### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

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# Submission Report

Submission Information

Submission ID : 25349398 Submission Name : 1H-Benzotriazole, 4or 5-methylAR Number :



## Chemical Information

CAS Number : 29385-43-1 Ninth Collective Index Name : 1H-Benzotriazole, 4(or 5)-methyl

## Submitter Information

Submitter's Name : EPA
Address : City : Washington
State : DC
ZIP/Postal Code : Country : Type of Submitter : Company 

## Technical Point of Contact Information

Technical Point of Contact :
Title :
Phone Number :
Fax Number :
E-Mail Address : 

## Additional Technical Point of Contact Information

Technical Point of Contact :
Title :
Phone Number :
Fax Number :
E-Mail Address :



## Sponsor Information
Sponsor's Name : EPA
Address : City :
State : ZIP/Postal Code : Country :
Phone Number :
Fax Number : Share

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] E-Mail Address : Type of Sponsor : Company 

## Technical Point of Contact Information

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## Acute Toxicity to Aquatic Vertebrates

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity To Aquatic Vertebrates

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Referred to as 5-Methyl-1H-Benzotriazole or 4-Methyl-1H-Benzotriazole (MeBT); Provided by Cincinnati Specialties (Cincinnati Ohio)

Sponsored Chemical Result Type: Measured

Test Substance Result Type: Measured



### Method - Acute Toxicity To Aquatic Vertebrates
Year Study Performed:

Method/Guideline Followed: Other

Other Method/Guideline: Methods for measuring the acute toxicity of effluents and receiving waters to freshwater and marine microorganisms

Deviations from Method/Guideline:
Species: Pimephales promelas

GLP: No Data

Analytical Monitoring: Yes

Test Type: Semi-Static

Test Vessel:

Water Media Type:

Test Concentrations: Both

Nominal and Measured Concentrations: nominal: 150.8 and 188.9 mg/L; measured high concentrations of MeBT were between 95.5% and 99.5% of nominal concentrations at a confidence interval of 95%.

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Exposure Period: 96 Hours

Vehicle Used:

Vehicle Name:

Vehicle Amount and Units:
Alkalinity:

Dissolved Oxygen:

pH Value:

Test Temperature and Units: Value/Lower Range: 20

Photo (Light/Dark): 16/8

Salinity:

TOC:

Water Hardness:

Method/Guideline Test Conditions Remarks: Fathead minnows were 7 days old at test initiation.

A dilution water control was included.

The toxicity of MeBT to fathead minnows was assessed in moderately hard water using acute tests according to U.S. Environmental Protection Agency procedures. Mortality was the endpoint. The nominal (predilution) test concentrations of MeBT were 150.8 and 188.9 mg/L. Test chambers were 500-mL glass beakers. All tests were conducted at 20 C with a light:dark cycle of 16:8 h. Fathead minnows were fed 0.2 mL of brine shrimp nauplii (Artemia sp.) at 48 h and were renewed two hours after feeding with freshly prepared dilutions. LC50 values were calculated using either the graphical, probit, Spearman-Karber, or trimmed Spearman-Karber methods according to U.S. Environmental Protection Agency established criteria.

Limit Test:



### Test Results - Acute Toxicity To Aquatic Vertebrates
NOEC Exposure Duration:

NOEC:

LOEC Exposure Duration:

LOEC:

NOELR Exposure Duration:

NOELR:

LOELR Exposure Duration:

LOELR:

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] LC/EC Mean Value : Exposure Duration Exposure Units LC/EC % Value Description Mean Value

or Lower Mean Value Upper Mean Value Units Effect Observed Basis for Concentration

96 Hours LC 50 % = 38 65 mg/L Mortality

Results Remarks: Measured high concentrations of MeBT were between 95.5 and 99.5% of nominal concentrations at a confidence interval of 95%. Lower test concentrations were calculated as a proportion of the 100% measured concentration. The LC50s were calculated based on measured 100% concentration. 

### Reliability/Data Quality - Acute Toxicity To Aquatic Vertebrates
Reliability: 2 - Valid With Restrictions

Reliability Remarks:
Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Vertebrates

Reference: Cornell, J.S. et al. 2000. Comparative measures of the toxicity of component chemicals in aircraft deicing fluid. Environmental Chemistry, 19(6): 1465-1472.



## Acute Toxicity to Aquatic Vertebrates

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity To Aquatic Vertebrates

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Referred to as Tolyltriazole-Cobratec TT-100, Sample 1090. Supplied as tan granules.

Sponsored Chemical Result Type: Measured

Test Substance Result Type: Measured



### Method - Acute Toxicity To Aquatic Vertebrates
Year Study Performed: 1974

Method/Guideline Followed: Other

Other Method/Guideline: Fish - Pesticide Acute Toxicity Test guideline; January 20, 1971.

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Deviations from Method/Guideline:
Species: Lepomis macrochirus

GLP: No Data

Analytical Monitoring: Yes

Test Type:

Test Vessel:

Water Media Type:

Test Concentrations: Nominal

Nominal and Measured Concentrations: 3.16, 3.55, 3.98, 4.47 ppm

Exposure Period: 96 Hours

Vehicle Used: Yes

Vehicle Name: Acetone

Vehicle Amount and Units:
Alkalinity:

Dissolved Oxygen:

pH Value:

Test Temperature and Units:

Photo (Light/Dark):
Salinity:

TOC:

Water Hardness:

Method/Guideline Test Conditions Remarks: Average length of fish was 42 mm; average weight of fish was 0.8 gm.

Fish were held in a plastic wading pool prior to bioassay. The water temperature of the pool was 72 ±2 F. Fish were held for 10 days before bioassay.

Test Medium - Container: 5 gallon glass water bucket lined with a clear polyethylene bag which was discarded and replaced after each use. Water Quantity: 16 liters per test vessel. Water Quality: Standard reconstituted distilled water. Water Temperature: 72 ±2 F.

Test Chemicals - Candidate: Tolyltriazole-Cobratec TT-100, Sample 1090, (technical grade chemical) supplied by Sherwin-Williams Chemicals.

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Solvent, acetone (0.3 mL/liter of test water) Reference: p, p'DDT, 74% Solvent, acetone (0.3 mL/liter of test water)

Determinations of Lethal Concentrations - Screening tests using 2 fish per dose level were conducted to locate the general level of toxicity of Tolyltriazole-Cobratec TT-100, Sample 1090, and 74% p, p'DDT to the particular lot of fish.

From the results of the screening tests, the following concentrations were chosen as the doses to use in the bioassay: Cobratec TT-100 (ppm): 3.16, 3.55, 3.98, 4.47 74% p, p'DDT (ppm): 0.016, 0.013, 0.010, 0.008

The test lasted for 96 hours with mortality readings taken every 24 hours.

Twenty fish were exposed to each of the above concentrations. The fish were loaded ten to a test vessel (0.5 gm fish/liter of water). Twenty fish were untreated and served as controls.

Reaction of fish: 1. Controls - there were no deaths and no signs of intoxication. 2. Tolyltriazole - Cobratec TT-100, Sample 1090 - the major symptom of toxicity was loss of equilibrium prior to death. 3. 74%, p'DDT - death was usually preceded by loss of equilibrium.

Litchfield and Wilcoxon's method was used to compute the confidence limits for the LC1, LC50, and LC99 (at 24, 48 and 96 hours).

Limit Test:



### Test Results - Acute Toxicity To Aquatic Vertebrates
NOEC Exposure Duration:

NOEC:

LOEC Exposure Duration:

LOEC:

NOELR Exposure Duration:

NOELR:

LOELR Exposure Duration:

LOELR:

LC/EC Mean Value : Exposure Duration Exposure Units LC/EC % Value Description Mean Value

or Lower Mean Value Upper Mean Value Units Effect Observed Basis for Concentration

96 Hours LC 50 % = 3.47 ppm Mortality

48 Hours LC 50 % = 3.55 ppm Mortality

24 Hours LC 50 % = 4.02 ppm Mortality

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Results Remarks: The positive control responded appropriately. It is concluded that Cobratec TT-100 is less toxic to the bluegill sunfish than is 74% p, p'DDT. 

### Reliability/Data Quality - Acute Toxicity To Aquatic Vertebrates
Reliability: 2 - Valid With Restrictions

Reliability Remarks:
Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Vertebrates

Reference: TSCATS submission (OTS0521121, Doc ID 86-890000613, received 11/06/74): Huntingdon Research Center, Sherwin Williams Company. 1974. Initial submission: The Acute Toxicity of Tolyltriazole-Cobratec TT-100 Sample 1090 to the Bluegill Sunfish (Lepomis macrochirus) with Attachments and Cover Letter Dated 061289.



## Acute Toxicity to Aquatic Vertebrates

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity To Aquatic Vertebrates

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: off-white particles

Sponsored Chemical Result Type: Measured

Test Substance Result Type: Nominal



### Method - Acute Toxicity To Aquatic Vertebrates
Year Study Performed: 1992

Method/Guideline Followed: OECD 203

Other Method/Guideline:

Deviations from Method/Guideline:
Species: Pimephales promelas

GLP: Yes

Analytical Monitoring: No Data

Test Type: Semi-Static

Test Vessel:

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Water Media Type: Freshwater

Test Concentrations: Nominal

Nominal and Measured Concentrations: 13, 22, 36, 60 and 100 mg/L

Exposure Period: 96 Hours

Vehicle Used:

Vehicle Name:

Vehicle Amount and Units:
Alkalinity: 184-188 mg/L CaCO3

Dissolved Oxygen: 8.4 to 9.2 mg/L

pH Value: Value/Lower Range: = 7.8 Upper Range: 8.5

Test Temperature and Units: Value/Lower Range: 21 Upper Range: 22.9 °C

Photo (Light/Dark):
Salinity:

TOC:

Water Hardness: 144 mg/L

Method/Guideline Test Conditions Remarks: Fathead minnows were exposed to a geometric series of five test concentrations and a negative (well water) control under static-renewal test conditions for 96 hours. Two replicate test chambers were maintained in each treatment and control group, with 10 fish in each test chamber. Nominal test concentrations were selected in consultation with the sponsor, and were based upon results of range finding tests. Nominal concentrations selected were 13, 22, 36, 60 and 100 mg/L. Immediately prior to initiating the test, fish were impartially removed from holding tanks in groups of two and distributed among the test chambers until each contained 10 fish. At 48 hours, new test solutions were prepared and fish were transferred to the new solutions. Observations of mortality and other clinical signs of toxicity were made at approximately 24, 48, 72, and 96 hours after test initiation. Cumulative percent mortality values in each treatment group were used to calculate LC50 values and 95% confidence limits at 24, 48, 72 and 96 hours. The no mortality concentration was determined by visually examining the mortality data.

Limit Test: Yes



### Test Results - Acute Toxicity To Aquatic Vertebrates
NOEC Exposure Duration:

NOEC:

LOEC Exposure Duration:

LOEC:

NOELR Exposure

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Duration:

NOELR:

LOELR Exposure Duration:

LOELR:

LC/EC Mean Value : Exposure Duration Exposure Units LC/EC % Value Description Mean Value

or Lower Mean Value Upper Mean Value Units Effect Observed Basis for Concentration

96 Hours LC 50 % = 43 mg/L Mortality

96 Hours LC 0 % = 22 mg/L Mortality

Results Remarks: Water temperatures taken daily with a hand-held thermometer ranged from 21.0 to 22.9
C. Dissolved oxygen and pH readings ranged from 8.4 to 9.2 mg/L and 7.8 to 8.5, respectively. Dissolved oxygen concentrations exceeded 60% of saturation throughout the test. Conductivity, alkalinity, and hardness measured at test initiation (0 hours) in the Negative Control Replicate A were 295 umhos/cm, 188 mg/L as CaCO3, and 144 mg/L as CaCO3, respectively. At test termination (96 hours) they were 325 umhos/cm, 184 mg/L as CaCO3, and 144 mg/L as CaCO3, respectively, in the Negative Control Replicate A.

After the test substance was added to the vortex, a clear colorless solution as formed in the test chambers. White particulate matter was noted on the bottom of one tank (Replicate A) of the 36 mg/L treatment group, approximately 24 hours after test initiation. The particulate matter remained until test termination.

Fish in the negative control, with the exception of one fish which appeared lethargic at 48 hours, appeared healthy and normal throughout the test. The 13 and 22 mg/L groups appeared normal and healthy throughout the test with no signs of disease or
stress. Overt signs of toxicity observed at the 36 mg/L concentration, beginning approximately 24 hours after test initiation, included lethargy, loss of equilibrium, and erratic swimming. At the 36 mg/L concentration, 15% mortality was observed at 24 hours with no further mortality occurring by 96 hours. At the 60 and 100 mg/L concentrations, 100% mortality was observed at 24 hours.



