SRC TR

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HPV4 CHEMICAL DATA A DEQUACY REVIEW

Benzotriazole , CAS No. 95

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Prepared by:

Julie Ramage , Ashlee Aldridge, Justine Von Runnen and Joan D. Garey

Environmental Science Center

SRC, Inc.

7502 Round Pond Road

North Syracuse , NY 13212

Contract No. EP

- W

- 09

- 027

Task FG007.0.009.002

Submitted to:

OPPT/RAD/HPVCB (7403M)

U.S. Environmental Protection Agency

1200 Pennsylvania Avenue

Washington, D.C. 20460

Elizabeth Margosches

, Alternate Project Officer

Meena Sonawane, Work Ass ignment Manager

David Brooks, Alternate Work Assignment Manager

July 23 , 2010HPV4 Chemical Data Adequacy Review

Benzotriazole , CAS No. 95

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- 7

July 2010

Submitted to:

High Production Volume Chemicals Branch

Risk Assessment Division

Office of Pollution Prevention and Toxics

Environmental Protection Agency

WAM: Meena Sonawane

Alternate WAM: David Brooks

1200 Pennsylvania Avenue, NW

Washington, DC 20460

- 0001

Prepared By:

SRC , Inc.

Environmental Science Center

2451 Crystal Drive, Suite 804

Arlington, VA 22202Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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1 A. PREPARATION OF THE DATA ADEQUACY ASSESSMENTS

A

- 1. Source material used for assessment

For this unsponsored chemical, a comprehensive literature search was performed to

identify available toxicity data for the follo wing SIDS endpoints:

Acute toxicity to fish

Acute toxicity to Daphnia

Toxicity to aquatic plants

Acute (oral, dermal, inhalation) toxicity

Repeated

- dose toxicity

Reproductive toxicity

Developmental toxicity

Gene mutation

Chromosomal aberrations

Studies r elevant to non

- SIDS endpoints (e.g., eye and skin irritation; neurotoxicity) conducted

via oral, inhalation, or dermal routes were also identified through literature searching.

The following sources were searched for toxicity data:

PubMed

- B ibliographic database that contains references to journal articles in the life

sciences with a concentration on biomedicine

T OXNET

- N ational L ibrary of M edicine databases on toxicology, hazardous chemicals,

environmental health, and toxic releases; includes:

**•** TOXLINE with BIOSIS and NTIS ;

**•** Development and Reproductive Toxicology File (DART) ;

**•** Chemical Carcinogenesis Research Information System (CCRIS)

- Carcinogenicity and mutagenicity test results for over 8000

**•** chemicals) ;

**•** Hazardous Substances Data Bank (HSDB)

- C omprehensive, peer

reviewed toxicology data for about 5000 chemicals ;

**•** Genetic Toxicology Data Bank (GENE

- TOX)

- Peer

- reviewed genetic

toxicology test data for over 3000 chemicals.

TSCATS

- U npublished technical reports submitted by industry to the EPA und er the

Toxic Substances Control Act]

Registry of toxic effects of chemical substances (RTECs)

Environmental Fate Database (EFDB)

ECOTOX

- EPA’s peer

- reviewed ecotoxicology databaseDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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2 EPA’s Integrated Risk Information System (IRIS) website

Agency for Toxi c Substances and Disease Registry (ATSDR) website

National Toxicology Program (NTP) website

International Program on Chemical Safety Concise International Chemical Assessment

Documents (IPCS INC HE M or CICADs)

European Chemical Substances Information Sy stem (ESIS)

- source of IUCLID Datasets

Any available IUCLID Datasets found through the ESIS website and robust summaries

submitted to EPA by former sponsors on the assigned chemicals were retrieved. Original

sources were obtained for any relevant toxici ty data found for the assigned chemicals.

While IUCLID D ataset s sometimes contained studies of interest, certain study

descriptions contained insufficient detail to assess adequacy. In these cases, the primary source

of the study, if available and acce ssible, was obtain ed and review ed.

A

- 2. Evaluating data adequacy

Using standard OECD testing guidelines and professional judgment as guides, available

toxicity studies for the unsponsored chemical were reviewed for adequacy. Summaries were

prepared for all relevant data found, even if little or no study detail was available. Each study

was summarized according to a standard format and assigned a Klimisch reliability code; all

study summaries for the chemical were combined into a single summary docume nt. For studies

already assigned Klimisch codes (e.g., studies summarized in IUCLID datasets), an explanation

was provided if the reviewer’s assessment of data adequacy differed from the pre

- assigned

Klimisch code. A table summarizing data availability/a dequacy for each SIDS endpoint is

included at the end of the summary document.

B. SPECIFIC INFORMATION ON THE AVAILABLE TOXICITY DATA

The available data for each of the SIDS toxicity endpoints are evaluated on the following

pages. Studies for a com mon 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 is determined to be inadequate, a basis for that

co nclusion is supplied.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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3 B

- 1. TOXICITY TO FISH

Summary of endpoint: A dequate

Study 1: Acute toxicity study in fathead minnows

Title: The effects of BT

- D on trout, bluegill and fathead minnow fingerlings with attachments

and cover letter dated 061289

T his study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Benzotriazole, CAS No. 95

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Remarks: The sample received on October 1, 1969, was contained in a bottle labeled “BT

- D

Sample No. SD

- 7414 , Date 9/2 6/69.”

METHOD

Method/guideline followed: Not stated

Test type: Acute toxicity, static and dynamic tests

GLP compliant?: No

Year study was performed: 1969

Test species: Trout, bluegill and fathead minnows

Analytical monitoring: Yes

Exposure durati on: 96 hours

Concentrations tested: between 10 and 34 ppm as appropriate for the species

Test method/conditions remarks: Trout were maintained for 96 hours prior to use at 55

- 60oF in

specially designed dynamic fresh water recirculating tanks of 100

- gall on capacity. The minnows

and bluegills were maintained in similar tanks for 96 hours at 68 oF. All species were fed a dry

commercial fish flake until 24 hours prior to initiation of testing.

In order to estimate the concentration to be used in the dynami c system, a series of static tests

were performed using 0.1, 1.0, 10, 30, 50 and 100 ppm BT

- D.

Th is assay used is described by the Federal Water Pollution Control Administration.

The water in both static and dynamic systems were circulated and that in the dynamic system

was passed through a bank of American Machine and Foundry Cuno Division Water Filters at a

low rate of five gallons per minute. The BT

- D concentration was monitored twice daily.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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4 RESULTS

Endpoint value(s): TL

m values using bluegills and mi nnows in static and dynamic test systems,

are respectively 27.5 and 30.2 ppm at 48 hours and 25.0 and 28.0 ppm at 96 hours. For the trout

tested solely under dynamic conditions the TL

m values were 15 and 12 ppm at 48 and 96 hours,

respectively.

(TL

m is t he medial tolerance limit)

Results remarks:

Static exposure of bluegills and fathead minnows to BT

- D

Concentration (ppm) No. of Fish Mortality

48 hours 96 hours

22 15 0 0

24 15 13.3 46.7

26 15 26.7 60.0

28 15 60.0 86.7

Control 15 0 0

Control + 0.1g

uranine 15 0 6.6

Dynamic exposure to bluegills and fathead minnows to BT

- D

Concentration (ppm) No. of Fish Mortality

48 hours 96 hours

26 15 20 35

28 15 32 50

30 15 41.9 66.1

32 15 52.3 80

34 15 63.0 95.2

Control 15 0 0

Dynamic exposure t o trout to BT

- D

Concentration (ppm) No. of Fish Mortality

48 hours 96 hours

10 30 3.3 3.3

12 30 25.0 50

14 30 41.6 90.0

16 30 55.0 100.0

Control 30 0 0

CONCLUSIONS

Static and dynamic systems were both carried out to determine the 48

- hour and 96

- hour LC50 of

BT

- D. The fish were exposed to levels of BT

- D between 10 and 34 ppm as appropriate for the

species. The concentration of test material was estimated by analysis of uranine dye added to theDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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5 system and provided evidence of correlation with th e calculated values.

The findings in both systems using bluegills and minnows in static and dynamic test systems, are

respectively 27.5 and 30.2 ppm at 48 hours and 25.0 and 28.0 ppm at 96 hours and indicate no

significant differences in toxicity between the two systems. For the trout tested solely under

dynamic conditions the LC50 values were 15 and 12 ppm at 48 and 96 hours, respectively.

It is concluded that trout are approximately twice as sensitive to BT

- D as bluegills and minnows

under the conditio ns of the dynamic test system.

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

The effects of BT

- D on trout, bluegill and fathead minnow fingerlings with attachments and

cover letter dated 061289. Submitting Organization: Sherwin Willia ms Company. Performing

Laboratory: Food and Drug Research Labs. OTS0520640Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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6 Study 2: Acute toxicity study in zebrafish

Title: Report on test for acute toxicity of REOMET BTA M to zebra fish ( Brac h ydanio rerio );

TO10637 with cover letter dated 072893

This study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: REOMET BTA M ; TK 10637

Remarks: The test substance was identified on the basis of the code number. The

characterization and analysis were not par t of the study and not within the responsibility of the

study director.

Yellow solid, batch no. EN 45689.

METHOD

Method/guideline followed: EEC

- Guideline L251 Vol. 27, 84/449/EEC: C

Test type: Acute s tatic

GLP compliant?: Not stated

Year study was performed: 1989

Test species: zebra fish ( Bracydanio rerio ); size 20

- 24 mm (average 23 mm) , weight 0.11 g

(0.07

– 0.015 g), 10 fishes per concentration and control, 10 fishes per aquarium.

Analytical monitoring: No t stated

Exposure duration: 96 hours

Concentrations tested: N ominal concentrations of 32, 58, 100, 180, 320, 580 and 1000 mg/L

Test method/conditions remarks: Glass aquaria of 20 liters filled with 15 liters (36/22/25 cm)

dechlorinated tap water (carbon filter), water hardness 168 mg CaCo3 /L, temperature 22 ± 1°C,

no aeration, fluorescent light for 16 hours daily, study duration 96 hours. The test substance was

added direction to the water in the tanks at nominal concentrations of 32, 58, 100, 180, 320, 580

and 1000 mg/L. Calculated amoun ts of test substance to produce the desired test concentrations

were given into the water and were homogeneously distributed. The fish were then transferred

into the aquaria. A slight deposit was observed at concentrations 100

- 1000 mg/L (nominal) from

th e start of the test.

The LC50

- values were calculated according to SPEARMAN

- KAERBER in: D.J. Finney 524

- 530, London (1964). LC

- values were graphically determined on gausso

- logarithmic probability

paper.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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7 RESULTS

Endpoint value(s): The 96

- hour LC50 value (calculated) for zebra fish was 130 mg/L. 95%

confidence limits: 110

– 140 mg/L. 96

- hour LC0 was 58 mg/L and the 96

- hour LC100 was 180

mg/L. The 72

- hour LC50 for zebra fish was 140 mg/L, 95% confidence limit: 130

- 160 mg/L.

The 48

- hour LC50 was 160, 95 % confidence limit: 130

- 190 mg/L. The 24

- hour LC%0 was 180

mg/L, 95% confidence limit: 150

- 220 mg/L.

Results remarks: Values are based on nominal concentrations (2 significant digits)

Mortalities:

Concentration

(nominal) mg/L Mortalities

– Number of d ead fish

24 h 48 h 72 h 96 h

Blank 0 0 0 0

32 0 0 0 0

58 0 0 0 0

100 0 0 0 1

180 5 7 9 10

320 10 10 10 10

580 10 10 10 10

1000 10 10 10 10

Symptoms observed at the different test concentrations:

Conc.

(nominal)

mg/L Swimming

behavior Loss of

eq uilibrium Respiratory

function Exophtalmus Pigmentation

24 48 72 96 24 48 72 96 24 48 72 96 24 48 72 96 24 48 72 96

Blank 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

32 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

58 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

10 0 1 2 2 2 1 1 1 2 1 1 1 2 0 0 0 0 0 0 1 1

180 3 3 3 3 3 3 1 1 1 0 0 0 0 0 1

320

580

1000

Effect: 0: normal; 1: slight; 2: moderate; 3 severe

CONCLUSIONS

96h LC50 = 130 mg/L

STUDY RE LIABILITY

2 = reliable with restrictionsDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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8 REFERENCE

Report on test for acute toxicity of REOMET BTA M to zebra fish ( Bracydanio rerio ); TO10637

with cover letter dated 072893. Submitting Organization Ciba

- Geigy Corporation.

OTS0538216.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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9 Study 3: Acu te toxicity study in several fish species

Title: Acute toxicity to rainbow trout, carp, catfish and bluegill of TK 10, 637 with cover letter

dated 072893

This study is: Adequate Not Adequate

TEST SUBSTANCE

Ide ntity: 1H

- benzotriazole, CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: Tested in accordance with the method reported by Bathe et al.

(1974) which is a modification of the procedure proposed by the United States Department of the

Interior Fish and Wildlife Service, reported in “Procedure for Evaluation of Acute Toxicity of

Pesticides to Fish and Wildlife” of December 14, 1964.

Test type: Not stated

GLP compliant?: Not stated

Year study was performed: 1978

Test species: rain bow trout ( Salmo gairdneri ), supplied by “Mandli”, Liestal, Swetzerland, fish

breeding station, average weight 2.6 g, average length 61 mm; carp ( Cyprinus carpio ), supplied

by “Riegger”, Ettenheim, W. Germany, fish breeding station, average weight 7.2 g, a verage

length 79 mm; catfish ( Ictalurus melas ), supplied by “Weber”, Wohlen Switzerland, average

weight 2.6 g, average length 57 mm; bluegill ( Lepomis macrochirus ),

Analytical monitoring: Not stated

Exposure duration: 96 hours

Concentrations tested: Ra inbow trout: 15, 21, 37, 65 ppm; Carp: 8.7, 21, 37, 49, 65 ppm;

Catfish:10, 49, 100 ppm; Bluegill: 10, 100 ppm; All concentrations were nominal.

Test method/conditions remarks: Prior to testing the food was withdrawn for 3 days. All fish

were in good con dition and free from disease.

The fish were tested in glass tanks, of 12 and 15 litre capacity. Testing was performed in

reconstituted water prepared of deionised water to which salts were added in the following

amount per litre: 30 mg calcium sulphate, 30 mg magnesium sulphate, 48 mg sodium

bicarbonate, 3 mg potassium chloride.

The fish were tested at 14 ± 2°C (trout, bluegill) and at 21 ± 1°C (carp, catfish). The water wasDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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10 not aerated during testing. Two to five fish were placed in each tank.

Various concentrations of TK 10637 were prepared by adding the test material diluted in aqua

dest. (stock solution) at the appropriate amount ensuring that the total volume in each vessel

remained the same.

RESULTS

Endpoint value(s): Rainbow trout = 23.9 (17.6

- 2 9.9) ppm ; Carp = 30.3 (23

- 39.3) ppm ; Catfish

= > 100 ppm ; Bluegill = > 100 ppm

Results remarks:

Rate of deaths for rainbow trout

Concentration

ppm Number of

fish tested Deaths after % Mortality

after 96 hours 24 h 48 h 72 h 96 h

15 10 0 0 0 0 0

2 1 10 0 0 5 5 50

37 10 1 9 9 9 90

65 10 10 10 10 10 100

Rate of deaths for carp

Concentration

ppm Number of

fish tested Deaths after % Mortality

after 96 hours 24 h 48 h 72 h 96 h

8.7 10 0 0 0 0 0

21 10 0 1 1 2 20

37 10 0 2 4 6 60

49 10 1 4 7 8 80

65 10 7 10 10 10 100

Rate of deaths for catfish

Concentration

ppm Number of

fish tested Deaths after % Mortality

after 96 hours 24 h 48 h 72 h 96 h

10 10 0 0 0 0 0

49 10 0 0 0 0 0

100 10 0 0 0 0 0

Rate of deaths for bluegill

Concentration

pp m Number of

fish tested Deaths after % Mortality

after 96 hours 24 h 48 h 72 h 96 h

10 10 0 0 0 0 0

100 10 0 1 2 4 40

CONCLUSIONS

The test material has a slight acute toxicity to tout and carp and is practically devoid of acuteDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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11 toxicity to catfi sh and bluegill when tested and evaluated according to the procedure of Bathe et

al (1974).

96

- hour LC50:

Rainbow trout = 23.9 (17.6

- 29.9) ppm, LC 50 including 95% confidence limits were

calculated by the logit model. Concentrations were expressed as wei ght of substance per

volume of water, in parts per million (ppm). These values based upon nominal

concentrations of TK 10637.

Carp = 30.3 (23

- 39.3) ppm

Catfish = > 100 ppm

Bluegill = > 100 ppm

STUDY RELIABILITY

2 = reliable with restrictions

REFERE NCE

Acute toxicity to rainbow trout, carp, catfish and bluegill of TK 10, 637 with cover letter dated

072893. Submitting Organization: Ciba

- Geigy Corporation. OTS0538215.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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12 Study 4: Acute toxicity study in Brachydanio rerio

Title: Acute/prolonged tox icity to fish

This study is: Adequate Not Adequate

[ Lacking number of fish tested, concentrations tested and study details .]

TEST SUBSTANCE

Identity: Benzotriazole, CAS No. 95

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Remarks: Substance type: or ganic

Physical status: solid

Purity: ca. 100 % w/w

METHOD

Method/guideline followed: other: "Letale Wirkung beim Zebrabaerbling

– Brachydanio rerio "

(LC 0, LC 50, LC 100; 48

- 96 Stunden), Verfahrensvorschlag Umweltbundesamt, Mai 1984

Test type: S tati c

GLP compliant?: N o data

Year study was performed: 1984

Test species: Zebrafish, Brachydanio rerio (Fish, fresh water)

Analytical monitoring: No

Exposure duration: 96 hours

Concentrations tested: No data

Test method/conditions remarks:

time ( hr) temp (degree C) O2 (mg/l) pH

0 19.6 9.8 6.5

24 23.0 9.3 6.7

48 22.0 9.7 6.8

72 22.5 9.3 6.9

96 22.6 8.0 7.0

RESULTS

Endpoint value(s): The 96

- hour LC50 value for zebrafish is >100 mg/L. T he 96

- hour LC0 isDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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13 100 mg/L.

Results remarks:

time (hr) Abnormal swimming habit Mortality

24 3/10 0/10

48 10/10 0/10

72 10/10 0/10

96 10/10 0/10

CONCLUSIONS

96h LC50 > 1000 mg/L

STUDY RELIABILITY

4 = not assigna ble . Lacking number of fish tested, concentrations tested and study details.

(2 = valid with restrictions Meets generally accepted scientific standards, well documented and

acceptable for assessment

– per IUCLID Dataset.)

REFERENCE

IUCLID (2001) . Bayer report Prof. Caspers in 1985

- 08

- 03

.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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14 Study 5: Acute toxicity study in rainbow trout

Title: Acute/prolonged toxicity to fish

This study is: Adequate Not Adequate

[Lacking number of fish tested, concentrations teste d and study details.]

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

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Remarks: Cobratec 99; purity >99%

METHOD

Method/guideline followed: Batelle Protocol

Test type: No data

GLP compliant?: N o data

Year study was performed: No data

Test species: Rainbow trout, Salmo gairdneri (Fish, estuary, fresh water)

Analytical monitoring: N o data

Exposure duration: 96 hours

Concentrations tested: No data

Test method/conditions remarks: No data

RESULTS

Endpoint value(s): The 96

- hour L C50 value for rainbow trout was 39 mg/L and the EC50 was

24.4 mg/L

.

Results remarks: LC50 generated by Binomial method; 95% confidence limits = 28.4

- 49.5

mg/L

EC50 generated by Binomial method; 95% confidence limits = 21.0

- 28.4 mg/L.

CONCLUSIONS

96 h LC50 = 39 mg/LDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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15 96h EC50 = 24.4 mg/L

STUDY RELIABILITY

4 = not assignable . Lacking number of fish tested, concentrations tested and study details.

(2 = valid with restrictions Meets generally accepted scientific standards, well documented and

acceptab le for assessment

– per IUCLID Dataset)

REFERENCE

IUCLID (2001) . Battelle Study. 10/1/85; PMC Specialties Group, Inc. MSDS.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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16 Study 6: Acute toxicity study in fathead minnows and killifish

Title: Acute/prolonged toxicity to fish

This study is: Adequate Not Adequate

[Lacking number of fish tested, concentrations tested and study details.]

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

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Remarks: P urity not noted

METHOD

Method/guideline followed: No data

Test type: F low through

GLP compliant?: N o data

Year study was performed: No data

Test species: Killifish, Lebistes reticulatus and fathead minnow, Pimephales promelas (Fish,

fresh water)

Analytical monitoring: No data

Exposure duration: 96 hours

Concentrations tested: No data

Test method/conditions remarks: No data

RESULTS

Endpoint value(s): The 96

- hour LC50 value for fathead minnows and killifish was 28 mg/L.

Results remarks:

CONCLUSIONS

96h LC50 = 28mg/LDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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17 STUDY RELIABILITY

4 = not assignable . Lacking number of fish tested, concentrations tested and study details.

(2 = valid with restrictions

– per IUCLID Dataset. )

REFERENCE

IUCLID (2001) . Davis, L.N. et al.: Investigation of Selected Potential Environmental

Contaminant s. Benzotriazoles. EPA 560/2

- 77

- 001, 137

- 138 (1977).Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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18 Study 7: Acute toxicity study in rainbow trout

Title: Acute/prolonged toxicity to fish

This study is: Adequate Not Adequate

[Lacking number of fish tested, conce ntrations tested and study details.]

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

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Remarks: purity not noted

METHOD

Method/guideline followed:

Test type: F low through

GLP compliant?: N o data

Year study was performed:

Test speci es: Rainbow trout, Salmo gairdneri (Fish, estuary, fresh water)

Analytical monitoring: No data

Exposure duration: 96 hours

Concentrations tested: No data

Test method/conditions remarks: fingerlings

RESULTS

Endpoint value(s): The 96

- hour LC50 value for rainbow trout was 12 mg/L.

Results remarks: No data

CONCLUSIONS

96h LC50 = 12 mg/LDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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19 STUDY RELIABILITY

4 = not assignable

REFERENCE

IUCLID (2001) . Davis, L.N. et al.: Investigation of Selected Potential Environmental

Contaminants. Benzotriazo les. EPA 560/2

- 77

- 001, 137

- 138 (1977).Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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20 Study 8: Acute toxicity study in several fish species

Title: Acute/prolonged toxicity to fish

This study is: Adequate Not Adequate

[Lacking number of fish tested, concentratio ns tested and study details.]

