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

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# Submission Report

Submission Information

Submission ID : 25349326 Submission Name : 2,5-Furandione, 3-dodecenyldihydroAR Number :



## Chemical Information

CAS Number : 25377-73-5 Ninth Collective Index Name : 2,5-Furandione, 3-(dodecenyl)dihydro

## 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 10:31:51 AM] E-Mail Address : Type of Sponsor : Company 

## Technical Point of Contact Information

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



## Acute Toxicity to Aquatic Vertebrates

CAS Number : 25377-73-5

Sponsored Chemical Name : 2,5-Furandione, 3-(dodecenyl)dihydro

### Test Substance - Acute Toxicity To Aquatic Vertebrates

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydroTest Substance Purity/Composition and Other Test Substance Comments: Purity not specified

Sponsored Chemical Result Type: Measured

Test Substance Result Type: Measured



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

Method/Guideline Followed: Other

Other Method/Guideline: Standard Methods for the Examination of Water and Wastewater (1971) American Public Health Association, Static acute toxicity test, Method No. 231

Deviations from Method/Guideline: Yes

Species: Carassius auratus

GLP: No Data

Analytical Monitoring: Yes

Test Type: Static

Test Vessel: Closed

Water Media Type: Freshwater

Test Concentrations: Measured

Nominal and Measured Concentrations: Not Specified

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Exposure Period: 96 Hours

Vehicle Used: No Data

Vehicle Name:

Vehicle Amount and Units:
Alkalinity:

Dissolved Oxygen:

pH Value: Value/Lower Range: = 6 Upper Range: 7

Test Temperature and Units: Value/Lower Range: 19 Upper Range: 21 °C

Photo (Light/Dark):
Salinity:

TOC:

Water Hardness:

Method/Guideline Test Conditions Remarks: Ten fish per test concentration were exposed at 20 ± 1 °C in 25 L of solution contained in all-glass tanks measuring 42 x 28 x 28 cm. The duration of the test was 24 or 96 hours and the solutions were aerated throughout the test period. The concentration of the chemical in solution was determined before and after each test, either with a total organic carbon analyzer or by extraction and subsequent GC analysis. Based on the average fish weight of 3.3 g and the testing of 10 fish/25L, the loading rate was 1.32 g fish/L. The latter value exceeds the OECD TG 203- recommended maximum loading of 1g fish/L for static tests.

Solutions were made in local tap water. The analysis was: Cl- = 65 mg/L NO2- = 0 mg/L NO3- = 4 mg/L SO42- = 35 mg/L PO43- = 0.15 mg/L HCO3- = 25 mg/L SiO2 = 25 mg/L NH4+ = 0 mg/L Fe = 0.05 mg/L Mn = 0 mg/L Ca2+ = 100 mg/L Mg2+ = 8 mg/L Na+ = 30 mg/L pH = 7.8

Limit Test: Unknown



### 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 10:31:51 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

24 Hours LC 50 % = 2 4 mg/L Mortality Calculated

96 Hours LC 50 % = 2 3 mg/L Mortality Calculated

Results Remarks: 24-hour LC50 (pH = 7.0) = 4 mg/L 96-hour LC50 (pH = 7.0) = 3 mg/L 24-hour LC50 (pH = 6.0) = 2 mg/L 96-hour LC50 (pH = 6.0) = 2 mg/L

The LC50 was obtained by interpolation from a graph of the logarithm of the concentration versus the percentage mortality.



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

Reliability Remarks: Non-preferred species and fish loading rate that exceeded guidline recommendations; missing study details included water hardness, total organic carbon, test substance purity, nominal and measured test concentrations, and mortality at each concentration
Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Vertebrates

Reference: Bridie et al. (1979) The acute toxicity of some petrochemicals to goldfish. Water Research 13, 623 - 626.

Bridie et al. (1974) Determination of Acute Toxicity to Fish of Shell Chemicals,
II. AMGR.0071.74.

TSCATS OTS0206206; Doc ID: 878210130. Shell Oil Company (12/10/82).



