SRC TR-10-127

HPV4 CHEMICAL DATA ADEQUACY REVIEW

Benzotriazole, CAS No. 95-14-7

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 Assignment Manager

David Brooks, Alternate Work Assignment Manager

July 23, 2010
HPV4 Chemical Data Adequacy Review

Benzotriazole, CAS No. 95-14-7

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 22202
Draft HPV4 Chemical Data Adequacy Review CAS No. 95-14-7

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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 following 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 relevant 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 - Bibliographic database that contains references to journal articles in the life 

sciences with a concentration on biomedicine

TOXNET -National Library of Medicine 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) - Comprehensive, peerreviewed toxicology data for about 5000 chemicals;

• Genetic Toxicology Data Bank (GENE-TOX) - Peer-reviewed genetic 

toxicology test data for over 3000 chemicals.

TSCATS - Unpublished technical reports submitted by industry to the EPA under the 

Toxic Substances Control Act]

Registry of toxic effects of chemical substances (RTECs)

Environmental Fate Database (EFDB)

ECOTOX -EPA’s peer-reviewed ecotoxicology database
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EPA’s Integrated Risk Information System (IRIS) website

Agency for Toxic Substances and Disease Registry (ATSDR) website

National Toxicology Program (NTP) website

International Program on Chemical Safety Concise International Chemical Assessment 

Documents (IPCS INCHEM or CICADs)

European Chemical Substances Information System (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 toxicity data found for the assigned chemicals. 

While IUCLID Datasets 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 accessible, was obtained and reviewed.

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 document. 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/adequacy 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 common endpoint are grouped together in the document and an endpoint 

adequacy assessment is provided near the top of the first page of the summaries for that 

endpoint. If the data available for the endpoint is determined to be inadequate, a basis for that 

conclusion is supplied. 
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B-1. TOXICITY TO FISH

Summary of endpoint: Adequate

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

This study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: Benzotriazole, CAS No. 95-14-7

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

Sample No. SD-7414 , Date 9/26/69.”

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 duration: 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-gallon 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 dynamic system, a series of static tests 

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

This 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.
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RESULTS

Endpoint value(s): TLm values 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. For the trout 

tested solely under dynamic conditions the TLm values were 15 and 12 ppm at 48 and 96 hours, 

respectively.

(TLm is the 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 to 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 the 
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system and provided evidence of correlation with the 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 conditions 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 Williams Company. Performing 

Laboratory: Food and Drug Research Labs. OTS0520640
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Study 2: Acute toxicity study in zebrafish

Title: Report on test for acute toxicity of REOMET BTA M to zebra fish (Brachydanio 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 part 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 static

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: Not stated

Exposure duration: 96 hours

Concentrations tested: Nominal 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 amounts 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 

the 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.
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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 dead 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 

equilibrium

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

100 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 RELIABILITY

2 = reliable with restrictions
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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. 
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Study 3: Acute 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 

Identity: 1H-benzotriazole, CAS No. 95-14-7

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: rainbow 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, average 

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: Rainbow 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 condition 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 was 
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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 (17.6-29.9) 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 

24 h 48 h 72 h 96 h after 96 hours

15 10 0 0 0 0 0

21 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 

24 h 48 h 72 h 96 h after 96 hours

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 

24 h 48 h 72 h 96 h after 96 hours

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 

ppm

Number of 

fish tested

Deaths after % Mortality 

24 h 48 h 72 h 96 h after 96 hours

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 acute 
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toxicity to catfish 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 weight 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

REFERENCE

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

072893. Submitting Organization: Ciba-Geigy Corporation. OTS0538215.
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Study 4: Acute toxicity study in Brachydanio rerio

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. 95-14-7

Remarks: Substance type: organic 

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: Static

GLP compliant?: No 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. The 96-hour LC0 is 
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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 assignable. 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.
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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 tested and study details.]

TEST SUBSTANCE 

Identity: benzotriazole, CAS No. 95-14-7

Remarks: Cobratec 99; purity >99%

METHOD

Method/guideline followed: Batelle Protocol

Test type: No data

GLP compliant?: No data

Year study was performed: No data

Test species: Rainbow trout, Salmo gairdneri (Fish, estuary, 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 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

96h LC50 = 39 mg/L
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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 

acceptable for assessment – per IUCLID Dataset)

REFERENCE

IUCLID (2001). Battelle Study. 10/1/85; PMC Specialties Group, Inc. MSDS.
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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. 95-14-7

Remarks: Purity not noted

METHOD

Method/guideline followed: No data

Test type: Flow through

GLP compliant?: No 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/L
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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).
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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, concentrations tested and study details.]

TEST SUBSTANCE 

Identity: benzotriazole, CAS No. 95-14-7

Remarks: purity not noted

METHOD

Method/guideline followed: 

Test type: Flow through

GLP compliant?: No data

Year study was performed:

Test species: 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/L
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STUDY RELIABILITY

4 = not assignable

REFERENCE

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

Contaminants. Benzotriazoles. EPA 560/2-77-001, 137-138 (1977).
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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, concentrations tested and study details.]