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

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Remarks: purity not noted

METHOD

Method/guideline followed: No data

Test type: S tatic

GLP compliant?: N o data

Year study was performed: No data

Test spe cies: Fathead minnows, Pimephales promelas and bluegill, Lepomis macrochirus (Fish,

fresh water)

Analytical monitoring: No data

Exposure duration: 96 hours

Concentrations tested: No data

Test method/conditions remarks: No data

RESULTS

Endpoint valu e(s): The 96

- hour LC50 value for fathead minnows and bluegill was 25 mg/L.

Results remarks: No data

CONCLUSIONS

96h LC50 = 25 mg/LDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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21 STUDY RELIABILITY

4 = not assignable . Lacking number of fish tested, concentrations tested and study details.

(2 = valid with restrictions

– per IUCLID Dataset.)

REFERENCE

IUCLID (2001) . Davis, L.N. et al.: Investigation of Selected Potential Environmental

Contaminants. Benzotriazoles. EPA 560/2

- 77

- 001, 137

- 138 (1977).Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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22 Study 9: Acute toxicity study in fathead minno ws

Title: Acute/prolonged toxicity to fish

This study is: Adequate Not Adequate

[Lacking number of fish tested, concentrations tested and study details.]

TEST SUBSTANCE

Identity: benzotriazole (BT

- D) , CAS No. 95

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Remarks: purity not noted

METHOD

Method/guideline followed:

Test type: flow through

GLP compliant?: N o

Year study was performed:

Test species: Fathead minnows, Pimephales promelas (Fish, fresh water)

Analytical monitoring: no data

Exposure duration: 96 hours

Concentrations tested: No data

Test method/conditions remarks: No data

RESULTS

Endpoint value(s): The 96

- hour LC50 value for fathead minnows was 2 8 mg/L.

Results remarks: Median Tolerance Limit ( LC50 ) = 30.2 ppm (48 hrs ); 28.0 ppm (96 hrs).

CONCLUSIONS

96h LC50 = 28 mg/LDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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23 STUDY RELIABILITY

4 = not assignable

REFERENCE

IUCLID (2001) . Food and Drug Research Laboratories, Study No. 90801,

December 1, 1969.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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24 Study10: Acute toxicity study in bluegill sunfish

Title: Acu te/prolonged toxicity to fish

This study is: Adequate Not Adequate

[Lacking number of fish tested, concentrations tested and study details.]

TEST SUBSTANCE

Identity: benzotriazole (BT

- D), CAS No. 95

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Rema rks: purity not noted

METHOD

Method/guideline followed:

Test type: flow through

GLP compliant?: N o

Year study was performed:

Test species: Bluegill, Lepomis macrochirus (Fish, fresh water)

Analytical monitoring: no data

Exposure duration: 96 hours

Concentrations tested: No data

Test method/conditions remarks: No data

RESULTS

Endpoint value(s): The 96

- hour LC50 value for bluegill was 25 mg/L.

Results remarks: LC50 = 27.5 ppm (48 hrs); 25.0 ppm (96

hrs).

CONCLUSIONS

96h LC50 = 25 mg/LDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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25 STUDY RELIABILITY

4 = not assignable

REFERENCE

IUCLID (2001) . Food and Drug Research Laboratories, Study No. 90801,

December 1, 1969.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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26 Study 11: Acute toxicity study in rainbow trout

Title: Acute/prolonged toxicity to fish

This study is: Adequate Not Adequate

[Lacking number of fish tested, concentrations tested and study details.]

TEST SUBSTANCE

Identity: benzotriazole (BT

- D), CAS No. 95

- 14

- 7

Remarks: purity not noted

METHOD

Method/guideline followe d: No data

Test type: F low through

GLP compliant?: N o

Year study was performed: No data

Test species: Rainbow trout, Salmo gairdneri (Fish, estuary, fresh water)

Analytical monitoring: N o data

Exposure duration: 96 hours

Concentrations tested: No data

Test method/conditions remarks: No data

RESULTS

Endpoint value(s): The 96

- hour LC50 value for rainbow trout was 12 mg/L.

Results remarks: LC50 = 15 ppm (48 hrs); 12 ppm (96 hrs).

CONCLUSIONS

96h LC50 = 12mg/LDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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27 STUDY RELIABILITY

4

- not as signable.

REFERENCE

IUCLID (2001) . Food and Drug Research Laboratories, Study No. 90801,

December 1, 1969.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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28 Study 12: Acute toxicity study in bluegill sunfish

Title: Acute/prolonged toxicity to fish

This study is: Adequate Not Adequate

[Lacking number of fish tested, concentrations tested and study details.]

TEST SUBSTANCE

Identity: benzotriazole (BT

- D), CAS No. 95

- 14

- 7

Remarks: purity not noted

METHOD

Method/guideline followed: EPA OTS 797.1400

Test t ype: S tatic

GLP compliant?: N o

Year study was performed: No data

Test species: Bluegill, Lepomis macrochirus (Fish, estuary, fresh water)

Analytical monitoring: N o data

Exposure duration: 96 hours

Concentrations tested: No data

Test method/cond itions remarks: No data

RESULTS

Endpoint value(s): The 96

- hour LC50 value for bluegill was 25 mg/L.

Results remarks: Median Tolerance Limit ( LC50 ) = 27.5 ppm (48 hrs); 25 ppm (96 hrs).

CONCLUSIONS

96h LC50 = 25 mg/LDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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29 STUDY RELIABILITY

4 = not ass ignable

REFERENCE

IUCLID (2001). Food and Drug Research Laboratories, Study No. 90801, December 1, 1969.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

30 Study 13: Acute toxicity study in fathead minnows

Title: Acute/prolonged toxicity to fish

This study is: Adequate Not Adequate

[Lacking number of fish tested, concentrations tested and study details.]

TEST SUBSTANCE

Identity: benzotriazole (BT

- D), CAS No. 95

- 14

- 7

Remarks: purity not noted

METHOD

Method/guideline followed: No data

Test type: S tati c

GLP compliant?: N o

Year study was performed: No data

Test species: Fathead minnow, Pimephales promelas (Fish, estuary, fresh water)

Analytical monitoring: N o data

Exposure duration: 96 hours

Concentrations tested: No data

Test method/condition s remarks: No data

RESULTS

Endpoint value(s): The 96

- hour LC50 value for fathead minnow was 28 mg/L.

Results remarks: Median Tolerance Limit ( LC50 ) = 30.2 ppm (48 hrs); 28 ppm (96 hrs).

CONCLUSIONS

LC50 = 30.2 ppmDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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31 STUDY RELIABILITY

4 = not assign able

REFERENCE

IUCLID (2001). Food and Drug Research Laboratories, Study No. 90801,

December 1, 1969.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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32 Study 14: Acute toxicity study in bluegill sunfish

Title: Acute toxicity to fish

This study is: Adequate Not Adequate

[Small sample size , non

- standard duration, lacking important study details .]

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

- 14

- 7

Remarks: purity not reported

METHOD

Method/guideline followed: Not stated

Test type: Static

GLP co mpliant?: Not stated

Year study was performed: 1957

Test species: B luegill sunfish ( Lepomis macrochirus ) , 2 fish used

, fingerlings, ≤ 4 inches

Analytical monitoring: No

Exposure duration: 24 hours

Concentrations tested: 5 ppm

Test method/conditions remarks: The test article was introduced at a one time concentration of

5000 μg/L. The test media was fresh water, tempe rature was kept at 55°F, and pH ranged from

7.5

- 8.2.

RESULTS

Endpoint value(s): Not reported

Results remarks: No effects were observed at 5 ppm.

CONCLUSIONS

No effects were observed at 5 ppm.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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33 STUDY RELIABILITY

4 = not assignable

REFERENCE

EC OTOX (2010). 638 Applegate, V.C., J.H. Howell, A.E. Hall Jr., and M.A. Smith, 1957.

Title: Toxicity of 4,346 Chemicals to Larval Lampreys and Fishes. Special Scientific Report

- -

F isheries No. 207. March 1957.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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34 Study 15: Acute toxicity study in Coho s alm on

Title: Lethal Effects of 1888 Chemicals upon Four Species of Fish from Western North

America

This study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

- 14

- 7

Remarks: None sta ted

METHOD

Method/guideline followed: Not stated

Test type: Static

GLP compliant?: Not stated

Year study was performed: Not stated

Test species: Coho s almon ( Oncorhynchus kisutch ), 5

- 10 cm

Analytical monitoring: No

Exposure duration: 24 hours

Concentrations tested: 10 ppm

Test method/conditions remarks: The test article was introduced at a concentration of 10,000

μg/L. The test media was fresh water, temperature was kept at 12°C, and pH at 7.2, the water

hardness was 7 mg/L CaCO3, and the alkalinity was 7 mg/L CaCO3. Polyethylene bags were

used as the t est vessel and aerated, 23 fish were used for testing.

RESULTS

Endpoint value(s): Not reported

Results remarks: Neither loss of equilibrium nor death occurred.

CONCLUSIONS

No effects were observed.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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35 STUDY RELIABILITY

2 = valid with restrictions

R EFERENCE

ECOTOX (2010). 15148 MacPhee, C., and R. Ruelle, 1969. Lethal Effects of 1888 Chemicals

Upon Four Species of Fish from Western North America. Bull. No. 3, Forest, WildI. And Range

Exp. Stn., Univ of Idaho, Moscow, ID: 112p.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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36 Study 16: Acute toxicity study in rainbow trout

Title: Toxicity of 4,346 Chemicals to Larval Lampreys and Fishes

This study is: Adequate Not Adequate

[Small sample size , non

- standard duration, missing important study details ]

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: Not stated

Test type: Static

GLP compliant?: Not stated

Year study was performed: Not stated

Test species: Oncorhynchus mykiss (Rainb ow Trout)

, fingerlings, ≤ 4 inches

Analytical monitoring: Not stated

Exposure duration: 24 hours

Concentrations tested: 5 ppm

Test method/conditions remarks: The test article was introduced at a concentration of 5,000

μg/L. The test media was fresh wa ter, temperature was kept at 55°F, and pH ranged from 7.5

- 8.2,

dissolved oxygen ranged from 8.6

- 13.7 ppm, 4 fish were used for testing.

RESULTS

Endpoint value(s): Not reported

Results remarks: No effects were observed at 5 ppm.

CONCLUSIONS

No effec ts were observed at 5 ppm.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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37 STUDY RELIABILITY

4 = not assignable

REFERENCE

ECOTOX (2010). 638 Applegate, V.C., J.H. Howell, A.E. Hall Jr., and M.A. Smith, 1957.

Title: Toxicity of 4,346 Chemicals to Larval Lampreys and Fishes. Special Scientific Report

- -

F isheries No. 207. March 1957.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

38 Study 17: Acute toxicity study in Chinook salmon

Title: Acute fish toxicity

This study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

- 14

- 7

R emarks: None stated

METHOD

Method/guideline followed: Not stated

Test type: Static

GLP compliant?: Not stated

Year study was performed:

Test species: Oncorhynchus tshawytscha (Chinook Salmon)

Analytical monitoring: No

Exposure duration: 24 hours

Concentrations tested: 10 ppm

Test method/conditions remarks: The test article was introduced at a concentration of 10,000

μg/L. The test media was fresh water, temperature was kept at 12°C with a pH of 7.2, hardness

of 0

- 17 mg/L CaCO3 and an alkalinity of 7 mg/L CaCO3. Polyethylene bags were used as the

test vessel, 22 fish were used for testing.

RESULTS

Endpoint va lue(s): Not reported

Results remarks: Neither loss of equilibrium nor death occurred.

CONCLUSIONS

No effects were observed.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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39 STUDY RELIABILITY

2 = valid with restrictions

REFERENCE

ECOTOX (2010). 15148 MacPhee, C., and R. Ruelle, 1969. Lethal E ffects of 1888 Chemicals

Upon Four Species of Fish from Western North America. Bull. No. 3, Forest, WildI. And Range

Exp. Stn., Univ of Idaho, Moscow, ID: 112p.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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40 B

- 2. TOXICITY TO AQUATIC INVERTEBRATES

Summary of endpoint: Adequate

Study 1: Acute tox icity study in Daphnia magna

Title: Report on test for acute toxicity of REOMET BTA M to Daphnia magna ; TK 10637 with

cover letter dated 072893

This study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: RE OMET BTA M, TK 10637, CAS No. 95

- 14

- 7

Remarks: The test substance was identified on the basis of the code number. The

characterization and analysis were not part of the study and not within the responsibility of the

study director.

METHOD

Method/guidel ine followed: EEC Guideline L251 Vol. 24 84/449/EEC: C.1

Test type: Static

GLP compliant?: Not stated

Year study was performed: 1989

Test species: Daphnia magna Straus 1820, 20 Daphnia per concentration and control, 4

replicates of 5 Daphnia each

An alytical monitoring: Not stated

Exposure duration: 24 hours

Concentrations tested: 0.58, 1.0, 1.8, 3.2, 5.8, 10, 18, 32, 58, 100, 180 mg/L

Test method/conditions remarks: Stock solution was prepared: 0.4 g TK 10647 were diluted

in and made up to 1000 ml with water. 20 Daphnia per concentration and control, 4 replicates of

5 Daphnia each.

Cultures of Daphnia are maintained in glass vessels containing approx. 2.5 L of reconstituted

water at 20 ± 1°C (water is renewed partially thrice weekly). At each r enewal the Dapnia are fed

a suspension of green algae ( Scenedesmus subspicatus ) supplemented by a suspension of

TETRAMIN

- extract in such quantities that food is consumed after 24 hours.

≤24 h before the start of the test reproductive Daphnia are separated from the young by sievingDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

41 all individuals through a 800 μm sieve. This operation is repeated 6 h before the start of the test

and the young (6

- ≤24h of age) are retained for the test.

Be akers covered with watch glasses, 100 mL solution per beaker were used. Reconstituted water

was produced by dissolving 65 mg NaHCO3, 294 mg CaCl2, 2 H20, 123 mg mgSO4, 7 H20 and

mg KCl in 1000 mL bidistilled water. Total hardness is 240 mg CaCO3/L, the water was aerated

with clean air for at least 24 h before use. Temperature was 20 ± 1°C, no aeration during

exposure, fluorescent light was used for 16 hours daily, approx. 1500 lux.

The test substance was added at nominal concentrations of 0.58, 1.0, 1.8 , 3.2, 5.8, 10, 18, 32, 58,

100 and 180 mg/L. Calculated amounts of stock solution to produce the desired test

concentrations were given into the water and were homogeneously distributed. The Daphnia

were then transferred into the beakers. The test subs tance was dissolved in the test vessels.

RESULTS

Endpoint value(s): The 24

- hour EC50 value (calculated) for daphnids was 20 mg/L. 95%

Confidence limits: 11

- 32 mg/L. The 24

- hour EC0 and EC100 were 3.2 and >180 mg/L,

respectively.

Results remarks: Value s are based on nominal concentrations (2 significant digits).

The EC50 values were calculated according to BERKSON, JASA 48 (1953), 569

- 599.

EC

- values were graphically determined on gausso

- logarithmic probability paper.

Immobilization:

Concentration

(nomin al)

mg/L Immobilization after 24 h in vessel Total

number Total % 1 2 3 4

Blank 0 0 0 0 0 0

0.58 0 0 0 0 0 0

1.0 0 0 0 0 0 0

1.80 0 0 0 0 0 0

3.2 0 0 0 0 0 0

5.8 0 0 1 0 1 5

10 2 0 2 3 7 35

18 4 4 3 4 15 75

32 5 5 3 4 17 85

58 4 4 4 3 15 75

100 3 5 4 3 15 75

180 4 3 3 5 15 75

CONCLUSIONS

24h EC50 = 20 mg/LDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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42 STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Report on test for acute toxicity of REOMET BTA M to Daphnia magna ; TK 10637 with cover

letter dated 072893. Submitting Organization Ciba

- Geigy Corporation. OTS0538217.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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43 Study 2: Acute toxicity study in Daphnia magna

Title: Acute toxicity to aquatic invertebrates

This study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: B enzotriazole, CAS No. 95

- 14

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Remarks: Substance type: organic

Physical status: solid

Purity: ca. 100 % w/w

METHOD

Method/guideline followed: OECD Guide

- line 202, part 1 "Daphnia sp., Acute Immobilisation

Test"

Test type: S tatic

GLP compliant?: Y es

Year study was performed: 1991

Test species: Daphnia magna (Crustacea)

Analytical monitoring: No

Exposure duration: 48 hours

Concentrations tested: 0, 63, 88, 125, 177, 250, 354, 500 mg/L (nominal)

Test method/conditions remarks: None stated

RESU LTS

Endpoint value(s): The 48

- hour EC50 value for daphnids was 91 mg/L. The EC0 and EC100

were 63 and 250 mg/L, respectively.

Results remarks:

Nominal Conc (mg/l) % immobilization

24 hr 48 hr

0 0 0

63 0 0

88 25 60Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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44 125 40 90

177 40 95

250 50 100

354 100

500 100

CONCLUSIONS

48h EC50 = 91 mg/L

STUDY RELIABILITY

1 = valid without restriction; GLP Guideline study

REFERENCE

IUCLID (2001). Bayer report 248 A/91 D; 1991

- 07

- 29.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

45 Study 3: Acute tox icity study in Daphnia magna

Title: Acute toxicity to aquatic invertebrates

This study is: Adequate Not Adequate

[Lacking study details.]

TEST SUBSTANCE

Identity: Cobratec 99, CAS No. 95

- 14

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Remarks: purity >99%

METHOD

Method/guideline followed: Battelle protocols

Test type: No data

GLP compliant?: N o data

Year study was performed: No data

Test species: Daphnia magna (Crustacea)

Analytical monitoring: N o data

Exposure duration: 48 hours

Concentrati ons tested: No data

Test method/conditions remarks: No data

RESULTS

Endpoint value(s): The 48

- hour LC50 value for daphnids was 141.6 mg/L.

Results remarks: LC50 generated by Probit method; 95% confidence limits = 127.7

- 173.5

mg/L.

CONCLUSIONS

48h EC50 = 141.6 mg/LDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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46 STUDY RELIABILITY

4 = not assignable. Lacking study details.

(2 valid with restrictions Meets generally accepted scientific standards, well documented and

acceptable for assessment

– per IUCLID dataset)

REFERENCE

IUCLD (2001). Batte lle Study. 10/1/85; PMC Specialties Group, Inc. MSDS.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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47 B

- 3. TOXICITY TO AQUATIC PLANTS

Summary of endpoint: Inadequate

Study 1: Aquatic toxicity study in green algae

Title: Acute toxicity to aquatic plants e.g. Algae

This study is: Adequate Not Adequate

[Lacking study details.]

TEST SUBSTANCE

Identity: Benzotriazole, CAS No. 95

- 14

- 7

Remarks: Substance type: organic

Physical status: solid

Purity: ca. 100 % w/w

METHOD

Method/guideline followed: DIN 38 412 L 9

Test type: No data

GLP compliant?: Y es

Year study was performed: 1992

Test species: Green algae, Scenedesmus subspicatus (Algae)

Analytical monitoring: No

Exposure duration: 72 hours

Concentrations tested: 0, 1, 3.2, 10, 32, 100, 320, 1000 m g/L (nominal)

Test method/conditions remarks: Endpoint

– growth rate

RESULTS

Endpoint value(s): The 72 hour EC50 for green algae was 231 mg/L, the EC10 was 58 mg/L.

Results remarks:Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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48 Nominal Conc (mg/l) % inhibition (72 hr)

0 0

1 0

3.2 0

10 0

32 0

100 8.3

320 91.7

1000 100.0

CONCLUSIONS

72 h EC50 = 231 mg/L

STUDY RELIABILITY

4 = not assignable. Lacking study details.

(1 = valid without restriction, GLP Guideline study

– per IUCLID Dataset)

REFERENCE

IUCLID (2001). Bayer report 248 A/91 A1; 1992

- 01

- 07.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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49 Study 2: Aquatic toxicity study in green algae

Title: Acute toxicity to aquatic plants e.g. Algae

This study is: Adequate Not Adequate

[Lacking study details.]

TEST SUBSTANCE

Identity: Benzotriazole, CAS No. 95

- 14

- 7

Remarks: Substance type: organic

Physical status: solid

Purity: ca. 100 % w/w

METHOD

Method/guideline followed: DIN 38412 L 9

Test type: No data

GLP compliant?: Y es

Year study was per formed: 1992

Test species: Green algae, Scenedesmus subspicatus (Algae)

Analytical monitoring: No

Exposure duration: 72 hours

Concentrations tested: 0, 1, 3.2, 10, 32, 100, 320, 1000 mg/L (nominal)

Test method/conditions remarks: Endpoint

– biomass

R ESULTS

Endpoint value(s): The 72 hour EC50 for green algae was 102 mg/L, the EC10 was 20 mg/L.

Results remarks:

Nominal Conc (mg/l) % inhibition (72 hr)

0 0

1 2.7

3.2 2.0

10 0.7Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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50 32 10.7

100 47.6

320 96.3

1000 100.0

CONCLUSIONS

72 h EC50 = 102 mg/L

STUDY RELIABILITY

4 = not assignable. Lacking study details. (1 = valid without restriction, GLP Guideline study

– per IUCLID Dataset)

REFERENCE

IUCLID (2001). Bayer report 248 A/91 A1; 1992

- 01

- 07.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

51 Stud y 3: Aquatic toxicity study in green algae

Title: Acute toxicity to aquatic plants e.g. Algae

This study is: Adequate Not Adequate

[Lacking study details.]

TEST SUBSTANCE

Identity: Cobratec 99; CAS No. 95

- 14

- 7

Remarks: purity >99%

METHOD

Method/guideline followed: other: Battelle protocol

Test type: No data

GLP compliant?: N o data

Year study was performed: No data

Test species: O ther aquatic plant: algae

Analytical monitoring: No data

Exposure durat ion: 96 hours

Concentrations tested: No data

Test method/conditions remarks: None stated

RESULTS

Endpoint value(s): The 96 hour EC50 for algae was 15.4 mg/L, the NOEC and LOEC were 4.1

and 8 mg/L, respectively.

Results remarks: EC50 generated by Probi t method; 95% confidence limits = 2.544.7

mg/L.

NOEC and LOEC generated by ANOVA, Bonferroni test.

CONCLUSIONS

96h EC50 = 15.4 mg/LDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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52 STUDY RELIABILITY

4 = not assignable. Lacking study details.

REFERENCE

IUCLID (2001). Battelle Study. 10/1/85; PMC Specialties Group, Inc. MSDS.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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53 B

- 4. CHRONIC TOXICITY STUDY IN AQUATIC INVERTEBRATES

S tudy 1 : Chronic toxicity study in Daphnia magna

Title: Chronic toxicity to aquatic invertebrates

This study is: Adequate Not Adeq uate

TEST SUBSTANCE

Identity: Benzotriazole, CAS No. 95

- 14

- 7

Remarks: Substance type: organic

Physical status: solid

Purity: ca. 100 % w/w

METHOD

Method/guideline followed: OECD Guide

- line 202, part 2 "Daphnia sp., Reproduction Test"

Test ty pe: Semi

- static

GLP compliant?: Y es

Year study was performed: 1994

Test species: Daphnia magna (Crustacea)

Analytical monitoring: Yes

Exposure duration: 21 day

Concentrations tested: No data

Test method/conditions remarks: To produce the stock solut ion, the test substance was

weighed into water and treated for 1 hour in an ultrasonic device and, subsequently stirred for 1

hour with a magnetic stirrer.

