toxicity is similar between the sexes, and there were ample data available within the same report to show that such is the case. However, this issue was never dealt with in the presentation regarding this as say.

Criteria for Positive Results: The frequencies of micronucleated PCEs m ust be significantly above the solvent control, with a significant dose

- response relationship

.

RESULTS

Genotoxicity result: Negative

Cytotoxic dose: No data

Results remarks: The test chemical did not induce a statistically significant increase in micronuclei in bone marrow PCEs under the conditions of this assay. The positive and vehicle controls gave appropriate responses. Only t he treated group that was injected three times with 400 mg/kg

- bw (i.e., the next to highest dose) gave any suggestion of an increase in the frequency of micronucleated PCEs, and for it the pairwise P

- value was 0.043. According to the statistical test used, this value would have had to be l ess than 0.006 to reach statistical significance. The numbers of micronucleated PCEs per 1000 PCEs in the solvent control and in the 400 mg/kg

- bw group were 1.2 ± 0.4 and 2.2 ± 0.7, respectively.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

32 CONCLUSIONS

The test chemical does not appear to have the potential to induce chromosomal aberrations in somatic cells in vivo

.

STUDY RELIABILITY

[ 2 ] Reliable with restriction s

. This was not a guideline study. The report does not provide data describing the effects of the test substance on the PCE:NCE ratio or evidence of systemic toxicity; therefore, it is unclear whether this study met the recommendations of OECD TG 474, which indicates that the highest dose tested should produce signs of toxicity. However, Caprino et al. (1976) reported an LD 50 of 727 mg/kg

- bw in mice administered benzophenone via the intraperitoneal route , and the highest dose tested in this micronucleus study ( 500 mg/kg

- bw) approached the reported LD 50 value. Therefore, this micronucleus study is considered acceptable.

REFERENCE

NTP. 2000. NTP technical report on the toxicity studies on benzophenone (CAS No. 119

- 61

- 9) administered in feed to F344/N rats and B6C3F1 mice. Toxicology Report Series No. 61. As summarized in HPVIS (summary available at http://www.regulations.gov/#!documentDetail; D=EPA

- HQ

- OPPT

- 2007

- 0531

- 0057 as of July 2, 2013)

.
Supporting I nformation: Caprino, L., Togna, G., and Mazzei, M. 1976. Toxicological studies of photosensitizer agents and photodegradable polyolefins. European Journal of Toxicology 9: 99

- 103. As summarized in NTP Report No. TR

- 533

, 2004

, “ Toxicological and carcinogenesis studies on benzophenone (CAS 119

- 61

- 9) in F344/N rats and B6C3F1 mice (feed studies) ”.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

33 D

- 8

. Reproductive /Developmental Toxicity

Summary of Reproductive Toxicity E ndpoint: Adequate Summary of Developmental Toxicity E ndpoint: Adequate

Study 1: Two

- generation reproductive toxicity study in rats ( Hoshino et al., 2005 )

Title : A two

- generation reproductive toxicity study of benzophenone in rats

Th is study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9 (referred to as benzophenone in study description)

Remarks: ≥ 99.98% purity; lot/batch no. 112D2013

METHOD

Method/guideline followed: OECD TG 416

Test type: Two

- G eneration Reproduction Toxicity

GLP compliant?: Not specified

Year study performed: 200 5

Test species: Crj:CD(SD) rats ; supplier: Charles River Japan, Inc. ; age at initiation: 5 weeks (P) or 3 weeks (F1)

Number of animals/sex/dose: 24

Route of administration: Oral feed

Vehicle: None

Exposure duration: P males were exposed for 10 weeks; P females were exposed for at least 10 weeks during pre

- mating, mating, gestation, lactation, and weaning of the F1 offspring (until postnatal day [PND] 21); F1 animals were exposed for the same duration as the P generation, starting at the time of weaning (three weeks old)

Doses/concentrations tested: 0, 100, 450, and 2000 ppm in diet (nominal concentrations) ; approximately equivalent to 0, 6.4, 29, and 130 mg/kg

- bw /day for the F0 male parents, 0, 8. 4

, 38, and 16 7 mg/kg

- bw/day for the F0 female parents, 0, 7.8, 3 5 , and 159 mg/kg

- bw /day for the F1 male parents, and 0, 8. 8 , 4 1 , and 179 mg/kg

- bw /day for the F1 female parents, respectivelyFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

34 Statistical methods used:

“ Data concerning effects on the offspring until their weaning were based on values calculated per litter as the specimen unit. Using weights of bilateral organs, the sums of the left and right organs were employed for statistical analysis. Metric data wer e analyzed for homogeneity of variance by Bartlett’s method (Bartlett, 1937). When the variance was homogeneous, one

- way ANOVA was carried out. When not homogeneous, on the other hand, a Kruskal

- Wallis’s test (Kruskal and Wallis, 1952) was performed. Wh en a significant inter

- group difference was found, Dunnett’s method (Dunnett, 1955) or a Dunnett type multiple

- comparison method (Dunnett, 1964) was applied. For some examination items, the Kruskal

- Wallis test was applied first, and when a significant int er

- group difference was found, Dunnett type multiple

- comparison method was conducted. Numerical data were analyzed by the Fisher’s exact probability method (Fisher, 1955). The level of statistical significance was basically set at 5%. ” [ ECHA ]

Test meth od/conditions remarks:

“ Test Animals

- Source: Charles River Japan, Inc. (Astugi breeding Center )

- Age at study initiation: (P) 5 wks; (F1) 3 wks

- Housing: Polycarbonate cages

- Diet: ad libitum

- Water: ad libitum

- Acclimation period: 5 days ” [ECHA]

“ Environmental Conditions

- Temperature (°C): 19.0

- 25.0

- Humidity (%): 35

- 75

- Air changes (per hr): 12

- Photoperiod: 12 hrs dark /12 hrs light ” [ECHA]

“ Details on Mating Procedure

- M/F ratio per cage: 1:1

- Length of cohabitation: 2 weeks

- Proof o f pregnancy: vaginal plug and /or sperm in vaginal smear referred to as day 0 of
pregnancy

- Further matings after two unsuccessful attempts: no

...

- Any other deviations from standard protocol: none known ” [ECHA]

“ Administration of F0 parental animals st arted from an age of 5 weeks and continued in males for 10 weeks. For females, administration lasted through 10 weeks or more of the pre

- mating, mating, gestational, [and] lactational [periods] and during weaning of the F1 offspri ng (PND 21). ” [ ECHA ]

“ Administration to F1 parental animals was started from the time of weaning (three weeks old); in F1 males it was continued until necropsy through 10 weeks or more of the pre

- mating and mating periods, and in F1 females until necropsy through 10 weeks or m ore of the pre

- mating,Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

35 mating, gestational, [and] lactational periods, and during weaning of the F2 offspring (PND 21). ” [ECHA]

Frequency of treatment: continuous through diet Control animals: plain diet

“ Dose selection rationale: B ased on the results of a preliminary dose

- range finding study over 4 weeks with dietary concentrations of 0, 600, 2000, 6000 or 20000 ppm, the highest dose for the definite study was set at 2000 ppm

. ” [ ECHA ]

“

PARENTAL ANIMALS: OBSERVATIONS AND EXAMINATIONS Cage

- side observations : Yes

- Time schedule: once daily D etailed clinical observations : No data B ody weight : Yes