### Reliability/Data Quality - Acute Toxicity To Aquatic Vertebrates
Reliability: 1 - Valid Without Restrictions

Reliability Remarks:
Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Vertebrates

Reference: TSCATS submission (OTS0557287, Doc ID 86940000877S, received 06/02/94): Wildlife International Ltd, CBI. 1993. Initial submission: A 96-hour Static Renewal ToxicityTest with the Fathead Minnow (Pimephales promelas) of Tolyltriazole - 1HBenzotriazole, Methyl with Cover Letter Dated 04/29/94 (SANITIZED).

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] 

## Acute Toxicity to Aquatic Vertebrates

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity To Aquatic Vertebrates

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Referred to as Tolyltriazole-Cobratec TT-100, Sample 1090. Supplied as tan granules.

Sponsored Chemical Result Type: Measured

Test Substance Result Type: Measured



### Method - Acute Toxicity To Aquatic Vertebrates
Year Study Performed: 1975

Method/Guideline Followed: Other

Other Method/Guideline: Fish - Pesticide Acute Toxicity Test guideline; January 20, 1971

Deviations from Method/Guideline:
Species: Lepomis macrochirus

GLP: No Data

Analytical Monitoring: Yes

Test Type:

Test Vessel:

Water Media Type: Freshwater

Test Concentrations: Nominal

Nominal and Measured Concentrations: 25.1, 28.2, 31.6, 35.5, 39.8 and 44.7 ppm

Exposure Period: 96 Hours

Vehicle Used: Yes

Vehicle Name: Acetone

Vehicle Amount and Units:
Alkalinity:

Dissolved Oxygen:

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] pH Value:

Test Temperature and Units:

Photo (Light/Dark):
Salinity:

TOC:

Water Hardness:

Method/Guideline Test Conditions Remarks: Average length of fish was 38 mm; average weight of fish was 0.9 gm.

Fish were held in a plastic wading pool prior to bioassay. The water temperature of the pool was 72 ±2 F. Fish were held for 10 days before bioassay.

Test Medium - Container: 5 gallon glass water bucket Water Quantity: 18 liters per test vessel. Water Quality: Standard reconstituted distilled water. Water Temperature: 72 ±2 F.

Test Chemicals - Candidate: Tolyltriazole-Cobratec TT-100, Sample 1090, (technical grade chemical) supplied by Sherwin-Williams Chemicals. Solvent, acetone < 0.3 mL/liter of test water

Determinations of Lethal Concentrations - Screening tests using 2 fish per dose level were conducted to locate the general level of toxicity of Tolyltriazole-Cobratec TT-100, Sample 1090 to the particular lot of fish.

From the results of the screening tests, the following concentrations were chosen as the doses to use in the bioassay: Cobratec TT-100 (ppm): 25.1, 28.2, 31.6, 35.5, 39.8 and 44.7 ppm

The test lasted for 96 hours with mortality readings taken every 24 hours.

Twenty fish were exposed to each of the above concentrations. The fish were loaded ten to a test vessel (0.5 gm fish/liter of water). Twenty fish were untreated and served as controls.

Reaction of fish: 1. Controls - there were no deaths and no signs of intoxication. 2. Tolyltriazole - Cobratec TT-100, Sample 1090 - the major symptom of toxicity was loss of equilibrium, sporadic swimming, and lying on their side prior to death

Litchfield and Wilcoxon's method was used to compute the confidence limits at 96 hours.

Limit Test:



### Test Results - Acute Toxicity To Aquatic Vertebrates
NOEC Exposure Duration:

NOEC:

LOEC Exposure Duration:

LOEC:

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] NOELR Exposure Duration:

NOELR:

LOELR Exposure Duration:

LOELR:

LC/EC Mean Value : Exposure Duration Exposure Units LC/EC % Value Description Mean Value

or Lower Mean Value Upper Mean Value Units Effect Observed Basis for Concentration

96 Hours LC 50 % = 31 ppm Mortality

Results Remarks: Cobratec TT-100 No. Dead/No. Dosed 35.5 20/20 31.6 10/20 28.2 2/20 25.1 1/20



### Reliability/Data Quality - Acute Toxicity To Aquatic Vertebrates
Reliability: 2 - Valid With Restrictions

Reliability Remarks:
Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Vertebrates

Reference: TSCATS submission (OTS0521122, Doc ID 86-890000614, received 09/29/75): Huntingdon Research Center, Sherwin Williams Company. 1975. Initial submission: The Acute Toxicity of Tolyltriazole-Cobratec TT-100 Sample 1090 to the Bluegill Sunfish (Lepomis macrochirus) with Attachments and Cover Letter Dated 061289.



## Acute Toxicity to Aquatic Vertebrates

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity To Aquatic Vertebrates

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Referred to as Tolyltriazole-Cobratec TT-100, Sample 1090. Supplied as tan granules.

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Sponsored Chemical Result Type: Measured

Test Substance Result Type: Measured



### Method - Acute Toxicity To Aquatic Vertebrates
Year Study Performed:

Method/Guideline Followed: Other

Other Method/Guideline: Fish - Pesticide Acute Toxicity Test guideline; January 20, 1971

Deviations from Method/Guideline:
Species: Salmo sp.

GLP: No Data

Analytical Monitoring: Yes

Test Type:

Test Vessel:

Water Media Type: Freshwater

Test Concentrations: Nominal

Nominal and Measured Concentrations: 25.1, 28.2, 31.6, 35.5, 39.8 and 44.7 ppm

Exposure Period: 96 Hours

Vehicle Used: Yes

Vehicle Name: Acetone

Vehicle Amount and Units:
Alkalinity:

Dissolved Oxygen:

pH Value:

Test Temperature and Units:

Photo (Light/Dark):
Salinity:

TOC:

Water Hardness:

Method/Guideline Average length of fish was 43 mm; average weight of fish was 0.35 gm.

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Test Conditions Remarks: Fish were held in an indoor tank prior to bioassay. The water temperature of the pool was 52 ±2 F. Fish were held for 10 days before bioassay.

Test Medium - Container: 5 gallon glass water bucket Water Quantity: 14 liters per test vessel. Water Quality: Standard reconstituted distilled water. Water Temperature: 52 ±2 F.

Test Chemicals - Candidate: Tolyltriazole-Cobratec TT-100, Sample 1090, (technical grade chemical) supplied by Sherwin-Williams Chemicals. Solvent, acetone (0.3 mL/liter of test water)

Reference: p, p'DDT, 74% Solvent, acetone (0/.3 mL/liter of test water)

Determinations of Lethal Concentrations - Screening tests using 5 fish per dose level were conducted to locate the general level of toxicity of Tolyltriazole-Cobratec TT-100, Sample 1090, and 74% p, p'DDT to the particular lot of fish.

From the results of the screening tests, the following concentrations were chosen as the doses to use in the bioassay: Cobratec TT-100 (ppm): 10.01, 14.10, 20.00, 28.20 74% p, p'DDT (ppm): 0.0200, 0.0251, 0.0316, 0.0308

The test lasted for 96 hours with mortality readings taken every 24 hours.

Twenty fish were exposed to each of the above concentrations. The fish were loaded ten to a test vessel (0.5 gm fish/liter of water). Twenty fish were untreated and served as controls.

Reaction of fish: 1. Controls - there were no deaths and no signs of intoxication. 2. Tolyltriazole - Cobratec TT-100, Sample 1090 - the major symptom of toxicity was loss of equilibrium prior to death. 3. 74% p, p'DDT - death was usually preceded by loss of equilibrium.

Litchfield and Wilcoxon's method was used to compute the confidence limits at 96 hours.

Limit Test:



### Test Results - Acute Toxicity To Aquatic Vertebrates
NOEC Exposure Duration:

NOEC:

LOEC Exposure Duration:

LOEC:

NOELR Exposure Duration:

NOELR:

LOELR Exposure Duration:

LOELR:

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] LC/EC Mean Value : Exposure Duration Exposure Units LC/EC % Value Description Mean Value

or Lower Mean Value Upper Mean Value Units Effect Observed Basis for Concentration

96 Hours LC 50 % = 21.4 ppm Mortality

Results Remarks:



### Reliability/Data Quality - Acute Toxicity To Aquatic Vertebrates
Reliability: 2 - Valid With Restrictions

Reliability Remarks:
Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Vertebrates

Reference: TSCATS submission (OTS0521123, Doc ID 86-890000615, received 06/13/89): Huntingdon Research Center, Sherwin Williams Company. 1974. Initial submission: The Acute Toxicity of Tolyltriazole-Cobratec TT-100 Sample 1090 to the Rainbow Trout (Salmo gairdneri) with Attachments and Cover Letter Dated 061289.



## Acute Toxicity to Aquatic Vertebrates

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity To Aquatic Vertebrates

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: 50% solution in water of the sodium salt of tolyltriazole. Referred to as Cobratec TT 50S. Described as dark amber, water-soluble liquid.

Sponsored Chemical Result Type: Measured

Test Substance Result Type: Nominal



### Method - Acute Toxicity To Aquatic Vertebrates
Year Study Performed: 1979

Method/Guideline Followed: Other

Other Method/Guideline: Committee on Methods for Toxicity Tests with Aquatic Organisms (1975)

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Deviations from Method/Guideline:
Species: Lepomis macrochirus

GLP: No Data

Analytical Monitoring: No

Test Type: Static

Test Vessel:

Water Media Type: Freshwater

Test Concentrations: Nominal

Nominal and Measured Concentrations: ~ 0, 16.0, 28.0, 50.0, 90.0 and 160.0 mg sodium tolyltriazole/L (0, 32.0, 56.0, 100.0, 180.0 and 320.0 mg Cobratec TT 50S / L)

Exposure Period: 96 Hours

Vehicle Used:

Vehicle Name:

Vehicle Amount and Units:
Alkalinity: 30 mg/L

Dissolved Oxygen: 5.2 to 9.3 mg/L

pH Value: Value/Lower Range: = 7.18 Upper Range: 9.4

Test Temperature and Units:

Photo (Light/Dark):
Salinity:

TOC:

Water Hardness: 42 mg/L

Method/Guideline Test Conditions Remarks: Dilution water for the test was obtained from a well on the Tarrytown site, treated with a Continental Model 3020 Reverse Osmosis Water System, and deionized; then, following procedures of Marking and Dawson (1973), it was reconstituted to a pH of 7.46, total hardness of 42 mg/L as CaCO3, total alkalinity of 30 mg/L as CaCO3 and specific conductance of 155 μmhos/cm. Hardness and alkalinity were determined according to standard analytical procedures (American Public Healthy Association, 1976), pH with an ORION pH Meter, conductivity with a YSI Conductivity Bridge and dissolved oxygen with a YSI Oxygen Meter. Test practices followed these recommended by the Committee on Methods for Toxicity Tests with Aquatic Organisms (1975), except that replicate concentrations were not used. Samples of Cobratec TT 50S were weighed to a precision of 0.1 mg and added directly to the test containers. Test vessels were 19.6 liter, chemically clean glass jars, each containing 15 liters of soft reconstituted dilution water and immersed in a constant temperature recirculating water bath. Testing was begun by adding measured quantities of test material to the water in

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] five test vessels, and mixing thoroughly, to yield nominal concentrations of 32.0, 56.0, 100.0, 180.0 and 320.0 mg/L. A sixth vessel, as control, contained 100% dilution water. Ten fish were introduced at random into each of the test and control vessels; these were approximately 4 months old with a mean (10 organisms) length of 37 mm and mean weight of 0.50 grams. Biological loading was 0.33 g/L. At the beginning of the test, and every 24 hours thereafter, dissolved oxygen and pH of the control and each test concentration, and temperature of the water bath, were determined. Mortalities among the test fish, and any observable abnormal behavioral responses, were noted and recorded every 24 hours. The no effect level was determined, by observation, at 96 hours. The concentration of test material lethal to 50% of the test population, and 95% confidence limits, were determined for the 72- and 96-hour exposure periods by the Spearman-Karber Estimator. LC50s could not be calculated for the 24 and 48 hour exposure periods due to the lack of sufficient mortalities. LC50 calculations are based on nominal concentrations of the test material in soft reconstituted water.

Limit Test:



### Test Results - Acute Toxicity To Aquatic Vertebrates
NOEC Exposure Duration:

NOEC:

LOEC Exposure Duration:

LOEC:

NOELR Exposure Duration:

NOELR:

LOELR Exposure Duration:

LOELR:

LC/EC Mean Value : Exposure Duration Exposure Units LC/EC % Value Description Mean Value

or Lower Mean Value Upper Mean Value Units Effect Observed Basis for Concentration

Results Remarks: The 96-hour LC50 was ~ 95.6 mg sodium tolyltriazole/L (191.2 mg Cobratec TT 50S / L). The 96-hour NOEC was ~ 28.0 mg sodium tolyltriazole/L (56.0 mg Cobratec TT 50S /L). pH ranged from 7.18 to 9.4. Dissolved oxygen ranged from 5.2 to 9.3 mg/L. 

### Reliability/Data Quality - Acute Toxicity To Aquatic Vertebrates
Reliability: 1 - Valid Without Restrictions

Reliability Remarks:

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Vertebrates

Reference: TSCATS submission (OTS0521126, Doc ID 86-890000618, received 06/13/89): Union Carbide Corp, Sherwin Williams Company. 1979. Initial submission: The Acute Toxicity of Cobratec TT 50S to the Bluegill Sunfish (Lepomis macrochirus) Rafinesque with Cover Letter Dated 061289.