Endpoint: immobilization

RESULTS

Endpoint value(s): The 21

- day EC50 value for daphnids was 76.9 m g/L. The 21

- day NOEC for

daphnids is 25.9 mg/L. Arithmetical mean of analytical values: EC50: 76.9 mg/l (test substance)

corresp. to 47.3 mg/l TOC.

Results remarks: No dataDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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54 CONCLUSIONS

21d EC50 = 76.9 mg/L

STUDY RELIABILITY

1 = valid without restri ction, GLP Guideline study

REFERENCE

IUCLID (2001). Bayer AG study: January 18, 1995.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

55 S tudy 2 : Acute toxicity study in Daphnia magna

Title: Chronic toxicity to aquatic invertebrates

This study is: Adequate Not Ade quate

TEST SUBSTANCE

Identity: Benzotriazole, CAS No. 95

- 14

- 7

Remarks: Substance type: organic

Physical status: solid

Purity: ca. 100 % w/w

METHOD

Method/guideline followed: OECD Guide

- line 202, part 2 "Daphnia sp., Reproduction Test"

.

Test type: Semi

- static

GLP compliant?: Y es

Year study was performed: 1993

Test species: Daphnia magna (Crustacea)

Analytical monitoring: Yes

Exposure duration: 21 day

Concentrations tested: No data

Test method/conditions remarks: To produce the stock so lution, the test substance was

weighed into water and treated for 1 hour in an ultrasonic device and, subsequently stirred for 1

hour with a magnetic stirrer

. Endpoint: reproduction rate

RESULTS

Endpoint value(s): The 21

- day EC50 value for daphnids was 25.9

- 76.9 mg/L. The 21

- day

NOEC and LOEC for daphnids is 25.9 and 76.9 mg/L, respectively. Arithmetical mean of

analytical values: EC50: 25.9 mg/l (test substance) corresp. to 15.3 mg/l TOC; 76.9 mg/l (test

substance) corresp. to 47.3 mg/l TOC; NOEC: 2 5.9 mg/l (test substance) corresp. to 15.3 mg/l

TOC LOEC: 76.9 mg/l (test substance) corresp. to 47.3 mg/l TOC.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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56 Results remarks: No data

CONCLUSIONS

21

- day EC50 = 25.9

- 76.9 mg/L

STUDY RELIABILITY

1 = valid without restriction, GLP Guideline study

REFERENCE

IUCLID (2001). Bayer AG study: January 18, 1995.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

57 B

- 5 . ACUTE TOXICITY

Summary of endpoint: Adequate

Study 1: Acute oral toxicity study in rats

Title: Acute oral toxicity

The study is: Adequate Not Adeq uate

TEST SUBSTANCE

Identity: Cobratec #99 (benzotriazole); CAS No. 95

- 14

- 7

Remarks: purity >99%

METHOD

Method/guideline followed: Other

Test type: LD50

GLP compliant?: N o data

Year study performed: No data

Test species: R at, no data on strain

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

Route of administration: O ral

Vehicle: N o data

Doses/concentrations tested: 398, 502, 632, 795 mg/kg

Statistical methods used: Not stated

Test method/conditions remarks: 5 rats/sex/dose, single application of 4 dose levels, post

exposure observation time: 14 d.

RESULTS

Endpoint value(s): The oral LD50 in male and female rats was 560 mg/kg

- bw.

Results remarks: 398 mg/kg; mortality: 0/10; 502 mg/kg, mortality: 4/10; 632 mg/kg bw,Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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58 mortality: 7/10; 795 mg/kg, m ortality: 10/10

CONCLUSIONS

The oral LD50 in male and female rats was 560 mg/kg

- bw

STUDY RELIABILITY

2 = valid with restrictions. Meets generally accepted scientific standards, well documented and

acceptable for assessment

REFERENCE

IUCLID (2001). Huntingdon Research CTR, Acute oral LD50 investigation in rats of Cobratec

99 (at the request of Sherwin Williams CO), OTS 0520632. June 5, 1972.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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59 Study 2: Acute oral toxicity study in rats

Title: Acute oral toxicity

The study is: Adequ ate Not Adequate

TEST SUBSTANCE

Identity: benzotriazole; CAS No. 95

- 14

- 7

Remarks: purity not noted

METHOD

Method/guideline followed: Other

Test type: LD50

GLP compliant?: N o data

Year study performed: No data

Test species: ra t, Greenacres strain

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

Route of administration: O ral

Vehicle: C orn oil

Doses/concentrations tested: 46.4, 100, 215 mg/kg as 10 % wt/vol; 1000, 150 mg/kg as 50%

wt/vol

Statistical methods used: Not stated

Test m ethod/conditions remarks: The test substance was administered by stomach tube to six

groups of 5 male albino rats (weight range 305

- 370 g). The sample was administered in corn oil

as a 10% or 50% wt/vol suspension. (Dosage levels of 46.4, 100, 215 mg/kg as 10 % wt/vol;

1000, 150 mg/kg as 50% wt/vol).

Food was withheld for 18 hrs prior to dosing, following which food and water was supplied ad

libitum. All animals observed several times during day of dosing for gross signs of systemic

toxicity and mortalit y and daily thereafter for 14 days. Gross autopsies were done on all animals.

Statistical analysis of mortality data was done by "moving average" method (Weil, CS.

Biometrics. 8:249. 1952)Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

60 RESULTS

Endpoint value(s): The oral LD50 in male Greenacres rats was 909 mg/kg

- bw

Results remarks: LD 50 = 909 mg/kg bw (95% conf. limit = 546

- 1510 mg/kg).

Time of death Dose (mg/kg)

46.4 100 215 464 1000 2150

1 hr 0/5 0/5 0/5 0/5 0/5 1/5

2 hr 0/5 0/5 0/5 0/5 1/5 2/5

4 hr 0/5 0/5 0/5 0/ 5 3/5 2/5

24 hr 0/5 0/5 0/5 0/5 4/5 4/5

2 days 0/5 0/5 0/5 0/5 4/5 4/5

3 days 0/5 0/5 0/5 0/5 4/5 4/5

4 days 0/5 0/5 0/5 0/5 4/5 4/5

5 days 0/5 0/5 0/5 0/5 4/5 4/5

6 days 0/5 0/5 0/5 0/5 4/5 4/5

7

- 14 days 0/5 0/5 0/5 0/5 4/5 4/5

CONCLUSIONS

LD50 = 909 mg/kg

- bw

STUDY RELIABILITY

2 = valid with restrictions. Meets generally accepted scientific standards, well documented and

acceptable for assessment

REFERENCE

IUCLID (2001). Hill Top Resea rch, Inc. Report# S

- 186A. June 12, 1968.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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61 Study 3: Acute oral toxicity study in rats

Title: Acute oral toxicity

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: benzotriazole residue (sample 81469

- B ); purity not noted, CAS No. 95

- 14

- 7

Remarks:

METHOD

Method/guideline followed: Other

Test type: LD50

GLP compliant?: N o data

Year study performed: No data

Test species: Sprague

- Dawley rat

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

Route of administra tion: O ral

Vehicle: None stated

Doses/concentrations tested: 316, 632, 892, 1260 mg/kg

Statistical methods used: Not stated

Test method/conditions remarks: Five male and five female rats/group were given a single oral

dose via blunt tipped canula. Dos es levels were 316, 632, 892, 1260 mg/kg. Animals were

observed for toxicity and mortality for 14 days. All rats were subjected to a macroscopic

examination of the viscera at necropsy.

RESULTS

Endpoint value(s): The oral LD50 for male and female Sprague

- Dawley rats was 720 mg/kg

bw.

Results remarks: The LD50 value was calculated according to Litchfield and Wilcoxon, 1949.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

62 CONCLUSIONS

LD50 = 720 mg/kg

- bw

STUDY RELIABILITY

2 = valid with restrictions. Meets generally accepted scientific standards, wel l documented and

acceptable for assessment

REFERENCE

IUCLID (2001). North American Science Associates, Inc. Lab No. 81

- 5510.

August 3, 1981.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

63 Study 4: Acute inhalation toxicity study in rats

Title: Acute inhalation toxicity

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: benzotriazole; purity not noted, CAS No. 95

- 14

- 7

Remarks: None stated

METHOD

Method/guideline followed: Other

Test type: LC50

GLP compliant?: N o data

Year study performed: No data

Test species: Sprague

- Dawley rats

Number of animals/sex/dose: 10 (number per sex not specified)

Route of administration: Inhalation

Vehicle: O ther: as a dust

Doses/concentrations tested: 1.5 mg/L

Statistical methods used: Not stated

Test met hod/conditions remarks: The exposure was conducted for 4 hours at a target level of

2.5 mg/l. Exposure levels were determined by gravimetric analysis during each hour of expos ure.

Particle size of the aeros olized dust was determined using a cascade impacto r. BT was

administered via inhalation as a dust, by whole body exposure. Physical observations for

abnormal signs were conducted on all animals as a group at 15 minute intervals during the first

hour of exposure, and hourly for the remainder of the exposur e. All animals received detailed

physical observations prior to exposure and upon removal from the chamber, hourly for two

hours post exposure, and daily for the 14 day post

- exposure period. All animals were sacrificed

and complete postmortem examinations were performed.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

64 RESULTS

Endpoint value(s): The inhalation LC50 for male and femal e rats was > 1.5 mg/L.

Results remarks: All animals survived the exposure and the 14

- day post

- exposure observation

period. Signs of treatment included slight labored breat hing and/or nasal discharge during the

exposure or the first week following exposure. These symptoms abated during the second post

exposure week. Slight decreases in weight were observed on the day after exposure with a

recovery thereafter. Postmortem exam inations were unremarkable. The mean gravimetric

exposure concentration for the 4

- hour exposure was determined to be 1.5 mg/L (considered to be

the maximum attainable concentration).

**1.5 mg/L for 4 hours is considered equivalent to 6.0 mg/L for 1 hour

CONCLUSIONS

L C 50 = > 1.5 mg/L

STUDY RELIABILITY

(2) valid with restrictions. Meets generally accepted scientific standards, well documented and

acceptable for assessment.

REFERENCE

IUCLID (2001). Pharmaco LSR Inc. Study No. 93

- 5163. July 29, 1994.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

65 Study 5: Acute inhalation toxicity study in rats

Title: Acute inhalation toxicity

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Benzotriazole

Remarks: Purity not noted

METHOD

Method/guideline followed: Other

Test type: LC50

GLP compliant?: N o data

Year study performed: No data

Test species: A lbino rat

Number of animals/sex/dose: 10 males/dose

Route of administration: Inhalation

Vehicle: None stated

Doses/concentrations tested: Not stated

Statistical methods used: Not stated

Test method/conditions remarks: Five groups of ten male albino rats were exposed for 3 hours

to different concentrations of benzotriazole as a fine aerosol.

RESULTS

Endpoint value(s): The LC50 was determined to be 1.91 mg/l (95% conf. limits = 1.59

- 2.29

mg/L).

Results remarks: Animals which survived the exposure were generally in very good health one

two days post exposure. The primary effect appeared to be pulmonary irritation which resulted

intra

- tracheal edema and a hemorrhagic type response in the lungs.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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66 CONCLUSIONS

LC50 = 1.91 mg/L

STUDY RELIABILITY

– 2 = valid with restrictions. Meets generally accepted scientific

standards, well documented and acceptable for assessment

REFERENCE

IUCLID (2001). Huntingd on Research Center. Project No. 745

- 080. August 8,

1974.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

67 Study 6: Acute dermal toxicity study in rabbits

Title: Acute dermal toxicity

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: BT

- D (benzotri azole), CAS No. 95

- 14

- 7

Remarks: sample no. SD

- 7413; purity not noted

METHOD

Method/guideline followed: EPA OTS 798.1100

Test type: LD50

GLP compliant?: no data

Year study performed: No data

Test species: A lbino rabbits

Number of animals/sex/dose: 2/ dose, both sexes used

Route of administration: dermal

Vehicle: P hysiol saline

Doses/concentrations tested: 1000, 2150, 4640, 10000 mg/kg bw

Statistical methods used: Not stated

Test method/conditions remarks: Dosage levels of 1000, 2150, 4640, 10000 mg/kg bw were

applied to intact (2 animals/group) or abraded (2 animals/group) of male and female albino

rabbits and occluded for 24 hours. Animals were observed for 14 days post exposure.

RESULTS

Endpoint value(s): The dermal LD50 for male and female ra bbits was > 10,000 mg/kg

- bw.

Results remarks: No mortalities occurred at any dosage level. With the exception of purulent

nasal discharge noted in occasional rabbits, all animals generally exhibited normal appearance

and behavior throughout the study.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

68 CO NCLUSIONS

LD50 > 10,000 mg/kg

- bw

STUDY RELIABILITY

– 2 = valid with restrictions. Guideline study

REFERENCE

IUCLID (2001). Hill Top Research, Inc. Report T

- 413. October 29, 1969.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

69 Study 7: Acute dermal toxicity study in rats

Title: Acute dermal toxic ity

The study is: Adequate Not Adequate

[Lacking study details, exposure duration, occlusive or non

- occlusive application and dose

concentrations.]

TEST SUBSTANCE

Identity: benzotriazole; CAS No. 95

- 14

- 7

Remar ks: Purity not noted

METHOD

Method/guideline followed: Not stated

Test type: LD50

GLP compliant?: N o data

Year study performed: 1984

Test species: R at

Number of animals/sex/dose: No data

Route of administration: Dermal

Vehicle: N o data

Doses/concen trations tested: Not stated

Statistical methods used: Not stated

Test method/conditions remarks: Acute dermal application and a 14 day post

- treatment

observation period (no further information)

RESULTS

Endpoint value(s): The dermal LD50 for male and f emale rats was >2,000 mg/kg

- bw.

Results remarks: Symptoms: Dyspnea, exophthalmus, ruffled fur and abnormal body position

are sym p toms commonly seen during observation time following administration of substances byDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

70 gavage. No other symptoms which could be related to the treatment, especially no cutaneous

changes, were observed.

CONCLUSIONS

L D5 > 2000 mg/kg

- bw

STUDY RELIABILITY

4 = not assignable. Lacking study details, exposure duration, occlusive or non

- occlusive

application and dose concentrations.

REFERENCE

IUCLID (2001 ). Ciba

- Geigy Corp., Toxicolog y studies with benzotriazole in rats, rabbits,

Chinese Hamsters, Zebra Fish and guinea pigs. OTS0510543

- 2 6/09/87.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

71 Study 8: Acute dermal toxicity study in rabbits

Title: Acute dermal toxicity

The study is: Adequate Not Adequate

[Lacking study details, exposure duration, occlusive or non

- occlusive application.]

TEST SUBSTANCE

Identity: Cobratec 99 (BT)

, CAS No. 95

- 14

- 7

Remarks: Sample no. 6023; purity > 99%

METHOD

Method/guideline followed: Other

Test type: LD50

GLP compliant?: Not stated

Year study performed: Not stated

Test species: R abbit

Number of animals/sex/dose: 10 animals used, no further details

Route of administration: D ermal

Vehicle: None stated

Doses/concentrations tested: 2000 mg/kg

Statistical methods used: Not stated

Test method/conditions remarks: Cobratec 99 was tested for Acute Dermal Toxicity in 10

rabbits at 2000 mg/kg.

RESULTS

Endpoint value(s): LD50 > 2000 mg/kg

Results r emarks: According to the code of Federal Regulations, the substance is not toxic.

CONCLUSIONSDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

72 LD50 > 2000 mg/kg

STUDY RELIABILITY

4 = not assignable. Lacking study details, exposure duration, occlusive or non

- occlusive

application.

REFERENCE

IUCLID ( 2001). Huntingdon Research Center. Study No. 725

- 254. June, 29, 1972.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

73 Study 9: Acute i.p. toxicity study in mice

Title: Acute toxicity, other routes

The study is: Adequate Not Adequate

[Lacking any study detail.]

T EST SUBSTANCE

Identity: benzotriazole, CAS No. 95

- 14

- 7

Remarks: None stated

METHOD

Method/guideline followed: None stated

Test type: LD50

GLP compliant?: N o data

Year study performed: Not stated

Test species: M ouse

Number of animals/sex/dose: Not stated

Route of administration: i.p.

Vehicle: None stated

Doses/concentrations tested: Not stated

Statistical methods used: Not stated

Test method/conditions remarks: None stated

RESULTS

Endpoint value(s): The intraperitoneal LD50 in mice was 1 000 mg/kg

- bw.

Results remarks: None stated

CONCLUSIONSDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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74 LD50 = 1000 mg/kg

- bw

STUDY RELIABILITY

4 = not assignable. Lacking study details.

REFERENCE

IUCLID (2001). NTP Chemical Repository, Catalog ID number: 000343.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

75 Study 10: Acute i.v. toxicity study in mice

Title: Acute toxicity, other routes

The study is: Adequate Not Adequate

[Lacking any study detail.]

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

- 14

- 7

Remarks: None stated

METHOD

Method/g uideline followed: Not stated

Test type: LD50

GLP compliant?: N o data

Year study performed: Not stated

Test species: M ouse

Number of animals/sex/dose: Not stated

Route of administration: i.v.

Vehicle: None stated

Doses/concentrations tested: Not stat ed

Statistical methods used: Not stated

Test method/conditions remarks: Not stated

RESULTS

Endpoint value(s): The intravenous LD50 in mice was 238 mg/kg

- bw.

Results remarks: Not stated

CONCLUSIONSDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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76 LD50 = 238 mg/kg

- bw

STUDY RELIABILITY

4 = not assignable. Lacking study details.

REFERENCE

IUCLID (2001). NTP Chemical Repository, Catalog ID number: 000343.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

77 Study 11: Acute oral toxicity study in rats

Title: Toxicity and health hazard summary, MSDS and basic toxicity of benzotriazole with cover

letter dated 041989

The study is: Adequate Not Adequate

[Lacking information on test strain, sex, number of animals tested, test duration, and method.]

TEST SUBSTANCE

Identity: 1,2,3

- benzotriazole, CAS No. 95

- 14

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Remarks: None stated

METHOD

Method/guideline followed: Not stated

Test type: LD50

GLP compliant?: Not stated

Year study performed:

~ 1983

Test species: R at, no strain specified

Number of animals/sex/dose: Not stated

Route of administration: Gav age

Vehicle: None stated

Doses/concentrations tested: Not stated

Statistical methods used: Not stated

Test method/conditions remarks: Not stated

RESULTS

Endpoint value(s): Males

– 673 mg/kg (479

- 945); Females

– 566 mg/kg (400

- 800)

Results remarks: moderate weakness, prostration, cyanosis, rapid breathing and slight twitching

were observedDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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78 CONCLUSIONS

The oral LD50 for male and female rats was 673 and 556 mg/kg, respectively. Clinical signs of

toxicity included moderate weakness, prostration, cya nosis, rapid breathing and slight twitching.

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Toxicity and health hazard summary, MSDS and basic toxicity of benzotriazole with cover letter

dated 041989. Submitting Organization: Eastman Ko dak Company. OTS0516745.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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79 Study 12: Acute oral toxicity study in rats

Title: Toxicity and health hazard summary, MSDS and basic toxicity of benzotriazole with cover

letter dated 041989

The study is: Adequate Not Adeq uate

[Lacking study details.]

TEST SUBSTANCE

Identity: 1,2,3

- benzotriazole, CAS No. 95

- 14

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Remarks: None stated

METHOD

Method/guideline followed: Not stated

Test type: LD50

GLP compliant?: Not stated

Year study performed: Not stated

Test spec ies: R at

Number of animals/sex/dose: 5

Route of administration: orally

Vehicle: 0.2% in water

Doses/concentrations tested: 5

- 40 cc/kg

Statistical methods used: Not stated

Test method/conditions remarks: None stated

RESULTS

Endpoint value(s): > 40 cc/kg

Results remarks: weakness, ataxia.

CONCLUSIONSDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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80 LD50 > 40 cc/kg

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Toxicity and health hazard summary, MSDS and basic toxicity of benzotriazole with cover letter

dated 041989. Submitt ing Organization: Eastman Kodak Company. OTS0516745.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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81 Study 13: Acute oral toxicity study in rats

Title: Acute oral administration of benzotriazole to rats with cover letter dated 061289

The study is: Adequate Not Adequate

[No LD50 determined.]

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

- 14

- 7

Remarks: The sample used in this study was received from Maumee Chemical Company on

May 7, 1968. Benzotriazole (SD

- 7176) is an off

- white powder with a slight odor.

METHOD

Method/guideline followed: Not stated

Test type: LD50

GLP compliant?: Not stated

Year study performed: 1968

Test species: M ale albino rat

Number of animals/sex/dose: 5 males/group; Greenacres strain, weight range 305 to 370 grams

Route of administration: O rally by stomach tube

Vehicle: Corn oil

Doses/concentrations tested: 46.5, 100 and 215 mg/kg were administered as a 10% suspension.

464, 1000 and 2150 mg/kg were administered as a 50% suspension

Statistical methods used: Not stated

Test method/conditions remarks: The test sample was administered orally by stomach tube to

six groups, each composed of five male albino rats (Greenacres strain, weight range 305 to 370

grams). The sample was administered as a 10% or 50% weight volume sus pension in corn oil

(Mazola). Dosage levels of 46.4, 100 and 215 mg/kg were administered as a 10% suspension.

Dosage levels of 464, 1000 and 2150 mg/kg were administered as a 50% suspension.