TEST SUBSTANCE 

Identity: benzotriazole, CAS No. 95-14-7

Remarks: purity not noted

METHOD

Method/guideline followed: No data

Test type: Static

GLP compliant?: No data

Year study was performed: No data

Test species: 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 value(s): The 96-hour LC50 value for fathead minnows and bluegill was 25 mg/L.

Results remarks: No data

CONCLUSIONS

96h LC50 = 25 mg/L
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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).
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Study 9: 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: 

Test type: flow through

GLP compliant?: No 

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 28 mg/L.

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

CONCLUSIONS

96h LC50 = 28 mg/L
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STUDY RELIABILITY

4 = not assignable

REFERENCE

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

December 1, 1969.
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Study10: 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: 

Test type: flow through

GLP compliant?: No 

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/L
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STUDY RELIABILITY

4 = not assignable

REFERENCE

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

December 1, 1969.
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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 followed: No data

Test type: Flow through

GLP compliant?: No 

Year study was performed: No data

Test species: Rainbow trout, Salmo gairdneri (Fish, estuary, 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 rainbow trout was 12 mg/L. 

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

CONCLUSIONS

96h LC50 = 12mg/L
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STUDY RELIABILITY

4-not assignable.

REFERENCE

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

December 1, 1969.
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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 type: Static

GLP compliant?: No 

Year study was performed: No data

Test species: Bluegill, Lepomis macrochirus (Fish, estuary, 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: Median Tolerance Limit (LC50) = 27.5 ppm (48 hrs); 25 ppm (96 hrs).

CONCLUSIONS

96h LC50 = 25 mg/L
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STUDY RELIABILITY

4 = not assignable

REFERENCE

IUCLID (2001). Food and Drug Research Laboratories, Study No. 90801, December 1, 1969.
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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: Static

GLP compliant?: No 

Year study was performed: No data

Test species: Fathead minnow, Pimephales promelas (Fish, estuary, 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 minnow was 28 mg/L. 

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

CONCLUSIONS

LC50 = 30.2 ppm
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STUDY RELIABILITY

4 = not assignable

REFERENCE

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

December 1, 1969.
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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 compliant?: Not stated

Year study was performed: 1957

Test species: Bluegill 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, temperature 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.
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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- -

Fisheries No. 207. March 1957.
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Study 15: Acute toxicity study in Coho salmon

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 stated

METHOD

Method/guideline followed: Not stated

Test type: Static

GLP compliant?: Not stated

Year study was performed: Not stated

Test species: Coho salmon (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 test 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. 
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STUDY RELIABILITY

2 = valid with restrictions

REFERENCE

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.
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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. 95-14-7

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 (Rainbow 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 water, 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 effects were observed at 5 ppm.
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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- -

Fisheries No. 207. March 1957.
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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

Remarks: 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 value(s): Not reported

Results remarks: Neither loss of equilibrium nor death occurred.

CONCLUSIONS

No effects were observed. 
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STUDY RELIABILITY

2 = valid with restrictions

REFERENCE

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.
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B-2. TOXICITY TO AQUATIC INVERTEBRATES

Summary of endpoint: Adequate

Study 1: Acute toxicity 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: REOMET 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/guideline 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

Analytical 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 renewal 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 sieving 
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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.

Beakers 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 substance 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: Values 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 

(nominal) 

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/L
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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.
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Study 2: Acute toxicity study in Daphnia magna

Title: Acute toxicity to aquatic invertebrates

This study is: Adequate Not Adequate

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 1 "Daphnia sp., Acute Immobilisation 

Test"

Test type: Static

GLP compliant?: Yes

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

RESULTS

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 60 
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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.
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Study 3: Acute toxicity 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. 95-14-7

Remarks: purity >99%

METHOD

Method/guideline followed: Battelle protocols

Test type: No data

GLP compliant?: No data

Year study was performed: No data

Test species: Daphnia magna (Crustacea)

Analytical monitoring: No data

Exposure duration: 48 hours

Concentrations 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/L
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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). Battelle Study. 10/1/85; PMC Specialties Group, Inc. MSDS.
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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 38412 L 9

Test type: No data

GLP compliant?: Yes

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 mg/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:
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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

72h 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.
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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?: Yes

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 mg/L (nominal)

Test method/conditions remarks: Endpoint – biomass

RESULTS

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 0.7 
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32 10.7

100 47.6 

320 96.3

1000 100.0

CONCLUSIONS

72h 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.
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Study 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?: No data

Year study was performed: No data

Test species: Other aquatic plant: algae

Analytical monitoring: No data

Exposure duration: 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 Probit method; 95% confidence limits = 2.544.7

mg/L.