## Acute Toxicity to Aquatic Invertebrates

CAS Number : 25377-73-5

Sponsored Chemical Name : 2,5-Furandione, 3-(dodecenyl)dihydro

### Test Substance - Acute Toxicity To Aquatic Invertebrates

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydro-

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Test Substance Purity/Composition and Other Test Substance Comments:
Sponsored Chemical Result Type:

Test Substance Result Type: 

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

Method/Guideline Followed:

Other Method/Guideline:

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

GLP:

Analytical Monitoring:
Test Type:

Test Vessel:

Water Media Type:

Test Concentrations:

Nominal and Measured Concentrations:
Exposure Period:

Vehicle Used:

Vehicle Name:

Vehicle Amount and Units:
Alkalinity:

Dissolved Oxygen:

pH Value:

Test Temperature and Units:

Photo (Light/Dark):
Salinity:

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] TOC:

Water Hardness:

Method/Guideline Test Conditions Remarks:

Limit Test:



### Test Results - Acute Toxicity To Aquatic Invertebrates
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:



### Reliability/Data Quality - Acute Toxicity To Aquatic Invertebrates
Reliability:

Reliability Remarks: As of February 2010, no data were available for this endpoint.

Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Invertebrates

Reference:

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] 

## Acute Toxicity to Aquatic Plants

CAS Number : 25377-73-5

Sponsored Chemical Name : 2,5-Furandione, 3-(dodecenyl)dihydro

### Test Substance - Acute Toxicity To Aquatic Plants

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydroTest Substance Purity/Composition and Other Test Substance Comments:
Sponsored Chemical Result Type:

Test Substance Result Type: 

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

Method/Guideline Followed:

Other Method/Guideline:

Deviations from Method/Guideline:
Species:

GLP:

Analytical Monitoring:
Test Type:

Test Vessel:

Water Media Type:

Test Concentrations:

Nominal and Measured Concentrations:
Exposure Period:

Vehicle Used:

Vehicle Name:

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 10:31:51 AM] pH Value:

Test Temperature and Units:

Photo (Light/Dark):
Salinity:

TOC:

Water Hardness:

Method/Guideline Test Conditions Remarks:

Limit Test:



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

NOEC:

LOEC Exposure Duration:

LOEC:

NOELR Exposure Duration:

NOELR:

LOELR Exposure 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

Results Remarks:



### Reliability/Data Quality - Acute Toxicity To Aquatic Plants
Reliability:

Reliability Remarks: As of February 2010, no data were available for this endpoint.

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Key Study Sponsor Indicator: 

### Reference - Acute Toxicity To Aquatic Plants

Reference:



## Acute Toxicity

CAS Number : 25377-73-5

Sponsored Chemical Name

: 2,5-Furandione, 3-(dodecenyl)dihydro

### Test Substance - Acute Toxicity

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydroTest Substance Purity/Composition and Other Test Substance Comments: Purity not specified; Appearance: Amber liquid; Sponsor's identification code: 127-2638, Dodecenyl Succinic Anhydride; FDRL identification code: 80-1168

Sponsored Chemical Result Type: Measured



### Method - Acute Toxicity

Route of Administration: Inhalation

Type of Exposure: Aerosol

Species: Rat

Mammalian Strain: Sprague-Dawley

Gender: Both M/F

Number of Animals per Dose: 10

Concentration: 5.3 mg/L (nominal); 1.22 mg/L (time-weighted gravimetric concentration)
Year Study Performed: 1981

Method/Guideline Followed: Other

GLP: Yes

Method/Guideline and Test Condition Remarks: Method/guideline followed: Similar to OECD limit test; FDA guidelines for Good Laboratory Practice were followed.

Test species: Sprague-Dawley rats obtained from Blue Spruce Farms, Altamont, NY. Group mean body weight (g) on day 1: 285 ± 5 (males) and 247 ± 6 (females)

The purpose of the study was to evaluate the acute inhalation toxicity of dodecenyl succinic anhydride (DDSAA) according to the OECD Acute Toxicity Screening Program, Acute Inhalation Studies Protocol, as modified and amplified by the

sponsor. The aerosol consisted of 90% DDSAA and 10% ethanol (w/w) with a geometric mean particle size of 2.3 μm (S.D. = 2.2 μm). To serve as a control, additional animals were

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] exposed to an atmosphere containing a liquid droplet aerosol of ethanol and observed a required by the protocol. The animals were exposed to an airborne aerosol of the test article for 4 hours. During exposure, the animals were individually housed in wire mesh cages and denied food and
water. Following exposure, the animals remained individually housed and food and water were provided ad libitum.