Food was withheld from the rats for approximately 18 hours prio r to dosage. Following dosage,

food, consisting of commercial pellets, and water were available ad libitum. The rats were

housed in groups in wire mesh cages suspended above the droppings. All animals were observedDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

82 closely for gross signs of systemic to xicity and mortality at frequent intervals during the day of

dosage, and at least once daily thereafter for a total of 14 days. Gross autopsies were performed

on the animals that died. At the end of the 14

- day observation period the surviving rats were

w eighed, sacrificed by cerebral concussion and gross autopsies were performed. Statistical

analysis of the mortality data was by the moving average method (Weil, C.S., Biometrics 8, 249,

1952)

RESULTS

Endpoint value(s): LD50 = 909 mg/kg

- bw

Results rema rks: Below values are presented as number of animals dead/number of and animals

tested, cumulative

Dose Concentration

of suspension Time of Death

Mg/kg % Hours Days

1 2 4 24 2 3 4 5 6 7

- 14

46.4 10 0/5 0/5 0/5 0/5 0/5 0/5 0/5 0/5 0/5 0/5

100 10 0/5 0/ 5 0/5 0/5 0/5 0/5 0/5 0/5 0/5 0/5

215 10 0/5 0/5 0/5 0/5 0/5 0/5 0/5 0/5 0/5 0/5

464 50 0/5 0/5 0/5 0/5 0/5 0/5 0/5 0/5 0/5 0/5

1000 50 0/5 1/5 3/5 4/5 4/5 4/5 4/5 4/5 4/5 4/5

2150 50 1/5 2/5 2/5 4/5 4/5 4/5 4/5 4/5 4/5 4/5

LD50 = 909 mg/kg

95% Confid ence limits = 546

- 1510 mg/kg

Two rats in the 46.4 mg/kg dosage level group exhibited slight depression on the day of dosage

only. Otherwise all animals exhibited normal appearance and behavior throughout the study.

The rats at the 100 mg/kg level general ly showed slight depression with slightly depressed

righting and placement reflexes within approximately one hour following dosage. These signs

diminished markedly in intensity during the next five hours. All rats appeared normal in

appearance and behavi or throughout the remainder of the study.

The rats at the 215 mg/kg level generally showed depression with slightly depressed righting and

placement reflexes within two minutes following dosage. These signs progressed to marked

depression with depressed o r absent pain, righting, and placement reflexes after approximately

one hour. There was a marked reduction in intensity of these signs over the next five hours. All

rats showed normal appearance and behavior from the first post

- dosage day through termina tion.

At dosage levels of 464, 1000 and 2150 mg/kg all rats generally showed marked depression with

depressed righting and placement reflexes within two minutes following dosage. These signs

generally became more severe with rats at the higher levels show ing absence of pain, righting

and placement reflexes and shallow respiration prior to death. The rats at the 464 mg/kg level

generally showed some improvement during the day of dosage and appeared normal from the

first post

- dosage day through termination. Except for transient slight emaciation in the single

survivor at the 1000 mg/kg level, the survivors from the 1000 and 2150 mg/kg groups exhibited

normal appearance and behavior from the first or second post

- dosage day through termination.

Dose Average b ody weightDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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83 Start Finish Gain

Mg/kg gm gm gm

46.4 329 399 70

100 342 407 65

215 331 400 69

464 338 397 59

1000 339 380 (1 rat) 41

2150 350 370 (1 rat) 20

Except for the two single surviving rats from the 1000 and 2150 mg/kg groups which showed

mo derate or marked growth suppression, the average body weight gains for the surviving rats

were within normal limits for rats of the age, sex and strain used in this study.

Gross autopsies performed on those rats that died generally showed paleness of the e xtremities;

slight to moderate congestion of the lungs, kidneys and adrenals; and slight irritation of the small

intestine. Occasional rats also exhibited mottled livers. Gross autopsies performed on the

survivors at termination generally showed no gross pathology.

CONCLUSIONS

The acute oral toxicity of benzotriazole (SD

- 7176) for male albino rats was found to be 909

mg/kg of body weight, with 95% confidence limits of 546 and 1510 mg/kg.

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Acute oral administration of benzotriazole to rats with cover letter dated 061289. Submitting

Organization: Sherwin Williams Company. Performing Laboratory: Hill Top Research

Incorporated. OTS0520631.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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84 Study 14: Acute oral toxicity study in rats

Tit le: Acute oral LD50 investigation in rats of Cobratec #99 with cover letter dated 061289

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: COBRATEC #99, benzotriazole, CAS No. 95

- 14

- 7

Remarks: Not s tated

METHOD

Method/guideline followed: Not stated

Test type: LD50

GLP compliant?: Not stated

Year study performed: Not stated

Test species: R ats

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

Route of administration: O rally

Vehicle: Not stated

Doses/conce ntrations tested:

Statistical methods used: Not stated

Test method/conditions remarks: Cobratec #99 was administered orally in graduated doses to

4 groups of 10 rats. Five males and five females were in each group. They were observed for 14

days.

RES ULTS

Endpoint value(s): LD50 = ~ 560 mg/kg

Results remarks:

Dose Day of Death Total Dead

Mg/kg 0 1 2 Total on test

195 4/10 6/10

- 10/10Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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85 632 0/10 7/10

- 7/10

502 0/10 2/10 2/10 4/10

398 0/10

- - 0/10

LD50 determinatio n: The dose response curve i s very steep and does not permit the

establishment of confidence limits. The LD50 is 560 mg/kg

CONCLUSIONS

The LD50 was estimated to be 560 mg/kg

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Acute oral LD50 investigation in rats of Cobratec #99 with cover letter dated 061289

. Submitting Organization: Sherwin

- Williams Company. Performing Laboratory: Huntingdon

Research Center. OTS0520632.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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86 Study 15: Acute dermal toxicity study in rabbits

Title: Acute dermal toxicity investigati on in rabbits of Cobratec 99 (BT) sample #6023 with

attachments and cover letter dated 061289

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Cobratec 99 (benzotriazole), CAS No. 95

- 14

- 7

Remarks: The substance to be tested was labeled Cobrtec 99 (BT) Sample No. 6023. The

material was a light yellow powder.

METHOD

Method/guideline followed: Not stated

Test type: LD50

GLP compliant?: Not stated

Year study performed: Not stated

Test species: R abbi ts

Number of animals/sex/dose: 10 females

Route of administration: De rmal

Vehicle: None stated

Doses/concentrations tested: 2000 mg/kg

Statistical methods used: Not stated

Test method/conditions remarks: The material was applied to the clipped skin of 5 female

rabbits and to the clipped and abraded skin of 5 female rabbits. The material was administered

and allowed to remain in contact with the skin for 24 hours. The trunk of each animal was

wrapped with a dam. After 24 hours the dams were removed . The residue was washed off with

water. All rabbits were observed for mortality and toxic effects on the day of application and

daily thereafter until death occurred or the 14 day observation period ended.

RESULTS

Endpoint value(s): LD50 > 2000 mg/k gDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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87 Results remarks: at 2000 mg/kg 0/10 animals died

Acute dermal toxicity in rabbits

Rabbit

No. sex reaction Day

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

*1015A F Erythema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

edema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

1016A F Erythema 0 2 1 1 0 0 0 0 0 0 0 0 0 0

edema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

1017A F Erythema 1 2 1 1 0 0 0 0 0 0 0 0 0 0

edema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

1018A F Erythema 1 2 1 1 0 0 0 0 0 0 0 0 0 0

edema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

1019A F Erythema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

edema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

**1020I F Erythema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

edema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

1021I F Erythema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

edema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

1022I F Erythema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

edema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

1023I F Erythema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

edema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

1024I F Erythema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

edema 0 0 0 0 0 0 0 0 0 0 0 0 0 0

*A = Abraded skin

**I

– Intact skin

Body weights (kg)

Dose level Rabbit No. Day

Mg/kg 0 3 7 10 14

2000 1015A 2.4 2.6 2.6 2.7 2.8

1016A 2.4 2.3 2.4 2.4 2.5

1017A 2.4 2.4 2.4 2.5 2.6

1018A 2.5 2.5 2.5 2.5 2.5

1019A 2.6 2.6 2.6 2.7 2.7

1020I 2.3 2.2 2.3 2.4 2.6

1021I 2.4 2.5 2.5 2.6 2.6

1022I 2.4 2.5 2.5 2.6 2.6

1023I 2.4 2.4 2.4 2.4 2.5

1024I 2.4 2.5 2.5 2.5 2.6

Evaluation of skin reaction Value

Erythema and eschar formationDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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88 No erythema 0

Very slight erythema (Barely perceptible) 1

Well

- defined erythema 2

Moderate to severe erythema 3

Severe erythema (beet redness) to slight eschar

formation (injuries in depth) 4

Edema formation

No edema 0

Very slight edema (barely perceptible) 1

Slight edema (edges of area well defined by

Definite raisin g) 2

Moderate edema (raised approximately 1 mm) 3

Severe edema (raised more than 1 mm and extend

beyond the area of exposure 4

CONCLUSIONS

According to the code of Federal Regulations, the substance is not toxic.

STUDY RELIABILITY

2 = reli able with restrictions

REFERENCE

Acute dermal toxicity investigation in rabbits of Cobratec 99 (BT) sample #6023 with

attachments and cover letter dated 061289. Submitting Organization: Sherwin

- Williams

Company. Performing Laboratory: Huntingdon Re search Center. OTS0520634.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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89 Study 16: Acute inhalation toxicity study in rats

Title: Evaluation of the three

- hour LC50 of aerosolized benzotriazole (final report) with

attachments and cover letter dated 061289

The study is: Adequate Not Adequate

[Only 3 hours duration

– acute inhalation should be 4.]

TEST SUBSTANCE

Identity: Benzotriazole, CAS No. 95

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Remarks: The experimental material was supplied by the sponsor. The compound was a white

powder and was identified as follows: Sample BT

- PG, benzotriazole photo grade, sample No.

1422

METHOD

Method/guideline followed: Not stated

Test type: LD50

GLP compliant?: Not stated

Year study performed: Not stated

Test species: Sprague

- Dawley rats

Number of anima ls/sex/dose: 10 males/group

Route of administration: Inhalation

- aerosol

Doses/concentrations tested: 0.78, 1.46, 2.03, 2.23, 2.71 mg/L

Statistical methods used: Not stated

Test method/conditions remarks: Five groups of ten male rats were used for th is study.

Groups I through V each received a single 3

- hour exposure to one of five different aerosol

concentrations of the experimental compound. All groups were held for fourteen days post

exposure for observation of latent effects. All animals were ob served daily for pharmacotoxic

signs. Body weights were recorded prior to exposure and after seven and fourteen days. All

animals that died or were sacrificed after the observation period were necropsied and all major

organs were observed for gross abnor malities.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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90 RESULTS

Endpoint value(s): LC50 = 1.91 mg/L

Results remarks: Group No. Exposure concentration mg/L Percent mortality

I 0.78 0

II 1.46 10

III 2.03 20

IV 2.23 50

V 2.71 100

Observations for pharmacotoxic signs: During exposures, the only pharmacotoxic sign observed

was deep abdominal breathing with open mouth gasping apparent during exposures to the two

highest concentrations. All but one death occurred during the exposures; and usually these

deaths occ urred during the final half

- hour of exposure. One animal in Group IV (2.23 mg/L)

was found dead on the morning after exposure. Generally, animals which survived the exposure

appeared normal and healthy by two or three days post

- exposure.

Body weights: All surviving animals appeared to gain weight normally throughout the 14

- day

observation period

.

Gross pathology: Animals that died during the exposure were necropsied immediately after the

3

- hour exposure. There were two consistent effects observed at these necropsies. First the cut

trachea exhibited a very severe accumulation of white frothy fluid; and second the lungs

exhibited a moderate to severe incidence of dark red hemorrhagic areas in all lobes. There were

no other gross abnormalities observed . The cut surface of the lungs did not indicated the

presence of pulmonary edema.

The primary effect appeared to be pulmonary irritation which resulted in tracheal edema and a

hemorrhagic type response in the lungs. Animals which survived the exposure w ere generally in

very good health one or two days post

- exposure.

CONCLUSIONS

The LC50 for benzotriazole, under the aerosol generation and exposure conditions used in this

study was 1.91 mg/L. The 95% confidence limits were 1.59 to 2.29 mg/L. The fac t that animals

which survived the exposure generally gained body weight normally and appeared to be in good

health would indicate that the effects of inhaled benzotriazole are primarily local respiratory tract

irritations.

STUDY RELIABILITY

2 = reliab le with restrictions

REFERENCE

Evaluation of the three

- hour LC50 of aerosolized benzotriazole (final report) with attachmentsDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

91 and cover letter dated 061289. Submitting Organization: Sherwin

- Williams. Performing

Laboratory: Huntingdon Research Center

. OTS0520637Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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92 Study 17: Acute inhalation toxicity study in rats

Title: Environmental and health related studies summary report of Cobratec 99 (1,2,3

- benzotriazole), Cobratec TT100 (tolyl triazole), and para cresidine with cover letter dated

09/16/94

Th e study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Cobratec 99, 1,2,3

- benzotriazole, CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: None stated

Test type: LC50

GLP compliant?: Not stated

Year study performed: Not stated

Test species: Sprague

- Dawley CD rats

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

Route of administration: Inhalation as a dust, whole

- body exposure

Vehicle: Not stated

Doses/concentrations tested: 1.5 mg/L for 4 hours

Statistical methods used: Not stated

Test method/conditions remarks: Exposure was conducted for 4 hours at a target level of 2.5

mg/L, which was considered equivalent to 10.0 mg/L for 1 hour. Physical observations for

abnormal signs were conducte d on all animals as a group, at fifteen minute intervals during the

first hour of exposure, and hourly for the remainder of exposure. All animals received detailed

physical observations just prior to exposure and 1,2,4,7 and 14 days after exposure. After the

post

- exposure observation period, all surviving animals were sacrificed. Complete post mortem

examinations were performed on all animals.

RESULTSDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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93 Endpoint value(s): LC50 = 6 mg/L

Results remarks: The mean gravimetric exposure concentration of CO BRATEC 99 for the 4

- hour exposure was determined to be 1.5 mg/L of air (equivalent) to 6.0 mg/L for 1 hour). This

was considered to have been the maximum attainable exposure level.

All animals survived the exposure and the 14

- day post

- exposure observation period. Signs of

treatment included slight respiratory (labored breathing) and/or secretory (nasal discharge)

responses during the exposure or for the first week following exposure. These generally abated

during the second week after the exposure.

Sligh t decreases in body weights were observed on the day after exposure with a recovery

thereafter. Postmortem findings were unremarkable.

CONCLUSIONS

In conclusion, the one

- hour LC50 of COBRATEC 99 was considered greater than 6.0 mg/L, the

maximum attainab le concentration.

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Environmental and health related studies summary report of Cobratec 99 (1,2,3

- benzotriazole),

Cobratec TT100 (tolyl triazole), and para cresidine with cover letter dated 09/1 6/94. Submitting

Organization: Cincinnati Specialties Incorporated. Performing Laboratory: Wildlife

International Limited. OTS0557484.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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94 Study 18: Acute oral toxicity study in rats

Title: Letter from Sherwin Williams Company to the USEPA regarding in formation on 1,2,3

- benzotriazole and tolytriazole with attachments.

The study is: Adequate Not Adequate

[Lacking study details and methods.]

TEST SUBSTANCE

Identity: 1H

- benzotriazole, CAS No. 95

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Remarks: Synonyms: 1,2,3

- benzotriazole, azimidobenzene, benzene azimide. Appearance:

white to off

- white free

- flowing powder

METHOD

Method/guideline followed: None stated

Test type: LD50

GLP compliant?: Not stated

Year study performed: Not stated

Test species: W hite rat

Number of animals/sex/dose: Not stated

Route of administration: O ral

Vehicle: None stated

Doses/concentrations tested: Not stated

Statistical methods used: Not stated

Test method/conditions remarks: Not stated

RESULTS

Endpoint value(s): The oral LD50 value in white rats is 560 mg/kg.

Results remarks: Not statedDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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95 CONCLUSIONS

Chemicals with values in this range are generally considered moderately toxic.

STUDY RELIABILITY

4 = not assignable

REFERENCE

Letter from Sherwin Williams Co mpany to the USEPA regarding information on 1,2,3

- benzotriazole and tolytriazole with attachments. Submitting Organization: Sherwin Williams

Company. OTS0520630.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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96 Study 19: Acute oral toxicity study in rats

Title: Acute oral test with cover sheet date d 061289

The study is: Adequate Not Adequate

[Study determined approximate lethal dose, but is lacking LD50, and details on test method.]

TEST SUBSTANCE

Identity: 1,2,3

- benzotriazole, CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: None stated

Test type: Acute toxicity

GLP compliant?: Not stated

Year study performed: Not stated

Test species: ChR

- Cd male rats

Number of animals/sex/dose: 1 male/dose

Route of administration: O ral

- gav age

Vehicle: C orn oil

Doses/concentrations tested: 130, 200, 300, 450, 670, 2250 mg/kg

Statistical methods used: Not stated

Test method/conditions remarks: The test material, as a suspension in corn oil, was

administered to young adult ChR

- CD male ra ts in single doses by intragastric intubation.

Survivors were sacrificed 14 days after dosing.

RESULTS

Endpoint value(s): ALD = 670 mg/kg

- bwDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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97 Results remarks:

Dose (mg/kg) Suspension (%) Fate Clinical signs ALD

2250 20 Dead

– 16 hours Lethal doses

– none

observed 670 mg/kg

670 20 Dead

– 16 hours

450 5 Sacrificed

– 14

days Sublethal doses

– temporary prostration

on day of dosing at 450

mg/kg. Lethargy at

200 and 300 mg/kg.

initial weight loss at

450 and 300 mg/kg. 300 5 Sacrificed

– 14

days

200 5 Sacrificed

– 14

days

130 5 Sacrificed

– 14

days

CONCLUSIONS

Benzotriazole is moderately toxic to young adult ChR

- CD male rats when administered in single

oral doses. Its Approximate Lethal Dose (ALD) is 670 mg/kg of body weight.

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Acute oral test with cover sheet dated 061289. Submitting Organization: E I Dupont De

Nemours and Company. Performing Laboratory: Haskell Laboratories. OTS0520954.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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98 B

- 6 . REPEATED

- DOSE TOXICI TY

Summary of endpoint: Adequate

Study 1: Repeated

- Dose toxicity study in rats

Title: Toxicity and health hazard summary, MSDS and basic toxicity of benzotriazole with cover

letter dated 041989

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: 1,2,3

- benzotriazole, CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: Not stated

Test type: Repeated

- dose toxicity

GLP compliant?: Not stated

Year study performed: Not stated

Test spec ies: R at

Number of animals/sex/dose: 5 male rats/group

Route of administration: O ral

– in diet

Vehicle: 2% corn oil

Doses/concentrations tested: 0.0, 0.1 or 1.0% of the compound, with 2.0% corn oil

Statistical methods used: Not stated

Test method/co nditions remarks: Groups of 5 male rats were fed diets containing 0.0, 0.1 or

1.0% of the compound, with 2.0% corn oil for 13 days. These animals consumed 103 and 978

mg/kg/day of the test compound on the low and high dose diets, respectively. Number of

treatments = 13, number of days of experiment = 14

RESULTSDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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99 Endpoint value(s): NOEL ≤ 978 mg/kg/day

Results remarks: The high dose animals had reduced feed intake during the first 3 days of the

experiment, but feed consumption returned to normal for the remainder of the experiment. Feed

intake and weight gain by the low

- dose animal s was normal. Mean body weight of the high dose

animals lagged behind the other animals throughout the experiment as a result of their initial

slightly reduced feed intake. A slight amount of hair loss was noted on the necks and backs of 2

high dose anim als; no other clinical signs were noted. No differences between treated and

control animals were seen in the following hematological parameters: red blood cell count,

hemoglobin and hematocrit, and total differential white cell counts. A slight reductio n in platelet

count and MCV was noted in the blood from high dose animals, but only 2 samples were

available for study. With the exception of a slight reduction in absolute liver weights in the high

dose animals, absolute and relative liver and kidney wei ghts were unaffected by treatment. The

liver weight decrease in the high dose animals reflected the lower terminal body weight of these

animals. No differences between treated and control animals were seen in the following clinical

chemistries: alanine aminotransferase, alkaline phosphatase, creatinine, urea nitrogen and

glucose. One high dose animal had slightly elevated aspartate aminotransferase and moderately

elevated lactate dehydrogenase; these clinical chemistries in all other animals were normal . No

gross or histopathologic lesions related to the compound were identified.

CONCLUSIONS

No site of toxic action was identified, and the no

- effect dose was estimated to be near but

somewhat below 978 mg/kg/day.

STUDY RELIABILITY

2 = reliable wi th restrictions

REFERENCE

Toxicity and health hazard summary, MSDS and basic toxicity of benzotriazole with cover letter

dated 041989. Submitting Organization: Eastman Kodak Company. OTS0516745.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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100 Study 2: Repeated i.p. injections in rats

Title: Eval uation of benzotriazole as a copper and iron chelator after intraperitoneal injection in

the rat with attachments and cover letter dated 061289.

The study is: Adequate Not Adequate

[Purpose of study is not appropriate for repeated

- dose toxicity; not a standard SIDS endpoint

study; non standard route]

TEST SUBSTANCE

Identity: Benzotriazole, Photo Grade X19BT

- 5589, Sample No. 75109, CAS No. 95

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Remarks: Fine white material, test solutions were prepared by in corporating the com p ound into

a mixture of 75% propylene glycol and 25% distilled water.

METHOD

Method/guideline followed: Not stated

Test type: Intraperitoneal study

GLP compliant?: Not stated

Year study performed: 1975

Test species: Sprague

- Dawley al bino rats, from the Charles River Breeding Laboratories.

Number of animals/sex/dose: 5/sex

Route of administration: I ntraperitoneal injection

Vehicle: Mixture of 75% propylene glycol and 25% distilled water

Exposure duration: Single injection for five consecutive days

Doses/concentrations tested: 2 ml/kg

- bw

Statistical methods used: Not stated

Test method/conditions remarks: Benzotriazole was injected intraperitoneally at 150 mg/kg of

body weight to a group of five female and five male rats. A si ngle injection was given for 5

consecutive days and the animals were euthanized. Blood and liver samples were analyzed for

copper and iron levels to determine the chelating affect, if any, of the test compounds. Body

weight and hematological observations were also made.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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101 Following a two

- week acclimation period, 5 males and 5 females were randomly assigned to a

treatment group and the remaining 10 rats to a vehicle control group. The test rats received a

single intraperitoneal injection of the test solutio n for 5 consecutive days at the rate of 2 ml/kg of

body weight. The control animals received a similar daily injection of the vehicle alone.

All rats were weighed on one occasion prior to treatment, each day prior to injection and at

termination.

Observa tions of appearance and behavior were made frequently after dose.

After 5 consecutive daily doses, t he animals were anesthetized with CO2. The maximum

amount of whole blood was collected via the descending aorta and analyzed for iron and copper

leve l s. H ematocrit, er yth rocyte count and hemoglobin values were also determined on all

animals.

A gross examination of the major organs was made and the liver was removed in toto, weighed,

cut into 40 mm blocks and analyzed for iron and copper levels.

RESULTS

E ndpoint value(s): None stated

Results remarks: No animals died during the five day study. Body weight gain appears

affected by the daily treatment of benzotriazole. Hematological parameters as well as copper

and iron levels in the blood and liver were within acceptable limits or comparable between

treated and control animals.

A dose level of 150 mg/kg was selected based on mortality and lethargy observations noted at

doses of 185 mg/kg/day. All treated rats seem to tolerate the dosing well and there w ere no

deaths or adverse reactions.