NOEC and LOEC generated by ANOVA, Bonferroni test.

CONCLUSIONS

96h EC50 = 15.4 mg/L
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STUDY RELIABILITY

4 = not assignable. Lacking study details.

REFERENCE

IUCLID (2001). Battelle Study. 10/1/85; PMC Specialties Group, Inc. MSDS.
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B-4. CHRONIC TOXICITY STUDY IN AQUATIC INVERTEBRATES

Study 1: Chronic toxicity study in Daphnia magna

Title: Chronic toxicity to aquatic invertebrates

This study is: Adequate Not Adequate

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?: Yes

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 solution, 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 mg/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 data
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CONCLUSIONS

21d EC50 = 76.9 mg/L

STUDY RELIABILITY

1 = valid without restriction, GLP Guideline study

REFERENCE

IUCLID (2001). Bayer AG study: January 18, 1995.
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Study 2: Acute toxicity study in Daphnia magna

Title: Chronic toxicity to aquatic invertebrates

This study is: Adequate Not Adequate

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?: Yes

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 solution, 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: 25.9 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.
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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.
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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 Adequate

TEST SUBSTANCE 

Identity: Cobratec #99 (benzotriazole); CAS No. 95-14-7

Remarks: purity >99%

METHOD

Method/guideline followed: Other

Test type: LD50

GLP compliant?: No data

Year study performed: No data

Test species: Rat, no data on strain

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

Route of administration: Oral

Vehicle: No 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, 
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mortality: 7/10; 795 mg/kg, mortality: 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.
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Study 2: Acute oral toxicity study in rats

Title: Acute oral toxicity

The study is: Adequate 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?: No data

Year study performed: No data

Test species: rat, Greenacres strain

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

Route of administration: Oral

Vehicle: Corn 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 method/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 mortality 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)
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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 Research, Inc. Report# S-186A. June 12, 1968.
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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?: No data

Year study performed: No data

Test species: Sprague-Dawley rat

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

Route of administration: Oral

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. Doses 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/kgbw.

Results remarks: The LD50 value was calculated according to Litchfield and Wilcoxon, 1949.
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CONCLUSIONS

LD50 = 720 mg/kg-bw

STUDY RELIABILITY

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

acceptable for assessment

REFERENCE

IUCLID (2001). North American Science Associates, Inc. Lab No. 81-5510.

August 3, 1981.
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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?: No 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: Other: as a dust

Doses/concentrations tested: 1.5 mg/L

Statistical methods used: Not stated

Test method/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 exposure. 

Particle size of the aerosolized dust was determined using a cascade impactor. 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 exposure. 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.
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RESULTS

Endpoint value(s): The inhalation LC50 for male and female 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 breathing and/or nasal discharge during the 

exposure or the first week following exposure. These symptoms abated during the second postexposure week. Slight decreases in weight were observed on the day after exposure with a 

recovery thereafter. Postmortem examinations 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

LC50 = > 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.
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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?: No data

Year study performed: No data

Test species: Albino 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 onetwo days post exposure. The primary effect appeared to be pulmonary irritation which resulted 

intra-tracheal edema and a hemorrhagic type response in the lungs.
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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). Huntingdon Research Center. Project No. 745-080. August 8,

1974.
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Study 6: Acute dermal toxicity study in rabbits

Title: Acute dermal toxicity

The study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: BT-D (benzotriazole), 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: Albino rabbits

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

Route of administration: dermal

Vehicle: Physiol 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 rabbits 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.
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CONCLUSIONS

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.
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Study 7: Acute dermal toxicity study in rats

Title: Acute dermal toxicity

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

Remarks: Purity not noted

METHOD

Method/guideline followed: Not stated

Test type: LD50

GLP compliant?: No data

Year study performed: 1984

Test species: Rat

Number of animals/sex/dose: No data

Route of administration: Dermal

Vehicle: No data

Doses/concentrations 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 female rats was >2,000 mg/kg-bw.

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

are symptoms commonly seen during observation time following administration of substances by 
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gavage. No other symptoms which could be related to the treatment, especially no cutaneous 

changes, were observed.

CONCLUSIONS

LD5 > 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., Toxicology studies with benzotriazole in rats, rabbits, 

Chinese Hamsters, Zebra Fish and guinea pigs. OTS0510543-2 6/09/87.
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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: Rabbit

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

Route of administration: Dermal

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

CONCLUSIONS
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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.
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Study 9: Acute i.p. 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/guideline followed: None stated

Test type: LD50

GLP compliant?: No data

Year study performed: Not stated

Test species: Mouse

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 1000 mg/kg-bw.

Results remarks: None stated

CONCLUSIONS
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LD50 = 1000 mg/kg-bw

STUDY RELIABILITY

4 = not assignable. Lacking study details.