- Time schedule for examinations: on gestation days 0, 7, 14 and 20, on lactation days 0, 4, 7, 14 and at necropsy F ood consumption and compound intake (if feeding study) :

- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

- Compound intake calculated as time

- weighted averages from the consumption and body weight gain data: Yes ” [E CHA]

“ Estrous cyclicity (Parental animals)

- Vaginal smears were collected from female animals everyday in the morning to examine the
estr o us cycle during the two weeks before mating, starting from 13 weeks of age for the F0 parents and from 11 weeks for the F1 parents. ” [ ECHA ]

“ Sperm parameters (Parental animals) Parameters examined in F0 and F1 male parental generations:

- In 10 animals of the control and 2000 ppm group, the number of homogenization

- resistant spermatids in the testis and number of sperms in the cauda epididymal were counted. ” [ ECHA ]

“ LITTER OBSERVATIONS Standardi s ation o f Litters

- Performed on day 4 postpartum: yes

- Maximum of 8 pups/litter (4)/sex/litter as nearly as possible; excess pups were kille d and discarded. Parameters Examined The following parameters were examined in [F0 / F1 / F2] offspring:

- number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalitie s, other: reflex tests Gross Examination Of Dead Pups : yes ” [ECHA]Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

36 “

POSTMORTEM EXAMINATIONS (PARENTAL ANIMALS) Sacrifice

- Male animals: All surviving animals as soon as possible after the last litters in each generation were produced

- Maternal animals: All surviving animals after the last litter of each generation was weaned Gross Necropsy

- Gross necropsy consisted of: no data Histopathology / Organ Weights

- Organ weights in all F0 and F1 parental animals: brain, pituitary gland, t hyroid including parathyroid, liver, kidneys, adrenal glands, spleen, testes, epididymes, prostate, semin al vesicle, ovaries and uterus

- Histopathological examinations were conducted for the following organs from all males and females of the control and 2 000 ppm groups in the F0 and F1 parents. The brain, pituitary gland, thyroid and parathyroid, liver, kidneys, adrenal glands, spleen, testes, epididymes, seminal vesicles (including coagulating glands), prostate (ventral lobe), ovaries, uterus (including the cervical region), vagina, and mammary glands. Furthermore, the liver and kidneys in the 100 and 450 ppm groups in the F0 and F1 parent animals and any macroscopically abnormal site s were also histopathologically examined. Also , any animals that died during the course of the study or were sacrificed upon becoming moribund w ere examined to investigate the causes. ” [ ECHA ]

“

POSTMORTEM EXAMINATIONS (OFFSPRING) Sacrifice

- Excluding the pups that died before selection on PND 4 or the pups not selected on PND 4 (when their numbers were adjusted), all the remaining pups were necropsied when they were sacrificed or were found dead. Gross Necropsy

- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal v iscera. Organ weights (at weaning)

...

- The brain, thymus, and spleen of one F1 and F2 male and female pups each selected from each litter were weighed on PND 21. ” [ ECHA ]

“ REPRODUCTIVE INDICES No. of days until copulation Mating index (%) = (Number of ma les and females showing evidence of copulation / number of males and females used for mating) × 100. Fertility index (%) = (Number of pregnant females / number of copulated females) × 100. Gestation length (days) = Number of days from GD 0 until the delive ry day. Gestation index (%) = (Number of females with normal delivery / number of pregnant females) × 100. Sex ratio = Total number of male pups delivered / total number of the pups delivered. ” [ECHA]

“

OFFSPRING VIABILITY INDICES Birth index (%) = (Number of live pups delivered / number of implantations) × 100. Viability rate on PND 21 (weaning rate) (%) = (Number of live pups on PND 21 / number of live pups on PND 4) × 100.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

37 Viability on PND 4 (the survival rate on day 4) (%) = (Number of live pups on PND 4 / number of live pups on PND 0) × 100. ” [ECHA]

RESULTS

Endpoint value(s): The LOAEL for parental toxicity was 100 ppm ( 6.4

- 7.8 and 8.4

- 8.8 mg/kg

bw /day for males and females, respectively; lowest dose tested) , based on hypertrophy of centrilobular hepatocytes in males and females and increased relative liver weights in females

. The NOAEL for parental toxicity was not established . The N OAEL for reproductive toxicity

was 2000 ppm (130

- 159 and 167

- 179 mg/kg

- bw /day for males and females, respectively; highest dose tested), based on no effects at the highest dose tested. The LOAEL for developmental toxicity in F1 and F2 pups was 2000 ppm (130

- 179 mg/kg

- bw /day), based on reductions in pup body weights and body weight gain during lactation

, compared to controls

. The NOAEL for developmental toxicity was 450 ppm (29

- 41 mg/kg

- bw /day)

.

Results remarks:

“

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS) No effects ” [ ECHA ]

“ BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS) Treatment

... at 450 ppm or 2000 ppm inhibited body weight gain and food consumption in both F0 and F1 males and females

. ” [ ECHA ]

“ TEST SUBSTANCE INTAKE (PARENTAL ANIMALS) The mean daily intakes of the test substance in the group receiving the concentrations (doses) of 0, 1 00, 450, and 2000 ppm were equivalent to 0, 6.445, 29.01, and 130.0 mg/kg for the F0 male parents, 0, 8.379, 38.15, and 166.5 mg/kg for the F0 female parents, 0, 7.785, 34.60, and 159.4 mg/kg for the F1 male parents, and 0, 8.776, 40.52, and 179.2 mg/kg fo r the F1 female parents, respectively. ” [ ECHA ]

“ REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS) Effects

... were not found in any dose group of the F0 and F1 females

. ” [ ECHA ]

“ REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS) In both F0 and F1 male parents, no effects of

... treatment were found, even in the 2000 ppm groups, on the sperm motility rate, number of homogenization

- resistant spermatids in the testis and number of sperm in the cauda epididymis, or on the frequency of morphologically a bnormal
sperm

. ” [ ECHA ]

“ REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS) There were no effects of

... treatment to the reproduct ive performance in any of the t ested
groups

. ” [ ECHA ]

“

ORGAN WEIGHTS (PARENTAL ANIMALS) 100 ppm: the relative hepatic weights increased significantly in the F0 females, but there was noFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

38 difference in the absolute weights nor was there a significant difference in absolute hepatic weights found in the F1 females. ” [ ECHA ]

“

450 ppm: a significa nt increase was also evident for both the absolute and relative hepatic weights and the relative renal weights in both F0 males and females; this was also the case for the absolute renal weights in F0 males, and the relative liver and kidney weights in bot h the F1 males and females. ” [ ECHA ]

“

2000 ppm group: significantly elevated values were found for both the absolute and relative weights of the liver and kidneys in F0 and F1 males and females. ” [ ECHA ]

“ GROSS PATHOLOGY (PARENTAL ANIMALS) No changes re lated to

... treatment were found in either F0 or F1 males or females. ” [ ECHA ]

“ HISTOPATHOLOGY (PARENTAL ANIMALS) 100 ppm group:

- hypertrophy of centrilobular hepatocytes in the parental F0/F1 males and females

. ” [ ECHA ]

“

450 ppm or 2000 ppm groups:

- dilation of renal proximal tubules was observed in both F0 and F1 males and females. Regeneration of the proximal tubular epithelium was also apparent in all groups of F0 and F1 males and females, including the control group. The incidence of the renal c hange was higher in the males receiving 450 ppm or 2000 ppm and in the females given 2000 ppm as compared with the control, and cases with moderate or marked change were included only in the former groups. Regeneration of tubular epithelium was more remar kable in males than females, and was pathologically prevalent around the dilated tubules

. ” [ ECHA ]

“

OTHER FINDINGS (PARENTAL ANIMALS) Serum hormone levels 100 ppm group (or higher):

- testosterone levels in the F0 males showed a tendency to be elevated, w hile values for F1 males were not significantly different from the control, so that no consistent trend was apparent. In the F0 and F1 males, no effects were found with regard to the mating index or the fertility index

. Furthermore, when testosterone lev els were compared between the F0 and F1 males, the treatment was found to have had no biologically significant effect. The change observed in F0 males was judged to be due to the low value in one control male. With serum levels of FSH, LH and estradiol, no change related to BZP treatment was found in any dose groups of F0 and F1 animals of either sex. In addition, no abnormalities were found in the hormone levels measured in three F1 control and one 100 ppm F1 animals , all of which were non

- copulating or non

pregnant. ” [ ECHA ]

“ VIABILITY (OFFSPRING) No changes related to

... treatment were found in the number of F1 and F2 pups delivered, the viabilities on PNDs 0, 4 and 21, and the other parameter s examined. ” [ ECHA ]Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

39 “

CLINICAL SIGNS (OFFSPRING) In the F1 and F2 offspring, no changes related to

... treatment were found in clinical signs in any dose group. ” [ ECHA ]

“ BODY WEIGHT (OFFSPRING) 100 ppm group: N ormal body weight changes as shown in the control were observed in both F1 and F2 male and female offsprin g. ” [ ECHA ]

“

450 ppm group: T he F2 male offspring showed significantly elevated body weights on PND 0. ” [ ECHA ]

“

2000 ppm group: A significant elevation was noted for body weights on PND 0 in F1 male and F2 male/female offspring, while significantly l ow values were found as follows: for body weights on PNDs 14

- 21 in the F1 and F2 male/female offspring and for body weight gain on PNDs 7

- 21 in F1 male/female and F2 female offspring. ” [ ECHA ]

“ SEXUAL MATURATION (OFFSPRING) No eff ec ts noted in sexual matu ration ” [ ECHA ]

“

ORGAN WEIGHTS (OFFSPRING) In the 2000 ppm group, significantly elevated relative brain weights were observed in both male and female F1/F2 pups, and lower absolute spleen weights were found in both F1 sexes. These changes were considered due to inhibition of the body weight gain induced by

... [ treatment ]

. In the groups treated with 450 ppm or less, no substance related changes were found. ” [ ECHA ]

“ GROSS PATHOLOGY (OFFSPRING) No changes related to

... treatment were observed in any dose group of F1 and F2 pups. ” [ ECHA ]

“ HISTOPATH OLOGY (OFFSPRING) [Not] performed

, as no macroscopic changes were observed

. ” [ ECHA ]

CONCLUSIONS

“

With regard to the effects of benzophenone on the F0 and F1 parental animals, inhibition of body weight gain and food consumption, significantly elevated renal weights, dilatation of the renal proximal tubules, and regeneration of the proximal tubular epithelium were recognized at doses of 450 ppm and 2000 ppm, along with an increase in hepatic weight and centrilobular hepatocytic hypertrophy, considered as vital adaptive changes, at the doses of 100 ppm or more. Obvious effects on the endocrine system and reproductive toxicological effects were no t found even at the dose of 2000 ppm in the F0 or F1 parent. There were no test substance

- related changes in the estrous cycle, reproductive capability, delivery and lactation, sperm parameters, serum hormone levels, or necropsy findings. As for effects on the offspring, inhibition of body weight gain was observed in both the F1 and F2 males and females of the 2000 ppm group. No effects of treatment were recognized in the number of male and female F1 or F2 pups delivered,Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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40 viability, anogenital distance, physical development, the results of reflex and response tests, or on the observation results of external abnormalities. ” [ ECHA ]

STUDY RELIABILITY

[1] Reliable without restriction.

REFERENCE

Hoshino, N., Tani , E., Wako, Y., and Takahashi, K. 2005. A two

- generation reproductive toxicity study of benzophenone in rats. The Journal of Toxicological Sciences 30 : 5

- 20

. As summarized at http://echa.europa.eu/ (Exp Key Toxicity to reproduction .00 1 for CASRN 119

- 61

- 9

, fourth row of the “Search results” table resulting from a “Chemical Substance Search” for CASRN 119

- 61

- 9 ).

Copyright Notice: © European Chemicals Agency, 2007

- 2011. Reproduction is authorised provided the source i s 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).

Text in quotes is taken directly from ECHA ( no date provided ) study summary (available online
at http://apps.echa.europa.eu/registered/data/dossiers/DISS

- b6b4 ef07

- 75ea

- 7379

- e044

- 00144f67d031/DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031_DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031.html as of June 28, 2013).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

41 Study 2 : Developmental toxicity study in rabbit s ( NTP, 2004 )

Title: Final study report developmental toxicity evaluation for benzophenone (CAS No. 119

- 61

- 9) administered by gavage to New Zealand White rabbits on gestational days 6 through 29

Th is study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks: > 99% pure

METHOD

Method/guideline followed: OPPTS 870.3700

Test type: Prenatal Developmental Toxicity Study

GLP compliant?: Yes

Year study performed: 2004

Test species: New Zealand White rabbits

Number of animals/sex/dose: 24 females /dose

Route of administration: Oral gavage

Exposure duration: Daily on gestation days 6

- 29 (total of 24 days)

Doses/concentrations tested: 0, 5, 25, and 45 mg/kg

- bw /day

Vehicle: 0.5% Ethylcellulose

S tatistical methods used: No data

Test method/conditions remarks: Groups of 24 naturally mated New Zealand W hite rabbits were administered 0, 5, 25, or 45 mg/kg

- bw /day in 0.5% ethylcellulose by gavage on gestation day s (GD) 6

- 29. Animals were observed daily for general condition and 1

- 2 hours post

- dosing for clinical signs of toxicity. Body weight was recorded on GD 0, 3, and 6

- 30. Food consumption was measured on GD 3, 6, 9, 12, 15, 18, 21, 24, 27, 29, and 30.

Anim als were sacrificed on GD 30 and subjected to gross necropsy. The liver, pair ed kidneys, and gravid uterus were weighed. The number of corpora lutea on each ovary was counted. Uterine contents were examined for number of implants, resorptions, dead fetu ses, and liveFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

42 fetuses. Live fetuses were weighed and examined for external abnormalities, including cleft palate. Approximately one

- half of the fetuses were decapitated and the heads fixed in Bouin's solution for examination by free

- hand sectioning. All carcasses were sexed and examined for visceral abnormalities using fresh dissection. The carcasses were eviscerated and stained for skeletal examination.