## Acute Toxicity to Aquatic Vertebrates

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity To Aquatic Vertebrates

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: 50% solution in water of the sodium salt of tolyltriazole. Referred to as Cobratec TT 50S. Described as dark amber, water soluble liquid.

Sponsored Chemical Result Type: Measured

Test Substance Result Type: Nominal



### Method - Acute Toxicity To Aquatic Vertebrates
Year Study Performed: 1979

Method/Guideline Followed: Other

Other Method/Guideline: Committee on Methods for Toxicity Tests with Aquatic Organisms (1975)

Deviations from Method/Guideline:
Species: Salmo sp.

GLP: No Data

Analytical Monitoring: No

Test Type: Static

Test Vessel:

Water Media Type: Freshwater

Test Concentrations: Nominal

Nominal and Measured Concentrations: ~ 0, 2.8, 5.0, 9.0, 16.0 and 28.0 mg sodium tolyltriazole / L (0, 5.6, 10.0, 18.0, 32.0 and 56.0 mg Cobratec TT 50S/L)

Exposure Period: 96 Hours

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Vehicle Used:

Vehicle Name:

Vehicle Amount and Units:
Alkalinity: 28 mg/L

Dissolved Oxygen: 6.2 to 9.4 mg/L

pH Value: Value/Lower Range: = 7.03 Upper Range: 8.42

Test Temperature and Units:

Photo (Light/Dark):
Salinity:

TOC:

Water Hardness: 42 mg/L

Method/Guideline Test Conditions Remarks: Dilution water for the test was obtained from a well on the Tarrytown site, treated with a Continental Model 3020 Reverse Osmosis Water System, and deionized; then, following procedures of Marking and Dawson (1973), it was reconstituted to a pH of 7.47, total hardness of 42 mg/L as CaCO3, total alkalinity of 28 mg/L as CaCO3 and specific conductance of 180 umhos/cm. Hardness and alkalinity were determined according to standard analytical procedures (American Public Healthy Association, 1976), pH with an ORION pH Meter, conductivity with a YSI Conductivity Bridge and dissolved oxygen with a YSI Oxygen Meter. Test practices followed these recommended by the Committee on Methods for Toxicity Tests with Aquatic Organisms (1975), except that replicate concentrations were not used. A fresh working stock solution of the test material in deionized water was prepared by weight to a precision of 0.1 mg. Five concentrations were tested, with a control. Test vessels were 19.6 liter, chemically clean glass jars, each containing 15 liters of soft reconstituted dilution water and immersed in a constant temperature recirculating water bath. Testing was begun by adding measured volumes of stock solution to the water in five test vessels, and mixing thoroughly, to yield nominal test material concentrations of 5.6, 10.0, 18.0, 32.0 and 56.0 mg/L. A sixth vessel, as control, contained 100% dilution water. Ten fish were introduced at random into each of the test and control vessels; these were approximately 3 months old with a mean (10 organisms) length of 54 mm and mean weight of 1.27 grams. Biological loading was 0.85 g/L. At the beginning of the test, and every 24 hours thereafter, dissolved oxygen and pH of the control and each test concentration, and temperature of the water bath, were determined. Mortalities among the test fish, and any observable abnormal behavioral responses were noted and recorded every 24 hours. The no effect level was determined, by observation, at 96 hours. The concentration of test material lethal to 50% of the test population was determined for the 24, 48, 72 and 96 hour exposure periods by the Spearman-Karber Estimator. Confidence limits for the 48 hour exposure period were also calculated. LC50 calculations are based on nominal concentrations of the test material in soft reconstituted water.

Limit Test:



### Test Results - Acute Toxicity To Aquatic Vertebrates
NOEC Exposure Duration:

NOEC:

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] LOEC Exposure Duration:

LOEC:

NOELR Exposure Duration:

NOELR:

LOELR Exposure Duration:

LOELR:

LC/EC Mean Value : Exposure Duration Exposure Units LC/EC % Value Description Mean Value

or Lower Mean Value Upper Mean Value Units Effect Observed Basis for Concentration

Results Remarks: The 96-hour LC50 was ~ 11.9 mg sodium tolyltriazole/L (23.7 mg Cobratec TT 50S /L). The 96-NOEC was ~ 5.0 mg sodium tolyltriazole/L (10.0 mg Cobratec TT 50S /L). pH ranged from 7.03 to 8.42. Dissolved oxygen ranged from 6.2 to 9.4 mg/L. 

### Reliability/Data Quality - Acute Toxicity To Aquatic Vertebrates
Reliability: 1 - Valid Without Restrictions

Reliability Remarks:
Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Vertebrates

Reference: TSCATS submission (OTS0521128, Doc ID 86-890000620, received 06/13/89): Union Carbide Corp, Sherwin Williams Company. 1979. Initial submission: The Acute Toxicity of Cobratec TT50S to the Rainbow Trout (Salmo gairdneri) Richardson with Cover Letter Dated 061289.



## Acute Toxicity to Aquatic Vertebrates

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity To Aquatic Vertebrates

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Referred to as Tolyltriazole-Cobratec TT-100, Sample 1090. Supplied as tan granules.

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Comments:
Sponsored Chemical Result Type: Measured

Test Substance Result Type: Nominal



### Method - Acute Toxicity To Aquatic Vertebrates
Year Study Performed: 1974

Method/Guideline Followed: Other

Other Method/Guideline: Fish - Pesticide Acute Toxicity Test guideline. January 20, 1971

Deviations from Method/Guideline:
Species: Pimephales promelas

GLP: No Data

Analytical Monitoring: Yes

Test Type:

Test Vessel:

Water Media Type:

Test Concentrations: Nominal

Nominal and Measured Concentrations: 17.80, 22.40, 28.20, 35.50 ppm

Exposure Period: 96 Hours

Vehicle Used: Yes

Vehicle Name: Acetone

Vehicle Amount and Units:
Alkalinity:

Dissolved Oxygen:

pH Value:

Test Temperature and Units:

Photo (Light/Dark):
Salinity:

TOC:

Water Hardness:

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Method/Guideline Test Conditions Remarks: Average length of fish was 65mm; average weight of fish was 1.2 gm.

Fish were held in a plastic wading pool prior to bioassay. The water temperature of the pool was 68 ±2 deg F. Fish were held for 10 days before bioassay.

Test Medium - Container: 5 gallon glass water bucket lined with a clear polyethylene bag which was discarded and replaced after each use. Water Quantity: 12 liters per test vessel. Water Quality: Standard reconstituted distilled water. Water Temperature: 68 ±2 deg F.

Test Chemicals - Candidate: Tolyltriazole-Cobratec TT-100, Sample 1090, (technical grade chemical) supplied by Sherwin-Williams Chemicals. Solvent, acetone (0.3 mL/liter of test water) Reference: p, p'DDT, 74% Solvent, acetone (0.3 mL/liter of test water)

Determinations of Lethal Concentrations - Screening tests using 2 fish per dose level were conducted to locate the general level of toxicity of Tolyltriazole-Cobratec TT-100, Sample 1090, and 74% p, p'DDT to the particular lot of fish.

From the results of the screening tests, the following concentrations were chosen as the doses to use in the bioassay: Cobratec TT-100 (ppm): 17.80, 22.40, 28.20, 35.50 74% p, p'DDT (ppm): 0.0563, 0.0709, 0.0892, 0.1120, 0.1410

The test lasted for 96 hours with mortality readings taken every 24 hours.

Twenty fish were exposed to each of the above concentrations. The fish were loaded five to a test vessel (0.5 gm fish/liter of water). Twenty fish were untreated and served as controls.

Reaction of fish: 1. Controls - there were no deaths and no signs of intoxication. 2. Tolyltriazole - Cobratec TT-100, Sample 1090 - the major symptom of toxicity was loss of equilibrium prior to death. 3. 74%, p'DDT - death was usually preceded by loss of equilibrium.

Litchfield and Wilcoxon's method was used to compute the confidence limits for the LC1, LC50, and LC99 (at 24, 48 and 96 hours).

Limit Test:



### Test Results - Acute Toxicity To Aquatic Vertebrates
NOEC Exposure Duration:

NOEC:

LOEC Exposure Duration:

LOEC:

NOELR Exposure Duration:

NOELR:

LOELR Exposure Duration:

LOELR:

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] LC/EC Mean Value : Exposure Duration Exposure Units LC/EC % Value Description Mean Value

or Lower Mean Value Upper Mean Value Units Effect Observed Basis for Concentration

96 Hours LC 50 % = 25.5 ppm Mortality

48 Hours LC 50 % = 29.8 ppm Mortality

24 Hours LC 50 % = 36.9 ppm Mortality

Results Remarks: The positive control responded appropriately. It is concluded that Cobratec TT-100 is less toxic to the fathead minnow than is 74% p, p'DDT. 

### Reliability/Data Quality - Acute Toxicity To Aquatic Vertebrates
Reliability: 2 - Valid With Restrictions

Reliability Remarks:
Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Vertebrates

Reference: TSCATS submission (OTS0521120, Doc ID 86-890000612; received 11/06/74): Huntingdon Research Center, Sherwin Williams Company. 1974. Initial submission: The Acute Toxicity of Tolyltriazole-Cobratec TT-100 Sample 1090 to the Fathead Minnow (Pimephales promelas) with Attachments and Cover Letter dated 061289.



## Acute Toxicity to Aquatic Invertebrates

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity To Aquatic Invertebrates

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: 5-Methyl-1H-Benzotriazole or 4-Methyl-1H-Benzotriazole (MeBT), CAS No. 29385-43-1

Sponsored Chemical Result Type: Measured

Test Substance Result Type: Measured



### Method - Acute Toxicity To Aquatic Invertebrates
Year Study Performed:

Method/Guideline Followed: Other

Other Method/Guideline: Methods for measuring the acute toxicity of effluents and receiving waters to freshwater and marine microorganisms

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Deviations from Method/Guideline:
Species/In Vitro System: Ceriodaphnia dubia

GLP: No Data

Analytical Monitoring: No Data

Test Type:

Test Vessel:

Water Media Type:

Test Concentrations: Both

Nominal and Measured Concentrations: nominal: 152.1 and 183.8 mg/L; measured: Measured high concentrations of MeBT were between 95.5 and 99.5% of nominal concentrations at a confidence interval of 95%.

Exposure Period: 48 Hours

Vehicle Used:

Vehicle Name:

Vehicle Amount and Units:
Alkalinity:

Dissolved Oxygen:

pH Value:

Test Temperature and Units: Value/Lower Range: 20 °C

Photo (Light/Dark): 16/8

Salinity:

TOC:

Water Hardness:

Method/Guideline Test Conditions Remarks: All C. dubia used in the tests were <24 (±8) h old at test initiation. The toxicity of MeBT to C. dubia was assessed in moderately hard water using acute tests according to U.S. Environmental Protection Agency procedures. Mortality was the endpoint. The nominal (predilution) test concentrations of MeBT were 152.1 and 183.8 mg/L. A dilution water control was also included. Test chambers were 30-mL plastic cups. All tests were conducted at 20 C with a light:dark cycle of 16:8 h. Testing was conducted under static conditions for a 48 hour duration. The LC50 values werecalculated using either the graphical, probit, Spearman-Karber, or trimmed SpearmanKarber methods according to U.S. Environmental Protection Agency-established criteria.

Limit Test:



### Test Results - Acute Toxicity To Aquatic Invertebrates

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] NOEC Exposure Duration:

NOEC:

LOEC Exposure Duration:

LOEC:

NOELR Exposure Duration:

NOELR:

LOELR Exposure Duration:

LOELR:

LC/EC Mean Value : Exposure Duration Exposure Units LC/EC % Value Description Mean Value

or Lower Mean Value Upper Mean Value Units Effect Observed Basis for Concentration

48 Days EC 50 % = 102 108 mg/L

Results Remarks: Measured high concentrations of MeBT were between 95.5 and 99.5% of nominal concentrations at a confidence interval of 95%. Lower test concentrations were calculated as a proportion of the 100% measured concentration. The LC50s were calculated based on measured 100% concentration. 

### Reliability/Data Quality - Acute Toxicity To Aquatic Invertebrates
Reliability: 2 - Valid With Restrictions

Reliability Remarks:
Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Invertebrates

Reference: Cornell, J.S. et al. 2000. Comparative measures of the toxicity of component chemicals in aircraft deicing fluid. Environmental Chemistry, 19(6): 1465-1472.



## Acute Toxicity to Aquatic Invertebrates

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity To Aquatic Invertebrates

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methyl-

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Test Substance Purity/Composition and Other Test Substance Comments: Referred to as RM 27513 (EnviroSystems Sample Number 2694E). Delivered on November 15,
1990. RM 27513 was supplied by 3M Company, 935 Bush Avenue, St. Paul, Minnesota.