The animals were observed for pharmacotoxic signs during exposure (day 1), subsequent to exposure on day one, and daily for 14 additional days (days 2 - 15). Body weights were also recorded from day 1 to 15 of the study. All animals were subject to gross necropsy. During necropsy, all major organs were examined for any signs of abnormalities and the following tissues excised and preserved in 10% neutral buffered formalin: whole head with nasal passages, trachea, bronchi, lungs, liver, and kidneys.

The actual exposure concentrations were determined by gravimetric analysis; a known volume of chamber air was drawn through a preweighed filter and the actual concentration was calculated by dividing the filter weight gain by the sample volume, and then correcting for dilution (by multiplying by 0.90).



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

LC 50 % > 1.2 mg/L

Number of Deaths ( Male ): 2

Number of Deaths ( Female ): 2

Number of Deaths ( Total ): 4

Results Remarks: The observed mortality was 2/5 for females by the second day and 2/5 males by the fifth day of post-exposure observation. No further deaths were observed by the 15th day post-exposure. Thus, the LC50 for 4-hour inhalation exposure is greater than 1.2 mg/L.

Significant observations included labored breathing, nasal discharge, urinary incontinence, and eye irritation. Labored breathing was present in all animals and continued throughout the study. Bloody nasal discharge was also observed in all animals during exposure and generally persisted throughout the observation period. Eye irritation, closing of the eyes (due to swelling of periorbital tissues) and a cloud or film-covered appearance of the eye was evident. Apparent blood around the eyes may have been porphyrin-containing secretions of the Harderian glands and is often seen in debilitated animals. Urinary incontinence was noted in all males on days two through four, and alopecia (hair loss) was evident in two males and one

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] female subsequent to day seven. Following exposure to the test article, body weight loss occurred in all animals. This was due to dodecenyl succinic anhydride exposure, as ethanol treatment caused no comparable effect. By day 15, the three surviving males and one female had regained their original body weights.

No findings attributable to test article exposure were evident upon gross necropsy. Various color alterations of the lung and other organs (dark, pale, spotting) are routine post mortem observations in the laboratory rat and results from post mortem congestion within the tissues.

Conclusion: Four-hour exposure to liquid droplet aerosol of dodecenyl succinic anhydride (90% w/w solution in ethanol) produced mortality in two of five male and two of five female
rats. Labored breathing, transient urinary incontinence, alopecia, eye irritation, and body weight loss were observed following exposure to the test article. 

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

Reliability Remarks: Mortalities were observed in the limit test. Further acute toxicity testing is recommended.
Key Study Sponsor Indicator: 

### Reference - Acute Toxicity

Reference: This summary is based on a study from a TSCATS submission OTS0505452 (Document ID: 88-8100369) conducted by Food and Drug Research Laboratories, for Buffalo Color Corporation: Acute Inhalation Toxicity of Dodecenyl Succinic Anhydride in Sprague-Dawley Rats with Cover Letter Dated 04/08/82. (Study produced on March 24, 1981)



## Acute Toxicity

CAS Number : 25377-73-5

Sponsored Chemical Name

: 2,5-Furandione, 3-(dodecenyl)dihydro

### Test Substance - Acute Toxicity

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydroTest Substance Purity/Composition and Other Test Substance Comments: Purity not specified

Sponsored Chemical Result Type: Measured



### Method - Acute Toxicity

Route of Administration: Oral

Type of Exposure: Gavage

Species: Rat

Mammalian Strain: Other

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Other Strain:

Gender: Male

Number of Animals per Dose: 5

Dose: 1, 2, 4, 8, 16 g/kg

Year Study Performed: 1978

Method/Guideline Followed: OECD 401

GLP: Not Applicable

Method/Guideline and Test Condition Remarks: Single administration of the test material was given intragastrically to five fasted male rats at each dose level. The animals were observed for signs of toxicity or behavioral changes on the day of treatment and throughout the 14-day observation period. Individual weights were recorded immediately prior to dosing and prior to termination. The surviving animals were euthanized at the conclusion of the observation period. Gross autopsies were performed on all animals.