REFERENCE

IUCLID (2001). NTP Chemical Repository, Catalog ID number: 000343.
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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/guideline followed: Not stated

Test type: LD50

GLP compliant?: No data

Year study performed: Not stated

Test species: Mouse

Number of animals/sex/dose: Not stated

Route of administration: i.v.

Vehicle: None stated

Doses/concentrations tested: Not stated

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

CONCLUSIONS
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LD50 = 238 mg/kg-bw

STUDY RELIABILITY

4 = not assignable. Lacking study details.

REFERENCE

IUCLID (2001). NTP Chemical Repository, Catalog ID number: 000343.
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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. 95-14-7

Remarks: None stated

METHOD

Method/guideline followed: Not stated

Test type: LD50

GLP compliant?: Not stated

Year study performed: ~ 1983

Test species: Rat, no strain specified

Number of animals/sex/dose: Not stated

Route of administration: Gavage

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 observed
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CONCLUSIONS

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

toxicity included moderate weakness, prostration, cyanosis, 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 Kodak Company. OTS0516745.
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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 Adequate

[Lacking study details.]

TEST SUBSTANCE 

Identity: 1,2,3-benzotriazole, CAS No. 95-14-7

Remarks: None stated

METHOD

Method/guideline followed: Not stated

Test type: LD50

GLP compliant?: Not stated

Year study performed: Not stated

Test species: Rat

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. 

CONCLUSIONS
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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. Submitting Organization: Eastman Kodak Company. OTS0516745.
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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: Male albino rat

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

Route of administration: Orally 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 suspension 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 prior 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 observed 
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closely for gross signs of systemic toxicity 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 

weighed, 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 remarks: 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% Confidence 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 generally 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 behavior 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 or 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 termination.

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 showing 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 body weight
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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 

moderate 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 extremities; 

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.
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Study 14: Acute oral toxicity study in rats

Title: 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 stated

METHOD

Method/guideline followed: Not stated

Test type: LD50

GLP compliant?: Not stated

Year study performed: Not stated

Test species: Rats

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

Route of administration: Orally

Vehicle: Not stated

Doses/concentrations 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.

RESULTS

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 - 10/10
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632 0/10 7/10 - 7/10

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

398 0/10 - - 0/10

LD50 determination: The dose response curve is 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.
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Study 15: Acute dermal toxicity study in rabbits

Title: Acute dermal toxicity investigation 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: Rabbits

Number of animals/sex/dose: 10 females

Route of administration: Dermal

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/kg
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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 formation
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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 raising) 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 = reliable 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 Research Center. OTS0520634.
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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. 95-14-7

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 animals/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 this 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 postexposure for observation of latent effects. All animals were observed 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 abnormalities.
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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 occurred 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 were 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 fact 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 = reliable with restrictions

REFERENCE 

Evaluation of the three-hour LC50 of aerosolized benzotriazole (final report) with attachments 
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and cover letter dated 061289. Submitting Organization: Sherwin-Williams. Performing 

Laboratory: Huntingdon Research Center. OTS0520637
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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

The study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: Cobratec 99, 1,2,3-benzotriazole, CAS No. 95-14-7

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 conducted 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.

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

Results remarks: The mean gravimetric exposure concentration of COBRATEC 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.

Slight 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 attainable 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 09/16/94. Submitting 

Organization: Cincinnati Specialties Incorporated. Performing Laboratory: Wildlife 

International Limited. OTS0557484.
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Study 18: Acute oral toxicity study in rats

Title: Letter from Sherwin Williams Company to the USEPA regarding information 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. 95-14-7

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: White rat

Number of animals/sex/dose: Not stated

Route of administration: Oral

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 stated
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CONCLUSIONS

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

STUDY RELIABILITY

4 = not assignable

REFERENCE

Letter from Sherwin Williams Company to the USEPA regarding information on 1,2,3-

benzotriazole and tolytriazole with attachments. Submitting Organization: Sherwin Williams 

Company. OTS0520630.
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Study 19: Acute oral toxicity study in rats

Title: Acute oral test with cover sheet dated 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. 95-14-7

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: Oral - gavage

Vehicle: Corn 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 rats in single doses by intragastric intubation. 

Survivors were sacrificed 14 days after dosing.

RESULTS

Endpoint value(s): ALD = 670 mg/kg-bw
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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.
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B-6. REPEATED-DOSE TOXICITY

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. 95-14-7

Remarks: None stated

METHOD

Method/guideline followed: Not stated

Test type: Repeated-dose toxicity

GLP compliant?: Not stated

Year study performed: Not stated

Test species: Rat

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

Route of administration: Oral – 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/conditions 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

RESULTS
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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 animals 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 animals; 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 reduction 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 weights 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 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.
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Study 2: Repeated i.p. injections in rats

Title: Evaluation 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. 95-14-7

Remarks: Fine white material, test solutions were prepared by incorporating the compound 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 albino rats, from the Charles River Breeding Laboratories.