Sponsored Chemical Result Type: Measured

Test Substance Result Type: Nominal



### Method - Acute Toxicity To Aquatic Invertebrates
Year Study Performed: 1991

Method/Guideline Followed: Other

Other Method/Guideline: based on procedures of the OECD (1984)

Deviations from Method/Guideline:
Species/In Vitro System: Daphnia magna

GLP: Yes

Analytical Monitoring: No

Test Type: Static

Test Vessel:

Water Media Type: Freshwater

Test Concentrations: Nominal

Nominal and Measured Concentrations: 0 mg/L (control), 15 mg/L, 25 mg/L, 40 mg/L, 60 mg/L, and 100 mg/L

Exposure Period: 48 Hours

Vehicle Used: No

Vehicle Name:

Vehicle Amount and Units:
Alkalinity:

Dissolved Oxygen: 8.3 to 9.7 mg/L

pH Value: Value/Lower Range: = 7.7 Upper Range: 8.4

Test Temperature and Units: Value/Lower Range: 19 Upper Range: 20.7 °C

Photo (Light/Dark): 16/8

Salinity:

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] TOC:

Water Hardness:

Method/Guideline Test Conditions Remarks: Dilution water - Water used for acclimation of test organisms and for all toxicity testing was collected from wells at EnviroSystems in Hampton, New Hampshire. Water was stored in 500-gallon polyethylene tanks where it was aerated.

Test organism - Juvenile daphnids employed as test organisms were from a single source and were identified using an appropriate taxonomic key. Daphnids used in the test were produced from an in-house culture and were less than 24 hours old at the start of the
test. Control daphnids were weighed at the conclusion of the toxicity test. Prior to testing, daphnids were maintained in 100% dilution water under static conditions in 4 liter glass jars. During acclimation daphnids were not treated for disease and they were free of apparent sickness, injuries, and abnormalities at the beginning of the
test. During the acclimation period 24 hours prior to the test initiation the temperature was 19.0 - 19.8 C. Daphnids were fed yeast, trout chow, and/or the freshwater alga Selenastrum capricornutum once or twice daily before the test.

Toxicity testing - A screening test with the test substance was conducted during November 27 to 29, 1990. Nominal concentrations of test substance were: 0.1, 1.0, 10, 100, and 1000 mg/L. After 48 hours of exposure there was 0% survival at 1000 mg/L, 75% survival at 100 mg/L, and 100% survival at all other tested concentrations. The definitive toxicity test was performed during December 12 to 14, 1990, according to EnviroSystems Test Protocol 90169-3 (Static, Acute Toxicity Test With RM 27513 And The Daphnid, Daphnia magna), which was signed by the Study Director on November 16, 1990. It is based on procedures of the OECD (1984). The test was conducted at a target temperature of 20 ± 1 C with five concentrations of test substance and a dilution water control. A 1000 mg/L stock solution was prepared for combining 1.0 g of test substance with dilution water and adjusting the final concentrations to 1000 ml without the use of a solvent. Nominal concentrations of the test material were: 0 mg/L (control), 15 mg/L, 25 mg/L, 40 mg/L, 60 mg/L, and 100 mg/L RM
27513. Twenty daphnids were randomly and equally distributed among four replicates of each
treatment. The test was performed in 250 ml glass beakers that contained 200 ml of test solution (water depth was approximately 6 cm). Test vessels were randomly arranged in an incubator during the 48 hour test (a random numbers table was used to select the location of each vessel). A 16 hour light and 8 hour dark photoperiod was automaticallymaintained with cool-white fluorescent lights that provided a light intensity of 30 uEs1m-2. Aeration was not required to maintain dissolved oxygen concentrations above acceptable levels. Daphnids were not fed during the test. The number of surviving organisms and the occurrence of sublethal effects and immobilization (inability to swim within 15 seconds after gentle agitation) or other sublethal effects (loss of equilibrium, erratic swimming, loss of reflex, excitability, discoloration, or change in behavior) were determined visually and recorded initially after 24 and 48 hours. Dead test organisms were removed when first observed. Dissolved oxygen, pH, conductivity and temperature were measured and recorded daily in each test chamber that contained live animals. Results of the toxicity test were interpreted by standard statistical techniques (Stephan, 1983). Nominal concentrations of test substance the number of daphnids killed or immobilized were used to calculate the EC50 value.

Limit Test:



### Test Results - Acute Toxicity To Aquatic Invertebrates
NOEC Exposure Duration:

NOEC:

LOEC Exposure Duration:

LOEC:

NOELR Exposure

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Duration:

NOELR:

LOELR Exposure Duration:

LOELR:

LC/EC Mean Value : Exposure Duration Exposure Units LC/EC % Value Description Mean Value

or Lower Mean Value Upper Mean Value Units Effect Observed Basis for Concentration

48 Hours EC 50 % = 19 mg/L Immobilization Nominal

24 Minutes EC 50 % = 34 mg/L Immobilization Nominal

Results Remarks: Test vessels containing RM 27513 appeared clear initially and remained free of insoluble material throughout the test. Ninety-five percent survival occurred in the control

exposure. Control daphnids had an average wet weight (blotted dry) of 0.001 g at the end of the test. Loading rate during the toxicity test was approximately 0.02 g/L. Exposure of daphnids to the reference toxicant sodium dodecyl sulfate resulted in a 48 hour median lethal concentration (LC50) of 13.8 mg/L.

pH ranged from 7.7 to 8.4 throughout testing. Conductivity ranged from 12000 to 13000 umho/cm. Temperature ranged from 19.0 to 20.7 C. Dissolved oxygen ranged from 8.3 to 9.7 mg/L.



### Reliability/Data Quality - Acute Toxicity To Aquatic Invertebrates
Reliability: 2 - Valid With Restrictions

Reliability Remarks:
Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Invertebrates

Reference: TSCATS submission (OTS0533466, Doc ID 86-920000103S, received 10/31/91): Resource Analysts, Inc., 3M Company. 1991. Initial submission: Environmental Effects of Tolyltriazole with Attachments and Cover Letter 102891 (SANITIZED). pp. 75-91.



## Acute Toxicity to Aquatic Plants

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity To Aquatic Plants

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Referred to as RM 27513 (EnviroSystems Sample Number 2694E). Delivered on November 15, 1990. RM 27513 (white granules) was supplied by 3M Company, 935 Bush

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Substance Comments: Avenue, St. Paul, Minnesota.

Sponsored Chemical Result Type: Measured

Test Substance Result Type: Nominal



### Method - Acute Toxicity To Aquatic Plants
Year Study Performed: 1991

Method/Guideline Followed: Other

Other Method/Guideline: Based on procedures of the OECD (1984)

Deviations from Method/Guideline:
Species: Pseudokirchnerella subcapitata

GLP: No Data

Analytical Monitoring: No

Test Type: Static

Test Vessel: Closed

Water Media Type: Freshwater

Test Concentrations: Nominal

Nominal and Measured Concentrations: 0 mg/L (control), 62.5 mg/L, 125 mg/L, 250 mg/L, 500 mg/L, and 1000 mg/L

Exposure Period: 96 Hours

Vehicle Used:

Vehicle Name:

Vehicle Amount and Units:
Alkalinity:

Dissolved Oxygen:

pH Value: Value/Lower Range: = 6.7 Upper Range: 9.4

Test Temperature and Units: Value/Lower Range: 23.1 Upper Range: 23.5 °C

Photo (Light/Dark): 24/0

Salinity:

TOC:

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Water Hardness:

Method/Guideline Test Conditions Remarks: Dilution water - Water used for acclimation of test organisms and for all toxicity testing was sterile enriched media (U.S. EPA, 1978) with a pH of approximately
8.0.

Test organism - Algae used for the test was from a culture originally procured from the Culture Collection of Algae at the University of Texas at Austin and delivered to Envirosystems on April 12, 1990. The identity of the culture was verified using an appropriate taxonomic key. The culture was transferred to sterile enriched media identical to media used for this test and maintained at test conditions.

Toxicity testing - The definitive toxicity test was performed during April 30 to May 4, 1991. It is based on procedures of the OECD (1984). The test was conducted at a target temperature of 22 ± 2 C with five concentrations of test substance and a dilution water control. A 1,000 mg/L stock solution was prepared in dilution water. The stock solution was added directly to dilution water contained in the test vessels without the use of a solvent. Nominal concentrations of the test material were: 0 mg/L (control), 62.5 mg/L, 125 mg/L, 250 mg/L, 500 mg/L, and 1000 mg/L. Algae was randomly and equally distributed among three replicates of each treatment at the rate of 10,000 cells/mL. The test was performed in 250 mL glass Erlenmeyer flasks that contained 100 mL of test solution. The flasks were capped with inverted glass beakers. Test vessels were randomly arranged on the rotary shaker in an incubatory during the 96 hour test (a random numbers table was used to select the location of each vessel). A 24 hour light and 0 hour dark photoperiod was maintained with cool-white fluorescent lights that provided a light intensity of 35 uEs-1m-2. The number of algal cells/mL in each test vessel was determined visually every 24 hours by means of direct microscopic counts with a hemacytometer. The pH (Beckman model pHI 12 meter) was determined in each test flask at the beginning and end of the test, and temperature (ASTM mercury thermometer) was measured and recorded daily in the incubator

Statistical methods - Results of the toxicity test were interpreted by standard statistical techniques (Stephan, 1983). The binomial or moving average method was used by the author to calculate EC50 values using nominal concentrations of test substance. The NOEC was calculated using Dunnett's test, which includes a parametric one-way analysis of variance (ANOVA). The average specific growth rate was calculated as the natural log of the number of cells per ml at 96 hours minus the natural log of the number of cells per ml at 0 hours divided by the exposure period. The percent change from control is calculated by subtracting the sample average specific growth rate from the control average specific growth rate, dividing that value by the average specific growth rate in the control and multiplying that value by 100. The percent change from control was used to compute EC50 and NOEC values.

Limit Test:



### Test Results - Acute Toxicity To Aquatic Plants
NOEC Exposure Duration: 96 Hours

NOEC: = 250 mg/L Nominal

LOEC Exposure Duration:

LOEC:

NOELR Exposure Duration:

NOELR:

LOELR Exposure

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Duration:

LOELR:

Effect : Exposure Duration Exposure Units Type % Value Description Mean Value

or Lower Mean Value Upper Mean Value Units Basis for Effect Basis for Concentration

72 Hours EC 50 % < 62.5 mg/L Growth Rate
96 Hours EC 50 % = 87 mg/L Growth Rate

Results Remarks: Temperature varied from 23.1 to 23.5 C throughout testing. pH varied from 6.7 to 9.4 throughout testing. 

### Reliability/Data Quality - Acute Toxicity To Aquatic Plants
Reliability: 2 - Valid With Restrictions

Reliability Remarks:
Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Plants

Reference: TSCATS submission (OTS0533466, Doc ID 86-920000103S, received 10/31/91): Resource Analysts, Inc., 3M Company. 1991. Initial submission: Environmental Effects of Tolyltriazole with Attachments and Cover Letter 102891 (SANITIZED). pp. 30-44



## Other

CAS Number : 29385-43-1

Sponsored Chemical Name

: 1H-Benzotriazole, 4(or 5)-methylTest Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: 5-Methyl-1H-Benzotriazole or 4-Methyl-1H-Benzotriazole (MeBT), CAS No. 29385-43-1

Other End Point Name: Microtox assay

Other End Point Description:

Reference: Cornell, J.S. et al. 2000. Comparative measures of the toxicity of component chemicals in aircraft deicing fluid. Environmental Chemistry, 19(6): 1465-1472.
Description: Microtox studies measured the luminescence of the marine bacterium Vibrio fischeri (formerly known as Photobacterium phosphoreum) according to the basic method described by Beckman Instruments. The nominal (predilution) test concentration was 47.8 mg/L. Two replicates were

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] tested. Five- and 15-min EC50s were calculated using methods described by AZUR Environmental.

Results: 15-min EC50 = 7 mg/L

Measured high concentrations of MeBT were between 95.5 and 99.5% of nominal concentrations at a confidence interval of
95%. Lower test concentrations were calculated as a proportion of the 100% measured concentration. The LC50s were calculated based on measured 100% concentration.



## Acute Toxicity

CAS Number : 29385-43-1

Sponsored Chemical Name

: 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Referred to as Cobratec TT 100 Sample No. 5459

Sponsored Chemical Result Type: Measured



### Method - Acute Toxicity

Route of Administration: Oral

Type of Exposure: No Data

Species: Rat

Mammalian Strain: No Data

Gender: Both M/F

Number of Animals per Dose: 10

Dose: 502, 632, 795 and 1001 mg/kg

Year Study Performed:

Method/Guideline Followed: No Data

GLP: No Data

Method/Guideline and Test Condition Remarks: Cobratec TT 100 Sample No. 5459 was administered orally in graduated doses to 4 groups of 10 rats. Five males and five females in each group. They were observed for 14 days.