RESULTS

Endpoint value(s): LOAEL (maternal toxicity) = 25 mg/kg

- bw/day based on mortality, reduced body weight gain and food consumption, and early termination of preganacy ; NOAEL (maternal toxicity) = 5 mg/kg

- bw/day ; LOAEL (developmental toxicity) = 45 mg/kg

- bw/day based on reduced fetal body weight ; N OAEL (developmental toxicity) = 25 mg/kg

- bw/day

Results remarks: Maternal Effects: No adverse effects were noted in low

- dose animals. Two mid

- dose and five high

- dose animals were found dead or sacrificed moribund between GDs 24

- 30. “ At 25 mg/kg/day, one doe aborted (gd 24) and two delivered early (gd 29). At 45 mg/kg/day, seven does delivered early (gd 27

- 30). In addition, two does in each of the mid and high dose groups were removed due to dosing errors or animal husbandry deviations. Thus, the total number of confirmed pregnant does evaluated at necropsy was 24, 21, 17 and 10 for the control through high

- dose groups, respectively. The confirmed pregnancy rate at termination was 88

- 100% per
group. ” [NTP (online summary)]

Body weight gain during the treatment interval was significantly less for the mid

- (39% of control) and high

- dose (net loss of 4.8 g) groups compared with controls (+320.8 g). Overall food consumption by the mid

- and high

- dose groups was 86 and 83%, respectively, of the control group level during the trea tment period. “ Gravid uterine weight, maternal liver and paired kidney weights (absolute or relative) were not affected at any dose. ” [NTP (online summary)]

Fetal Effects: Number of corpora lutea/doe, number of implantations/doe, percent of pre

- and
po st

- implantation losses, number of live fetuses/litter, and fetal sex ratio were similar between the treated and control groups. “ Average fetal body weight per litter exhibited decreasing dose

response trends (males, females and both sexes combined). At t he high dose, significant reductions were noted for male fetal body weight (81% of control weight), female fetal body weight (85% of control weight) and both sexes (83% of control weight). Benzophenone had no adverse effect on prenatal viability or incide nces of fetal morphological anomalies among litters evaluated on gd 30. ” [NTP (online summary)]

No treatment

- related external, visceral, or skeletal malformations were observed.

CONCLUSIONS

“

In summary, benzophenone was administered to timed

- pregnant NZW rabbits by gavage on gd 6 through 29. The maternal toxicity NOAEL was 5 mg/kg/day. The LOAEL for maternal toxicity was 25 mg/kg/day based on reduced maternal body weight and weight change, a s well asFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

43 dose

- related incidences of maternal mortality and early termination of pregnancy (i.e., abortion or early delivery). The developmental toxicity NOAEL was 25 mg/kg/day, and the LOAEL was 45 mg/kg/day based on reduced fetal body weight. ” [NTP (on line summary)]

STUDY RELIABILITY

[ 1 ] Reliable with out restrictions.

REFERENCES

NTP (National Toxicology Program). 2004. Final study report developmental toxicity evaluation for benzophenone (CAS No. 119

- 61

- 9) administered by gavage to New Zealand White rabbits on gestational days 6 through 29. NTIS PB2004

- 105387. 95 pp. As summarized in abstract ( http://ntp.niehs.nih.gov/index.cfm?objectid=07313273

- FB82

- 4032

- AAE7B779BD397F5D ) and HPVIS (summary available at http://www.regulations.gov/ #!documentDetail;D=EPA

- HQ

- OPPT

- 2007

- 0531

- 0057 as of July 2, 2013)

. Supporting study: NTP (National Toxicology Program). 2002. Final study report developmental toxicity evaluation for benzophenone ( CAS No. 119

- 61

- 9) administered by gavage to Sprague

- Dawley (CD) rats on gestational days 6 through 19. NTIS PB2003

- 101281. 132 pp.

Text in quotes is taken directly from the NTP online summary ( http://ntp.niehs.nih.gov/ index.cfm?objectid=07313273

- FB82

- 4032

- AAE7B779BD397F5D )

.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

44 E . SUPPLEMENTAL DATA (NON

- SIDS ENDPOINTS)

E

- 1. Skin Irritation

Study 1: Dermal irritation study in rabbits ( From ECHA website, Exp Key Skin irritation/corrosion.001 )

Title: Not provided

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks: White to pale yellowish crystals; lot/batch no. 50391499; expiration date: December 1999; stored in dark refrigerator; test substance ID: HR 98/660032 ; purity not provided

METHOD

Method/guideline followed: OECD TG 404 and EU Method B.4

Test type: Acute Dermal Irritation/Corrosion

GLP compliant?: Yes

Year study performed: 19 98

- 1999

Test species: MOL:Russian rabbit

Number of animals/sex/dose: 4 animals total , sex not indicated

Route of administration: Dermal

Vehicle: Ethanol 96% and Diethyl phthalate in a ratio of 1:1

Doses/concentrations tested: 0.5 mL of 2, 5, 10, 25 and 100%

Statistical methods used: None

Test method/conditions remarks:

“ TEST ANIMALS

- Source: M&B, Ejby, DK

- 4623 Lille Skensved, Denmark

- Weight at study initiation: 2.2

- 2.4 kg

- Housing: indi vidual in PPO/HIPS cages

- Diet : ad libitum

- Water: ad libitumFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

45 - Acclimation period: 5 days ” [ECHA]

“ ENVIRONMENTAL CONDITIONS

- Temperature (°C): 20 ± 3°C

- Humidity (%): 55 ± 15%

- Air changes (per hr): 10/hour

- Photoperiod (hrs dark / hrs light): 12/ 12 ” [ECHA]

“

IN

- LIFE DATES: 14 Dec 1998 t o 01 January 1999 ” [ECHA]

“ TEST SYSTEM Type of coverage : semiocclusive Preparation of test site : shaved Vehicle : Ethanol 96% and Diethyl phthalate in a ratio of 1:1 Test Material

- Amount (s) applied (volume or weight with unit) : 0.5 g of the test item was moistened with 0.5 mL vehicle, or 0.5 mL of the test article soluti on (if diluted), were applied

- Concentration (if solution) : 2, 5, 10, 25 and 100% Vehicle

- A mount (s) applied (volume or weight with unit) : 0.5 mL Duration of treatment/exposure : 4 hours Observation period : 1 hour after exposure period as well as 24, 48 and 72 hours after exposure period Number of animals: 4 ” [ECHA]

“ Test site

- Area of exposure: 10x10 cm, 6 sit es were selected on this area

- Type of wrap if used: gauze patches secured with 2.5 cm adhesive tapes Removal of test substance

- W ashing (if done): yes, wi th lukewarm water and mild soap

- Time after start of exposure: 4 hours after ex posure Scoring system : Erythema and Eschar formation: Score 0 to 4; Oedema formation: Score 0 to 4 ” [ECHA]

RESULTS

Endpoint value(s): Not irritating; scores at 24, 48

, and 72 hours were 0.

Results remarks: All treated skin sites were scored with 0 ; no signs of irritation were observed at any of the tested concentrations.

CONCLUSIONS

The test substance was not irritating to rabbit skin.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

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46 STUDY RELIABILITY

[ 2 ] Reliable with restrictions

. The primary source of the study is not identified and the purity of the test substance is not provided.

REFERENCE

Study available from ECHA website, http://echa.europa.eu

, fourth row of the “Search results” table resulting from a “Chemical Substance Search” for CASRN 119

- 61

- 9 . Study indicated as Exp Key Skin irritation/corrosion.001

. The data source is referred to as a “study report”, with a report date of 1999

- 01

- 07. No other reference information is provided.

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).