Number of animals/sex/dose: 5/sex

Route of administration: Intraperitoneal 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 single 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.
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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 solution 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.

Observations of appearance and behavior were made frequently after dose.

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

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

levels. Hematocrit, erythrocyte 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

Endpoint 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 were 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.

Copper 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 increase 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 does not appear to be a copper or iron chelator.
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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 dated 061289. Submitting Organization: Sherwin Williams 

Company. Performing Laboratory: Huntingdon Research Center. OTS0520638.
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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. 95-14-7

Remarks: Substance type: organic 

Physical status: solid 

Purity: ca. 100 % w/w

METHOD

Method/guideline followed: OECD Guide-line 476 "Genetic Toxicology: In vitro Mammalian 

Cell Gene Mutation Tests"

Test type: HGPRT assay

GLP compliant?: Yes

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 %
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Results remarks: None stated

CONCLUSIONS

The test substance was not considered to be mutagenic.

STUDY RELIABILITY

1 = valid without restriction. GLP Guideline study

REFERENCE

IUCLID (2001). Hazleton Biotechnologies, The Netherlands, at the request of Bayer AG, 1987.
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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. 95-14-7

Remarks: None stated

METHOD

Method/guideline followed: Not stated

Test type: Ames test

GLP compliant?: No data

Year study performed: Not stated

Test species: Salmonella typhimurium

Metabolic activation: No data

Concentrations tested: Not stated

Statistical methods used: Not stated

Test method/conditions remarks: Not stated

RESULTS

Genotoxicity result: Positive

Cytotoxic concentration (if applicable): No data

Results remarks: Not stated

CONCLUSIONS

The test substance was considered to be mutagenic.
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STUDY RELIABILITY

4 = not assignable. Lacking study details.

REFERENCE

IUCLID (2001). Dunkel, v.c.; Simmon, V.F. in: IARC Publication vol. 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).
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Study 3: Mouse micronucleus tests (in vivo)

Title: Genetic toxicity ‘in vivo’

The 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: OECD Guide-line 474 "Genetic Toxicology: Micronucleus Test"

Test type: Micronucleus assay

GLP compliant?: Yes

Year study performed: 1988

Test species: NMRI mice

Metabolic activation: Not stated

Concentrations tested: 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 remarks: Not stated

CONCLUSIONS
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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.
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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 equivocal]

TEST SUBSTANCE 

Identity: 1H-benzotriazole, 95-14-7

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 histidineauxotrophic mutants of Salmonella typhimurium. 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 with 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 was 
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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 buffer; 2) for strain TA100: 4-

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

bacteria in the treated and control cultures that have undergone back-mutation to histidineprototrophism. 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 cofactors)

RESULTS

Genotoxicity result: Weakly positive

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 

effect 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 the 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.
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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.
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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 equivocal]

TEST SUBSTANCE 

Identity: 1H-benzotriazole, 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

GLP compliant?: Not stated

Year 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 histidineauxotrophic 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, 1500 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 0.5mM. In the experiments in 
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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.

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 TA1535: nmethyl-n’-nitro-n-nitrosoguanidine, 3 and 5 μg/0.1 ml phosphate buffer; 4) for strain TA1537: 

9(5)aminoacridine 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 of the numbers of 

bacteria in the treated and control cultures that have undergone back-mutation to histidineprototrophism. 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 (rat liver microsomes and co-factors).

RESULTS

Genotoxicity result: Weak positive

Cytotoxic concentration (if applicable):

Results remarks: 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 at the concentrations of 1500 and 2025 μg/0.1 ml.

CONCLUSIONS 

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

STUDY RELIABILITY

3 = not reliable

REFERENCE
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Salmonella/mammalian-microsome mutagenicity test with TK 11’237 with cover letter dated 

072893. Submitting Organization: Ciba-Geigy Corporation. OTS0538210.
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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 study 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 confirmation assay

Statistical methods used: Not stated

Test method/conditions remarks: TK 10 637 was tested for mutagenic effects on histidineauxotrophic mutants of Salmonella typhimurium. The investigations were performed on strains 

TA 98, TA 100, TA 1535 and TA 1537 with the following concentrations of the 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 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 0.5mM. In the experiments in 
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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.

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 TA1535: nmethyl-n’-nitro-n-nitrosoguanidine, 3 and 5 μg/0.1 ml phosphate buffer; 4) for strain TA1537: 

9(5)aminoacridine 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 of the numbers of 

bacteria in the treated and control cultures that have undergone back-mutation to histidineprototrophism. 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 (rat liver microsomes and co-factors).