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

LC 50 % = 2900 mg/kgbw

Number of Deaths ( Male ): 15

Number of Deaths ( Female ): 0

Number of Deaths ( Total ): 15

Results Remarks: During the first three days of study all animals treated at the 4, 8 and 16 g/kg dose levels died. No deaths were observed at the 1 and 2 g/kg dose levels. No clinical signs of toxicity were observed at 1 g/kg. At 2 g/kg the animals were lethargic and had an oily appearance for up to 48 hours post dosing. All animals at the 4, 8 and 16 g/kg dose levels were severely depressed prior to death. No body weight effects occurred at 1 or 2 g/kg. Body weight data were not available at higher dose levels due to the observed mortality. No test material related macroscopic findings were evident.

Conclusion: LD50 (males) = 2.9 g/kg



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

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Reliability Remarks: Purity not specified

Key Study Sponsor Indicator: 

### Reference - Acute Toxicity

Reference: HPV submission for Alkenyl Succinic Anhydrides category, received by EPA from Petroleum Additives Panel Health & Environmental Research Task Group on November 22,2002; HPV submission references "Unpublished confidential business information" updated 2/23/01 (RTA-074).



## Repeated-Dose Toxicity

CAS Number : 25377-73-5

Sponsored Chemical Name : 2,5-Furandione, 3-(dodecenyl)dihydro

### Test Substance - Repeated-Dose Toxicity

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydroTest Substance Purity/Composition and Other Test Substance Comments:
Sponsored Chemical Result Type: 

### Method - Repeated-Dose Toxicity

Route of Administration:
Type of Exposure:

Species:

Mammalian Strain:

Gender:

Number of Animals per Dose:

Dose:

Year Study Performed:

Method/Guideline Followed:

GLP:

Exposure Period:

Frequency of Treatment:
Post-Exposure Period:

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Method/Guideline and Test Condition Remarks: 

### Test Results - Repeated-Dose Toxicity
Concentration (LOAEL/ LOAEC/ NOAEL/ NOAEC) : LOAEL/ LOAEC/ NOAEL/ NOAEC Population Value Description Value/Lower Concentration Upper Concentration Units

Results Remarks:

Conclusion:



### Reliability/Data Quality - Repeated-Dose Toxicity
Reliability:

Reliability Remarks: As of February 2010, no data were available for this endpoint.

Key Study Sponsor Indicator: 

### Reference - Repeated-Dose Toxicity

Reference:



## Genetic Toxicity in vivo

CAS Number : 25377-73-5

Sponsored Chemical Name

: 2,5-Furandione, 3-(dodecenyl)dihydro

### Test Substance - Genetic Toxicity in vivo

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydroTest Substance Purity/Composition and Other Test Substance Comments:
Sponsored Chemical Result Type: 

### Method - Genetic Toxicity in vivo
Type of Study:

Type of Test:

Route of Administration:

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Type of Exposure:

Species:

Strain:

Gender:

Dose:

Year Study Performed:

Method/Guideline Followed:

GLP:

Duration of Treatment/ Exposure Period and Units:
Frequency of Treatment:

Positive, Negative, and Solvent Control Substance(s):
Number of Animals per Sex and Dose Group:

Method/Guideline and Test Condition Remarks: 

### Test Results - Genetic Toxicity in vivo
Systemic Toxicity:

Genotoxic Effect:

Results Remarks:

Conclusion:



### Reliability/Data Quality - Genetic Toxicity in vivo
Reliability:

Reliability Remarks: As of February 2010, no data were available for this endpoint.
Key Study Sponsor Indicator: 

### Reference - Genetic Toxicity in vivo

Reference:



## Genetic Toxicity in vitro

CAS Number : 25377-73-5

Sponsored Chemical Name : 2,5-Furandione, 3-(dodecenyl)dihydro-

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Test Substance - Genetic Toxicity in vitro

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydroTest Substance Purity/Composition and Other Test Substance Comments: Purity not specified

Sponsored Chemical Result Type: Measured



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

Concentrations: 0.3, 1, 3.3, 10, 100 μg/plate without activation and 100, 333, 1000, 3333, 10,000 μg/plate with activation
Year Study Performed:

Method/Guideline Followed: Other

GLP: No Data

Postive, Negative, and Solvent Control Substance(s): Concurrent solvent control substance: DMSO Positive control substances were not specified

Method/Guideline and Test Condition Remarks: Method/guideline followed: Ames et al. 1975, with and without metabolic activation. Plate incorporation method.

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. For each plate receiving microsomal enzymes, 0.5 mL of S9 mix was added.