Text in quotes is taken directly from ECHA (no date provided) study summary (available online
at http://apps.echa.europa.eu/registered/data/dossiers/DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031/DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031_ DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031.html as of June 28, 2013).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

47 Study 2: Dermal irritation study in rabbits ( Calas et al., 1977 )

Title: Not provided

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks: Purity not provided

METHOD

Method/guideline followed: Draize (1944) method

Test type: Acute Dermal Irritation/Corrosion

GLP compliant?: Not applicable (pre

- GLP)

Year study performed: 1977

Test species: New Zealand White rabbit

Number of animals/sex/dose: 6 animals total , sex not indicated

Route of administration: Dermal

Vehicle: Olive oil

Doses/concentrations tested: 0.5 mL of 2 0% solution

Statistical methods used: None

Test method/conditions remarks: The test substance was applied to scratched and non

scratched test sites. The vehicle was olive oil. 0.5 mL of a 200 g/L (20%) test solution was applied to each test site. Treated sites were scored for erythema and edema at 24 and 72 hours post

- administration using the Draize (1944) method.

RESU LTS

Endpoint value(s): Moderately irritating; primary dermal irritation index (PDII) = 2.0

.

Results remarks: Erythema and edema were most often slight, but were moderate in 1/6 animals and intense in 1/6 animals. Histolo patholo g ical changes consisted of focal necrosis with and without dyskeratosis.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

48 CONCLUSIONS

“ Benzophenone, applied to the scratched or unscratched rabbit skin, had a medium irritating
response. The primary irritation index was at 2.0

. ” [ ECHA ]

STUDY RELIABILITY

[2] Reliable with restrictions. Test details are limited; missing details include test substance purity, description of test sites and wrap used to cover sites, and individual irritation scores

.

REFERENCE

Calas

, E

. , Castelain

, P

. Y

. , Lapointe

, H

. R

. , Duco s , P

. , Cavelier

, C

. , Duprat

, P ., and Pottou

, P

. 1977

. Allergic contact dermatitis to a photom erizable resin used in printing. Contact Dermatitis 3: 186

- 194. As summarized at http://echa.europa.eu/ ( Exp Supporting Skin irritation/corrosion.00 2

, fourth row of the “Search results” table resulting from a “Chemical Substance Search” for CASRN 119

- 61

- 9 ).

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 Communicat ion Unit (info (at) echa.europa.eu).

Text in quotes is taken directly from ECHA ( no date provided ) study summary (available online
at http://apps.echa.europa.eu/registered/data/dossiers/DISS

- b6b4 ef07

- 75ea

- 7379

- e044

- 00144f67d031/DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031_DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031.html as of June 28, 2013).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

49 E

- 2

. Eye Irritation

Study 1: Eye irritation study in rabbits ( Eastman Kodak Co., 1984)

Title: Letter from Eastman Kodak to USEPA RE: Chemicals listed in the '83 list of chemicals selected for review by TSCA ITC (48 FR 5152) w/attachments

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks: Purity not provided

METHOD

Method/guideline followed: Not specified

Test type: Acute Eye Irritation

GLP compliant?: Not specified

Year study performed: 19 84

Test species: R abbits (strain and sex not specified)

Number of animals/sex/dose: 6 total , sex not indicated

Route of administration: Ocular

Vehicle: None

Doses/concentrations tested: Several crystals (not further specified)

Statistical methods used: None specified

Test method/conditions remarks:

“ Several crystals of the compound were placed into the conjunctival sac of 6 rabbit eyes (3 rinsed/not rinsed each). ” [ ECHA ]

RESULTS

Endpoint value(s): Slightly irritating

Results remarks:

“ The treatment caused only slight eye irritation in all animals. ” [ ECHA ]Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

5 0 CONCLUSIONS

The test substance was slightly irritating to rabbit eyes.

STUDY RELIABILITY

[ 4 ] Not assignable

. Many s tudy details were not provided. Missing details include test substance purity, volume of test material administered, duration of observation period, individual irritation scores and observations , and primary irritation index.

REFERENCE

Eastman Kodak Co. 1984. Initial submission: Letter from Eastman Kodak to USEPA RE: Chemicals listed in the '83 list of chemicals selected for review by TSCA ITC (48 FR 5152) w/attachments, dated 07/04/84. NTIS OTS0001156, FYI

- OTS

- 0794

- 1156. As summariz ed at http://echa.europa.eu/ (Exp WoE Eye irritation.00 2

, fourth row of the “Search results” table resulting from a “Chemical Substance Search” for CASRN 119

- 61

- 9 ).

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 pr ovided written notification is given to the ECHA Communication Unit (info (at) echa.europa.eu).

Text in quotes is taken directly from ECHA ( no date provided ) study summary (available online
at http://apps.echa.europa.eu/registered/data/dossiers/DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031/DISS

- b6b4ef07

- 75ea

- 7379

- e04 4

- 00144f67d031_DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031.html as of June 28, 2013).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

51 E

- 3

. Skin Sensitization

Study 1: Dermal sensitization study in guinea pigs (Sharp, 1978 )

Title: The sensitization potential of some perfume ingredients tested using a modified Draize procedure

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks: Purity not specified; the test item used was from commercial batches which had passed quality control checks

METHOD

Method/guideline followed: Similar to OECD TG 406 (modified Draize test)

Test type: Skin sensitization

GLP compliant?: No

Year study performed: 1978

Test species: Dunkin

- Hartley g uinea p igs

Number of animals/sex/dose: 10 test animals and 4 challenge control animals (sex distribution not specified , both males and females used )

Route of administration: Induction: intradermal; Challenge: intradermal and epicutaneous

Vehicle: Not specified

Doses/concentrations tested: Intradermal injection induction concentration = 1%; Topical challenge concentration = 20%; Intradermal i njection challenge concentration = 0.25%

Test method/conditions remarks: Test animals were obtained from an in

- house inb red colony and weigh ed 350 g at study initiation.

RANGE

- FINDING TESTS

“

Intradermal injection: Four animals of the same sex were each injected intradermally on the shaved flanks with 0.1 mL aliquots of a range of concentrations of test item in a suitable solvent. The reactions were examined for size, erythema and oedema 24 hours later and the concentration giving slight but perceptible irritation with no oede ma was selected as the injection challenge concentration (IIC). ” [ ECHA ]Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

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52 “ Topical application: Aliquots (0.1 mL) of a range of concent r a tions of the test item in a suitable solvent were applied in small circular areas to the shaved flanks of 4 guinea pigs of the same sex. The reactions were examined 24 hours later for erythema and the highest concentration which caused no irritation was se lected as the application challenge conc entration (AAC). ” [ ECHA ]

MAIN STUDY

“

Induction Exposure : At Day 0, in 10 animals, 0.1 mL aliquots of the test item at 1% ( 2.5 times the IIC ) were injected intradermally at 4 sites which overlie the 2 a u xilla ry and inguinal lymph nodes. ” [ ECHA ]

Challenge Exposure :

“

Fourteen days after induction, each animal was challenged intradermally in one flank and topically in the other with 0.1 mL aliquots of the test item at the respective ICC (0.25%) and ACC (20%)

. T he topical chall e nge was made by spreading 0.1 mL of the test substance onto the shaved flank in a small circular area which was not covered. Twenty