### Test Results - Acute Toxicity
Concentration (LC/LD) : LC/LD % Value Description Value/Lower Concentration Upper Concentration Units

LD 50 % = 675 mg/kgbw

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http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Number of Deaths ( Male ):
Number of Deaths ( Female ):
Number of Deaths ( Total ): 23

Results Remarks: LD50 = 675 mg/kg (95% confidence limits were 535-851 mg/kg)

Mortalities were as follows: 502 mg/kg - 3 deaths on day 0 632 mg/kg - 1 death on day 0; 2 deaths on day 1 795 mg/kg - 6 deaths on day 0; 1 death on day 1 1001 mg/kg - 10 deaths on day 0

Conclusion: None stated



### Reliability/Data Quality - Acute Toxicity
Reliability: 2 - Valid With Restrictions

Reliability Remarks:

Key Study Sponsor Indicator: 

### Reference - Acute Toxicity

Reference: TSCATS submission (Doc ID 86-890000263, received 06/05/89): Huntingdon Research Center, Union Carbide Corp. 1989. Initial submission: Health and Safety Studies for 19 Chemicals with Cover Letter Date 060289. pp. 32-34.



## Acute Toxicity

CAS Number : 29385-43-1

Sponsored Chemical Name

: 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Referred to as Cobratec RTT 100, Sample No. 3000. The compound was a fine powder, off-white in color. The melting point was reported to be 76  ̈¬C, with rapid decomposition occurring at temperatures ¡Ã 300  ̈¬C.

Sponsored Chemical Result Type: Measured



### Method - Acute Toxicity

Route of Administration: Inhalation

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http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Type of Exposure: Aerosol

Species: Rat

Mammalian Strain: Sprague-Dawley

Gender: Both M/F

Number of Animals per Dose: 20

Concentration: 1.73 mg/L

Year Study Performed:

Method/Guideline Followed: Other

GLP: No Data

Method/Guideline and Test Condition Remarks: : One group of ten male and ten female Sprague-Dawley rats (Charles River) was exposed for one hour to a concentration of 1.73 +/- 0.67 mg/L of an aerosol of tolyltriazole. Vapor phase compound was also monitored. Following the exposure, all animals were maintained for an additional fourteen days; with food and water ad libitum. Body weights were takenprior to exposure and at seven and fourteen days postexposure. All animals were observed for signs of toxicity during the exposure and daily during the post-exposure observation period. All animals that died or were sacrificed after fourteen days were necropsied. At this time, all organs were observed for macroscopic abnormalities. The following tissues were stored in 10% neutral buffered formalin in the event that histopathology might be required: lungs and trachea, heart, liver, kidneys, spleen, stomach, intestines, gonads, whole head, abnormal tissue.

Animals were allowed 7 days to accommodate to the laboratory conditions before being assigned to the study.

Aerosol generation system: Approximately one hundred grams of tolyltriazole is placed in the round bottom three necked flask and melted. The temperature of the flask and delivery tube is maintained at 200  ̈¬C by the heating mantle, air heater and heating tape. A controlled air flow of ten L/min is provided to the vapor generating flask. The tolyltriazole vapor-laden air is taken from the top of the generating flask, diluted and cooled by admixture of a controlled flow of clean dry air. This system provided a very fine white aerosol. Chamber concentration was controlled by varying the air flow rate through the generator.

Exposure concentrations were determined by sampling the chamber atmosphere. Actual exposure concentrations were determined by drawing (at 0.82 L/min) a known volume of chamber air through a pre-weighted glass fiber filter in a closed face filter holder. The sample time was three minutes. The filter was then re-weighed and the chamber concentration calculated.

Vapor phase compound was determined by sampling the chamber atmosphere. The sampling rate was 0.82 L/min for six minutes. The sample was drawn through two 25 mL fritted gas bubblers in series, each containing 10 mL of 0.5N sodium hydroxide. After sample collection the contents of each gas bubbler was transferred to a 25 mL volumetric flask, brought

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] to volume with additional 0.5N sodium hydroxide and mixed well. The absorbance was determined at 274 nm using a UV spectrophotometer and matched 1 cm. silica cells. The tolyltriazole content was obtained by reference to a standard

curve.

Particle size analysis was also conducted during the

exposure. An Anderson cascade impactor was used for this determination. Pre-weighted glass fiber filters, specially cut to fit the impactor, were used for collecting the aerosol on each stage. The filter discs were re-weighed immediately after sampling. A particle size distribution graph was then constructed from the weight of material collected on each stage. The aerosol was found to have an Equivalent Aerodynamic Diameter (EAD) of 0.35 µm with a geometric standard deviation of 2.62. The sample flow rate was 1 CFM for ten minutes.



### Test Results - Acute Toxicity
Concentration (LC/LD) : LC/LD % Value Description Value/Lower Concentration Upper Concentration Units

LC 50 % > 1.73 mg/L

Number of Deaths ( Male ): 0

Number of Deaths ( Female ): 0

Number of Deaths ( Total ): 0

Results Remarks: During the exposure, the only pharmacologic or toxicologic sign observed was a decrease in general activity. By 24 hours post-exposure, all animals were apparently normal and remained so during the entire 14-day observation period. All surviving animals appeared to gain weight normally throughout the 14 day observation period. Since there were no deaths during the exposure or 14-day post-exposure observation period, all animals were necropsied following the 14-day observation period. All organs were observed for gross abnormalities. Respiratory tract involvement, characterized by slight to moderate incidence of hemorrhage or hepatization, was found in all animals. Some incidence of pale liver and kidneys was also noted in males and females.

Conclusion: None specified



### Reliability/Data Quality - Acute Toxicity
Reliability: 1 - Valid Without Restrictions

Reliability Remarks:

Key Study Sponsor Indicator:

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Reference - Acute Toxicity

Reference: TSCATS submission (OTS0521119, Doc ID 86-890000610, received 06/13/89): Huntingdon Research Center, Sherwin Williams
Co. 1976. Initial submission: Evaluation of the Acute Inhalation Toxicity of Tolyltriazole (Final Report) with Attachments and Cover Letter Dated 061289.



## Acute Toxicity

CAS Number : 29385-43-1

Sponsored Chemical Name

: 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Substance was labeled Cobratec TT-100 (Tolyltriazole) Sample
No. 6022. The material was light tan granular in nature.

Sponsored Chemical Result Type: Measured



### Method - Acute Toxicity

Route of Administration: Dermal

Type of Exposure: Occlusive

Species: Rabbit

Mammalian Strain: No Data

Gender: Female

Number of Animals per Dose: 10

Dose: 2000 mg/kg

Year Study Performed:

Method/Guideline Followed: No Data

GLP: No Data

Method/Guideline and Test Condition 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.



### Test Results - Acute Toxicity
Concentration (LC/LD) : LC/LD % Value Description Value/Lower Concentration Upper Concentration Units

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http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] LD 50 % > 2000 mg/kgbw

Number of Deaths ( Male ):
Number of Deaths ( Female ): 0

Number of Deaths ( Total ): 0

Results Remarks: No mortalities were observed. Body weights remained stable throughout the study (varying from 2.8 to 3.1 kg). Very slight erythema (grade 1) was observed in three rabbits, each from the abraded group; these effects subsided after the third day of observation. No additional signs of skin irritation were observed.

Conclusion: None stated



### Reliability/Data Quality - Acute Toxicity
Reliability: 2 - Valid With Restrictions

Reliability Remarks:

Key Study Sponsor Indicator: 

### Reference - Acute Toxicity

Reference: TSCATS submission (OTS0516797, Doc ID 86-890000263, received 06/05/89): Huntingdon Research Center, Union Carbide Corp. 1989. Initial submission: Health and Safety Studies for 19 Chemicals with Cover Letter Date 060289. pp. 10-15.



## Acute Toxicity

CAS Number : 29385-43-1

Sponsored Chemical Name

: 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Referred to as Olin 58734. Pinkish grey granular material

Sponsored Chemical Result Type: Measured



### Method - Acute Toxicity

Route of Administration: Dermal

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Type of Exposure:

Species: Rabbit

Mammalian Strain: No Data

Gender: Both M/F

Number of Animals per Dose: 4

Dose: 500, 1000, 2000 and 4000 mg/kg-bw

Year Study Performed:

Method/Guideline Followed: Other

GLP: No Data

Method/Guideline and Test Condition Remarks: Method/guideline followed: Method in Appraisal of the Safety of Chemicals in Foods, Drugs and (illegible) Association of Food and Drug Officials of the U.S. 1959

Animals - Four albino rabbits per four levels in weight range of 2.2 to 3.6 kg depilated over the trunk.

Maintenance conditions - Trunk encased in sleeve with animal restrained in stock during application. Thereafter individual housing with food and water ad libitum.

Dosage - 24 hour continuous contact with the skin. Test substance was administered as a 40% aqueous paste.

Observations - Body weight, appearance, behavior, skin irritation and mortality over a 14-day period



### Test Results - Acute Toxicity
Concentration (LC/LD) : LC/LD % Value Description Value/Lower Concentration Upper Concentration Units

LD 50 % > 4000 mg/kgbw

Number of Deaths ( Male ): 0

Number of Deaths ( Female ): 0

Number of Deaths ( Total ): 0

Results Remarks: No deaths occurred at any level in 14 days. The primary irritation scores were 1.4, 1.5, 1.1 and 1.4, for the 0.5, 1, 2 and 4 g/kg levels, respectively. No other noteworthy findings were observed.

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http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Conclusion: The acute dermal toxicity of the tested substance is greater than 4 g/kg-bw and this product evokes no irritation. 

### Reliability/Data Quality - Acute Toxicity
Reliability: 3 - Not Valid

Reliability Remarks: Lack of study details; study is outdated

Key Study Sponsor Indicator: 

### Reference - Acute Toxicity

Reference: TSCATS submission (OTS0520643, Doc ID 86-89000060A, received 06/13/89): Huntingdon Research Center, Sherwin Williams
Co. 1968. Initial submission: Summary of Mucuous Membrane (Eye) Irritations in Rabbits Treated with Olin 58734 with Attachment and Cover Letter Dated 061289.



## Acute Toxicity

CAS Number : 29385-43-1

Sponsored Chemical Name

: 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Acute Toxicity

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Referred to as Olin 58734. Pinkish grey granular material

Sponsored Chemical Result Type: Measured



### Method - Acute Toxicity

Route of Administration: Oral

Type of Exposure: Gavage

Species: Rat

Mammalian Strain: Other

Other Strain:

Gender: Both M/F

Number of Animals per Dose: 5

Dose: 500, 1000, 2000, 4000 and 8000 mg/kg-bw

Year Study Performed:

Method/Guideline Followed: Other

GLP: No Data

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Method/Guideline and Test Condition Remarks: Method/guideline followed: Method in Appraisal of the Safety of Chemicals in Food, Drugs, and Cosmetics, Association of Food and Drug Officials of the U.S. 1959

Animals - Adult albino rats, FDRL strain, 5 rats per dose
level

Observations - Body weight, behavior, appearance, mortality, gross changes at autopsy of rats that died and of representative survivors sacrificed at 14 days

Test substance was administered intragastrically as a 25% suspension in 0.5% (illegible).

Groups were comprised of three males and two females.



### Test Results - Acute Toxicity
Concentration (LC/LD) : LC/LD % Value Description Value/Lower Concentration Upper Concentration Units

LD 50 % = 1830 mg/kgbw

Number of Deaths ( Male ):
Number of Deaths ( Female ):
Number of Deaths ( Total ): 14

Results Remarks: Deaths occurred in 3 hours to 5 minutes, following CNS depression, shallow breathing, lacrimation, and salivation. Liver congestion and pale stomach mucosae were seen at autopsy.

One mortality was observed at 0.5 g/kg; 0 mortalities at 1.0 g/kg; 3 mortalities at 2.0 g/kg; 5 mortalities at 4.0 g/kg; 5 mortalities at 8.0 g/kg.

Conclusion: The acute oral LD50 for rats is 1.83 g per kg body weight; confidence limits = 1.20 to 2.81 g per kg body weight. 

### Reliability/Data Quality - Acute Toxicity
Reliability: 3 - Not Valid

Reliability Remarks: Study is dated; lack information on test substance

Key Study Sponsor Indicator: 

### Reference - Acute Toxicity

Reference: TSCATS submission (OTS0520643, Doc ID 86-89000060A, received 06/13/89): Huntingdon Research Center, Sherwin Williams
Co. 1968. Initial submission: Summary of Mucuous Membrane (Eye) Irritations in Rabbits Treated with Olin 58734 with Attachment and Cover Letter Dated 061289.

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] 

## Genetic Toxicity in vitro

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Genetic Toxicity in vitro

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Referred to as TK 10 637/1

Sponsored Chemical Result Type: Measured



### Method - Genetic Toxicity in vitro
Type of Study: Bacterial reverse mutation assay

Concentrations: 25, 75, 675, 2025 μg/0.1 mL without activation and 500, 1000, 2000, 4000, 8000 μg/0.1 mL with activation.