For testing in the absence of S9 mix, 100 μL of the tester strain and 50 μL of the solvent or test chemical were added to 2.5 mL of molten selective top agar at 45 ± 2 oC. When S9 was used, 0.5 mL of S9 mix, 50 μL of tester strain, and 50 μL of solvent or test chemical were added to 2.0 mL of molten selective top agar at 45
± 2 oC. 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
oC. Five doses of test chemical, together with the appropriate concurrent solvent (DMSO) and positive controls (not specified), 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.



### Test Results - Genetic Toxicity in vitro

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] 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 Negative Negative

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 Negative Negative

Results Remarks: Positive and negative controls responded appropriately; Cytotoxic concentration: None specified

Conclusion: The test substance did not cause an increase in mutation frequency in Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 or TA1538 with or without metabolic activation. 

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

Reliability Remarks: Purity not specified. Although the findings from this study were presented in a review article and a citation to a previous publication of the data was not provided, the primary data from the study were made available in a "Supporting Information" source at http://pubs.acs.org.
Key Study Sponsor Indicator: 

### Reference - Genetic Toxicity in vitro

Reference: Seifried et al. (2006) A compilation of two decades of mutagenicity test results with the Ames Salmonella typhimurium and L5178Y mouse lymphoma cell mutation assays. Chemical Research in Toxicology 19, 627-644.



## Genetic Toxicity in vitro

CAS Number : 25377-73-5

Sponsored Chemical Name : 2,5-Furandione, 3-(dodecenyl)dihydro

### Test Substance - Genetic Toxicity in vitro

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydroTest Substance Purity/Composition and Other Test Substance Comments: Purity not specified

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Sponsored Chemical Result Type: Measured



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

Concentrations: 0.017, 0.022, 0.026, 0.031, 0.035 μg/mL without activation and 0.035, 0.08, 0.13, 0.17, 0.22, 0.26, 0.31 μg/mL with activation

Year Study Performed:

Method/Guideline Followed: Other

GLP: No Data

Postive, Negative, and Solvent Control Substance(s): Solvent control: DMSO Positive Control (without metabolic activation): ethyl methylsulfonate at 4.7 x 10-6 M (or methyl methanesulfonate at 10 - 20 μg/mL) Positive Control (with metabolic activation): 3-methylcholanthrene at 1.86 x 10-5 M (or dimethylbenz[a]-anthracene at 0.5 - 4 μg/mL)

Method/Guideline and Test Condition Remarks: 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. (1979). 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 10,000 μg/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 (DMSO) controls, a positive control of ethyl methylsulfonate at 4.7 x10-6 M (or methyl methanesulfonate at 10 - 20 μg/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 μg/mL) for the test with metabolic activation.

The mutagenicity assay was performed according to the procedure described by Clive and Spector (1975). 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 μg/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 ~ 0.2 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 adoubling of the mutant frequency over the concurrent solventtreated control value as an indication of a positive effect,

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] 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

Mammalian Cell Line Mouse Lymphoma

L5178Y

Cells With and Without Negative Negative

Results Remarks: Positive and negative controls responded appropriately

Genotoxicity result: Negative with and without metabolic activation using the evaluation criteria that are described above. The Seifried et al. (2006) publication also presented mutagenicity conclusions that were stated to be "new" evaluations based on 'current international "harmonization" recommendations'. However, these "new" evaluations were considered to be uninterpretable and are not considered further because this publication neither presented the criteria used for the "new" evaluations nor cited literature that present such criteria.

Cytotoxic concentration: ≥ 0.031 μg/mL without activation, ≥ 0.13 μg/mL with activation

Conclusion: The test substance was negative for mutagenicity in mouse lymphoma cells with and without metabolic activation. 

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

Reliability Remarks: Test substance purity was not specified. Although the findings from this study were presented in a review article and a citation to a previous publication of the data was not provided, the primary data from the study were presented in "Supporting Information" documents at http://pubs.acs.org.
Key Study Sponsor Indicator: 

### Reference - Genetic Toxicity in vitro

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Reference: Seifried et al. (2006) A compilation of two decades of mutagenicity test results with the Ames Salmonella typhimurium and L5178Y mouse lymphoma cell mutation assays. Chemical Research in Toxicology 19,
627-644.