RESULTS

Genotoxicity result: Weak positive

Cytotoxic concentration (if applicable):

Results remarks: In the experiments performed without 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 back-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 the 
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substance on the growth of the bacteria. In the experiment with activation this effect was only 

observed with strains 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

REFERENCE

Salmonella/mammalian-microsome mutagenicity test with TK 10 637 with cover letter dated 

072893. Submitting Organization: Ciba-Geigy Corporation. OTS 538213.
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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 specified

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 specified

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

Genotoxicity result: Positive (with activation), Negative (without activation)

Cytotoxic concentration (if applicable): Not specified

Results remarks: None stated
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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.
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Study 8: Sister chromatid exchange assay

Title: Sister Chromatid Exchange

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: Sister Chromatid Exchange

GLP compliant?: Not specified

Year study performed: 1985

Test species: 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 specified

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 positive 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 activation)

Cytotoxic concentration (if applicable): Not specified

Results remarks: None stated
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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.
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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, concurrent no treatment

Statistical methods used: Not stated

Test methods/conditions remarks: Groups of 50 rats of each sex were administered IHbenzotriazole at one of two time-weighted average doses, either 6,700 or 12,100 ppm, for 78 

weeks. Except for five 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. All rats 

surviving to weeks 104-106 were then killed.
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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 reproductive 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 of 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 RELIABILITY

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 carcinogenicity (1978).
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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 IHbenzotriazole 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. All mice surviving to weeks 106-109 were then 

killed.

RESULTS

Endpoint value(s): The parenteral NOAEL was > 23,500 ppm.
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Results remarks: The 104 week oral study of 1H-benzotriazole in mice did not find any 

evidence of pathology in the reproductive 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 ovaries.

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, well documented and 

acceptable for assessment

REFERENCE

IUCLID (2001). NTIS # PB285202/AS. US National Cancer Institute : Bioassay

of 1-H-Benzotriazole for possible carcinogenicity (1978).
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B-9. CARCINOGENICITY

Study 1: Chronic carcinogenicity study in 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

Statistical methods used: Not stated

Test method/conditions remarks: Groups of 50 rats of each sex were administered 1hbenzotriazole 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 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. All rats surviving 

to weeks 104-106 were then killed.
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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 groups 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 historical-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 occurred 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 incidence 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 

when 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 rats 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.

CONCLUSIONS

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 carcinogenicity (CAS No. 95-14-7). U.S. 

Department of Health, Education, and Welfare. National Institutes of Health. NCI-CG0TR-88.
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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 were 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 were administered IHbenzotriazole 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. All mice surviving to weeks 106-109 were then 

killed.

RESULTS

Endpoint value(s): Not stated
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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. Survival 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 possible 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 conditions 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, Education, and Welfare. National Institutes of Health. NCICG0TR-88.
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B-10. EYE IRRITATION

Study 1: Eye 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 letter 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 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: Eye irritation study

GLP compliant?: Not stated

Year study performed: 1969

Test species: Rabbits, albino

Number of animals/sex/dose: 6

Route of administration: Eye

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 of 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 weighted 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 total 
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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 observation period.

CONCLUSIONS 
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The acute dermal toxicity and the acute eye irritative potential of Sample BT-D, Sample No. SD7413, 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.
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Study 2: Eye irritation study in rabbits

Title: Results of range finding toxicological tests on benzotriazole (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: Eye irritation

GLP compliant?: Not stated

Year study performed: ~ 1960

Test species: Rabbit

Number of animals/sex/dose: Not stated

Route of administration: Eye 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 irritation
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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 likelihood 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 formulations irritating.

STUDY RELIABILITY

2 = reliable with restrictions

REFERENCE

Results of range finding toxicological tests on benzotriazole (purified). Submitting 

Organization: Dow Corning Corporation. OTS0520472.
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Study 3: Eye 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. 95-14-7

Remarks: None stated

METHOD

Method/guideline followed: None stated

Test type: Eye irritation

GLP compliant?: Not stated

Year study performed: Not stated

Test species: Rabbit

Number of animals/sex/dose: 3

Route of administration: Eye

Vehicle: 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 examined up to 2 weeks 

following administration.

RESULTS

Endpoint value(s): None stated

Results remarks: Moderate eye irritation
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CONCLUSIONS

The 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.
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Study 4: Eye 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. 95-14-7

Remarks: None stated

METHOD

Method/guideline followed: 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: Eye

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 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 reactions was facilitated by 

the use of an opthalmoscope. After the recording 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”, section 191.12 of the Federal Hazardous Substances labeling Act 

Regulations.
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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 visible

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.
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Study 5: Eye irritation study in rabbits

Title: Eye irritation study in the rabbit of benzotriazole (Cobratec 99) sample #1088 with 

attachments and cover letter dated 061289

The study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: benzotriazole, CAS No. 95-14-4

Remarks: Cobratec 99, Sample # 1088

METHOD

Method/guideline followed: 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: Eye

Vehicle: None stated

Doses/concentrations 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 ophthalmoscope 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 ophthalmoscope. After the 

recording of observations at 24 hours, the eyes were stained with fluorescein and examined with 

an ultraviolet lamp to detect corneal damage of present.
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The reactions were scored from: “Illustrated guide for grading eye irritation by hazardous 

substances”, Section 191.12 of the Federal Hazardous Substances 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 – 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