- four hours later

, the reactions were scored and apparent senitization reactions confirmed 7 days later by a s econd challenge with controls included. In the absence of sensitization reactions at first challenge, the induction and challenge procedures were repeated on days 35 and 42 , but this time a

co n firmatory challenge with controls was included irrespective of any apparent sensitization reactions at the previous challenge. ” [ ECHA ]

Challenge controls :

“ At each challenge with controls, 4 previously untreated animals of the same sex and similar weight to the test animals were treated intradermally and topically on opposite flanks with 0.1 mL aliquots of test substance at the ICC and ACC respectively. ” [ ECHA ]

Positive control substance(s) : no

Scoring:

“ Each injection reaction was given a total score based on size (2 largest diameters), erythema and oedema. Individual reactions were considered positive when their total score was significantly greater than the average for total score for control reactions. Application reactions were scored on a 0 to +++ scale and individual reactions wer e considered positive if they were a) + or greater and b) there were no erythema reactions in controls. ” [ ECHA ]

RESULTS

Endpoint value(s): Not sensitizing

Results remarks: None specified

CONCLUSIONS

The test article was not sensitizing in guinea pigs.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

53 STUDY RELIABILITY

[ 2 ] Reliable with restrictions

. P ositive control s were not used. Missing test details include purity of test substance and individual scores. Although 4 (instead of 10) intradermal injections were used in the induction phase in this modified Draize test, the test modification as compared to the original Draize method was validated by the performing laboratory

.

REFERENCE

Sharp, D.W. 1978. The sensitization potential of some perfume ingredients tested using a modified Draize procedure. Toxicology 9: 261

- 271. As summarized at http://echa.europa.eu/ (Exp Key Skin sens i tisation.001 for CASRN 119

- 61

- 9

, fourth row of the “Search results” table resulting from a “Chemical Substance Search” for CASRN 119

- 61

- 9 ).

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).

Text in quotes is taken directly from ECHA ( no date provided ) study summary (available online
at http://apps.echa.europa.eu/registered/data/dossiers/DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031/DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031_DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031.html as of June 28, 2013).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

54 Study 2 : D ermal sensitization study in guinea pigs ( Calas et al. , 197 7 )

Title: Allergic contact dermatitis to a photomerizable resin used in printing

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks: Purity not specified

METHOD

Method/guideline followed : OECD TG 406

Test type: Skin sensitization

GLP compliant?: Not applicable (pre

- GLP)

Year study performed: 1977

Test species: Hartley guinea pigs

Number of animals/sex/dose: 2 0 females

Route of administration: Induction: intradermal and epicutaneous ; Challenge: epicutaneous (open)

Vehicle: Olive oil, Freund’s adjuvant

Doses/concentrations tested: Intradermal injection induction concentration = 1%; Topical induction concentration = 1 0%; Intradermal injection challenge concentration s = 1 and 5 %

Test method/conditions remarks: Test animals weigh ed 400 g at study initiation.

MAIN STUDY

“

Induction Exposure :

- No. of exposures: 2 [ intra ] dermal injections in olive oil (1%); 2 [ intra ] dermal injections of Freund's complete adjuvant (50% in distilled water) and 2 [ intra ] dermal injections of suspended test item in Freund's adjuvant (1%)

- Exposure period: After one week, a topical application was applied at a 10% concentration for 48 hour s

- Test groups: 1

- Control group: none ” [ECHA]

“

Challenge Exposure :

- No. of exposures: 1Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

55 - Day(s) of challenge: 2 weeks after dermal induction

- Exposure period: 48 hours

- Test groups: 1

- Control group: none

- Site: same as induction

- Concentration s: 1 and 5%

- Evaluation (hr after challenge): 24 and 48 hours ” [ECHA]

Positive control substance(s) : no

RESULTS

Endpoint value(s): Not sensitizing

Results remarks: At 24 hours after the challenge exposure, 0/20 animals displayed positive reactions.

CONCLUSIONS

The test article was not sensitizing in guinea pigs.

STUDY RELIABILITY

[2] Reliable with restrictions. Positive controls were not used. Missing test details include purity of test substance and individual scores.

REFERENCE

Calas, E., Castelain, P.Y., Lapointe, H.R., Ducos, P., Cavelier, C., Duprat, P., and Pottou, P. 1977. Allergic contact dermatitis to a photomerizable resin used in printing. Contact Dermatitis 3: 186

- 194. As summarized at http://echa.europa.eu/ (Exp Supporting Skin sensitisation .00 3

, fourth row of the “Search results” table resulting from a “Chemical Substance Search” for CASRN 119

- 61

- 9 ).

Copyright Notice: © European Chemicals Agency, 200 7

- 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 C ommunication Unit (info (at) echa.europa.eu).

Text in quotes is taken directly from ECHA (no date provided) study summary (available online
at http://apps.echa.europa.eu/registered/data/dossiers/DISS

- b6b4 ef07

- 75ea

- 7379

- e044

- 00144f67d031/DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031_DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031.html as of June 28, 2013).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

56 E

- 4 . Carcinogenicity

Study 1 : Carcinogenicity study in rats (NTP, 200 6 )

Title: Toxicology and carcinogenesis studies of benzophenone (CAS 119

- 61

- 9) in F344/N rats and B6C3F1 mice (feed studies)

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks: > 99% pure

METHOD

Method/guideline followed: NTP study equivalent to EPA 870.4300

Test type: 2

- Year Oral Toxicity in Rodents (dietary)

GLP compliant?: Yes

Year study performed: 200 6

Test species: Fischer 344 rats (male and female)

Number of animals/sex/dose: 5 0

Route of administration: Oral

– dietary

Vehicle: None

Exposure duration: 2 years

Doses/concentrations tested: 0, 312, 625, and 1250 ppm (equivalent to average daily doses of approximately 15, 30, and 60 mg /kg

- bw to males and 15, 30, and 65 mg/kg

- bw to females)

Test method/conditions remarks:

“ Groups of 50 male and 50 female rats were fed diets containing 0, 312, 625, or 1250 ppm benzophenone (equivalent to average daily doses of approximately 15, 30, and 60 mg benzophenone/kg body weight to males and 15, 30, and 65 mg/kg to females) for 105 weeks. ” [NTP 2006, p. 5]

Carcinogenic effect: YesFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

57 Results remarks:

“ Survival of 1250 ppm males was significantly less than that of controls. Mean body weights of 1250 ppm males were markedly less than those of the controls during year 2 of the study, and weights of exposed females were consistently less than controls throughout the study. Feed consumption by 1250 ppm males was less than that b y the controls after week 70; feed consumption by 1250 ppm females was generally less than that by the controls throughout the study. ” [NTP 2006, p. 5]

“ There was a positive trend in the incidences of renal tubule adenoma in males, and the incidences in 625 and 1250 ppm males exceeded the historical control range for all routes; these neoplasms were accompanied by significantly increased incidences of renal tubule hyperplasia. Due to these findings, additional kidney sections were evaluated; results indi cated additional renal tubule adenomas in all groups of males and renal tubule hyperplasia in all groups of males and females. The incidences of pelvic transitional epithelium hyperplasia and the severity of nephropathy were significantly increased in all exposed groups of male rats. ” [NTP 2006, p. 5]