## Reproductive Toxicity

CAS Number : 25377-73-5

Sponsored Chemical Name : 2,5-Furandione, 3-(dodecenyl)dihydro

### Test Substance - Reproductive Toxicity

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydroTest Substance Purity/Composition and Other Test Substance Comments:
Sponsored Chemical Result Type: 

### Method - Reproductive Toxicity

Route of Administration:
Type of Exposure:

Species:

Mammalian Strain:

Gender:

Number of Animals per Dose:

Dose:

Year Study Performed:

Method/Guideline Followed:

GLP:

Exposure Period:

Frequency of Treatment:
Post-Exposure Period:

Method/Guideline and Test Condition Remarks:
Pre-Mating Exposure / Males:
Pre-Mating Exposure / Females:

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Test Results - Reproductive Toxicity
Concentration (LOAEL/ LOAEC/ NOAEL/ NOAEC) : LOAEL/ LOAEC/ NOAEL/ NOAEC Population Value Description Value/Lower Concentration Upper Concentration Units

Results:

Results Remarks:

Conclusion:



### Reliability/Data Quality - Reproductive Toxicity
Reliability:

Reliability Remarks: As of February 2010, no data were available for this endpoint.

Key Study Sponsor Indicator: 

### Reference - Reproductive Toxicity

Reference:



## Developmental Toxicity/Teratogenicity

CAS Number : 25377-73-5

Sponsored Chemical Name : 2,5-Furandione, 3-(dodecenyl)dihydro

### Test Substance - Developmental Toxicity/Teratogenicity

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydroTest Substance Purity/Composition and Other Test Substance Comments:
Sponsored Chemical Result Type: 

### Method - Developmental Toxicity/Teratogenicity

Route of Administration:
Type of Exposure:

Species:

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Mammalian Strain:

Gender:

Number of Animals per Dose:

Dose:

Year Study Performed:

Method/Guideline Followed:

GLP:

Exposure Period:

Frequency of Treatment:
Post-Exposure Period:

Method/Guideline and Test Condition Remarks: 

### Test Results - Developmental Toxicity/Teratogenicity
Concentration (LOAEL/ LOAEC/ NOAEL/ NOAEC) : LOAEL/ LOAEC/ NOAEL/ NOAEC Population Value Description Value/Lower Concentration Upper Concentration Units

Results Remarks:

Conclusion:



### Reliability/Data Quality - Developmental Toxicity/Teratogenicity
Reliability:

Reliability Remarks: As of February 2010, no data were available for this endpoint.

Key Study Sponsor Indicator: 

### Reference - Developmental Toxicity/Teratogenicity

Reference:



## Skin Irritation

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] CAS Number : 25377-73-5

Sponsored Chemical Name

: 2,5-Furandione, 3-(dodecenyl)dihydro

### Test Substance - Skin Irritation

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydroTest Substance Purity/Composition and Other Test Substance Comments: Light yellow, clear, viscous liquid; MW: 266; Solubility: reacts with water and alcohols, soluble in certain oils and industrial solvents

Sponsored Chemical Result Type: Measured



### Method - Skin Irritation
Species: Other

Other Species: Guinea Pig

Mammalian Strain: Other

Other Strain: Albino

Type of Coverage: Occlusive

Preparation of Test Site: Shaved

Gender: Female

Number of Animals per Dose:

Amount/Concentration Applied: pure compound (400 mg) or 10% solution in corn oil

Year Study Performed: 1964

Method/Guideline Followed: No Data

GLP: No Data

Exposure Period: 24 Hours

Total Volume applied and Units: 0.4 ml

Control Group Type: No Data

Vehicle Used: Yes

Vehicle Name: Corn Oil

Vehicle Amount and Units:

Post-Exposure Period: 14 Days

Grading Scale:

Method/Guideline and Patch tests were performed on female, albino guinea pigs

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Test Condition Remarks: weighing 387 to 395 g, using 0.4 mL of the indicated concentrations of the test solution.

Undiluted material was applied to closely clipped abdominal skin of albino guinea pigs by means of cotton-gauze patches (2 x 2 cm) that were held in contact with the skin by means of a 3-in. Elastoplastic bandage. The patches were removed 24 hours after application, and the animals were observed for a period of at least 14 days; when damage occurred, observations were continued until the damaged areas healed. When a response was elicited with the undiluted material, tests were repeated using 10% and 1% solutions.