Body weight gain was not remarkable but within acceptable limits. The slight depression of

weight gain among the treated rats is compound related.

Hematological observations are comparable among treated and control.

C opper analysis of the blood and liver are comparable among treated and control groups.

Iron analysis of the blood and liver are within acceptable limits and show no decrease among

treated animals as compared to the control.

Liver weights show a slight inc rease among the treated males as percent of body weight. This

increase is due mainly to less weight gain in the treated animals as compared to the controls.

The females remain comparable.

CONCLUSIONS

Based on the results of this study, benzotriazole d oes not appear to be a copper or iron chelator.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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102 STUDY RELIABILITY

(4) Not a standard SIDS endpoint study

REFERENCE

Evaluation of benzotriazole as a copper and iron chelator after intraperitoneal injections in the rat

with attachments and cover letter d ated 061289. Submitting Organization: Sherwin Williams

Company. Performing Laboratory: Huntingdon Research Center. OTS0520638.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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103 B

- 7

. GENETIC TOXICITY

Summary of endpoint: Adequate

Study 1: Mammalian gene mutation assay (in vitro)

Title: Genetic toxicity ‘in vitro’

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Benzotriazole, CAS No. 95

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Remarks: Substance type: organic

Physical status: solid

Purity: ca. 100 % w/w

METHOD

Method/gui deline followed: OECD Guide

- line 476 "Genetic Toxicology: In vitro Mammalian

Cell Gene Mutation Tests"

Test type: HGPRT assay

GLP compliant?: Y es

Year study performed: 1987

Test species: CHO cells

Metabolic activation: With and without

Concentrations tested: without: 150, 300, 400, 500, 600, 800, 1000 μg/ml; with: 50, 100, 200,

350, 500, 700, 1000 μg/ml

Statistical methods used: Not stated

Test method/conditions remarks: Not tested

RESULTS

Genotoxicity result: Negative

Cytotoxic concentration (if applicable): without: 1000 μg/ml rel. survival: 82.4 % with: 500

μg/ml_ 19.1 %, 1000 μg/ml: 0.0 %Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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104 Results remarks: None stated

CONCLUSIONS

The test substance was not considered to be mutagenic.

STUDY RELIABILITY

1 = valid without restriction. GLP Gui deline study

REFERENCE

IUCLID (2001). Hazleton Biotechnologies , The Netherlands, at the request of Bayer AG, 1987.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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105 Study 2: Bacterial reverse mutation assay

Title: Genetic toxicity ‘in vitro’

The study is: Adequate Not Adequate

[Lacking study details.]

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: Not stated

Test type: Ames test

GLP compliant?: N o data

Year study performed: Not stated

Te st species: Salmonella typhimurium

Metabolic activation: N o data

Concentrations tested: Not stated

Statistical methods used: Not stated

Test method/conditions remarks: Not stated

RESULTS

Genotoxicity result: Positive

Cytotoxic concentration (if app licable): N o data

Results remarks: Not stated

CONCLUSIONS

The test substance was considered to be mutagenic.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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106 STUDY RELIABILITY

4 = not assignable. Lacking study details.

REFERENCE

IUCLID (2001). Dunkel, v.c.; Simmon, V.F. in: IARC Publication vo l. 27 (1980), S. 283

- 302.

IUCLID (2001). Zeiger, Errol in: Cancer Research 47 (1987), S. 1287

- 1296 (zitiert in : Ashby,

John; Tennart, Raymond in: Mutation Research 204 (1988), S. 17

- 115).Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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107 Study 3: Mouse micronucleus tests ( in vivo )

Title: Genetic tox icity ‘in vivo’

The study is: Adequate Not Adequate

[Lacking study details.]

TEST SUBSTANCE

Identity: Benzotriazole, CAS No. 95

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Remarks: Substance type: organic

Physical status: solid

Purity: ca. 100 % w/w

METHOD

Method/guideline followed: OECD Guide

- line 474 "Genetic Toxicology: Micronucleus Test"

Test type: Micronucleus assay

GLP compliant?: Y es

Year study performed: 1988

Test species: NMRI mice

Metabolic activation: Not stated

Concentrations tes ted: 0, 800 mg/kg bw

Statistical methods used: Not stated

Test method/conditions remarks: Route of administration

– gavage, exposure period

- once

RESULTS

Genotoxicity result: Negative

Cytotoxic concentration (if applicable): Not stated

Results rema rks: Not stated

CONCLUSIONSDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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108 The test substance was not considered to be mutagenic.

STUDY RELIABILITY

4 = not assignable. Lacking study details (1

– valid with restriction. GLP Guideline study

– per

IUCLID Dataset.]

REFERENCE

IUCLID (2001). Bayer AG data, 1988.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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109 Study 4: Bacterial reverse mutation assay

Title: Salmonella/mammalian

- microsome mutagenicity test with TK 11’237 with cover letter

dated 072893

The study is: Adequate Not Adequate

[Findings were eq uivocal]

TEST SUBSTANCE

Identity: 1H

- benzotriazole, 95

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Remarks: Not stated

METHOD

Method/guideline followed: The tests were carried out in accordance with the method

described by Ames et al.

Test type: Ames

GLP compliant?: Not stated

Year study performed: 1982

Test species: Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538

Metabolic activation: With and without

Concentrations tested: 444, 667, 1000, 1500 and 2250 μg/0.1 ml and 500, 1000, 2000, 4000

and 8000 μg/0.1 ml.

Statistical methods used: Not stated

Test method/conditions remarks: TK 11 237 was tested for mutagenic effects on histidine

auxotrophic mutants of Salmonella typhim urium . The investigations were performed with the

following concentrations of the trial substance without and with microsomal activation: 444,

667, 1000, 1500 and 2250 μg/0.1 ml. In order to confirm the results, the experiments on strains

TA 100, TA1535 and TA 1537 were repeated with the concentrations of 500, 1000, 2000, 4000

and 8000 μg/0.1 ml. The substance was dissolved in acetone. Acetone alone was used for the

negative controls (the substances and vehicles used for positive controls are indicated below).

Each Petri dish contained: 1) approx. 20 ml of minimum agar, 2) 0.1 ml of the solution of the test

substance or the vehicle and 0.1 ml of a bacterial culture in 2.0 ml of soft agar. The soft agar

was composed of: 100 ml of 0.6% agar solution wit h 0.6% NaCl and 10 ml of a solution of 1

- histidine, 0.5 mM and +biotin 0.5mM. In the experiments in which the substance wasDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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110 metabolically activated, 0.5 ml of an activation mixture was added also. 1 ml activation mixture

contained: 0.3 ml S9 fraction of liver from Chinese hamsters induced with Aroclor 1254 and 0.7

ml of a solution of co

- factors.

Positive control experiments were carried out simultaneously with the following substances: 1)

for strain TA98: daunorubicin

- HCl, 5 and 10 μg/0.1 ml phosphate buf fer; 2) for strain TA100: 4

- nitroquinoline

- n

- oxide, 0.125 and 0.25 μg/0.1 ml phosphate buffer; 3) for strain TA1535: n

methyl

- n’

- nitro

- n

- nitrosoquanidine, 3 and 5 μg/0.1 ml phosphate buffer; 4) for strain TA1537:

9(5)animoacridine hydrochloride monohydrate , 50 and 100 μg/0.1 ml DMSO; 5) for strain

TA1538: 2

- nitrofluorene, 5 and 10 μg/0.1 ml DMSO. The activation mixture was tested with

strain TA1535 and cyclophosphamide, 250 μg/0.1 ml phosphate buffer.

In the experiments without and with the addition of mic rosomal activation mixture three Petri

dishes were prepared per strain and per group (i.e. per concentration or per control group).

The plates were incubated for about 48 hours at 37°C in darkness.

When the colonies had been counted, the arithmetic mean wa s calculated. The test substance is

generally considered to be non

- mutagenic of the colony count in relation to the negative control

is not doubled at any concentration.

These tests permit the detection of point mutations in bacteria induced by chemical s ubstances.

Any mutagenic effects of the substances are demonstrable on comparison of the numbers of

bacteria in the treated and control cultures that have undergone back

- mutation to histidine

prototrophism. To ensure that mutagenic effects of metabolites of the test substances formed in

mammals would also be detected, experiments were performed in which the cultures were

additionally treated with and activation mixture (Chinese hamster liver microsomes and co

factors)

RESULTS

Genotoxicity result: Weakly p ositive

Cytotoxic concentration (if applicable): None stated

Results remarks: In the experiments performed without microsomal activation, treatment with

TK 11237 led to a light increase in the number of back

- mutant colonies of strain TA1535. This

eff ect was observed at the concentrations of 1000 and 1500 μg/0.1 ml.

In the experiments carried out with microsomal activation, treatment with TK 11237 led to a

slight increase in the number of back

- mutant colonies of strain TA1535 at the concentrations of

444 to 2250 μg/0.1 ml.

After exposure to TK 11237 in th e concentrations of 4000 and 8000 μg/0.1 ml in the experiments

on strains TA100, TA 1535 and TA1537, the number of histidine

- prototrophic mutants was

reduced, as a result of the inhibitory effect of the substance on the growth of the bacteria.

CONCLUSIONS

In the experiments performed without and with microsomal activation on strain TA 1535

treatment with TK 11237 led to a slight increase in the number of back

- mutant colonies.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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111 TK 11237 and its metabolites formed as a result of microsomal activation thus exerted an albeit

weak mutagenic action in this test system.

STUDY RELIABILITY

3 = not reliable

REFERENCE

Salmonella/mammalian

- microsome mutagenicity test with TK 11’237 with cover letter dated

072893. Submitting Organization: Ciba

- Geigy Corporation . OTS0538209.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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112 Study 5: Bacterial reverse mutation assay

Title: Salmonella/mammalian

- microsome mutagenicity test with TK 11’237 with cover letter

dated 072893

The study is: Adequate Not Adequate

[Findings were e quivocal]

TEST SUBSTANCE

Identity: 1H

- benzotriazole, 95

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Remarks: None stated

METHOD

Method/guideline followed: The tests were carried out in accordance with the method

described by Ames et al.

Test type: Ames

GLP compliant?: Not stated

Yea r study performed: Not stated

Test species: Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538

Metabolic activation: With and without

Concentrations tested: 25, 75, 225, 675 and 2025 ug/0.1 ml. Results were confirmed with

testing at concentrations of 444, 667, 1000, 1500 and 2250 ug/0.1

ml

Statistical methods used: Not stated

Test method/conditions remarks: TK 11 237 was tested for mutagenic effects on histidine

auxotrophic mutants of Salmonella typhimurium . The investigations were performed with the

following concentrations of the trial substance without and with microsomal activation: 25, 75,

225, 375 and 2025 μg/0.1 ml. In order to confirm the results, the experiments were repeated with

the concentrations of 444, 667, 1000, 150 0 and 2250 μg/0.1 ml. The substance was dissolved in

acetone. Acetone alone was used for the negative controls (the substances and vehicles used for

positive controls are indicated below). Each Petri dish contained: 1) approx. 20 ml of minimum

agar, 2) 0.1 ml of the solution of the test substance or the vehicle and 0.1 ml of a bacterial culture

in 2.0 ml of soft agar. The soft agar was composed of: 100 ml of 0.6% agar solution with 0.6%

NaCl and 10 ml of a solution of 1

- histidine, 0.5 mM and +biotin 0.5 mM. In the experiments inDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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113 which the substance was metabolically activated, 0.5 ml of an activation mixture was added also.

1 ml activation mixture contained: 0.3 ml S9 fraction of liver from rats induced with Aroclor

1254 and 0.7 ml of a solution of co

- f actors.

Positive control experiments were carried out simultaneously with the following substances: 1)

for strain TA98: daunorubicin

- HCl, 5 and 10 μg/0.1 ml phosphate buffer; 2) for strain TA100: 4

- nitroquinoline

- n

- oxide, 0.125 and 0.25 μg/0.1 ml phosphate buffer; 3) for strain TA15 35: n

methyl

- n’

- nitro

- n

- nitrosog uanidine, 3 and 5 μg/0.1 ml phosphate buffer ; 4) for strain TA1537:

9(5)amin oacridine hydrochloride monohydrate, 50 and 100 μg/0.1 ml DMSO; 5) for strain

TA1538: 2

- nitrofluorene, 5 and 10 μg/0.1 ml DMSO. The activation mixture was tested with

strain TA1535 and cyclophosphamide, 250 μg/0.1 ml phosphate buffer.

In the experiments without and with the addition of microsomal activation mixture three Petri

dishes were prepared per strain and per group ( i.e. per concentration or per control group).

The plates were incubated for about 48 hours at 37°C in darkness.

When the colonies had been counted, the arithmetic mean was calculated. The test substance is

generally considered to be non

- mutagenic of the colony count in relation to the negative control

is not doubled at any concentration.

These tests permit the detection of point mutations in bacteria induced by chemical substances.

Any mutagenic effects of the substances are demonstrable on comparison o f the numbers of

bacteria in the treated and control cultures that have undergone back

- mutation to histidine

prototrophism. To ensure that mutagenic effects of metabolites of the test substances formed in

mammals would also be detected, experiments were p erformed in which the cultures were

additionally treated with and activation mixture (rat liver microsomes and co

- factors).

RESULTS

Genotoxicity result: Weak p ositive

Cytotoxic concentration (if applicable):

Results remarks: In the experiments performe d without and with microsomal activation on

strain TA 1535 treatment with TK 11237 led to a slight increase in the number of back

- mutant

colonies at the concentrations of 1500 and 2025 μg/0.1 ml.

CONCLUSIONS

TK 11237 thus exerted an albeit very weak mu tagenic action in this test system.

STUDY RELIABILITY

3 = not reliable

REFERENCEDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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114 Salmonella/mammalian

- microsome mutagenicity test with TK 11’237 with cover letter dated

072893. Submitting Organization: Ciba

- Geigy Corporation. OTS0538210.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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115 Study 6: Bacterial reverse mutation assay

Title: Salmonella/mammalian

- microsome mutagenicity test with TK 10 637 with cover letter

dated 072893

The study is: Adequate Not Adequate

[Findings were equivocal]

TEST SUBSTANCE

Identity: TK 10 637, benzotriazole, CAS No. 95

- 14

- 7

Remarks: None stated

METHOD

Method/guideline followed: The tests were carried out in accordance with the method

described by Ames et al.

Test type: Ames assay

GLP compliant?: Not stated

Year stu dy performed: 1982

Test species: Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 1538

Metabolic activation: With and without

Concentrations tested: 25, 75, 225, 675 and 2025 ug/0.1 ml; 50, 150, 450, 1350 and 4050

ug/0.1 ml in the c onfirmation assay

Statistical methods used: Not stated

Test method/conditions remarks: TK 10 637 was tested for mutagenic effects on histidine

auxotrophic mutants of Salmonella typhimurium . The investigations were performed on strains

TA 98, TA 100, TA 1535 and TA 1537 with the following concentrations of th e trial substance

without and with microsomal activation: 25, 75, 225, 675 and 2025 μg/0.1 ml. In order to

confirm the results, additional experiments were performed on all strains including TA 1538 with

the concentrations of 50, 150, 450, 1350 and 4050 μ g/0.1 ml. The substance was dissolved in

acetone. Acetone alone was used for the negative controls (the substances and vehicles used for

positive controls are indicated below). Each Petri dish contained: 1) approx. 20 ml of minimum

agar, 2) 0.1 ml of th e solution of the test substance or the vehicle and 0.1 ml of a bacterial culture

in 2.0 ml of soft agar. The soft agar was composed of: 100 ml of 0.6% agar solution with 0.6%

NaCl and 10 ml of a solution of 1

- histidine, 0.5 mM and +biotin 0.5mM. In the experiments inDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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116 which the substance was metabolically activated, 0.5 ml of an activation mixture was added also.

1 ml activation mixture contained: 0.3 ml S9 fraction of liver from rats induced with Aroclor

1254 and 0.7 ml of a solution of co

- factors.

Pos itive control experiments were carried out simultaneously with the following substances: 1)

for strain TA98: daunorubicin

- HCl, 5 and 10 μg/0.1 ml phosphate buffer; 2) for strain TA100: 4

- nitroquinoline

- n

- oxide, 0.125 and 0.25 μg/0.1 ml phosphate buffer; 3) for strain TA15 35: n

methyl

- n’

- nitro

- n

- nitrosog uanidine, 3 and 5 μg/0.1 ml phosphate buff er; 4) for strain TA1537:

9(5)amin oacridine hydrochloride monohydrate, 50 and 100 μg/0.1 ml DMSO; 5) for strain

TA1538: 2

- nitrofluorene, 5 and 10 μg/0.1 ml DMSO. The activation mixture was tested with

strain TA1535 and cyclophosphamide, 250 μg/0.1 ml phosphate buffer.

In the experiments without and with the addition of microsomal activation mixture three Petri

dishes were prepared per strain and per group (i.e. per c oncentration or per control group).

The plates were incubated for about 48 hours at 37°C in darkness.

When the colonies had been counted, the arithmetic mean was calculated. The test substance is

generally considered to be non

- mutagenic of the colony cou nt in relation to the negative control

is not doubled at any concentration.

These tests permit the detection of point mutations in bacteria induced by chemical substances.

Any mutagenic effects of the substances are demonstrable on comparison of the numb ers of

bacteria in the treated and control cultures that have undergone back

- mutation to histidine

prototrophism. To ensure that mutagenic effects of metabolites of the test substances formed in

mammals would also be detected, experiments were performed i n which the cultures were

additionally treated with and activation mixture (rat liver microsomes and co

- factors).

RESULTS

Genotoxicity result: Weak p ositive

Cytotoxic concentration (if applicable):

Results remarks: In the experiments performed withou t and with microsomal activation

treatment with TK 10 637 led to an increase in the number of back

- mutant colonies of strains TA

98, TA 1535, TA 1537 and TA 1538 at the concentrations of 25 or 50 μg/0.1 ml and above. A

slight increase in the number of bac k

- mutants was also observed in the experiment on strain TA

1535 at the concentration of 2025 μg/0.1 ml.

In the experiments carried out with microsomal activation, this effect was less pronounced. An

increase in the number of back

- mutant colonies occurred in the experiments on strains TA 98,

TA 1535 and TA 1538 at the concentrations of 675 μg/0.1 ml and above. With strain TA 1538

this effect was observed at the concentrations of 150 μg/0.1 ml and above.

After exposure to TK 10 637 in the concentration of 4050 μg/0.1 ml in the experiments without

activation no histidine

- prototrophic mutants were found, as a result of the inhibitory effect of theDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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117 substance on the growth of the bacteria. In the experiment with activation this effect was only

observed with s trains TA 100 and TA 1535.

An inhibitory effect of TK 10 637 on the growth of the bacteria was observed at the

concentration of 4050 μg/0.1 ml.

CONCLUSIONS

The test substance was considered to be mutagenic.

STUDY RELIABILITY

3 = not reliable

REFEREN CE

Salmonella/mammalian

- microsome mutagenicity test with TK 10 637 with cover letter dated

072893. Submitting Organization: Ciba

- Geigy Corporation. OTS 538213.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

118 Study 7: Chromosomal aberrations assay

Title: Chromosomal Aberrations

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: 1,2,3,

- Benzotriazole, CAS # 95

- 14

- 7

Remarks: Not specified

METHOD

Method/guideline followed: Not specified

Test type: Chromosome Aberrations

GLP compliant?: Not spec ified

Year study performed: 1985

Test species: CHO cells

Metabolic activation: With and without

Concentrations tested: 1000, 1500, 2000, 2500 ug/mL (without activation)

503, 1257, 2513, 3750 ug/mL (with activation)

Statistical methods used: Not specifie d

Test method/conditions remarks: Two chromosomal aberrations tests were conducted; one

without activation and one with activation. The study without activation included dimethyl

sulfoxide as the vehicle control, and mitomycin

- c as the positive control . The study with

activation induced in rat liver S9 included dimethyl sulfoxide as the vehicle control, and

cyclophosphamide as the positive control. The harvest time without activation was 21.5 hours,

and 12.5 hours with activation.

RESULTS

Genotox icity result: Positive (with activation), Negative (without activation)

Cytotoxic concentration (if applicable): Not specified

Results remarks: None statedDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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119 CONCLUSIONS

The test substance was considered to be mutagenic.

STUDY RELIABILITY

[1] Reliable without restrictions. The study is acceptable and within necessary guidelines.

REFERENCE

NTP Study #770824, 1985.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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120 Study 8: Sister chromatid exchange assay

Title: Sister Chromatid Exchange

The study is: Adequate N ot Adequate

TEST SUBSTANCE

Identity: 1,2,3,

- Benzotriazole, CAS # 95

- 14

- 7

Remarks: Not specified

METHOD

Method/guideline followed: Not specified

Test type: Sister Chromatid Exchange

GLP compliant?: Not specified

Year study performed: 1985

Test spe cies: CHO cells

Metabolic activation: With and without

Concentrations tested: 50, 167, 500, 502, 1005, 1500, 1670, 2000 ug/mL (without activation)

167, 500, 1670, 2000, 2500, 3000, 3500, 5000 ug/mL (with activation)

Statistical methods used: Not specifie d

Test method/conditions remarks: Four separate sister chromatid exchange tests were

conducted; two without activation and two with activation. The studies without activation

included dimethyl sulfoxide as the vehicle control, and mitomycin

- c as the p ositive control. The

studies with activation induced in rat liver S9 included dimethyl sulfoxide as the vehicle

control, and cyclophosphamide as the positive control.

RESULTS

Genotoxicity result: Positive (without activation), Negative (with activat ion)

Cytotoxic concentration (if applicable): Not specified

Results remarks: None statedDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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121 CONCLUSIONS

The test substance was considered to be mutagenic.

STUDY RELIABILITY

[1] Reliable without restrictions. The study is acceptable and within necessary guidelines.

REFERENCE

NTP Study #770824, 1985.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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122 B

- 8

. REPRODUCTIVE TOXICITY

Summary of endpoint: Adequate

Study 1: Chronic reproductive toxicity study in rats

Title: Toxicity to reproduction

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: 1H

- benzotriazole; purity > 99%, CAS No. 95

- 14

- 7

Remarks: Lot No. 030737

METHOD

Methods /guideline followed: EPA OTS 798.3300

Test type: Chronic dietary carcinogenicity

GLP compliant?: Not stated

Year study performed: 1978

Test species: Fischer 344 rat, males and females

Number of animals/sex/dose: 50/sex

Route of administration: Oral feed

Exposure duration: 78 weeks

Doses/concentrations tested: 0, 6,700, 12,100 mg/kg

- bw

Control group: yes, concu rrent no treatment

Statistical methods used: Not stated

Test methods/conditions remarks: Groups of 50 rats of each sex were administered IH

benzotriazole at one of two time

- weighted average doses, either 6,700 or 12,100 ppm, for 78

weeks. Except for fiv e control and five high

- dose rats of each sex, which were killed at week 78,

all animals surviving at that time were observed for 26

- 27 additional weeks. Controls consisted

of groups of 50 untreated rats of each sex and were observed for 105

- 106 weeks. Al l rats

surviving to weeks 104

- 106 were then killed.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

123 RESULTS

Endpoint value(s): The NOAEL was > 12,100 ppm.