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 +

Conjuctiva

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 +
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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 deepened folds, congestion, swelling, moderate circumcorneal 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 all

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 reaction)

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 half 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 restrictions
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REFERENCE

Eye irritation study in the rabbit of benzotriazole (Cobratec 99) sample #1088 with attachments 

and cover letter dated 061289. Submitting Organization: Sherwin-Williams. Performing 

Laboratory: Huntingdon Research Center. OTS0520636.
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Study 6: Eye irritation study in rabbits

Title: Letter from Polaroid 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-benzotriazole, CAS No. 95-14-7

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: Eye

Vehicle: None stated

Doses/concentrations tested: 100 mg

Statistical methods used: Not stated

Test method/conditions remarks: 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 with 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 fluid. After an exposure period of five 
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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, sucrose 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 reexamined 7 days after treatment. Mild conjunctivitis was observed in all instances immediately 

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 etiology 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 enclosed results of the acute toxicity, corneal and 

dermal irritation studies on 1h-benzotriazole with attachments. Submitting Organization: 

Polaroid Corporation. OTS0520182. 
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Study 7: Eye 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. 95-14-7

Remarks: purity not noted

METHOD

Method/guideline followed: Draize test

Test type: Eye irritation

GLP compliant?: No data

Year study performed: Not stated

Test species: rabbit

Number of animals/sex/dose: Not stated

Route of administration: Eye

Vehicle: None – substance delivered undiluted

Doses/concentrations tested: 100 mg (undiluted)

Statistical methods used: Not stated

Test method/conditions remarks: Exposure 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.
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CONCLUSIONS

Highly irritating

STUDY RELIABILITY

4 = not assignable. Lacking study details

REFERENCE

IUCLID (2001). Hill Top Research, Inc. Report T-413. October 29, 1969.

IUCLID (2001). PMC Specialties Group, Inc. MSDS.
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B-11. SKIN IRRITATION

Study 1: Skin 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 letter dated 061289

The study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: benzotriazole, CAS No. 95-14-7

Remarks: The sample used in this study 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 irritation

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: Dermal

Vehicle: Physiological saline

Doses/concentrations 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 one 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.
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The sample was moistened 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 above 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 frequent 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 mortalities 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 -25
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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 persisted 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. SD7413, 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 Incorporated. OTS0520633.
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Study 2: Skin irritation study in rabbits

Title: Results of range finding toxicological tests on benzotriazole (purified)

The study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: benzotriazole, CAS No. 95-14-7

Remarks: None stated

METHOD

Method/guideline followed: Not stated

Test type: Skin irritation

GLP compliant?: Not stated

Year study performed: 1960

Test species: Rabbit

Number of animals/sex/dose: Not stated

Route of administration: Dermal

Vehicle: None or water

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 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.

RESULTS

Endpoint value(s): 

Results remarks:

Material Condition of skin No. of Appl. Site Response-remarks
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Undiluted intact 10 Belly Essentially no irritation observed

Undiluted abraded 2 Belly Slight hyperemia on 2nd and 3rd day 

of exposure, slight edema on 1st, 2nd and 3rd days of exposure, slight necrosis after 1st application 

and moderate necrosis after 2nd 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 Belly 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 dilute 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 abraded 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 

material. 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 restrictions

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 

Organization: Dow Chemical Company. OTS0520708.
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Study 3: Skin 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: Adequate Not Adequate

TEST SUBSTANCE 

Identity: benzotriazole, CAS No. 95-14-7

Remarks: None stated

METHOD

Method/guideline followed: Cobratex 99 (BT) Sample No. 6023 was tested for primary skin 

irritation on the intact and abraded skin 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 female

Route of administration: Dermal

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 backs 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.
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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 awarded 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 scores 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.
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Study 4: Skin irritation study in rabbits

Title: Letter from Polaroid 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-benzotriazole, CAS No. 95-14-7

Remarks: None stated

METHOD

Method/guideline followed: None stated

Test type: Skin irritation

GLP compliant?: Not stated

Year study performed: Not stated

Test species: New Zealand albino rabbits, weighing approximately 2 kg

Number of animals/sex/dose: 4/group (both sexes)

Route of administration: Dermal

Vehicle: Distilled 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 material 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 the 
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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 observations.