### Test Results - Skin Irritation

Grade:

Primary Irritation Index:

Lesions:

Erythema:

Edema:

Results Remarks: With the pure compound, there was a slight edema which persisted for 3 days, a scab which was evident on the 8th day, and complete healing on the 14th day. The 10% solution (corn oil) produced no effects.
Interpretation of Results:

Conclusion:



### Reliability/Data Quality - Skin Irritation
Reliability:

Reliability Remarks:

Key Study Sponsor Indicator:



### Reference - Skin Irritation

Reference: Longley et al. 1964; AMRL (1964) Toxic Hazard Study of Selected Missile Propellants Document Number(s): AD 441 640.
AMRL-TDR-64-28.



## Eye Irritation

CAS Number : 25377-73-5

Sponsored Chemical Name

: 2,5-Furandione, 3-(dodecenyl)dihydro

### Test Substance - Eye Irritation

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydroTest Substance Purity/Composition and Other Test Substance Comments: Light yellow, clear, viscous liquid; MW: 266; Solubility: Reacts with eater and alcohols, soluble in certain oils and industrial solvents

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Sponsored Chemical Result Type: Measured



### Method - Eye Irritation
Species: Other

Other Species: Guinea Pig

Mammalian Strain: Other

Other Strain: Albino

Gender: No Data

Number of Animals per Dose:

Amount/Concentration Applied: 50 mg

Year Study Performed: 1964

Method/Guideline Followed: No Data

GLP: No Data

Exposure Period:

Total Volume applied and Units: 50 mg

Control Group Type: No Data

Vehicle Used: No

Vehicle Name:

Vehicle Amount and Units:

Post-Exposure Period:

Grading Scale:

Tool:

Method/Guideline and Test Condition Remarks: Undiluted material was instilled into the conjunctival sac of albino guinea pigs. Effects were noted at 1 min, 5 min, 1 hour and 24 hours after exposure, and daily thereafter for a total of at least 14 days. When damage occurred, observations were continued until healing was completed.



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

Overall Irritation Score:

Primary Irritation Index:

Lesions:

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Cornea:

Iris:

Conjunctivae (Chemosis):

Conjunctivae (Redness):

Results Remarks: The pure compound produced slight swelling, with a very slight opacity in the cornea and a moderate discharge which lasted for 1 day. There appeared to be no effects persisting beyond the 2nd post-exposure day.
Interpretation of Results:

Conclusion:



### Reliability/Data Quality - Eye Irritation
Reliability:

Reliability Remarks:

Key Study Sponsor Indicator: 

### Reference - Eye Irritation

Reference: Longley et al. 1964; AMRL (1964) Toxic Hazard Study of Selected Missile Propellants Document Number(s): AD 441 640.
AMRL-TDR-64-28.



## Skin Sensitization

CAS Number : 25377-73-5

Sponsored Chemical Name

: 2,5-Furandione, 3-(dodecenyl)dihydro

### Test Substance - Skin Sensitization

Test Substance: (25377-73-5) 2,5-Furandione, 3-(dodecenyl)dihydroTest Substance Purity/Composition and Other Test Substance Comments: 98% purity; test substance referred to as "C12 ASA"

Sponsored Chemical Result Type: Measured



### Method - Skin Sensitization
Test Type: In Vivo

Study Type: Other

Species: Guinea pig

Mammalian Strain: Hartley

Route of Induction: Dermal

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] Route of Challenage Exposure: Dermal

Gender: Both M/F

Number of Animals per Dose: 20

Concentration: Irritation test: 1%, 2.5%, 5%, 10%, 25%, 50% solution in acetone or undiluted Induction: 25% w/v in acetone Primary challenge: 5% w/v in acetone Rechallenge: 3% w/v in acetone
Year Study Performed: 1986

Method/Guideline Followed: Other

GLP: Yes

Exposure Period: 6 Hours

Induction Frequency of Treatment: once/week for 3 weeks

Challenge Exposure Period: 6 Hours

Challenge Frequency of Treatment: once

Total Volume applied and Units: 0.3 ml

Control Group Type: Other

Vehicle Used: Yes

Vehicle Name: Acetone

Vehicle Amount and Units:

Positive Control Substance:
Post-Exposure Period: 48

Negative Control Substance: Hours

Method/Guideline and Test Condition Remarks: Method/Guideline Followed: Ritz and Buehler, 1980

No. of animals/dose: 4 male, 4 female for irritation study; 10 male, 10 female for test article; 5 male, 5 female for two naive control groups

In the irritation test, a single administration of the test material was applied dermally to sites clipped of hair on four male and four female guinea pigs. Each animal had 4 sites exposed to a different concentration of test article on a gauze patch. Sites were covered with a Hill Top chamber, which was covered with tape and a rubber dam. After 6 hours of application, the occlusion was removed, and the animals returned to their cages. On the day following application, the clipped sites were depilated with chemical hair remover, and the sites scored for severity of response at 24 hours and 48 hours. The animals were observed for signs of erythema and edema and scored either 0 (no reaction), +/- (slight patchy

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] erythema), 1 (slight confluent or moderate patchy edema), 2 (moderate erythema), and 3 (severe erythema with or without edema).

For induction of sensitization, the upper left quadrant of the backs of guinea pigs were clipped of hair. On the following day, moistened patches were applied, to the test group, the animals restrained as previously described, and the animals returned to their cages. Patches moistened with test article were applied to the skin in the same manner once a week for three applications. The same site was clipped on the day before application, and the restraint periods were six hours on each occasion.

For primary challenge two weeks after the last of the induction applications, a fresh application site was prepared by clipping the lower left quadrant of the backs of test and naïve control animals. The next day, challenge patch was applied to each guinea pig in the test and control groups. Each animal was restrained for 6 hours as before, and the animals returned to their cages. On the next day, sites were depilated and scored for severity of response at least two hours later and for a 48 hour reading. Scores of "1" or greater in the test group were considered to be indicative of sensitization providing grades of less than 1 were found in the naïve control group. If the naïve group had grades of "1" or greater, the reactions in the test group that exceeded the most severe reactions in the control group were presumed to be indicative of sensitization.

For rechallenge, eight days after the primary challenge, all of the original animals were single patch rechallenged. Ten previously unexposed naïve animals were identically treated to serve as a new naïve control group. The right rear quadrant was used for rechallenge. Depilation and observation procedures were the same as described for primary challenge.

Individual weights were recorded immediately prior to dosing and prior to termination. The surviving animals were euthanized at the conclusion of the observation period.



### Test Results - Skin Sensitization

Measurement Period and Units:

Percent Sensitized Test Substance:

Percent Sensitized Positive Control:

Percent Sensitized Negative Control:

Sensitization Score:

Results Remarks: No deaths were observed in the test. The incidence of grade "2" responses or greater in the test group exposed to 25% for induction, and 5% at primary challenge was greater for the test animals (13 of 20) compared to the naïve challenge group (0 of 10). The incidence and severity of responses in the test group were more pronounced than the responses of the naïve group, suggesting sensitization had occurred.

At rechallenge with 3% test article in acetone, the incidence of grade "1" responses or greater in the test group (17 of 20) was greater than that in the naïve control group (0 of 10). The incidence and severity of responses were more pronounced in the test group indicating that a sensitization response had been elicited.

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

http://iaspub.epa.gov/hpvis/ServSubmissionPrint [6/15/2011 10:31:51 AM] EPA Home Privacy and Security Notice Contact Us
http://iaspub.epa.gov/hpvis/ServSubmissionPrint Print As-Is
Last updated on Wednesday, June 15, 2011 Interpretation of Results: Sensitizing

Conclusion: The test article, when administered to guinea pigs according to the method of Ritz and Buehler, caused delayed contact hypersensitivity of the skin. 

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

Reliability Remarks:

Key Study Sponsor Indicator:



### Reference - Skin Sensitization

Reference: HPV submission for Higher Alkenyl Succinic Anhydrides category, received by EPA from Albemarle Corporation on April 5, 2006; HPV submission references the following:

Buehler, E.V., "Delayed Contact Hypersensitivity Study in Guinea Pigs of Dodecenyl Succinic Anhydride C12 ASA" for Ethyl Corporation, Hill Top Research Project No. 86-0873-21, Hill Top Research, Inc., 1986.

Ritz, H.L, and Buehler, E.V., (1980), In Current Concepts in Cutaneous Toxicity, (V.A. Drill and T.Lazar, eds.) pp. 25-40, Academic Press, New York.