Results remarks: The 78 week oral study of 1H

- benzotriazole in Fischer 344 rats did not find

any evidence of dose

- related pathology in the reprod uctive organs: prostate/testis/epididymis of

males and uterus/ovaries of females. Findings, not significantly different from controls, included

acute inflammation of the prostate, hyperplasia of the testis, acute inflammation, hydrometra, and

hyperplasia o f the uterus, inflammation and cyst formation of the ovaries.

CONCLUSIONS

In conclusion, under conditions of this study, 1H

- benzotriazole in Fischer 344 rats did not find

any evidence of dose

- related pathology in the reproductive organs.

STUDY RELIABILI TY

2 = valid with restrictions. Meets generally accepted scientific standards, well documented and

acceptable for assessment

REFERENCE

IUCLID (2001). NTIS # PB285202/AS. US National Cancer Institute : Bioassay

of 1

- H

- Benzotriazole for possible carcino genicity (1978).Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

124 Study 2: Chronic reproductive toxicity study in mice

Title: Toxicity to reproduction

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: 1H

- benzotriazole; purity > 99%, CAS No. 95

- 14

- 7

Remarks: Lot No. 030737

METHOD

Methods /guideline followed: EPA OTS 798.3300

Test type: Chronic dietary carcinogenicity

GLP compliant?: No data

Year study performed: 1978

Test species: B6C3F1 mice, both males and females

Number of animals/sex/dose: 50/sex

Route of administration: Oral feed

Exposure duration: 104 weeks

Doses/concentrations tested: 0, 11700, 23500 ppm

Control group: yes, concurrent no treatment

Statistical methods used: Not stated

Test methods/conditions remarks: Groups of 50 mice of each sex were administered IH

benzotriazole at one of two time

- weighted average doses, either 11,700 or 23,500 ppm, for 104

weeks, then observed for 2 additional weeks. Controls consisted of groups of 50 untreated mice

of each sex and were observe d for 109 weeks. All mice surviving to weeks 106

- 109 were then

killed.

RESULTS

Endpoint value(s): The parenteral NOAEL was > 23,500 ppm.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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125 Results remarks: The 104 week oral study of 1H

- benzotriazole in mice did not find any

evidence of pathology in the r eproductive organs: prostate/testis/epididymis of males and

uterus/ovaries of females. Findings, not significantly different from controls, included

hyperplasia of the testis, hydrometra and cystic hyperplasia of the uterus, and cyst formation of

the ovari es.

CONCLUSIONS

In conclusion, under conditions of this study, 1H

- benzotriazole in mice did not find any evidence

of pathology in the reproductive organs

.

STUDY RELIABILITY

2 = valid with restrictions. Meets generally accepted scientific standards, w ell documented and

acceptable for assessment

REFERENCE

IUCLID (2001). NTIS # PB285202/AS. US National Cancer Institute : Bioassay

of 1

- H

- Benzotriazole for possible carcinogenicity (1978).Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

126 B

- 9

. CARCINOGENICITY

Study 1: Chronic carcinogenicity study i n rats

Title: Bioassay of 1h

- benzotriazole for possible carcinogenicity (CAS No. 95

- 14

- 7)

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: 1H

- benzotriazole, (CAS No. 95

- 14

- 7)

Remarks: Two lots of IH

- benzotriazole were obtained from Aldrich Chemical Company,

Milwaukee, Wisconsin, and were stored at 4°C.

METHOD

Method/guideline followed: NTP

Test type: Chronic

GLP compliant?: Not stated

Year study performed: 1978

Test species: Fischer 344 rats, obtained from Charles River Breeding Laboratories, Inc.,

Wilmington, Massachusetts.

Number of animals/sex/dose: 50/sex/dose

Route of administration: oral

– in food

Exposure duration: 78 weeks

Doses/concentrations tested: 6,700 or 12,100 ppm

Statist ical methods used: Not stated

Test method/conditions remarks: Groups of 50 rats of each sex were administered 1h

benzotriazole at one of two time

- weighted doses, either 6,700 or 12,100 ppm, for 78 weeks.

Except for five control and five high

- dose rats o f each sex, which were killed at week 78, all

animals surviving at that time were observed for 26

- 27 additional weeks. Controls consisted of

groups of 50 untreated rats of each sex and were observed for 105

- 106 weeks. All rats surviving

to weeks 104

- 106 w ere then killed.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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127 RESULTS

Endpoint value(s): Not stated

Results remarks: Mean body weights of the dosed male and female rats were lower than those

of the corresponding controls throughout most of the bioassay. Survival of animals in dosed and

control gr oups of rats was at least 60%, and sufficient numbers of animals were at risk for

development of late

- appearing tumors.

In male rats, neoplastic nodules of the liver occurred at a statistically significant incidence (P =

0.024) in the high

- dose group when compared with the control group (controls 0/48, low

- dose

0/46, high

- dose 5/45 [11%]). The incidence of this tumor in control Fischer 344 rats used in

similar bioassays of other test chemicals at the same laboratory has varied from 0 to 11%, with

2/13 hist orical

- control groups having incidences of 10

- 11%. Since the incidence in the high

- dose

group is no higher than has been observed in some control groups, these tumors cannot be clearly

associated with administration of the test chemical.

Brain tumors occu rred in three dosed male rats, in one dosed female rat, and in none of the

controls. The occurrence of this rare tumor in dosed animals of each sex is suggestive of, but not

considered as sufficient evidence of, carcinogenicity.

In female rats, the incide nce of endometrial stromal polyps in the low

- dose group was

significantly higher (P = 0.010) than that in the corresponding controls (controls 2/48, low

- dose

10/45, high

- dose 8/50). However, the incidence in the high

- dose group was not significant, and

whe n the incidences of endometrial stromal polyps and endometrial stromal sarcomas were

combined, they were not significant in either the low

- or high

- dose groups. Thus, these tumors

cannot be associated with administration of the chemical.

In Fischer 344 r ats there was an increased incidence of brain tumors, suggesting a possible

carcinogenic effect. However, there was no convincing evidence that under the conditions of this

bioassay IH

- benzotriazole was carcinogenic in Fischer 344 rats of either sex.

CONC LUSIONS

There is no convincing evidence that under the conditions of this bioassay 1H

- benzotriazole was

carcinogenic.

STUDY RELIABILITY

1 = reliable without restrictions

REFERENCE

NTP. 1978. Bioassay of 1h

- benzotriazole for possible carcinogeni city (CAS No. 95

- 14

- 7). U.S.

Department of Health, Education, and Welfare. National Institutes of Health. NCI

- CG0TR

- 88.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

128 Study 2: Chronic carcinogenicity study in mice

Title: Bioassay of 1h

- benzotriazole for possible carcinogenicity (CAS No. 95

- 14

- 7)

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: 1H

- benzotriazole, (CAS No. 95

- 14

- 7)

Remarks: Two lots of IH

- benzotriazole were obtained from Aldrich Chemical Company,

Milwaukee, Wisconsin, and we re stored at 4°C.

METHOD

Method/guideline followed: NTP

Test type: Chronic

GLP compliant?: Not stated

Year study performed: 1978

Test species: B6C3F1 mice, obtained from Charles River Breeding Laboratories, Inc.,

Wilmington, Massachusetts.

Number of animals/sex/dose: 50/sex/dose

Route of administration: oral

– in food

Exposure duration: 104 weeks

Doses/concentrations tested: 11,700 or 23,500 ppm

Statistical methods used: Not stated

Test method/conditions remarks: Groups of 50 mice of each sex w ere administered IH

benzotriazole at one of two time

- weighted average doses, either 11,700 or 23,500 ppm, for 104

weeks, then observed for 2 additional weeks. Controls consisted of groups of 50 untreated mice

of each sex and were observed for 109 weeks. A ll mice surviving to weeks 106

- 109 were then

killed.

RESULTS

Endpoint value(s): Not statedDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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129 Results remarks: Mean body weights of the dosed male and female mice were lower than those

of the corresponding controls throughout most of the bioassay. Surviva l of animals in dosed and

control groups of mice was at least 60%, and sufficient numbers of animals were at risk for

development of late

- appearing tumors.

In male mice, no tumors occurred in dosed groups at incidences that were significantly higher

than those in controls. In female mice, alveolar/bronchiolar carcinomas occurred at a statistically

significant incidence (P = 0.001) only in the low

- dose group when compared with the control

group (controls 0/49, low

- dose 9/49 [18%], high

- dose 3/49 [6%]). The incidence in the high

- dose

group was not significant, and the data did not show a dose

- related trend. It should be noted that

the incidence of these tumors in control B6C3F1 female mice from other bioassays at this

laboratory has varied from 0 to 7%, with a mean of 4%. Therefore, the occurrence of this tumor

in the female mice cannot be clearly related to the administration of the test chemical.

In female B6C3F1 mice there was an increased incidence of alveolar/bronchiolar carcinomas,

suggesting a possibl e carcinogenic effect of iH

- benzotriazole. However, there was no convincing

evidence that under the conditions of this bioassay IH

- benzotriazole was carcinogenic in

B6C3F1 mice of either sex.

CONCLUSIONS

There is no convincing evidence that under the co nditions of this bioassay 1H

- benzotriazole was

carcinogenic.

STUDY RELIABILITY

1 = reliable without restrictions

REFERENCE

NTP. 1978. Bioassay of 1h

- benzotriazole for possible carcinogenicity (CAS No. 95

- 14

- 7).

U.S. Department of Health, Educatio n, and Welfare. National Institutes of Health. NCI

CG0TR

- 88.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

- 7

130 B

- 10

. EYE IRRITATION

Study 1: E ye irritation study in rabbits

Title: Acute dermal toxicity and eye irritation studies on sample BT

- D, sample #SD

- 7413 in

rabbits with attachment and cover l etter dated 061289

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity:

Remarks: The sample used in this study was received from Maumee Chemical Company on

September 29, 1969, and was labeled Sample B T

- D, Sample No. SD

- 7413. The test material is a

fine, yellowish

- tan powder with a slight odor.

METHOD

Method/guideline followed: Not stated

Test type: Eye irritation study

GLP compliant?: Not stated

Year study performed: 1969

Test species: Rabbits, alb ino

Number of animals/sex/dose: 6

Route of administration: E ye

Doses/concentrations tested: 100 mg

Statistical methods used: Not stated

Test method/conditions remarks: One hundred milligrams of the sample, as received, was

applied to the right eye o f each of six albino rabbits. The left eye was untreated and served as a

control. Examinations for gross signs of eye irritation were made at 24, 48, and 72 hours

following application. Scoring of irritative effects was according to the method of Draize

(Lehman, A.J., et al Appraisal of the safety of chemicals in foods, drugs, and cosmetics,

Association of Food and Drug Officials of the U.S., Austin, Texas, 1959) in which corneal, iris,

and conjunctival effects are scored separately. These scores are we ighted and combined into a

total composite score. In this scoring system, injuries to the cornea and iris are heavily weighted

due to the role of these organs in vision and may represent as much as 80 per cent of the totalDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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131 score.

RESULTS

Endpoint value( s): Irritating to rabbit eye

Results remarks: The results following application of Sample BT

- D, Sample No. SD

- 7413, to

the eyes of albino rabbits are shown in table

Rabbit

number Time Cornea Iris

Conjunctivae** Total

score*

Hours Opacity Area Erythema Swelling Discharge

17 24 1 3 1 1 2 2 30

48 1 3 1 1 2 2 30

72 1 3 1 1 1 1 26

18 24 1 4 1 1 3 2 37

48 1 4 1 1 2 2 35

72 1 4 1 1 1 1 31

19 24 2 4 1 1 2 2 55

48 2 4 1 1 2 2 55

72 2 4 1 1 1 1 51

20 24 2 4 1 1 2 2 55

48 2 4 1 1 2 2 55

72 2 4 1 1 1 1 51

21 24 4 2*** 1 1 2 2 55

minimum

48 4 2*** 1 1 2 3 57

minimum

72 4 2*** 1 1 2 2 55

minimum

22 24 2 4 1 1 3 3 59

48 2 4 1 1 3 3 59

72 4 2*** 1 1 2 3 57

minimum

* Total score is the sum of the following three sub

- totals

a) degree of opacity x area involved x 5

b) iris score x 5

c)sum of scores for erythema, swelling, and discharge x 2

Total possible score = 110

** Blanching of upper and/or lower conjunctivae noted in each rabbit throughout the study

*** One

- half of eye exhibited opacity of Grade 2 intensity

Irritative effects included moderate or marked conjunctivitis and corneal opacity, mild iritis, and

conjunctival blanching in each rabbit. These signs remained relatively unchanged throughout the

72

- hour observatio n period.

CONCLUSIONSDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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132 The acute dermal toxicity and the acute eye irritative potential of Sample BT

- D, Sample No. SD

7413, were evaluated in accordance with the techniques specified in the Regulations (Federal

Register, August 12, 1961 et seq) under the Federal Hazardous Substances Act.

Acute eye application produced corneal opacity, iritis, conjunctivitis, and conjunctival blanching

in each rabbit.

Based on these results Sample BT

- D, Sample No. SD

- 7413, is classified as non

- toxic by dermal

application and is an eye irritant as these terms are defined in the above

- cited Regulations.

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Acute dermal toxicity and eye irritation studies on sample BT

- D, sample #SD

- 7413 in rabbits

with attachment and cover letter dated 061289. Submitting Organization: Sherwin

- Williams

Company. Performing Laboratory: Hill Top Research Incorporated. OTS0520633.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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133 Study 2: E ye irritation study in rabbits

Title: Results of range finding toxicological tests on be nzotriazole (purified)

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

- 14

- 7

Remarks: These conclusions are based upon range finding toxicological tests and are limited to

precautions for industrial handling of the material. Development of specific uses will require

consideration of the health problems presented and of the need for further toxicological studies.

METHOD

Method/guideline followed: None stated

Test type: Ey e irritation

GLP compliant?: Not stated

Year study performed: ~ 1960

Test species: R abbit

Number of animals/sex/dose: Not stated

Route of administration: E ye contact

Doses/concentrations tested: undiluted or 0.03% solution in water

Statistical methods used: Not stated

Test method/conditions remarks: None stated

RESULTS

Endpoint value(s):

Results remarks: Material Treatment Response remarks

Undiluted unwashed moderated conjunctival

swelling, moderate corneal injury subsided in one week

Undiluted washed with water moderated conjunctival

swelling, superficial corneal injury subsiding in 48 hours

.03% sol. in water washed & unwashed essentially no irritationDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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134 observed

CONCLUSIONS

The undiluted material is moderately irritating to the eye. Direct contact would likely result in

moderated conjunctival swelling and moderate corneal injury which should subside in a week or

two. It is recommended that safety glasses with side shields be used while handling this material

whenever the likeli hood of eye contact exists. It is important to note that immediate and

thorough washing of contaminated eyes should reduce the degree of corneal injury appreciably.

The 0.03% solution in water studied in the laboratory is essentially nonirritating to the eye.

Special precautions should be unnecessary for safe industrial handling.

Due to the low eye and skin irritating potential of dilute solutions of this material, it would seem

most unlikely that a concentration of 0.03% in DC 2 would render the formula tions irritating.

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Results of range finding toxicological tests on benzotriazole (purified). Submitting

Organization: Dow Corning Corporation. OTS0520472.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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135 Study 3: E ye irritation study in rabbits

Title: Toxicity and health hazard summary, MSDS and basic toxicity of benzotriazole with cover

letter dated 041989

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: 1,2,3

- benzotriazole, CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: None stated

Test type: Eye irritation

GLP compliant?: Not stated

Year study performed: Not stated

Test species: R abbit

Number of animals/sex/dose: 3

Route of administration: E ye

Veh icle: None stated

Doses/concentrations tested: Not stated

Statistical methods used: Not stated

Test method/conditions remarks: Test item was administered as crystals placed in the

conjunctival sacs of rabbit eyes (3 washed, 3 unwashed). Animals were e xamined up to 2 weeks

following administration.

RESULTS

Endpoint value(s): None stated

Results remarks: M oderate eye irritationDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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136 CONCLUSIONS

T he compound was a moderate eye irritant when several crystals were placed in the conjunctival

sacs of rabbit eyes (3 washed, 3 unwashed). All 3 unwashed eyes developed slight to moderate

erythema and edema of the adnexa, slight to moderate corneal opacity, and injection of the irides.

Immediate irrigation of the treated eyes was not palliative, with 2/3 washed eyes having a

purulent discharge at a 24

- hour examination point. This discharge was not present by the 2

- week

examination point.

STUDY RELIABILITY

[2] Reliable with restrictions

REFERENCE

Toxicity and health hazard summary, MSDS and basic toxicity of benzotriazole with cover letter

dated 041989. Submitting Organization: Eastman Kodak Company. OTS0516745.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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137 Study 4: E ye irritation study in rabbits

Title: Eye irritation study in the rabbit of benzotriazole (Cobratec® 99) sample #1088

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Benzotriazole (Cobratec 99), CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: None stated

Test type: Eye irritation

GLP compliant?: Not stat ed

Year study performed: Not stated

Test species: albino New Zealand rabbits

Number of animals/sex/dose: 6 female

Route of administration: E ye

Vehicle: None stated

Doses/concentrations tested: 100 mg

Statistical methods used: Not stated

Test method /conditions remarks: 100 mg of test material was placed in one eye of each animal

by gently pulling the lower lid away from the eyeball to form a cup into which the test substance

was dropped. The lids were gently held together for one second and the anim al released. The

other eye, remaining untreated served as a control. The eyes were examined and the grade of

ocular reaction recorded at 24, 48, 72 hours and 14 days. Reading of reactions was facilitated by

the use of an opthalmoscope. After the record ing of observations at 24 hours, the eyes were

stained with fluorescein and examined with an ultra

- violet lamp to detect corneal damage if

present. The reactions were scored from : “Illustrated guide for grading eye irritation by

hazardous substances”, s ection 191.12 of the Federal Hazardous Substances labeling Act

Regulations.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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138 RESULTS

Endpoint value(s): None stated

Results remarks: Non

- irrigated No. Obs./No. Exposed

Cornea 1/6

– translucent areas of opacity

1/6

– no details of iris vis ible

4/6

– complete corneal opacity

6/6

– fluorescein examination positive

Iris 5/6

– moderate circumcorneal

Conjunctival redness 4/6

– diffuse crimson red

2/6

– diffuse beefy red

Chemosis 2/6

– swelling with lids about half closed

4/6

– swelling w/lids half closed or greater

CONCLUSIONS

Benzotriazole is considered to be an eye irritant.

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Eye irritation study in the rabbit of benzotriazole (Cobratec ® 99) sample #1088 . Submitting

Organization: Sherwin

- Williams. Performing Laboratory: Huntingdon Research Center.

OTS0516797.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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139 Study 5: E ye irritation study in rabbits

Title: Eye irritation study in the rabbit of benzotriazole (Cobratec 99) sample #1 088 with

attachments and cover letter dated 061289

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

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Remarks: Cobratec 99, Sample # 1088

METHOD

Method/guideline follo wed: None stated

Test type: Eye irritation

GLP compliant?: Not stated

Year study performed: Not stated

Test species: albino New Zealand rabbits

Number of animals/sex/dose: 6 female

Route of administration: E ye

Vehicle: None stated

Doses/concentration s tested: 100 mg

Statistical methods used: Not stated

Test method/conditions remarks: Six female albino New Zealand rabbits were used. Both

eyes of each animal were examined with the aid of an op h thalmoscope before testing. 100 mg of

test material was placed in one eye of each animal by gently pulling the lower lid away from the

eyeball to form a cup into which the test substance was dropped. The lids were gently held

together for one second and the animal released. The other eye, remaining untreated served as a

control.

The eyes were examined and the grade of ocular reaction recorded at 24, 48, 72 hours and 14

days. Reading of the reactions was facilitated by the use of an op h thalmoscope. After the

recording of observations at 24 hours, the eyes w ere stained with fluorescein and examined with

an ultraviolet lamp to detect corneal damage of present.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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140 The reactions were scored from: “Illustrated guide for grading eye irritation by hazardous

substances”, Section 191.12 of the Federal Hazardous Substanc es labeling Act

RESULTS

Endpoint value(s): None stated

Results remarks: Non

- irrigated No. Obs./No. Exposed

Cornea 1/6

– translucent area of opacity

1/6

– no details of iris visible

4/6

– complete corneal opacity

6/6

– fluor escein examination positive

Iris 5/6

– moderate circumcorneal

Conjunctival redness 4/6

– diffuse crimson red

2/6

– diffuse beefy red

Chemosis 2/6

– swelling with lids about half closed

4/6

– swelling w/lids half closed or great er

Ocular reactions elicited by benzotriazole

Rabbit

No. Day Positive

(+)

1 2 3 14 Negative

(

- )

1178 Cornea 2 2 2 3 +

Iris 1 1 0 0 +

Conjuctiva Redness 2 2 2 2 +

Chemosis 4 3 3 3 +

Fluor. exam + + + +

1179 Cornea 3 3 3 4 +

Iris 1 1 1 1 +

Conjuctiva Redness 3 2 2 1 +

Chemosis 3 3 3 2 +

Fluor. exam + +

- +

1180 Cornea 2 2 3 4 +

Iris 1 1 1 1 +

Conjuctiva Redness 2 2 2 0 +

Chemosis 3 3 4 1 +

Fluor. exam + + + +

1181 Cornea 1 2 2 2 +

Iris 1 1 1 0 +

Conjuctiv a Redness 1 2 2 0 +

Chemosis 3 3 3 1 +

Fluor. exam + +

- +

1182 Cornea 4 4 4 3 +

Iris Could not read because of

opacity 0

- Conjuctiva Redness 3 3 2 0 +Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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141 Chemosis 4 4 3 1 +

Fluor. exam + + + +

1183 Cornea 3 3 2 4 +

Iris 1 1 1 0 +

Conjuctiva Redness 2 2 1 1 +

Chemosis 4 4 3 2 +

Fluor. exam + + + +

Cornea

No ulceration or opacity 0

Scattered or diffuse areas of opacity (other than slight dulling

of normal luster), details of iris clearly visible 1 (positive reaction)

Easily discernible translucent areas, details of iris slightly

obscured 2

Nacreous areas, no details of iris visible, size of pupil barely

discernible 3

Complete corneal opacity, iris not discernible 4

Iris

Normal 0

Markedly deepene d folds, congestion, swelling, moderate circum

corneal injection (any of these combination of any thereof),

iris still reacting to light (sluggish reaction is positive) 1 (positive reaction)

No reaction to light, hemorrhage, gross destruction (any or al l

of these) 2

Conjunctivae

Redness (refers to palpebral and bulbar conjunctivae excluding

cornea and iris)

Vessels normal 0

Some vessels definitely injected 1

Diffuse, crimson red, individual vessels not easily discernible 2 (positive reac tion)

Diffuse beefy red 3

Chemosis

No swelling 0

Any swelling above normal (included nictitating membrane) 1

Obvious swelling with partial eversion of lids 2 (positive reaction)

Swelling with lids about half closed 3

Swelling with lids h alf closed to completely closed 4

CONCLUSIONS

According to the results of this study benzotriazole is considered to be an eye irritant.