Year Study Performed: 1982

Method/Guideline Followed: Other

GLP:

Postive, Negative, and Solvent Control Substance(s): negative control: acetone; Positive controls: 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 ug/0.1 mLphosphate buffer; 3) for strain TA1535: N-methyl-N'-nitro-Nnitrosoguanidine, 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.

Method/Guideline and Test Condition Remarks: Method/guideline followed: Ames, B.N., F.D. Lee, and W.E. Durston (1973), An Improved Bacterial Test System for the Detection and Classification of Mutagens and Carcinogens. Proc. Natl. Acad. Sci. USA 70, 782-786.

The substance was dissolved in acetone. Acetone alone was used for the negative controls. 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 l-histidine, 0.5 mM and +biotin 0.5 mM. In the experiments in 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 ratsinduced with Aroclor 1254 and 0.7 mL of a solution of cofactors.

Positive control experiments were carried out simultaneously with

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] 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 ug/0.1 mL phosphate buffer; 3) for strain TA1535: N-methyl-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 and with strain TA98 and 2-aminoanthracene, 5 μg/0.1 mL DMSO.

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 and 37 oC in darkness.

When the colonies bad been counted, the arithmetic mean was calculated. The test substance is generally considered to be nonmutagenic if the colony count in relation to the negative control is not doubled at any concentration.



### Test Results - Genetic Toxicity in vitro
Details on Cytogenetic Assay:

Statistics:

Effect : Species Other Species Strain Other Strain Metabolic Activation Genotoxic Effect Conclusion

Bacteria S. typhimurium TA 98 With and Without Positive Positive With Metabolic Activation
Bacteria S. typhimurium TA 100 With and Without Negative

Bacteria S. typhimurium

TA 1535 With and Without Negative

Bacteria S. typhimurium

TA 1537 With and Without Negative

Bacteria S. typhimurium

TA 1538 With and Without Positive Positive With Metabolic Activation

Results Remarks: In the experiments performed without microsomal activation, none of the tested concentrations of TK 10 637/1 led to an increase in the incidence of histidine-prototrophic mutants comparison with the negative controls.

In the experiments with microsomal activation treatment with TK 10 637/1 led to an increase in the number of back-mutant colonies of strains TA98 and TA1538 at the concentrations of 2000 ug/0.1 mL and above.

Conclusion:



### Reliability/Data Quality - Genetic Toxicity in vitro
Reliability: 1 - Valid Without Restrictions

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Reliability Remarks:

Key Study Sponsor Indicator: 

### Reference - Genetic Toxicity in vitro

Reference: TSCATS submission (OTS0519041, Doc ID 86-890000272, received 06/07/89): CIBA GEIGY Corp. 1982. Initial submission: Salmonella/Mammalian - Microsome Mutagenicity Test with TK 10 637/1 with Cover Letter Dated 060589.



## Genetic Toxicity in vitro

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Genetic Toxicity in vitro

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Code Identification - TT-100; Sample Number - X18BT7565 Cobratec 78669A; Description - Tan round crystals

Sponsored Chemical Result Type: Measured



### Method - Genetic Toxicity in vitro
Type of Study: Bacterial reverse mutation assay

Concentrations: 0, 80 μg, 400 μg, 2 mg, 10 mg

Year Study Performed:

Method/Guideline Followed: No Data

GLP: No Data

Postive, Negative, and Solvent Control Substance(s): Positive control/strain/concentration (μg/plate)

Nonactivation Sodium azide / TA1535 / 10 Quinacrine / TA1537 / 200 2-nitrofluorene / TA1538, TA98, TA100 / 10

Activation 2 amino anthracene / TA1535, TA1537 / 10 2 amino fluorine / TA1538, TA98, TA100 / 10

Method/Guideline and Test Condition Remarks: Dimethylsulfoxide (DMSO) was used to prepare stock dilutions of test materials and positive control chemicals. Approximately 108 cells from an overnight culture of each indicator strain (TA1535, TA1537, TA1538, TA98, TA100) were added to separate test tubes containing 2.0 mL of molten agar supplemented with a trace of histidine and biotin. For nonactivation tests, three dose levels of the extract were added to the content of the appropriate tubes and poured over the surface of selective agar plates. In activation tests, the extract

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] concentrations were likewise added to the appropriate tubes with cells. Then, just prior to pouring, an aliquot of the S-9 mix containing the rat liver microsomes was added to each of the top agar tubes which were then mixed and the contents poured over the surface of a minimal agar plate and allowed to solidify. The plates were incubated for 48 hours at 37 oC, and scored for the number of colonies growing on each plate.

Positive control/strain/concentration (μg/plate)

Nonactivation Sodium azide / TA1535 / 10 Quinacrine / TA1537 / 200 2-nitrofluorene / TA1538, TA98, TA100 / 10

Activation 2 amino anthracene / TA1535, TA1537 / 10 2 amino fluorine / TA1538, TA98, TA100 / 10



### Test Results - Genetic Toxicity in vitro
Details on Cytogenetic Assay:

Statistics:

Effect : Species Other Species Strain Other Strain Metabolic Activation Genotoxic Effect Conclusion

Bacteria S. typhimurium TA 98 With and Without Equivocal Equivocal With Metabolic Activation
Bacteria S. typhimurium TA 100 With and Without Negative Negative

Bacteria S. typhimurium

TA 1535 With and Without Negative Negative

Bacteria S. typhimurium

TA 1537 With and Without Negative Negative

Bacteria S. typhimurium

TA 1538 With and Without Equivocal Equivocal With Metabolic Activation

Results Remarks: The initial data obtained by the mutagenic bioassay of compound TT-100 suggested a possible response with tester strains TA1538 and TA98. Confirmatory assays were carried out two additional times. The results do not clearly indicate that TT-100 is a frameshift mutagen. The revertant response elicited at the highest concentration tested indicated a slight elevation of the revertant rate but did not produce a response of at least an excess of two times background. Higher concentrations of the test compound indicated a toxic response. It appears that the TT-100 may possess some very weak mutagenic activity in this system when enzymatically activated; however, confirmation of this activity is negated by the toxicity of the compound in this system.

For the initial assay, in the absence of metabolic activation, toxicity was observed at 10 mg in all strains tested. In the presence of metabolic activation, toxicity was observed at 10 mg in TA1535 and TA1537.

For the confirmation assay, toxicity was observed with metabolic activation at 10 mg for TA1538 and TA98.

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Positive and negative controls responded appropriately.

Conclusion:



### Reliability/Data Quality - Genetic Toxicity in vitro
Reliability: 2 - Valid With Restrictions

Reliability Remarks:

Key Study Sponsor Indicator: 

### Reference - Genetic Toxicity in vitro

Reference: TSCATS submission (OTS0516797, Doc ID 86-890000263, received 06/05/89): Huntingdon Research Center, Union Carbide Corp. 1989. Initial submission: Health and Safety Studies for 19 Chemicals with Cover Letter Date 060289. pp. 72-91.



## Genetic Toxicity in vitro

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Genetic Toxicity in vitro

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments:
Sponsored Chemical Result Type: Measured



### Method - Genetic Toxicity in vitro
Type of Study: Mammalian cell gene mutation assay

Concentrations: 12.5 - 750 ìg/mL

Year Study Performed:

Method/Guideline Followed: No Data

GLP: No Data

Postive, Negative, and Solvent Control Substance(s): For each assay, there were 2-4 solvent controls, a positive control of ethyl methylsulfonate at 4.7 x 10-6 M (or methyl methanesulfonate at 10-20 ìg/mL) for the test without metabolic activation, and a positive control of 3-methylcholanthrene at 1.86 x 10-5 M (or dimethylbenz[a]-anthracene at 0.5-4 ìg/mL) for the test with metabolic activation.

Method/Guideline and Test Condition Remarks: L5178Y TK+/- 3.7.C mouse lymphoma cells were Originally obtained from Dr. Donald Clive, Burroughs Wellcome Co. (Research Triangle Park, NC). The cells were grown in Fischer's medium for leukemic cells of mice (Gibco, Grand Island, NY, or Quality Biological, Gaithersburg, MD) supplemented with 10%

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] horse serum (Gibco or Hyclone, Logan, UT) and 0.02% pluronic F-68 (BASF Wyandotte Corp., Wyandotte, MI). Cells were screened for the presence of mycoplasma after cryopreservation. New cultures were initiated at approximately 3 month intervals from cells stored in liquid N2. The toxicity of each chemical was determined both with and without liver S9 prepared from Aroclor 1254-induced male Sprague-Dawley rats. S9 mix was prepared according to the procedure of Clive et al. Cells at a concentration of 6 x 105/mL (6 x 106 cells total) were exposed for 4 h to a range of concentrations from 0.0005 to 10000 ìg/mL. The cells were then washed, resuspended in growth medium, and incubated at 37 ±1 °C for 48 h. The rate of cell growth was determined for each of the treated cultures and compared to the rate of growth of the solvent controls. The doses of chemical selected for testing were within the range yielding approximately 0-90% cytotoxicity. For each assay, there were 2-4 solvent controls, a positive control of ethyl methylsulfonate at 4.7 x 10-6 M (or methyl methanesulfonate at 10- 20 ìg/mL) for the test without metabolic activation, and a positive control of 3-methylcholanthrene at 1.86 x 10-5 M (or dimethylbenz[a]-anthracene at 0.5-4 ìg/mL) for the test with metabolic activation. The mutagenicity assay was performed according to the procedure described by Clive and Spector. A total of 1.2 x 107 cells in duplicate cultures were exposed to the test chemical, positive control, and solvent control for 4 h at 37 ± 1 °C, washed twicewith growth medium, and maintained at 37 ±1 °C for 48 h in logphase growth to allow recovery and mutant expression. Cells in the cultures were adjusted to 3 x 105/mL at 24 h intervals. They were then cloned (1 x 106 cells/plate for mutant selection and 200 cells/plate for viable count determinations) in soft agar medium containing Fischer's medium, 20% horse serum, 2 mM sodium pyruvate, 0.02% pluronic F-68, and 0.23% granulated agar (BBL, Inc., Cockeysville, MD). Resistance to trifluorothymidine (TFT) was determined by adding TFT (final concentration, 3 ìg/mL) to the cloning medium for mutant selection. The 100x stock solution of TFT in saline was stored at -70 °C and was thawed immediately before use. Plates were incubated at 37 ±1 °C in 5% CO2 in air for 10-12 days and then counted with an Artek automated colony counter (Artek 982, DynaTech) or ProtoCol colony counter (Synbiosis, Frederick, MD). Only colonies larger than x0.2 mm in diameter were counted. Mutant frequencies were expressed as mutants per 106 surviving cells. Although there are several different methods for evaluating mouse lymphoma data, results from this study were interpreted using a doubling of the mutant frequency over the concurrent solvent-treated control value as an indication of a positive effect, together with evidence of a dose-related increase. Doubling of the mutant frequency was previously reported as representing a positive effect. Only doses yielding total growth values of 10% were used in the analysis of induced mutant frequency. Doses yielding less than 10% total growth were used in determining dose response. The size of mutant mouse lymphoma colonies was also determined using an Artek 982 colony counter/sizer or the ProtoCol colony counter. An internal discriminator was set to step sequentially to exclude increasingly larger colonies in approximate increments of 0.1 mm in colony diameter. The size range used was from 0.2 to 1.1

mm.



### Test Results - Genetic Toxicity in vitro

Details on Cytogenetic Assay:

Statistics:

Effect : Species Other Species Strain Other Strain Metabolic Activation Genotoxic Effect Conclusion

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http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Mammalian Cell Line Mouse Lymphoma

L5178Y

Cells With and Without Positive Positive With Metabolic Activation

Results Remarks: In the nonactivated test, the old evaluation was Positive at a concentration of 750 ìg/mL, and the new evaluation was inconclusive. In the activated test, the old evaluation was positive at 25 ìg/mL, and the new evaluation was positive at 50 ìg/mL.

Conclusion: Positive in the presence of metabolic activation. Inconclusive without metabolic activation. 

### Reliability/Data Quality - Genetic Toxicity in vitro
Reliability: 2 - Valid With Restrictions

Reliability Remarks:

Key Study Sponsor Indicator: 

### Reference - Genetic Toxicity in vitro

Reference: Seifried et al. 2006. Compilation of two decades of mutagenicity test results with the Ames Salmonella typhimurium and L5178Y mouse lymphoma cell mutation assays. Chem. Res. Toxicol. 19: 627-644.



## Genetic Toxicity in vitro

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Genetic Toxicity in vitro

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Code Identification - TT-100 Sample number - X18BT7565 Cobratec 78669A Description - Tan round crystals

Sponsored Chemical Result Type: Measured



### Method - Genetic Toxicity in vitro
Type of Study: in vitro mammalian cell transformation assay

Concentrations: 6, 60 and 600 μg

Year Study Performed:

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http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Method/Guideline Followed: No Data

GLP: No Data

Postive, Negative, and Solvent Control Substance(s): positive control: dimethylbenzanthracene

Method/Guideline and Test Condition Remarks: The C3H10T1/2 line is a fibroblastic cell line, derived from C3H mouse embryos which are highly sensitive to postconfluence inhibition of cell division. When C3H10T1/2 cells are exposed to a variety of chemical carcinogens morphologically transformed foci develop with a frequency that is dose dependent. The foci have been shown to produce fibrosarcomas when injected into syngeneic mice. Therefore, the C3H10T1/2 cell line can be used to assay for chemical carcinogens.