“

Increased incidences of mononuclear cell leukemia in all exposed groups of females exceeded the historical control range from feed studies, and the incidence in 625 ppm females was significantly greater than that in the controls. Male rats exposed to 312 or 625 ppm had significantly increased incidences of mononuclear cell leukemia. One 625 ppm female and two 1250 ppm females had histiocytic sarcomas, and the incidence in the 1250 ppm group exceeded the range in the historical controls. ” [NTP 2006, p p

. 5

- 6]

“ Liver lesions included significantly increased incidences of hepatocytic centrilobular hypertrophy in all exposed groups of males and females, cystic degeneration in 625 and 1250 ppm males, and bile duct hyperplasia in all exposed groups of females. ” [NTP 2006, p. 6 ]

“

Incidences of mammary gland fibroadenoma in females exposed to 625 or 1250 ppm were lower than expected

... ” [NTP 2006, p. 6 ]

CONCLUSIONS

“ Under the conditions of th is 2

- year stud y , there was some evidence of carcinogenic activity of benzophenone in male F344/N rats based on increased incidences of renal tubule adenoma; mononuclear cell leukemia in male F344/N rats may have been related to benzophenone
exposure. There was equivocal evidence of carcinogenic activity of benzophenone in female F344/N rats based on the marginally increased incidences of mononuclear cell leukemia and histiocytic sarcoma

. ” [NTP 2006, p. 6 ]

“ Administration of benzophenone in feed resulted in increased incidences and/or severities of nonneoplastic lesi ons in the kidney and liver of male and female rats . D ecreased incidences of mammary gland fibroadenoma in female rats were related to b enzophenone exposure. ” [NTP 2006, p p

. 6

- 7 ]Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

58 STUDY RELIABILITY

[ 1 ] Reliable with out restrictions.

REFERENCE

NTP Report No. TR

- 533. 200 6

. Toxicolog y and carcinogen e sis studies of benzophenone (CAS 119

- 61

- 9) in F344/N rats and B6C3F1 mice (feed studies).

Text in quotes is taken directly from NTP (2006).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

59 Study 2 : Carcinogenicity study in mice (NTP, 200 6 )

Title: Toxicology and carcinogenesis studies of benzophenone (CAS 119

- 61

- 9) in F344/N rats and B6C3F1 mice (feed studies)

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks: > 99% pure

METHOD

Method/guideline followed: NTP study e quivalent to EPA 870.4300

Test type: 2

- Year Oral Toxicity in Rodents (dietary)

GLP compliant?: Yes

Year study performed: 2006

Test species: B6C3F1 mice (male and female)

Number of animals/sex/dose: 50

Route of administration: Oral

– dietary

Vehicle: None

Exposure duration: 2 years

Doses/concentrations tested: 0, 312, 625, and 1250 ppm (equivalent to average daily doses of approximately 40, 80, and 160 mg/kg

- bw to males and 35, 70, and 150 mg/kg

- bw to females)

Test method/conditions rema rks:

“ Groups of 50 male and 50 female mice were fed diets containing 0, 312, 625, or 1250 ppm benzophenone (equivalent to average daily doses of approximately 40, 80, and 160 mg/kg body weight to males and 35, 70, and 150 mg/kg to females) for 105 weeks. Survival of all exposed groups of mice was generally similar to that of the control groups. Mean body weights of exposed females were less than vehicle controls. Feed consumption by exposed males and females was similar to that by the controls. ” [NTP 2006, p. 6 ]

Carcinogenic effect : YesFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

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60 Results remarks:

“

In male mice, there were significantly increased incidences of hepatoce llular adenoma in the 625 and 1 250 ppm groups, and these incidences exceeded the historical control range. All hepatocellular neoplasms combined occurred with a positive trend. In female mice, the incidences of hepatocellular adenoma in the 625 and 1250 ppm groups were higher than expected

... Incidences of centrilobular hepatocyte hypertrophy were significantly inc reased in all exposed groups of males and females. All exposed groups of male mice had significant increases in the incidences of multinucleated hepatocytes and chronic active inflammation. The incidences of cystic degeneration of hepatocytes in 625 and 1250 ppm males were significantly increased. The incidence of histiocytic sarcoma in 625 ppm females was significantly increased and exceeded the historical control range. ” [NTP 2006, p. 6 ]

“ The incidences of kidney nephropathy and mineralization in exposed groups of females and the severity of nephropathy in exposed groups of males were significantly increased. ” [NTP 2006,
p. 6 ]

“ The incidences of metaplasia of the olfactory epithelium were significantly increased in 1250 ppm males and females. The incidences of hyperplasia of lymphoid follicles in the spleen were significantly increased in all exposed groups of males and in 312 and 625 ppm females. ” [NTP 2006, p. 6 ]

CONCLUSIONS

“[ Under the conditions of this 2

- year study, ] there was some evi dence of carcinogenic activity of benzophenone in male B6C3F 1 mice based on increased incidences of hepatocellular neoplasms, primarily adenoma. There was some evidence of carcinogenic activity of benzophenone in female B6C3F 1 mice based on increased incidences of histiocytic sarcoma; the incidences of hepatocellular adenoma in female B6C3F 1 mice may have been related to benzophenone
exposure. ” [NTP 2006, p. 6 ]

“ Administration of benzophenone in feed resulted in increased i ncidences and/or severities of nonneoplastic lesions

... in the liver, kidney, nose, and spleen of male and female mice. ” [NTP 2006, p p

. 6

- 7 ]

STUDY RELIABILITY

[1] Reliable without restrictions.

REFERENCE

NTP Report No. TR

- 533. 2006. Toxicology and carcinogenesis studies of benzophenone (CAS 119

- 61

- 9) in F344/N rats and B6C3F1 mice (feed studies).

Text in quotes is taken directly from NTP (2006).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

61 E

- 5. Genetic Toxicity

– Other

Study 1: DNA Damage and Repair Study in Esch erichia coli ( Fluck et al., 19 7 6)

Title: Evaluation of a DNA polymerase

- deficient mutant of E. coli for the rapid detection of carcinogens

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks : Purity not provided

METHOD

Method/guideline followed: None specified

Test type: DNA damage and/or repair ( in vitro )

GLP compliant?: Not applicable (pre

- GLP)

Year study performed: 19 7 6

Test species: Escherichia coli : W3110 (Pol A+) and p3478 (Pol A

- )

Metabolic activation: With and without (rat liver extract)

Concentrations tested: 500 μg

Statistical methods used: None specified

Test method/conditions remarks:

“ T he procedure is based on the finding that cells exposed to agents that modify their DNA tend to protect themselves by excising the altered DNA portion and then resynthesizing the correct
sequence. DNA polymerase has been implicated in this repair process. Bacterial mutants deficient in DNA polymerase (pol A

- ) have been isolated. These strains are exceedingly sensitive to UV radiations, radiomimetic agents, and agents known to react with cellular DNA; moreover, they exhibit an increased rate of mutation. Advantage was taken of these properties of pol A

- strains to devise a screening procedure for detecting agents that modify cellular DNA. ” [ ECHA ]

“

Portions (0.1 m L

) of exponentially growing bacterial cultures [ Escherichia coli W3110 (pol A+) or its pol A

- derivative p3478)] were added to 3 mL of soft (0.75%) agar, which were then poured onto the surface of a hard (1.5%) agar plate (25 mL). For the growth of bacteria, solid as well as liquid media contained Medium HA (18) supplemented with 5 μg of thymi ne/mL. When the soft agar had solidified, a sterile disc impregnated with the test substance, dissolvedFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

62 preferably in water, was placed at the center of each plate. (Alternatively, 0.1 mL of the bacterial cultures was applied directly to the surface of t he hard agar plates.) After incubation at 37°C for 16 hrs the diameters of the zones of growth inhibition were measured. ” [ ECHA ]

Vehicle: W ater or acetone (if unsoluble) Negative controls: A mpicillin and colistin Positive control: D imethyl sulfate So lvent control: N o data

“ Evaluation criteria: Zones of inhibition were measured on inoculated plates. ” [ ECHA ]

RESULTS

Genotoxicity result: Negative with and without metabolic activation

Cytotoxic concentration (if applicable): No data

Results remarks: Positive and negative controls were valid.