STUDY RELIABILITY

2 = reliable with restrictionsDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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142 REFERENCE

Eye irritation study in the rabbit of benzotriazo le (Cobratec 99) sample #1088 with attachments

and cover letter dated 061289. Submitting Organization: Sherwin

- Williams. Performing

Laboratory: Huntingdon Research Center. OTS0520636.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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143 Study 6: E ye irritation study in rabbits

Title: Letter from Polaro id Corporation regarding the enclosed results of the acute toxicity,

corneal and dermal irritation studies on 1h

- benzotriazole with attachments

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: 1h

- ben zotriazole, CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: None stated

Test type: Eye irritation

GLP compliant?: Not stated

Year study performed: ~ 1965

Test species: New Zealand albino rabbits, weighing between 2.0 and 2.5 kg

Number of animals/sex/dose: Both sexes used, number per sex not specified. Total of 6 eyes

exposed.

Route of administration: E ye

Vehicle: None stated

Doses/concentrations tested: 100 mg

Statistical methods used: Not stated

Test method/conditions r emarks: The method consists in dusting 100 mg of either a test or

control substance (sodium chloride, sucrose, clean sand) into the eye of the rabbit.

Young adult New Zealand albino rabbits of both sexes weighing between 2.0 and 2.5 kg were

anesthetized w ith intravenously administered pentobarbital sodium 50 mg/ml (25 to 30 mg/kg).

A total of six eyes were used per compound. The eyelids were held open by gently pulling

hemostats placed on the margins of each lid using appropriate precautions to minimize the

degree of mechanical trauma. Each powdered (100 mesh) sample was introduced into the eye

over the corneal surface and followed immediately by the instillation of 0.2 ml of distilled water

to moisten the powder and simulate the effects of lacrimal flui d. After an exposure period of fiveDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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144 minutes the test material was removed from the eye by copious rinsing (50 ml minimum volume)

with sterile physiological saline.

The extent of injury to the cornea, iris and bulbar and palpebral conjunctivae were scored

individually and also pooled as suggested by Draize.

RESULTS

Endpoint value(s):

Results remarks: Corneal irritation studies were carried out on powdered samples. In addition,

eyes were exposed to powdered (100 mesh) samples of sodium chloride, sucros e and clean sand.

The latter substance served as a control for any irritation which might result from the mechanical

effects of a foreign body in the eye.

Corneal irritation was noted among the resulting samples in varying degrees of severity at the

end of the 24 hours. This irritation persisted for 7 days.

Powdered sodium chloride, sucrose and sand were similarly instilled into the eyes of animals in

which the eyelids were held open and found to produce no corneal irritation after 24 hours and

7 days. Circumcorneal injection was observed 24 hours after exposure in one of two animals

receiving sand and in one animal exposed to sodium chloride. It was not found when re

examined 7 days after treatment. Mild conjunctivitis was observed in all instances i mmediately

after treatment. After 24 hours one of the two eyes exposed to sand exhibited a mild

conjunctivitis. After 7 days both eyes showed mild conjunctivitis. In view of the delay in the

appearance of this conjunctival lesion in one case, the etiolo gy of the lesion and its significance

are in doubt.

Severe staining of the ocular tissues was noted, nasal secretions were also stained. In view of

these observations the risk of accidental poisoning, without awareness on the part of the subject,

appears to be rather small.

CONCLUSIONS

The test substance was highly irritating

STUDY RELIABILITY

2 = reliable with restrictions (multiple materials were tested; data is often all inclusive)

REFERENCE

Letter from Polaroid Corporation regarding the enclose d results of the acute toxicity, corneal and

dermal irritation studies on 1h

- benzotriazole with attachments . Submitting Organization:

Polaroid Corporation. OTS0520182.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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145 Study 7: E ye irritation study in rabbits

Title: Eye irritation

The study is: Adequate Not Adequate

[Lacking important study details.]

TEST SUBSTANCE

Identity: BT

- D (benzotriazole) sample no. SD

- 7413 (Sherwin Williams Co.); CAS No. 95

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Remarks: purity not noted

METHOD

Method/guideline followed: Draize test

Test type: E ye irritation

GLP compliant?: No data

Year study performed: Not stated

Test species: rabbit

Number of animals/sex/dose: Not stated

Route of administration: E ye

Vehicle: None

– substance delivered undiluted

Doses/con centrations tested: 100 mg (undiluted)

Statistical methods used: Not stated

Test method/conditions remarks: E xposure time: 72 hours

RESULTS

Endpoint value(s):

Results remarks: Irritative effects included moderate or marked conjunctivitis and corneal

opacity, mild iritis, and conjunctival blanching in each rabbit. These signs remained relatively

unchanged throughout the 72 hour observation period.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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146 CONCLUSIONS

Highly irritating

STUDY RELIABILITY

4 = not assignable. Lacking study details

REFERENC E

IUCLID (2001). Hill Top Research, Inc. Report T

- 413. October 29, 1969.

IUCLID (2001). PMC Specialties Group, Inc. MSDS.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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147 B

- 11 . SKIN IRRITATION

Study 1: S kin irritation study in rabbits

Title: Acute dermal toxicity and eye irritation studies on sam ple BT

- D, sample #SD

- 7413 in

rabbits with attachment and cover letter dated 061289

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

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Remarks: The sample used in this s tudy was received from Maumee Chemical Company on

September 29, 1969, and was labeled Sample BT

- D, Sample No. SD

- 7413. The test material is a

fine, yellowish

- tan powder with a slight odor.

METHOD

Method/guideline followed: Not stated

Test type: Dermal i rritation

GLP compliant?: Not stated

Year study performed: 1969

Test species: rabbit, albino; weight range 2319 to 2833 grams

Number of animals/sex/dose: 4/dose; sex not specified

Route of administration: D ermal

Vehicle: Physiological saline

Doses/con centrations tested: 1.00, 2.15, 4.64 and 10.0 gm/kg

- bw

Statistical methods used: Not stated

Test method/conditions remarks: The test material was applied to the skin of four groups, each

composed of four albino rabbits (weight range 2319 to 2833 grams). The dose was applied to the

abdominal skin area from which the fur had been previously removed with electric clippers. The

abdominal skin area of two rabbits in each group was abraded by making a series of longitudinal

minor epidermal incisions spaced o ne to two centimeters apart, using a hypodermic needle as a

cutting tool. The abrasions were sufficiently deep to penetrate the epidermis but not to induce

bleeding. The skin of the remaining two rabbits in each group remained intact.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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148 The sample was mois tened with sufficient physiological saline solution to form a paste and

applied at dosage levels of 1.00, 2.15, 4.64 and 10.0 gm/kg of body weight.

The sample was placed on a sleeve of freezer wrapping paper which was then wrapped around

the trunk of the animal and secured with staples. An outer layer of gauze and tape was placed

around the trunk of the animal. The rabbit was immobilized for 24 hours in a wooden restraining

stock.

At the end of the 24

- hour exposure period the binder was removed and any unabsorbed sample

remaining on the skin was removed by gentle sponging with a moistened towel. Each rabbit was

examined thoroughly for gross signs of systemic toxicity and dermal irritation . The rabbits were

placed in individual metal cages elevated abov e the droppings. Commercial pellets and water

were available at all times. All rabbits were maintained for 14 days following completion of the

exposure period. Examinations for gross signs of systemic toxicity and dermal irritation were

carried out at f requent intervals during this period.

At the end of the 14

- day observation period the rabbits were weighed, sacrificed by air

embolism, and a gross necropsy was performed on each animal.

RESULTS

Endpoint value(s): None stated

Results remarks: No mortal ities occurred at any dosage level tested. Therefore, the acute

dermal LD50 for Sample BT

- D, Sample No. SD

- 7413, for albino rabbits is greater than 10.0

gm/kg of body weight. With the exception of purulent nasal discharge noted in occasional

rabbits, all animals generally exhibited normal appearance and behavior throughout the study.

Rabbit

No. Sex Dose Skin Body weight

gm/kg Intact Abraded Start (gm) Finish

(gm) Change

(gm)

1 F 1.0 X 2808 2983 175

2 F 1.0 X 2833 2971 138

3 M 1.0 X 2686 2986 300

4 M 1.0 X 2361 2552 191

5 M 2.15 X 2442 2742 300

6 M 2.15 X 2590 2838 248

7 F 2.15 X 2764 2729

- 35

8 M 2.15 X 2770 2833 63

9 M 4.64 X 2319 2457 138

10 M 4.64 X 2326 2434 108

11 F 4.64 X 2535 2744 209

12 M 4.64 X 2495 2603 108

13 F 10.0 X 2423 2493 70

14 M 10.0 X 2718 2746 28

15 M 10.0 X 2818 2793

- 25Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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149 16 M 10.0 X 2348 2469 121

As shown above, the majority of rabbits showed substantial weight gains during the study.

On removal of the binders at the end of the 24

- hour exposure, dry sample was present on the

binders and abdomens of the rabbits, indicating incomplete dermal absorption of the applied

material.

Irritative effects noted at completion of the exposure period were confined to mild or spotted

erythema in all rabbits. This pe rsisted for one to five days. Desquamation, generally beginning

on the third or fourth post

- exposure day and persisting for two to 12 days was noted in the

majority of the animals.

Gross necropsy findings at termination included congested kidneys in four rabbits, markedly

cystic kidneys in one rabbit, and depleted body fat stores in two rabbits. No other significant

pathological alterations were noted.

CONCLUSIONS

The acute dermal toxicity and the acute eye irritative potential of Sample BT

- D, Sample No . SD

7413, were evaluated in accordance with the techniques specified in the Regulations (Federal

Register, August 12, 1961 et seq) under the Federal Hazardous Substances Act. The acute dermal

LD50 for albino rabbits was found to be greater than 10.0 gm/kg of body weight.

Based on these results Sample BT

- D, Sample No. SD

- 7413, is classified as non

- toxic by dermal

application and is an eye irritant as these terms are defined in the above

- cited Regulations.

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Acute dermal toxicity and eye irritation studies on sample BT

- D, sample #SD

- 7413 in rabbits

with attachment and cover letter dated 061289. Submitting Organization: Sherwin

- Williams

Company. Performing Laboratory: Hill Top Research Incorpo rated. OTS0520633.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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150 Study 2: S kin irritation study in rabbits

Title: Results of range finding toxicological tests on benzotriazole (purified)

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: benz otriazole, CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: Not stated

Test type: S kin irritation

GLP compliant?: Not stated

Year study performed: 1960

Test species: R abbit

Number of animals/sex/dose: Not stated

Route of admini stration: D ermal

Vehicle: None or w ater

Doses/concentrations tested: undiluted and 0.3%

Statistical methods used: Not stated

Test method/conditions remarks: These conclusions are based upon range finding

toxicological tests and are limited to precaut ions for industrial handling of the material.

Development of specific uses will require consideration of the health problems presented and of

the need for further toxicological studies.

RESULTS

Endpoint value(s):

Results remarks:

Material Condition of skin No. of Appl. Site Response

- remarksDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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151 Undiluted intact 10 Belly Essentially no irritation observed

Undiluted abraded 2 Belly Slight hyperemia on 2 nd and 3 rd day

of exposure, slight edema on 1 st , 2 nd and 3 rd days of exposure, slight necrosis afte r 1 st application

and moderate necrosis after 2 nd application. Skin essentially normal in 21 days.

.03% sol. in water intact 10 Ear Essentially no irritation observed

.03% sol. in water intact 10 Belly Same as above

.03% sol. in water abraded 3 Bel ly Very slight hyperemia thru four days

of exposure

CONCLUSIONS

There is no indication, from the skin irritation tests conducted, that this material is absorbed

through the skin in toxic amounts.

Due to the low eye and skin irritating potential of dilu te solutions of this material, it would seem

most unlikely that a concentration of 0.03% in DC 2 would render the formulations irritating.

The undiluted material is essentially nonirritating to the intact skin, moderately irritating to

abraded skin. To ab raded skin prolonged contact might produce a slight burn and repeated

prolonged contact, a serious burn, particularly if confined under clothing. It is recommended

that such exposure be avoided. Abraded skin should be protected while handling undiluted

m aterial. The 0.03% solution in water studied in the laboratory is essentially non

- irritating to

intact and abraded skin. Reasonable care and cleanliness should be sufficient for safe industrial

handling.

STUDY RELIABILITY

2 = reliable with restriction s

REFERENCE

Results of range finding toxicological tests on benzotriazole (purified). Submitting

Organization: Dow Corning Corporation. OTS0520472.

Results of range finding toxicological tests on benzotriazole (purified) (sanitized). Submitting

Org anization: Dow Chemical Company. OTS0520708.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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152 Study 3: S kin irritation study in rabbits

Title: Primary skin irritation study in rabbit of Cobratec 99 (BT) sample #6023 with attachments

and cover letter dated 061289

The study is: Adequ ate Not Adequate

TEST SUBSTANCE

Identity: benzotriazole, CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: Cobratex 99 (BT) Sample No. 6023 was tested for primary skin

irritation on the intact and abraded s kin of rabbits according to the methods described in the

Federal Hazardous Substances Act.

Test type: Skin irritation

GLP compliant?: Not stated

Year study performed: Not stated

Test species: New Zealand albino rabbits

Number of animals/sex/dose: 12 fem ale

Route of administration: D ermal

Vehicle: None stated

Doses/concentrations tested: 0.5 g

Statistical methods used: Not stated

Test method/conditions remarks: Twelve adult female New Zealand albino rabbits were used.

Prior to application the back s were clipped free of hair. The skin was left intact on six of the

rabbits and was abraded on the other six. One half gram of the sample was applied to an area of

skin approximately one inch square on each rabbit. Patches of surgical gauze measuring 1 inch

square, two single layers thick, were applied over the samples and covered with a non

- reactive

tape. The rabbits were then wrapped with a binder.

After the 24

- hour exposure, the binders and the tape were removed and the skin reactions

evaluated. An evaluation was also made at 72 hours after application.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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153 The reactions were scored according to the method described in the Federal Hazardous

Substances Act.

RESULTS

Endpoint value(s): Primary skin index was 0.0

Results remarks: Numerical scores were aw arded to the dermal reaction elicited by Cobratec 99

(BT) Sample No. 6023 on abraded skin

– all scores were 0 (no erythema/no edema).

Numerical scores were awarded to the dermal reaction elicited by Cobratec 99 (BT) Sample No.

6023 on intact skin

– all sc ores were 0 (no erythema/no edema)

CONCLUSIONS

The primary irritation index was 0.0. There was no skin irritation

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Primary skin irritation study in rabbit of Cobratec 99 (BT) sample #6023 with attachments and

cover letter dated 061289. Submitting Organization: Sherwin

- Williams Company. Performing

Laboratory: Huntingdon Research Center. OTS0520635.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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154 Study 4: S kin irritation study in rabbits

Title: Letter from Polaroid Corporation re garding the enclosed results of the acute toxicity,

corneal and dermal irritation studies on 1h

- benzotriazole with attachments

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: 1h

- benzotriazole, CAS N o. 95

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Remarks: None stated

METHOD

Method/guideline followed: None stated

Test type: S kin irritation

GLP compliant?: Not stated

Year study performed: Not stated

Test species: New Zealand albino rabbits, weighing approximately 2 kg

Number of anima ls/sex/dose: 4/group (both sexes)

Route of administration: D ermal

Vehicle: D istilled water

Doses/concentrations tested: 1 gram

Test method/conditions remarks: A total of 30 adult New Zealand albino rabbits of both sexes

weighing approximately 2 kg were divided into seven groups of four animals each. In addition,

two animals were employed as controls and exposed to sodium chloride and calcium carbonate

powder. Animals were prepared by shaving the test area.

One gram of powdered (100 mesh) test materia l was placed on skin and enough distilled water

added to make a paste of uniform consistency. It was then spread over an area of approximately

4”x4” and covered with an occlusive dressing which consisted of a double thickness of Saran

Wrap. The edges of the dressing were fixed to the skin with strips of waterproof adhesive tape.

Patches were not replaced during the exposure period. The test areas were afforded additional

protection by wrapping gauze sheeting around each animal’s trunk. After the third day theDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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155 patches were removed and the test areas examined. Results were recorded immediately, after 11

days and after 26 days.

Body weights were recorded before and 2 weeks after treatment.

RESULTS

Endpoint value(s): Draize scores were 0 at all observat ions.

Results remarks: Neither the animal exposed to sodium chloride nor the animal exposed to

calcium carbonate showed areas of focal necrosis. However, some erythema was noted after 11

days in the animal whose skin was exposed to sodium chloride powder

.

CONCLUSIONS

Draize scores for all observation times and groups being tested with benzotriazole were 0. 0 out

of 4 animals died during testing, dose 0.46 gm/kg, mean % weight change aver 14 days was

- 0.2.

STUDY RELIABILITY

2 = reliable with restric tions

REFERENCE

Letter from Polaroid Corporation regarding the enclosed results of the acute toxicity, corneal and

dermal irritation studies on 1h

- benzotriazole with attachments . Submitting Organization:

Polaroid Corporation. OTS0520182Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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156 Study 5: S kin irritation study in rabbits

Title: Report on skin irritation in the rabbit after single application of TK 11’237 with cover

letter dated 072893

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: 1H

- b enzotriazole, 95

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Remarks: physical appearance: solid; batch no.: SRF OA/3183 (1977). Received: January 11,

1982

METHOD

Method/guideline followed: The procedure used is described in the Proposed Guidelines of the

United States Environmental Agency (EPA) 163.81

- 5 “Primary dermal irritation study”, Federal

Register, Vol. 43, No. 163, August 22, 1978

Test type: Skin irritation

GLP compliant?: Not stated

Year study performed: 1982

Test species: New Zealand white rabbits

Number of animals/sex/dose: 3/ dose, both sexes, weighing 2 to 3 kg

Route of administration: Dermal

Vehicle: Polyethylene glycol (PEG 400) + saline (70:30 parts)

Doses/concentrations tested: 0.5 g

Statistical methods used: Not stated

Test method/conditions remarks: The test was p erformed on 3 male and 3 female adult New

Zealand White rabbits bred and raised at UFA Versuchsbetrieb Buhl, CH

- 5604 Hendschiken,

weighing 2 to 3 kg . They were housed individually in metal cages numbered by ear tags, were

kept at a constant room temperatu re of 22 ± 2°C, at a relative humidity of 55 ± 10 and on a 12

hours light cycle day. The animals received ad libitum standard rabbit food, NAFAG, No. 814,

Gossau SG

– and water. Prior to treatment they were adapted to our laboratories for a minimum

of 4 days.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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157 Before treatment the entire back and the flank of rabbits were shaved with an electric clipper and

immediately before treatment the shaven skin on one side was slightly scarified with the help of

a “Schropfschnapper”. Gauze patches of 2.5 x 2.5 cm laden with 0.5 g of the test material were

applied to the prepared abraded and intact skin.

The patches were covered with an impermeable material and were fastened to the body of the

rabbit with adhesive tape. The dressings were removed after a 24 hour application.

The skin reaction was appraised upon removal and during an observation period of 7 days on the

basis of the following evaluation scheme.

Score for skin irritation in rabbits

Erythema and eschar formation

No erythema 0

Very slight eryth ema (barely perceptible) 1

Well defined erythema 2

Moderate to severe erythema 3

Severe erythema (beet redness) to slight eschar

formation (injuries in depth) 4

Total possible erythema score 4

Edema formation

No edema 0

Very sligh t edema (barely perceptible) 1

Slight edema (edges of area well defined by

definite raising) 2

Moderate edema (raised approximately 1 mm) 3

Sever edema (raised more than 1 mm and extending

beyond area of exposure) 4

Total possible edema score 4

RESULTS

Endpoint value(s): Primary irritation index = 1.0

Results remarks:

Time after

exposure in

hours Mean reaction score

erythema edema

Intact skin Abraded skin Intact skin Abraded skin

24 0.7 1.5 0.3 0.7

72 0 0.3 0 0.3

Total 0.7 1.8 0.3 1.0Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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158 Total = 0.7 + 1.8 + 0.3 + 1.0 = 3.8

Primary irritation index = 3.8:4 = 1.0

CONCLUSIONS

Under the conditions of the present experiment TK 11’237 was found to cause a minimal

irritation when applied to intact and abraded rabbit skin.

The calcu lated primary irritation index was 1.0

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Report on skin irritation in the rabbit after single application of TK 11’237 with cover letter

dated 072893. Submitting Organization: Ciba

- Geigy Corpo ration. OTS0538206.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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159 Study 6: S kin irritation study in rabbits

Title: Report on skin irritation in the rabbit after single application of TK 10’367 and with cover

letter dated 072893

The study is: Adequate Not Adeq uate

TEST SUBSTANCE

Identity: 1H

- benzotriazole, CAS No. 95

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Remarks: Physical appearance: solid; batch no.: Op. 3173/74 (07/79); Received: January 11,

1982

METHOD

Method/guideline followed: The procedure used is described in the Proposed Gui delines of the

United States Environmental Protection Agency (EPA) 163.81

- 5 “Primary dermal irritation

study”, Federal Register Vol. 43, No. 163, August 22, 1978.

Test type: S kin irritation

GLP compliant?: Not stated

Year study performed: 1982

Test spec ies: New Zealand white rabbits bred and raised at UFA Versuchsbetrieb Buhl, CH

5604 Hendschiken, weighing 2 to 3 kg

Number of animals/sex/dose: 3/sex/dose

Route of administration: D ermal

Vehicle: P olyethylene glycol (PEG 400) + saline (70:30 parts)

Dos es/concentrations tested: 50%

Statistical methods used: Not stated

Test method/conditions remarks: The test was performed on 3 male and 3 female adult New

Zealand white rabbits. They were housed individually, identified with individual ear tags, kept

at a constant room temperature of 22 ± 2°C, at a relative humidity of 55 ± 10% and on a 12 hours

light cycle day. The animals received ad libitum standard rabbit food

– NAFAG, No. 814 and

water. Prior to treatment they were adapted to our laboratories fo r a minimum of 4 days.