Two sets of 60-mm culture dishes are seeded with cells. One set of dishes, 12 x replicates per dose, is seeded with 1000 cells. This set is used for the transformation assay. The second set was seeded with 200 cells per dish, 3 x per dose, and is utilized for cytotoxicity determinations.

Both sets of cells are treated in an identical manner with solutions of the test material. Following a 24-hour exposure period, the toxicant is washed from all the plates and the cells refed with normal growth media. Ten days following the initial seeding, the plates designated for the cytotoxicity assay are washed, fixed with methanol, and stained. The clones on all of the plates are counted and the plating efficiency determined. This plating efficiency is expressed as the number of colonies formed as a percentage of the number of cells seeded.

The transformation set are incubated for 6 weeks with weekly medium changes. During this time, the clones grow to form a confluent monolayer of cells on the bottom of the plates. At the termination of the incubation period, all remaining plates are washed, fixed, stained and scored for the transformed foci: Type I Foci: A Type I focus is composed of tightly packed cells Type II Foci: The cells display massive piling into opaque multilayers. Type III Foci: These foci are composed of highly polar, densely stained cells, which form a criss-crossed multilayer.

Type II and Type III foci are considered to be malignantly transformed. The transformation frequency is calculated as the percentage of cells that give rise to transformed foci corrected for plating efficiency which is determined by the cytotoxicity assay.



### Test Results - Genetic Toxicity in vitro

Details on Cytogenetic Assay:

Statistics:

Effect : Species Other Species Strain Other Strain Metabolic Activation Genotoxic Effect Conclusion

Mammalian Cell Line Other

- C3H10T1/2

Mouse Fibroblast Not Applicable Negative Negative

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http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Results Remarks: For the 6, 60, 600 μg and control groups, the transformation frequency was 0.32, 0.15, 0.07 and 0.07, respectively.

None of the sample chemicals at the dose levels tested produced significant increases in transformation above control cell population levels. As a positive control the cells did respond in a dose-dependent manner, to the chemical carcinogen dimethylbenzanthracene (DMBA).

Conclusion:



### Reliability/Data Quality - Genetic Toxicity in vitro
Reliability: 2 - Valid With Restrictions

Reliability Remarks:

Key Study Sponsor Indicator: 

### Reference - Genetic Toxicity in vitro

Reference: TSCATS submission (OTS0516797, Doc ID 86-890000263, received 06/05/89): Huntingdon Research Center, Union Carbide Corp. 1989. Initial submission: Health and Safety Studies for 19 Chemicals with Cover Letter Date 060289. P 72-91



## Genetic Toxicity in vitro

CAS Number : 29385-43-1

Sponsored Chemical Name

: 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Genetic Toxicity in vitro

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments:
Sponsored Chemical Result Type: Measured



### Method - Genetic Toxicity in vitro
Type of Study: Bacterial reverse mutation assay

Concentrations: 100 - 10,000 ug/plate

Year Study Performed:

Method/Guideline Followed: Other

GLP: No Data

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Postive, Negative, and Solvent Control Substance(s):

Method/Guideline and Test Condition Remarks: The experimental materials, methods, and procedures as explained below are based on those described by Ames et al. For initial mutagenicity testing, the plate incorporation assay was used; some compounds were also tested using the flavin mononucleotide (FMN)-dependent reducing conditions described by Prival and Mitchell to break the diazo bonds of the condensed dyes, forming the individual constituents. S9 Preparation. Liver S9 homogenate was prepared from male Sprague-Dawley rats and Syrian golden hamsters that had been injected with Aroclor 1254 at 500 mg/kg body weight. The postmitochondrial (microsomal) enzyme fractions were prepared as described by Ames et al. (26). The components of the S9 mix were 8 mM MgCl2, 33 mM KCl, 5 mM glucose-6-phosphate, 4 mM NADP, 100 mM sodium phosphate (pH 7.4), and the appropriate S9 homogenate at a concentration of 0.1 mL/mL of mix. For each plate receiving microsomal enzymes, 0.5 mL of S9 mix was added. Plate Incorporation Methodology. For testing in the absence of S9 mix, 100 ìL of the tester strain and 50 ìL of the solvent or test chemical were added to 2.5 mL of molten selective top agar at 45 ±2 °C. When S9 was used, 0.5 mL of S9 mix, 50 ìL of tester strain, and 50 ìL of solvent or test chemical were added to 2.0 mL of molten selective top agar at 45 ±2 °C. After it was vortexed, the mixture was overlaid onto the surface of 25 mL of minimal bottom agar. After the overlay had solidified, the plates were incubated for 48 h at 37 ±2 °C. Five doses of test chemical, together with the appropriate concurrent solvent and positive controls, were tested in triplicate on each tester strain without metabolic activation and also with activation by induced rat and hamster liver S9 preparations. The doses that were tested in the mutagenicity assay were selected based on the levels of cytotoxicity observed in a preliminary dose range-finding study using strain TA100. Ten dose levels of the chemical, one plate per dose, were tested in both the presence and the absence of induced hamster S9. If no toxicity was observed, a total maximum dose of 10 mg of test chemical per plate was used. The criteria used to evaluate a test were as follows: for a test article to be considered positive, it had to induce at least a doubling (TA98, TA100, and TA1535) in the mean number of revertants per plate of at least one tester strain. This increase in the mean revertants per plate had to be accompanied by a dose response to increasing concentrations of the test chemical. If the study showed a dose response with a less than 3-fold increase on TA1537 or TA1538, the response had to be confirmed in a repeat experiment. Preincubation Methodology under Reductive Conditions. For tests using the FMN-modified assay, strains TA98 and TA100 were used. The bacteria, uninduced hamster liver S9 (30% v/v), cofactors (FMN, NADH, glucose-6-phosphate dehydrogenase, and glucose-6-phosphate), and test chemical were added, mixed, and incubated at 30 °C for 30 min without shaking. Nitrogen was blown over the preincubation tube to keep the atmosphere reduced. At the end of the incubation period, 2 mL of molten top agar was added to each sample tube and the mixture was poured on a minimal agar plate containing 0.5% glucose rather than the 2% glucose specified by Ames et al. The plates were then incubated at 37 °C for 48 h. The positive control in all FMN experiments was Congo red. All plates were counted with

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http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] an Artek automated colony counter (Artek 880, DynaTech, Chantilly, VA) or Minicount colony counter (Imaging Products International, Inc., Chantilly, VA), which was calibrated prior to use.



### Test Results - Genetic Toxicity in vitro
Details on Cytogenetic Assay:

Statistics:

Effect : Species Other Species Strain Other Strain Metabolic Activation Genotoxic Effect Conclusion

Bacteria With and Without Negative Negative

Results Remarks:

Conclusion:



### Reliability/Data Quality - Genetic Toxicity in vitro
Reliability: 2 - Valid With Restrictions

Reliability Remarks:

Key Study Sponsor Indicator: 

### Reference - Genetic Toxicity in vitro

Reference: Seifried et al. 2006. Compilation of two decades of mutagenicity test results with the Ames Salmonella typhimurium and L5178Y mouse lymphoma cell mutation assays. Chem. Res. Toxicol. 19: 627-644.



## Genetic Toxicity in vitro

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Genetic Toxicity in vitro

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: 99+ % pure

Sponsored Chemical Measured

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http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Result Type: 

### Method - Genetic Toxicity in vitro
Type of Study: Bacterial reverse mutation assay

Concentrations: 0, 33, 100, 333, 1000, 1666, 3333, 6666 ìg/plate

Year Study Performed: 1983

Method/Guideline Followed: Other

GLP: No Data

Postive, Negative, and Solvent Control Substance(s): vehicle control: dimethylsulfoxide; The positive controls in the absence of metabolic activation were sodium azide (TA1535 andTA100), 9-aminoacridine (TA97 and TA1537), and 4-nitro-ophenylenediamine (TA98). The positive control for metabolic activation with all strains was 2-aminoanthracene.

Method/Guideline and Test Condition Remarks: Preparation of Liver S-9 Fractions - The S-9 (9,000 gsupernatant) fractions of Aroclor 1254-induced, male SpragueDawley rat and male Syrian hamster livers were prepared as described previously [Haworth et al, 1983]. The S-9 mixes were prepared immediately prior to use and contained either 10% or 30% S-9; occasionally, other levels were used. All chemicals were tested in the absence of metabolic activation and with rat and hamster S-9 fractions.

Test Protocol - The preincubation assay was performed as described previously [Haworth et al., 1983], with some differences, as described below. The test chemical (0.05 mL), Salmonella culture (0.10 mL), and S-9 mix or buffer (0.50 mL) were incubated at 37 ºC, without shaking, for 20 min. Chemicals known or suspected to be volatile were incubated in capped tubes. The top agar was added and the contents of the tubes were mixed and poured onto the surface of petri dishes containing Vogel-Bonner medium [Vogel and Bonner, 1956]. The histidine-independent (his+) colonies arising on these plates were counted following two days incubation at 37 ºC. Plates were machine counted unless precipitate was present which interfered with the count, or the color of the test chemical on the plate reduced the contrast between the colonies and the background agar. At the discretion of the investigators, plates with low numbers of colonies were counted by hand. Variations in the protocol among the tested chemicals reflect the evolution of the protocol originally described by Haworth et al. [1983]. Four protocol variations are evident. 1) Testing in strains TA97, TA98, TA100 and TA1535, with some additional testing in strain TA1537; 10% S-9 was used. 2) The first test of a chemical was without activation and with 10% S-9 in the mix. If a positive result was obtained the test was repeated. If the tests were negative they were repeated without S-9 and with 30%
S-9. 3) The order of use of 10% and 30% S-9 was reversed. 4) Initial testing was in strains TA98 and TA100 without activation and with 30% rat and hamster S-9s. If a positive result was obtained in one of these two strains it was repeated and the other strains were not used. If the tests were negative, the other strains were used with 30% and 10% S-9. A chemical was not designated nonmutagenic unless it had been tested in strains TA98, TA100, TA1535, and TA97 and/or TA1537, without activation and with 10% and 30% rat and hamster S-9. Occasionally, 5% S-9 was also used in all protocol variations. All chemicals were tested initially in a toxicity assay to determine the appropriate dose range for the mutagenicity

assay. The toxicity assay was performed using TA100 or the system developed by Waleh et al. [1982]. Toxic concentrations were those

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] that produced a decrease in the number of his+ colonies, or a clearing in the density of the background lawn, or both. Each chemical was tested initially at half-log dose intervals up to a dose that elicited toxicity, or to a dose immediately below one which was toxic in the preliminary toxicity test. Subsequent trials occasionally used narrower dose increments and may not have included doses in the toxic range. Experiments were repeated at least one week following the initial trial. A maximum of 0.05 mL solvent was added to each plate. Concurrent solvent and positive controls were run with each trial. The positive controls in the absence of metabolic activation were sodium azide (TA1535 and TA100), 9-aminoacridine (TA97 and TA1537), and 4-nitro-o-phenylenediamine (TA98). The positive control for metabolic activation with all strains was 2- aminoanthracene. Occasionally, a laboratory used a single solvent and/or positive control for more than one chemical tested on the same day. Each laboratory made its own determination regarding positive control dose levels.

A chemical was judged mutagenic or weakly mutagenic if it produced a reproducible dose-related response over the solvent control in replicate trials. A chemical was judged questionable if the results of the individual trials were not reproducible, if increases in his+ revertants did not meet the criteria for a 'weakly mutagenic' response, or if only single doses produced increases in his+ revertants in repeat trials. Chemicals were judged nonmutagenic if they did not meet the criteria for a mutagenic or questionable response.



### Test Results - Genetic Toxicity in vitro
Details on Cytogenetic Assay:

Statistics:

Effect : Species Other Species Strain Other Strain Metabolic Activation Genotoxic Effect Conclusion

Bacteria S. typhimurium TA 97 With and Without

Bacteria S. typhimurium TA 98 With and Without

Bacteria S. typhimurium TA 100 With and Without

Bacteria S. typhimurium

TA 1535 With and Without

Bacteria S. typhimurium

TA 1537 With and Without

Results Remarks: Test substance is weakly positive.

Toxicity was observed at 6666 ìg/plate in TA97 with activation and in TA1535 with activation.

Conclusion: Weakly positive.