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 restrictions

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. OTS0520182
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Study 5: Skin 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-benzotriazole, 95-14-7

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 performed 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 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, 

Gossau SG – and water. Prior to treatment they were adapted to our laboratories for a minimum 

of 4 days.
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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 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

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 = 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 1.0
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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 calculated 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 Corporation. OTS0538206.
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Study 6: Skin 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 Adequate

TEST SUBSTANCE 

Identity: 1H-benzotriazole, CAS No. 95-14-7

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 Guidelines 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: Skin irritation

GLP compliant?: Not stated

Year study performed: 1982

Test species: New Zealand white rabbits bred and raised at UFA Versuchsbetrieb Buhl, CH5604 Hendschiken, weighing 2 to 3 kg

Number of animals/sex/dose: 3/sex/dose

Route of administration: Dermal

Vehicle: Polyethylene glycol (PEG 400) + saline (70:30 parts)

Doses/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 for 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 a 
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“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 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

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 = 0.7
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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 irritation 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. OTS0538211.
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Study 7: Skin 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. 95-14-7

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: Albino guinea pigs

Number of animals/sex/dose: 10/group

Route of administration: Dermal

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

Statistical 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 

material in dimethyl phthalate (DMP). Following a two-week rest period, the test animals were 
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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

Results 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 = not reliable

REFERENCE

Primary skin irritation and sensitization test on guinea pigs with cover sheet dated 061289. 

Submitting Organization: E I Dupont De Nemours and Company. Performing Laboratory: 

Haskell Laboratories. OTS0520956.
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Study 8: Skin irritation 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. 95-14-7

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: 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.
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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. Submitting Organization: Eastman Kodak Company. OTS0516745.
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Study 9: Skin irritation 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. 95-14-7

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 stated

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 moderate erythema, slight edema, 

minute vesicles and eschars, moderate desquamation, and staining were present on the treated 
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animals.

CONCLUSIONS

None stated

STUDY RELIABILITY

3 = not reliable

REFERENCE

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

dated 041989. Submitting Organization: Eastman Kodak Company. OTS0516745.
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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. 95-14-7

Remarks: Material 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 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: Dermal

Vehicle: 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 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 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 pig pellets – NAFAG 
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No. 830, and water, supplemented with fresh carrots. 

During the 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 (“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:adjuvant = 1:1)

Fourteen days after the last sensitizing injection, a challenge injection of 0.1 ml of a freshly 

prepared 0.1% TK 11’237 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 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 mean 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 animals in the control group (treated with the vehicle alone) 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 11’237 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): Not senxitizing

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

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/No. 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 0
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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 11’237.

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.
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Study 2: Skin 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: Dermal

Vehicle: Intradermal application: saline; epidermal applications: soft white petrolatum

Doses/concentrations tested: Intradermal induction: 1% TK 11237; epidermal induction 30% 

TK 11237; epidermal challenge: 30% TK 11237.

Statistical methods used: Not stated

Test method/conditions remarks: The test was performed on 10 male and 10 female guineapigs per group weighing between 310 and 430 g (~10 weeks old). 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 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 TK 
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11237 in saline and with 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).

Challenge: 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 compound 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 treated 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

Animal 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, therefore, 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

REFERENCE
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Report on skin sensitizing (contact allergenic) effect in guinea pigs of TK 11’23 (maximization 

test) with cover letter dated 072893. Submitting Organization: Ciba-Geigy Corporation. 

OTS0538208.
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Study 3: Skin 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. 95-14-7

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 stated

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: Dermal

Vehicle: Iintradermal: 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 Pirbright 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 guinea 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 (except 
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weekends) to a total of 10 intracutaneous injections 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:adjuvant = 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 first 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 animals in 

the control group (treated with the vehicle alone) 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 10’637 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 skin 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

Vehicle 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 321 322 323 324 325 326 327 328 329 330
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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 conditions 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 albino 

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. OTS0538212.
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Study4: Skin sensitization study in guinea pigs

Title: Sensitization

The study is: Adequate Not Adequate

[Lacking any 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: OECD Guide-line 406 "Skin Sensitization"

Test type: Guinea pig maximization test

GLP compliant?: No data

Year study performed: Not stated

Test species: Guinea pig

Number of animals/sex/dose: Not stated

Route of administration: Eye

Vehicle: None stated

Doses/concentrations tested: Not stated

Statistical methods used: Not stated

Test method/conditions remarks: Not stated

RESULTS

Endpoint value(s): Slightly irritating

Results remarks: None stated
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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.
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Study 5: Skin 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

[Lacking information on method, minimal study details, inconsistent reporting of the LD50.]

TEST SUBSTANCE 

Identity: 1,2,3-benzotriazole, CAS No. 95-14-7

Remarks: None stated

METHOD

Method/guideline followed: None stated

Test type: Skin sensitization

GLP compliant?: Not stated

Year study performed: Not stated

Test species: Guinea pig, no strain specified

Number of animals/sex/dose: 5, sex not stated

Route of administration: Dermal

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 sensitized
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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 toxicity of benzotriazole with cover letter 

dated 041989. Submitting Organization: Eastman Kodak Company. OTS0516745.
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CAS No. 95-14-7: Summary of Endpoints

Endpoint Study Information 

Available

Acceptable Data Gap

Aquatic Effects

Acute Toxicity to Fish 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