Before treatment the entire back and the flank of the rabbits were shaved and immediately before

treatment the shaven skin on one side was slightly scarified with the help of aDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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160 “Schropfschnapper”. Gauze patches of 2.5 x 2.5 cm laden with 0.5 g of the test material were

applied to the prepared abraded and intact skin.

The patches were covered with an impermeable material and were fastened to the body of the

rabbit with adhesive tape. The dressings were removed after a 24 hour applica tion.

The skin reaction was appraised upon removal and during an observation period of 7 days on the

basis of the following evaluation scheme.

Score for skin irritation in rabbits

Erythema and eschar formation

No erythema 0

Very slight erythema (bare ly perceptible) 1

Well defined erythema 2

Moderate to severe erythema 3

Severe erythema (beet redness) to slight eschar

formation (injuries in depth) 4

Total possible erythema score 4

Edema formation

No edema 0

Very slight edema ( barely perceptible) 1

Slight edema (edges of area well defined by

definite raising) 2

Moderate edema (raised approximately 1 mm) 3

Sever edema (raised more than 1 mm and extending

beyond area of exposure) 4

Total possible edema score 4

RESULTS

Endpoint value(s): Primary irritation index = 0.7

Results remarks:

Time after

exposure in

hours Mean reaction score

erythema edema

Intact skin Abraded skin Intact skin Abraded skin

24 0.8 1.5 0 0.5

72 0 0 0 0.

Total 0.8 1.5 0 0.5

Total = 0.8 + 1.5 + 0 + 0.5 = 2.8

Primary irritation index = 2.4:4 = 0.7Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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161 CONCLUSIONS

Under the conditions of the present experiment TK 10’637 was found to cause a minimal

irritation when applied to intact and abraded rabbit skin.

The calculated primary irrit ation index was 0.7.

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Report on skin irritation in the rabbit after single application of TK 10’367 and with cover letter

dated 072893. Submitting Organization: Ciba

- Geigy Corporation. OTS053 8211.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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162 Study 7: S kin irritation study in guinea pigs

Title: Primary skin irritation and sensitization test on guinea pigs with cover sheet dated

061289.

The study is: Adequate Not Adequate

[fiche is not legible]

TEST SUBSTANCE

Identity: Benzotriazole, CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: None stated

Test type: Primary irritation test

GLP compliant?: Not stated

Year study performed: Not stated

Test species: A lbino guinea pi gs

Number of animals/sex/dose: 10/group

Route of administration: D ermal

Vehicle: Ethanol for dermal exposure; dimethyl phthalate for intradermal injections

Doses/concentrations tested: Epidermal induction: one drop (~0.05 ml) of a 50% and 25%

solution (wt./wt.) of the test material in 99% ethanol ; Intradermal injection: 0.1 of a 1% solution

(wt./vol.) of test material in dimethyl phthalate ; Challenge exposure: one drop (~0.05 ml) of a

50% and 25% solution (wt./wt.) of test material in 99% ethanol

Stat istical methods used: Not stated

Test method/conditions remarks: The test for primary irritation was conducted by applying,

and lightly rubbing in, one drop (~0.05 ml) of a 50% and 25% solution (wt./wt.) of the test

material in 99% ethanol on the shaved, intact shoulder skin of ten male, albino guinea pigs. To

test for the sensitization potential, a series of four sacral intradermal injections was given, one

each week over a 3

- week period, which consisted of 0.1 of a 1% solution (wt./vol.) of test

materi al in dimethyl phthalate (DMP). Following a two

- week rest period, the test animals wereDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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163 challenged for sensitization by applying, and lightly rubbing in one drop (~0.05 ml) of a 50%

and 25% solution (wt./wt.) of test material in 99% ethanol on the shaved, intact shoulder skin. A

group of ten previously unexposed guinea pigs received similar applications at the time of

challenge to provide a direct comparison of the challenge reactions on skin of similar age.

RESULTS

Endpoint value(s): None stated

Resul ts remarks: None stated

CONCLUSIONS

Benzotriazole produced mild to no skin irritation when tested, at a concentration up to 50% in

99% ethanol, on the shaved, intact skin of male, albino guinea pigs. It did not produce skin

sensitization. Although no serious skin irritation was observed, good industrial hygiene practices

should be observed when handling this or any chemical.

STUDY RELIABILITY

3 = n ot reliable

REFERENCE

Primary skin irritation and sensitization test on guinea pigs with cover sheet d ated 061289.

Submitting Organization: E I Dupont De Nemours and Company. Performing Laboratory:

Haskell Laboratories. OTS0520956.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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164 S tudy 8 : S kin irritation study in guinea pigs

Title: Toxicity and Health Hazard Summary, MSDS and Basic Toxicity of B enzotriazole with

cover letter dated 041989

The study is: Adequate Not Adequate

[ Lack of study details ]

TEST SUBSTANCE

Identity: Benzotriazole, CAS No. 95

- 14

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Remarks: None stated

METHOD

Method/guideline fo llowed: Not stated

Test type: Skin irritation

GLP compliant?: Not stated

Year study performed: Not stated

Test species: Guinea pigs

Number of animals/sex/dose: Not stated

Route of administration: Dermal

Vehicle: Not stated

Doses/concentrations tested : Not stated

Statistical methods used: Not stated

Test method/conditions remarks: Applied to depilated abdomen under an occlusive wrap for

24 hours.

RESULTS

Endpoint value(s):

Results remarks: Slight irritation resulted. Percutaneous absorption was not evident, and the

dermal LD50 was greater than 1.0 g/kg.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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165 CONCLUSIONS

Slightly irritating

STUDY RELIABILITY

3 = not reliable

REFERENCE

Toxicity and health hazard summary, MSDS and basic toxicity of benzotriazole with cover letter

dated 041989. S ubmitting Organization: Eastman Kodak Company. OTS0516745.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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166 S tudy 9 : S kin irrita tion study in guinea pigs

Title: Toxicity and Health Hazard Summary, MSDS and Basic Toxicity of Benzotriazole with

cover letter dated 041989

The study is: Adequate Not Adequate

[ Lack of study details ]

TEST SUBSTANCE

Identity: Benzotriazole, CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: Not stated

Test type: Skin irritation

GLP compliant?: Not stated

Year study performed: Not stated

Test species: Guinea pigs

Number of animals/sex/dose: 5 (sex not specified)

Route of administration: Dermal

Vehicle: Not stated

Doses/concentrations tested: 0.5 ml of a 50% solution

Statistical methods used: Not sta ted

Test method/conditions remarks: Repeated, daily applications (10) of 0.5 ml of a 50% solution

of the compound to the clipped backs of guinea pigs.

RESULTS

Endpoint value(s): None stated

Results remarks: By the tenth day of application, slight to mo derate erythema, slight edema,

minute vesicles and eschars, moderate desquamation, and staining were present on the treatedDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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167 animals.

CONCLUSIONS

None stated

STUDY RELIABILITY

3 = not reliable

REFERENCE

Toxicity and health hazard summary, MSDS and ba sic toxicity of benzotriazole with cover letter

dated 041989. Submitting Organization: Eastman Kodak Company. OTS0516745.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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168 B

- 12 . SKIN SENSITIZATION

Study 1: skin sensitization study in guinea pigs

Title: Report on skin sensitizing (contact allergenic ) effect in guinea pigs of TK 11’237 with

cover letter dated 072893

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: TK 11’237. SRF OA/3183 (1977), CAS No. 95

- 14

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Remarks: Material received: 14.0 1.1982

METHOD

Method/guideline followed: The optimization test was used, an intracutaneous sensitization

procedure exceeding the sensitivity of the method recommended in the “Appraisal of the Safety

of Chemicals in Foods, Drugs and Cosmetics” (1959), the US Association of Food and Drug

Officials (AFDO).

Test type: Skin sensitization

GLP compliant?: Not stated

Year study performed: 1982

Test species: Pirbright white guinea pigs, weighing between 279 to 416 grams

Number of animals/sex/dose: 10/sex/dose

Route of administration: D ermal

Vehicle: I ntradermal

– physiological saline; epidermal

– Vaseline PhH VI

Doses/concentrations tested: 0.1% for intradermal application; 30% for epidermal application

Statistical methods used: Not stated

Test method/con ditions remarks: The test was performed on 10 male and 10 female guinea

pigs of the Pirbright white strain, bred on our premises and weighing between 279 to 416 grams

(~10 weeks old). The animals were housed individually, indentified with individual ear t ags,

kept at a constant room temperature of 20 ± 1°C, at a relative humidity of 50 ± 10% and on a 14

hours light cycle day. The animals received ad libitum standard guinea pig pellets

– NAFAGDraft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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169 No. 830, and water, supplemented with fresh carrots.

During th e induction period the animals received one injection every second day (except

weekends) to a total of 10 intracutaneous injections of a freshly prepared 0.1% TK 11’237 in

physiological saline. One control group was treated with the vehicle alone (“negati ve control”).

On the first day, injections of 0.1 ml were administered into the shaved skin of the right flank

and the back, while on the following days a single intracutaneous injection was given into the

back.

During the second and third week of the indu ction period the test material was incorporated in a

mixture of the normal vehicle with complete Bacto Adjuvant (vehicle:adjuvant = 1:1)

Fourteen days after the last sensitizing injection, a challenge injection of 0.1 ml of a freshly

prepared 0.1% TK 11’23 7 in physiological saline was administered into the skin of the left flank.

Twenty

- four hours after each injection during the first week of the induction period and 24 hours

after the challenge injection the reactions were recorded.

The two largest perpend icular diameters (in mm) and the increase in the skin

- fold thickness (in

mm) were measured and by multiplication of these values a “reaction volume” was obtained (in

μL) for each reading from each animal. The mean volume plus one standard deviation of the

induction react ions observed in the individual animal in the first week was taken as representing

the skin irritation “threshold” for each animal. Any challenge reaction greater than this

threshold value in the induction period was graded as an allergic reaction and the animal termed

“positive”. The number of “positive” animals in the test group was compared with the number

of animals in the control group (treated with the vehicle alone) that showed a non

- specific

reaction of at least the same magnitude (“negative contr ol”).

Ten days after the intracutaneous challenge injection a subirritant dose of the test compound

(30% TK 11’237 in Vaseline) was applied epicutaneously under occlusive dressings which were

left in place for 24 hours. The reactions were evaluated 24 hou rs after removing of the bandages

according the Draize scoring scale.

RESULTS

Endpoint value(s): Not senxitizing

Results remarks: Intradermal injections of the vehicle alone failed to induce sensitization.

A) incidence of positive animals pre group of i ntradermal challenge injection

No. of positive animals/No. of treated animals P

Vehicle control 2/20

TK 11’237 3/20 >0.01

B) incidence of positive animals per group after occlusive epicutaneous application

No. of positive animals/N o. of treated animals

Vehicle control 0/20

TK 11’237 0/20

C) challenge reactions after occlusive epicutaneous administration of the test material

Erythema score (Draize Score) 24 hours after removal of the dressing.

Animal No. m 341 342 343 344 345 346 347 348 349 350

Score 0 0 0 0 0 0 0 0 0 0

Animal No. f 351 352 353 354 355 356 357 358 359 360

0 0 0 0 0 0 0 0 0 0Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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170 CONCLUSIONS

Under the experimental conditions employed, no differences between the test group and the

vehicle

- treated controls were seen, after either intradermal or epidermal challenge application of

TK 11’2 37.

TK 11’237 was found to be devoid of skin

- sensitizing (contact allergenic) potency in albino

guinea

- pigs.

STUDY RELIABILITY

(2) Reliable with restrictions

REFERENCE

Report on skin sensitizing (contact allergenic) effect in guinea pigs of TK 11’237 with cover

letter dated 072893. Submitting Organization: Ciba

- Geigy Corporation. OTS0538207.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

- 14

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171 Study 2: S kin sensitization study in guinea pigs

Title: Report on skin sensitizing (contact allergenic) effect in guinea pigs of TK 11’23

(maximization test) with cover letter dated 072893

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: TK 11237; SRF O 3183

Remarks: Recevied: 26.05. 1982

METHOD

Method/guideline followed: The test was carried out by the maximization test of Magnuson

and Kligman (j. invest. Dermatol. 52, 268

- 276, 1969).

Test type: Skin sensitization

GLP compliant?: Not stated

Year study performed: 1982

Test species: Pirbright white guinea pigs, weighing between 310 and 430 g.

Number of animals/sex/dose: 10/sex/dose

Route of administration: D ermal

Vehicle: I ntradermal application: saline; epidermal applications: soft white petrolatum

Doses/concentrations tested: I ntradermal induction: 1% TK 11237; epidermal induction 30%

TK 1123 7; epidermal challenge: 30% TK 11237.

Statistical methods used: Not stated

Test method/conditions remarks: The test was performed on 10 male and 10 female guinea

pigs per group weighing between 310 and 430 g (~10 weeks old). The animals were housed

in dividually, identified with individual ear tags, kept at a constant room temperature of 20 ± 1°C,

at a relative humidity of 50 ± 10% and on a 14 hours light cycle day. The animals received ad

libitum standard guinea

- pigs pellets

– NAFAG No. 830, and fresh water, supplemented with

fresh carrots.

Inductin, intradermal application: Two intradermal injections (0.1 per injection) were made into

the neck of the guinea

- pigs with a mixture of adjuvant and saline, with the test compound TKDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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172 11237 in saline and wit h the test compound TK 11237 in the adjuvant saline mixture.

Induction, epidermal application: One week later TK 11237 was incorporated in Vaseline and

applied on a filterpapier patch to the neck of the animals (occlusive administration for 48 hours).

Cha llenge: Two weeks after the epidermal induction application the animals were tested on the

flank with 30% TK 11237 in vaseline (24h occlusive application). 24 hours after removing the

dressings the challenge reactions were graded according the Draize scoring scale. The

application sites were chemically depilated 3 hours before examination.

The concentrations of the test compound for induction and challenge periods were determined on

separate animals

.

A control group was treated with adjuvant and the vehicle during the induction period. During

the challenge period the group was treated with the vehicle as well as with the test compound to

control the maximal subirritant concentration of the test com pound in adjuvant treated animals.

RESULTS

Endpoint value(s): Classified as lowest sensitization grade

Results remarks:

A) Incidence of positive animals per group after occlusive epicutaneous challenge application

No. of positive animals/No. of tre ated animals Sensitization rate

Control group 0/20 0%

TK 11237 0/20 0%

B) Challenge reactions after occlusive epicutaneous challenge application of the test material

Erythema score (Draize score) 24 hours after removal of the dressing

Anima l No. m 221 222 223 224 225 226 227 228 229 230

Score 0 0 0 0 0 0 0 0 0 0

Animal No. f 231 232 233 234 235 236 237 238 239 240

Score 0 0 0 0 0 0 0 0 0 0

During the challenge period TK 11237 did neither elicit an allergic reaction in the test group nor

a nonspecific irritant reaction in the control group.

TK 11237 can, therefo re, be classified to the lowest sensitization grade.

CONCLUSIONS

None of the animals was sensitized by TK 11237 under the experimental conditions employed.

According to the maximization grading TK 11237 could be classified to the lowest skin

sensitizing (contact allergenic) class in albino guinea pigs.

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCEDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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173 Report on skin sensitizing (contact allergenic) effect in guinea pigs of TK 11’23 (maximization

test) with cover letter dated 072893. Submit ting Organization: Ciba

- Geigy Corporation.

OTS0538208.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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174 Study 3: S kin sensitization study in guinea pigs

Title: Report on skin sensitizing (contact allergenic) effect in guinea pigs of TK 10’637 with

cover letter dated 072893

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: TK 10’637, Op. 3173/74 07/79, CAS No. 95

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Remarks: Received: 14.01.1982

METHOD

Method/guideline followed: The optimization test was used, an intracutaneous sensitization

procedure exceeding the sensitivity of the method recommended in the “Appraisal of the Safety

of Chemicals in Foods, Drugs and Cosmetics” (1959), the US Association of Food and Drug

Officials (AFDO).

Test type: Skin sensitization

GLP compliant?: Not stat ed

Year study performed: 1982

Test species: Pirbright white guinea pigs, weighing between 270 and 424 grams (~10 weeks old)

Number of animals/sex/dose: 10/sex/dose

Route of administration: D ermal

Vehicle: I intradermal: physiological saline; epidermal: Vaseline PhH VI

Doses/concentrations tested: 0.1% for intradermal application; 30% for epidermal application.

Statistical methods used: Not stated

Test method/conditions remarks: The test was performed on 10 male and 10 female guinea

pigs of the Pirbri ght white strain. The animals were housed individually, identified with

individual ear tags, kept at a constant room temperature of 20 ± 1°C, at a relative humidity of 50

± 10% and on a 14 hours light cycle day. The animals received ad libitum standard g uinea pig

pellets

– NAFAG No. 830, and water, supplemented with fresh carrots. The animals were

acclimated for 11 days.

During the induction period the animals received one injection every second day (exceptDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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175 weekends) to a total of 10 intracutaneous inje ctions of a freshly prepared 0.1% TK 10’647 in

physiological saline. One control group was treated with the vehicle alone (“negative control”).

On the first day, injections of 0.1 ml were administered into the shaved skin of the right flank

and the back, while on the following days a single intracutaneous injection was given into the

back.

During the second and third week of the induction period the test material was incorporated in a

mixture of the normal vehicle with complete Bacto Adjuvant (vehicle:adj uvant = 1:1).

Fourteen days after the last sensitizing injection, a challenge injection of 0.1 ml of a freshly

prepared 01. % TK 10’637 in physiological saline was administered into the skin of the left

flank.

24 hours after each injection during the firs t week of the induction period and 24 hours after the

challenge injection the reactions were recorded.

The two largest perpendicular diameters ( in mm) and the increase in the skin

- fold thickness (in

mm) were measured and by multiplication of these values a “reaction volume” was obtained (in

μL) for each reading from each animal. The man volume plus one standard deviation of the

induction reactions observed in the individual animal in the first week was taken as representing

the skin irritation “threshold ” for each animal. Any challenge reaction greater than this threshold

value in the induction period was graded as an allergic reaction and the animal termed “positive”.

The number of “positive” animals in the test group was compared with the number of an imals in

the control group (treated with the vehicle alon e ) that showed a non

- specific reaction of at least

the same magnitude (“negative control”).

Ten days after the intracutaneous challenge injection a subirritant dose of the test compound

(30% TK 10’6 37 in vaseline) was applied epicutaneously under occlusive dressings which were

left in place for 24 hours. The reactions were evaluated 24 hours after removing of the bandages

according the Draize scoring scale.

RESULTS

Endpoint value(s): Devoid of ski n sensitizing potential

Results remarks: Intradermal injections of the vehicle alone failed to induce sensitization.

A) incidence of positive animals pre group of intradermal challenge injection

No. of positive animals/No. of treated animals P

Vehicl e control 2/20

TK 10’637 5/20 >0.01

B) incidence of positive animals per group after occlusive epicutaneous application

No. of positive animals/No. of treated animals

Vehicle control 0/20

TK 10’637 0/20

C) challenge reactions after occlusive epicutaneous administration of the test material

Erythema score (Draize Score) 24 hours after removal of the dressing.

Animal No. m 321 322 323 324 325 326 327 328 329 330Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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176 Score 0 0 0 0 0 0 0 0 0 0

Animal No. f 331 332 333 334 335 336 337 338 339 340

0 0 0 0 0 0 0 0 0 0

CONCLUSIONS

Under the experimental condition s employed, no differences between the test group and the

vehicle

- treated controls were seen, after either intradermal or ipidermal challenge application of

TK 10’637.

TK 10’637 was found to be devoid of skin

- sensitizing (contact allergenic) potency in al bino

guinea

- pigs.

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Report on skin sensitizing (contact allergenic) effect in guinea pigs of TK 10’637 with cover

letter dated 072893. Submitting Organization: Ciba

- Geigy Corporation. OTS05382 12.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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177 Study4: S kin sensitization study in guinea pigs

Title: Sensitization

The study is: Adequate Not Adequate

[Lacking any study details.]

TEST SUBSTANCE

Identity: Benzotriazole, CAS No. 95

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Remarks: Sub stance type: organic

Physical status: solid

Purity: ca. 100 % w/w

METHOD

Method/guideline followed: OECD Guide

- line 406 "Skin Sensitization"

Test type: Guinea pig maximization test

GLP compliant?: N o data

Year study performed: Not stated

Test species: G uinea pig

Number of animals/sex/dose: Not stated

Route of administration: E ye

Vehicle: None stated

Doses/concentrations tested: Not stated

Statistical methods used: Not stated

Test method/conditions remarks: Not stated

RESULTS

Endpoint value(s): S lightly irritating

Results remarks: None statedDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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178 CONCLUSIONS

None stated

STUDY RELIABILITY

4 = not assignable. Lacking study details (1

- valid without restriction. Guideline study

– according to IUCLID dataset)

REFERENCE

IUCLID (2001). Bayer AG data.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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179 Study 5: S kin sensitization study in guinea pigs

Title: Toxicity and health hazard summary, MSDS and basic toxicity of benzotriazole with cover

letter dated 041989

The study is: Adequate Not Adequate

[Lack ing information on method, minimal study details, inconsistent reporting of the LD50.]

TEST SUBSTANCE

Identity: 1,2,3

- benzotriazole, CAS No. 95

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Remarks: None stated

METHOD

Method/guideline followed: None stated

Test type: Skin sensitization

GLP compliant?: Not stated

Year study performed: Not stated

Test species: G uinea pig, no strain specified

Number of animals/sex/dose: 5, sex not state d

Route of administration: D ermal

Vehicle: None stated

Doses/concentrations tested: .1M in A:D:G.P. Fat, Solvent control

– A:D:G.P. Fat;

Phenylhydraxine

- .1M in D:G.P. Fat

Statistical methods used: Not stated

Test method/conditions remarks: Drop

- on exposure. Scored after 24 and 48 hours.

RESULTS

Endpoint value(s): Not a sensitizer

Results remarks : 0/5 animals sensitizedDraft HPV4 Chemical Data Adequacy Review CAS No. 95

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180 CONCLUSIONS

The compound produced no evidence of skin sensitization in ten guinea pigs tested by a

standardized procedure.

STUDY RELIABILITY

3 = not reliable

REFERENCE

Toxicity and health hazard summary, MSDS and basic toxi city of benzotriazole with cover letter

dated 041989. Submitting Organization: Eastman Kodak Company. OTS0516745.Draft HPV4 Chemical Data Adequacy Review CAS No. 95

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181 CAS No. 95

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Endpoint Study Information

Available Acceptable Data Gap

Aquatic Effects

Acute Toxicity to Fis h Yes Yes No

Acute Toxicity to Daphnia Yes Yes No

Toxicity to Aquatic Plants Yes No Yes

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 No No Yes