“ Zones of inhibition were 0 (for Pol A+) and 1 mm for Pol A

- . ” [ ECHA ]

CONCLUSIONS

“

In a DNA repair test, Benzophenone did not reveal a positive result. ” [ ECHA ]

STUDY RELIABILITY

[2] Reliable with restrictions. This was not a guideline study.

REFERENCE

Fluck, E.R., Poirier, L.A., and Ruelius, H.W. 1976. Evaluation of a DNA polymerase

- deficient mutant of E. coli for the rapid detection of carcinogens

. Chem Biol Interactions 15 : 219

- 231

. As summarized at http://echa.europa.eu/ (Exp Key Genetic toxicity in vitro.00 1

, fourth row of the “Search results” table resulting from a “Chemical Substance Search” for CASRN 119

- 61

- 9 ).

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).

Text in quotes is taken directly from ECHA ( no date provided ) study summary (available online
at http://apps.echa.europa.eu/registered/data/dossiers/DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031/DISS

- b6b4ef07

- 75ea

- 7379

- e 044

- 00144f67d031_DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031.html as of June 28, 2013).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

63 Study 2 : DNA Damage and Repair Study in Salmonella typhimurium ( Takemoto et al., 2002 )

Title: Genotoxic activation of benzophenone and its two metabolites by human cytochrome P450s in SOS/ umu assay

TEST SUBSTANCE

Identity: Methanone, diphenyl

- , CASRN 119

- 61

- 9

Remarks : > 99% pure; supplier: Wako Pure Chemicals

METHOD

Method/guideline followed: None specified

Test type: DNA damage and/or repair ( in vitro SOS/ umu assay )

GLP compliant?: No data

Year study performed: 2002

Test species: Salmonella typhimurium TA 1535 pSK1002

Metabolic activation: With ( liver microsomes )

Concentrations tested: 1

- 1000 μM

Statistical methods used: None specified

Test method/conditions remarks:

“ The genotoxicity of benzophenone was determined by measuring the induction of umu gene expression in Salmonella typhimurium TA1535/pSK1002 or O

- acetyltransferase

- overexpressing strain NM2009. Overnight cell cultures of tester strain were grown in 1% bactotryptone, 0.5% sodium chloride, 0.2% glucose containing 50 μ g/m L ampicillin until the OD600 of the bacterial culture was ~ 0.2. The resulting bacterial suspension was incubated with benzopheno ne and its metabolites dissolved in dimethylsulfoxide at 37°C for 2 h. The expressed beta

- galactosidase activity in the tester strain was determined using o

- nitrophenyl

- ß

- D

- galactopyranoside as a substrate. The P450

- dependent activation of benzophenone a nd its metabolites causing an induction of umu gene expression was carried out with liver microsomes or recombinant P450/NPR in insect cells or E. coli membranes expressing human P450 and NPR. Incubation mixtures (final volume 1.0 mL) consisted of P450 en zyme sources with chemicals in 0.25 m L of 200 mM potassium buffer containing an NADPH

- generating system and 0.75 m L of bacterial suspension. In separated experiments, liver 9000 *g supernatant fraction (20

– 60 μ g/m L ) was used instead of liver microsomes, o r liver cytosol (30 μ g/m L ) was added to the liver microsomes. For inactivation of the enzyme activities, heat treatment (at 100°C for 2 min) was carried out.Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

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64 The umu gene expression was monitored by measuring beta

- galactosidase activities as described ab ove and expressed as units/(min nmol) P450 or NPR. ” [ ECHA ]

“ The concentrations of P450 and protein were determined spectrally. NADPH

– cytochrome c reductase activities were determined using an assumed specific activity of 3.0 umol cytochrome reduced/(min nmol) NPR based on purified human NPR preparations. The molar ratios for P450 1A1/NPR, 1A2/NPR, 1B1/NPR, 2A6/NPR, 2B6/NPR, 2C8/NPR, 2C9/NPR, 2C19/NPR, 2D6/NPR, 2E1/NPR, 3A4/NPR, and 3A5/NPR membranes used in this study were 0.70, 0.28, 0.65, 1.94, 0.80, 0.14, 1.95, 1.40, 0.95, 1.05, 1.10, and 0.36, respectively. ” [ ECHA ]

“

Metabolites [ of benzophenone ] : benzhydrol and p

- benzoylphenol

- Supplier: Wako Pure Chemicals and Pfaltz & Bauer (Waterbury, CT) ” [ECHA]

“ Target gene: umu gene expression in Salmonel la typhimurium TA1535/pSK1002 or O

acetyltransferase

- overexpressing in NM2009 ” [ECHA]

Vehicle: DMSO

RESULTS

Genotoxicity result: Positive with activation

Cytotoxic concentration (if applicable): No data

Results remarks:

“ The umu gene expression of benzophenone catalyzed by P450 2A6/NPR membranes was increased at substrate concentrations up to 200μM. This activation was dependent on P450 concentrations. ” [ ECHA ]

CONCLUSIONS

“

In the present study, benzophenone, benzhydrol and p

- benzylphenol induced umu gene expression after metabolic activation by human P450s, including P450 2A6 and P450 family 1
enzymes. The bioactivation reaction was dependent on the substrate and P450

- concentrations. ” [ ECHA ]

STUDY RELIABILITY

[2] Reliable with restrictions. This was not a guideline study. No details are provided on the use of controls.

REFERENCE

Takemoto, K., Yamazaki, H., Nakajima, M., and Yokoi, T. 2002. Genotoxic activation of benzophenone and its two met abolites by hu man cytochrome P450s in SOS/ umu assay

. Mutat Res 519 : 199

- 204

. As summarized at http://echa.europa.eu/ (Exp Key Genetic toxicity inFinal HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

65 vitro.00 6

, fourth row of the “Search results” table resulting from a “Chemical Substa nce Search” for CASRN 119

- 61

- 9 ).

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).

Text in quotes is taken directly from ECHA ( no date provided ) study summary (avai lable online
at http://apps.echa.europa.eu/registered/data/dossiers/DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031/DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031_DISS

- b6b4ef07

- 75ea

- 7379

- e044

- 00144f67d031.html as of June 28, 2013).Final HPV2 Chemical Data Adequacy Review (July 2 013) CASRN 119

- 61

- 9

66 F

. DATA ADEQ U ACY SUMMARY TABLE

Endpoint Study Information Available Acceptable Data Gap

Aquatic Effects Acute Toxicity to Fish Yes Yes No Acute Toxicity to Aquatic Invertebrates Yes Yes No Toxicity to Aquatic Plants 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 Yes Yes No Developmental Toxicity Yes Yes No