### Reliability/Data Quality - Genetic Toxicity in vitro
Reliability: 1 - Valid Without Restrictions

Reliability Remarks:

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http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Key Study Sponsor Indicator: 

### Reference - Genetic Toxicity in vitro

Reference: NTP study 651117. Zeiger, E., Anderson, B., Haworth, S., Lawlor, T., and Mortelmans, K. Salmonella mutagenicity tests. IV. Results from the testing of 300 chemicals Environ. Molec. Mutagen. Vol. 11 (Suppl 12) (1988) 1-158



## Genetic Toxicity in vitro

CAS Number : 29385-43-1

Sponsored Chemical Name : 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Genetic Toxicity in vitro

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Code Identification - TT-100; Sample number - X18BT7565 Cobratec 78669A; Description - Tan round crystals

Sponsored Chemical Result Type: Measured



### Method - Genetic Toxicity in vitro
Type of Study: DNA damage or repair assay, unscheduled DNA synthesis in mammalian cells in vitro

Concentrations: not legible

Year Study Performed:

Method/Guideline Followed: No Data

GLP: No Data

Postive, Negative, and Solvent Control Substance(s):

Method/Guideline and Test Condition Remarks: This test system is utilized to determine the potential of compounds to damage DNA in mammalian cells. One mL of the cell suspension (2 x 105 cells/mL) is added to four mL of media in 60-mm tissue culture plates (6 x replicates per dose level). The cells are allowed to grow to confluency (5-7 days) then exposed to 10 mM hydroxyurea for 2 hours. The cells are then washed and exposed to a mixture of the test material and tritiated thymidine (3H-TdR) for 24 hours. After the exposure the plates are washed 3 X with phosphate buffered saline (pH 7.2) and lysed with 2 mL of 0.1M
NaOH. After neutralization with 0.2 mL of 1M HCl, 2.2 mL of cold 10 percent TCA is added, and the plates incubated at 4 oC for 1 hour. The lysate is then filtered through a GF/A filter which is subsequently washed 3 X with TCA. The filters are dried for 8 hours and counted in a Nuclear Chicago Mark III liquid scintillation counter. Dosage levels which have caused DNA damage will exhibit elevated counts due to the increased update of 3H-TdR are mediated by the DNA repair mechanism in the cells.

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http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Test Results - Genetic Toxicity in vitro

Details on Cytogenetic Assay:

Statistics:

Effect : Species Other Species Strain Other Strain Metabolic Activation Genotoxic Effect Conclusion

Mammalian Cell Line Mammalian Cell Line:

Other - WI-38 Human Embryonic Lung Fibroblasts Without Negative Negative

Results Remarks: The potential for damaging cellular DNA was monitored in WI-38 human embryonic lung fibroblasts by measuring the amount of 3H-TdR incorporated into cells exposed to subtoxic levels of the chemical samples. Samples of TT-100 did not elicit significant incorporation above control levels at any of the dose levels tested.

Conclusion: Samples of TT-100 did not elicit significant incorporation above control levels at any of the dose levels tested. 

### Reliability/Data Quality - Genetic Toxicity in vitro
Reliability: 2 - Valid With Restrictions

Reliability Remarks:
Key Study Sponsor Indicator: 

### Reference - Genetic Toxicity in vitro

Reference: TSCATS submission (OTS0516797, Doc ID 86-890000263, received 06/05/89): Huntingdon Research Center, Union Carbide Corp. 1989. Initial submission: Health and Safety Studies for 19 Chemicals with Cover Letter
Date 060289. P 72-91



## Skin Irritation

CAS Number : 29385-43-1

Sponsored Chemical Name

: 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Skin Irritation

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Referred to as Cobratec TT-100 Sample No. 6022

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Sponsored Chemical Result Type: Measured



### Method - Skin Irritation
Species: Rabbit

Mammalian Strain: New Zealand White

Type of Coverage: Occlusive

Preparation of Test Site: Shaved

Gender: Female

Number of Animals per Dose: 12

Amount/Concentration Applied: 0.5 grams

Year Study Performed:

Method/Guideline Followed: No Data

GLP: No Data

Exposure Period: 24 Hours

Total Volume applied and Units:
Control Group Type:

Vehicle Used:

Vehicle Name:

Vehicle Amount and Units:

Post-Exposure Period: 48 Hours

Grading Scale:

Method/Guideline and Test Condition 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, 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.

The reactions were scored according to the method described in the Federal Hazardous Substances Act.



### Test Results - Skin Irritation

Grade:

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Primary Irritation Index: 0

Lesions:

Erythema:

Edema:

Results Remarks: The primary irritation index was 0.0. There was no skin irritation.
Interpretation of Results: Not Irritating

Conclusion: Not irritating to rabbit skin.



### Reliability/Data Quality - Skin Irritation
Reliability: 2 - Valid With Restrictions

Reliability Remarks:

Key Study Sponsor Indicator:



### Reference - Skin Irritation

Reference: TSCATS submission (OTS0516797, Doc ID 86-890000263, received 06/05/89): Huntingdon Research Center, Union Carbide Corp. 1989. Initial submission: Health and Safety Studies for 19 Chemicals with Cover Letter Date 060289. pp. 51-57.



## Eye Irritation

CAS Number : 29385-43-1

Sponsored Chemical Name

: 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Eye Irritation

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Referred to as Olin 58734. Pinkish grey granular material

Sponsored Chemical Result Type: Measured



### Method - Eye Irritation
Species: Rabbit

Mammalian Strain: No Data

Gender: Both M/F

Number of Animals per Dose: 6

Amount/Concentration Applied: 10 mg

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Year Study Performed:

Method/Guideline Followed: Other

GLP: No Data

Exposure Period:

Total Volume applied and Units: 10 mg

Control Group Type:

Vehicle Used:

Vehicle Name:

Vehicle Amount and Units:

Post-Exposure Period: 72 Hours

Grading Scale:

Tool:

Method/Guideline and Test Condition Remarks: Method/guideline followed: Method in Appraisal of the Safety of Chemicals in Food, Drugs, and Cosmetics, Association of Food and Drug Officials of the U.S. 1959, p. 46

Dosage - 10 mg of powdered sample in right eye, without washout.

Maintenance conditions - Animals maintained in stocks for 6 hours after dosage. Thereafter, individual housing with food and water ad libitum.

Observations - Evaluation of degree of eye irritation at 24, 48, and 72 hours, as described in Regulations



### Test Results - Eye Irritation
Overall Grading Score:

Overall Irritation Score:

Primary Irritation Index:

Lesions:

Cornea:

Iris:

Conjunctivae (Chemosis):

Conjunctivae (Redness):

Results Remarks: Cornea - Moderate opacity in small area in 3 on day 1, in 2 on day 2, in 1 on day 3.

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Iris - Irritation in 1 on days 1, 2, and 3. Conjunctiva - Lid redness slight in 6 on day 1, very slight in 2 on day 3. Chemosis slight in 4 on day 1. Discharge slight in 6 on day 1, very slight in 1 on day (illegible).

Interpretation of Results: Slightly Irritating

Conclusion: The test substance is slightly and transiently irritating to the eyes of rabbits. 

### Reliability/Data Quality - Eye Irritation
Reliability: 2 - Valid With Restrictions

Reliability Remarks:

Key Study Sponsor Indicator: 

### Reference - Eye Irritation

Reference: TSCATS submission (OTS0520643, Doc ID 86-89000060A, received 06/13/89): Huntingdon Research Center, Sherwin Williams
Co. 1968. Initial submission: Summary of Mucuous Membrane (Eye) Irritations in Rabbits Treated with Olin 58734 with Attachment and Cover Letter Dated 061289.



## Eye Irritation

CAS Number : 29385-43-1

Sponsored Chemical Name

: 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Eye Irritation

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments:
Sponsored Chemical Result Type: Measured



### Method - Eye Irritation
Species: Rabbit

Mammalian Strain: New Zealand White

Gender: Female

Number of Animals per Dose: 6

Amount/Concentration Applied: 100 mg

Year Study Performed:

Method/Guideline Followed: No Data

GLP: No Data

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Exposure Period:

Total Volume applied and Units: 100 mg

Control Group Type: Negative

Vehicle Used:

Vehicle Name:

Vehicle Amount and Units:

Post-Exposure Period: 14 Days

Grading Scale:

Tool: Fluorescein

Method/Guideline and Test Condition 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 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 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.



### Test Results - Eye Irritation
Overall Grading Score:

Overall Irritation Score:

Primary Irritation Index:

Lesions:

Cornea:

Iris:

Conjunctivae (Chemosis):

Conjunctivae (Redness):

Results Remarks: The test substance was found to be irritating to rabbit

eyes. Conjunctival irritation scores ranged from 0 to 4

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] throughout the observation period. Corneal irritation scores ranged from 0 to 3. Iridial irritation scores ranged from 0
to 1.

One rabbit died prior to the 14 day reading. All rabbits exhibited treatment-related effects on the conjunctiva and

cornea. 4/6 rabbits exhibited treatment-related effects on the iris.

No. Observed / No. Exposed Cornea observations - 2/6 - a diffuse area of corneal opacity 3/6 - discernible translucent areas of opacity, details of iris slightly obscured 1/6 - no details of iris visible 6/6 - fluorescein examination positive

Iris observations - 4/6 - moderate circumcorneal injection

Conjunctival redness - 4/6 - diffuse crimson red 2/6 - diffuse beefy red

Chemosis - 1/6 - swelling with partial eversion of lids 2/6 - swelling with lids about half closed 3/6 - swelling with lids half closed or greater

Interpretation of Results: Irritating

Conclusion: Irritating to rabbit eyes



### Reliability/Data Quality - Eye Irritation
Reliability: 2 - Valid With Restrictions

Reliability Remarks:

Key Study Sponsor Indicator: 

### Reference - Eye Irritation

Reference: TSCATS submission (Doc ID 86-890000263, received 06/05/89): Huntingdon Research Center, Union Carbide Corp. 1989. Initial submission: Health and Safety Studies for 19 Chemicals with Cover Letter Date 060289. P 60-65.



## Skin Sensitization

CAS Number : 29385-43-1

Sponsored Chemical Name

: 1H-Benzotriazole, 4(or 5)-methyl

### Test Substance - Skin Sensitization

Test Substance: (29385-43-1) 1H-Benzotriazole, 4(or 5)-methylTest Substance Purity/Composition and Other Test Substance Comments: Referred to as TK 10 637/1, Lot 1179/1186, 97.8% pure

Sponsored Chemical Result Type: Measured

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Method - Skin Sensitization
Test Type: In Vivo

Study Type: Maurer Optimization Test

Species: Guinea pig

Mammalian Strain: Pirbright-Hartley

Route of Induction: Intradermal

Route of Challenage Exposure: Intradermal and Epicutaneous

Gender: Both M/F

Number of Animals per Dose: 20

Concentration: 0.1% for intradermal application 10% for epidermal application

Year Study Performed: 1982

Method/Guideline Followed: Other

GLP: No Data

Exposure Period:

Induction Frequency of Treatment: once every second day (except weekends) to a total of 10

Challenge Exposure Period: 24 Hours

Challenge Frequency of Treatment:
Total Volume applied and Units:
Control Group Type: Vehicle

Vehicle Used: Yes

Vehicle Name: Other

Other Vehicle Name: physiological saline

Vehicle Amount and Units:

Positive Control Substance:
Negative Control Substance: phsyiological saline

Post-Exposure Period: 24 Hours

Method/Guideline and Test Condition Remarks: The optimization test was used, an intracutaneous sensitization procedure exceeding the sensitivity of the method recommended in the "Appraisal of the Safety of

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:30:31 AM] Chemicals in Foods, Drugs and Cosmetics" (1959), the US Association of Food and Drug Officials (AFDO).

The test was performed on 10 male and 10 female guinea pigs of the Pirbright white strain, bred on the premises and weighing between 221 to 431 grams (~10 weeks old).

The animals were housed individually in Macrolon cages type 3, assigned to the different groups by means of random numbers generated by the random number generator incorporated in the Hewlett-Packard desk computer, 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. The animals received ad libitum standard guinea pig pellets and water, supplemented with fresh
carrots. The animals were acclimatized for 12 days.

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. The reactions were scored according to the method described in the Federal Hazardous Substances Act.

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% solution of TK 10 637/1 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% solution of TK 10 637/1 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 (10% TK 10 637/1 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 to Draize scoring scale.

### High Production Volume Information System (HPVIS) | High Production Volume (HPV) Challenge | OPPT | US EPA

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### Test Results - Skin Sensitization

Measurement Period and Units:

Percent Sensitized Test Substance:

Percent Sensitized Positive Control:

Percent Sensitized Negative Control:

Sensitization Score:

Results Remarks: 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 10 637/1. TK 10 637/1 was found to be devoid of skin-sensitizing (contact allergenic) potency in albino guinea-pigs.
Interpretation of Results: Not Sensitizing

Conclusion: Not a sensitizer



### Reliability/Data Quality - Skin Sensitization
Reliability: 1 - Valid Without Restrictions

Reliability Remarks:

Key Study Sponsor Indicator:



### Reference - Skin Sensitization

Reference: TSCATS submission (OTS0519042, Doc ID 86-890000273, received 06/07/89): Ciba Geigy Corp. 1982. Initial submission: Report on Skin Sensitizing Effect in Guinea Pigs of TK 10 637/1 with Cover Letter Dated 060589.