SRC TR-10-059 

HPV1 TEST RULE STUDY – DATA ADEQUACY ASSESSMENT 

Methyl Heptanone, CAS No. 409-02-9 

Acute Toxicity to Daphnia 

Prepared by: 

Kendra Moran, Ashlee Aldridge, and Joan D. Garey 

Environmental Science Center 

SRC, Inc. 

7502 Round Pond Road 

North Syracuse, NY 13212 

Contract No. EP-W-09-027 

Task FG007.0.009.004 

Submitted to: 

OPPT/RAD/HPVCB (7403M) 

U.S. Environmental Protection Agency 

1200 Pennsylvania Avenue 

Washington, D.C. 20460 

Elizabeth Margosches, Project Officer 

Meena Sonawane, Work Assignment Manager 

David Brooks, Alternate Work Assignment Manager 

March 5, 2010 
HPV1 Test Rule – Data Adequacy Assessment 

Methyl Heptanone, CAS No. 409-02-9 

Acute Toxicity to Daphnia 

March 2010 

Submitted to: 

High Production Volume Chemicals Branch 

Risk Assessment Division 

Office of Pollution Prevention and Toxics 

Environmental Protection Agency 

WAM: Meena Sonawane 

Alternate WAM: David Brooks 

1200 Pennsylvania Avenue, NW 

Washington, DC 20460-0001 

Prepared By: 

SRC, Inc. 

Environmental Science Center 

2451 Crystal Drive, Suite 804 

Arlington, VA 22202 
Draft HPV1 Test Rule – Data Adequacy – Acute Toxicity to Daphnia CAS No. 409-02-9

PREPARATION OF THE DATA ADEQUACY ASSESSMENT

This document contains the review of a toxicity study conducted in response to the first 

HPV TSCA Section 4 Test Rule (71 FR 13708, March 16, 2006). Using the applicable test 

guideline and professional judgment as guides, the study was reviewed for adequacy and 

evaluated for compliance with the test guideline. (The applicable test guideline is not always the 

test guideline required by the test rule, because some test sponsors requested and received 

approval from EPA to use a different guideline.) The study was summarized according to a 

standard format, assessed for adequacy, and assigned a Klimisch reliability code. If the study 

was determined to be inadequate, a basis for that conclusion is supplied. For studies already 

assigned Klimisch codes, an explanation is provided if the reviewer’s assessment of data 

adequacy differed from the pre-assigned Klimisch code. 

A detailed evaluation of the test report and study conduct is provided at the end of the 

summary document in a section entitled “Review of TSCA Section 4 Final Study Report”. Any 

deviations from the applicable test guideline are described. The study is also evaluated for 

compliance with Standard Operating Procedures, Good Laboratory Practice Standards, and 

Quality Assurance procedures, as well as the use of appropriate controls and test substances. 

Any deficiencies in the descriptions of the study method, test system, animals studied, physical 

conditions, dosage information, individual animal data, calculations, and statistical analyses are 

described, and internal inconsistencies within the study report are identified. An overall 

assessment is provided for each of the following: study conduct, data reliability, and data utility. 

1 
Draft HPV1 Test Rule – Data Adequacy – Acute Toxicity to Daphnia CAS No. 409-02-9

ACUTE TOXICITY TO DAPHNIA 

Title: Acute Immobilization Test 

This study is: Adequate Not Adequate 

TEST SUBSTANCE 

 

Identity: Methyl Heptanone, CAS No. 409-02-9 

Remarks: The test substance, Baton Rouge MEK Heavy Ketone Heart Cut C8 Ketone Fraction, 

was characterized as a complex mixture. 

METHOD

Method/guideline followed: OECD TG 202 (1984)

Test type: Static

GLP compliant?: Yes

Year study was performed: 2007

Test species: Daphnia magna Strauss. The daphnids were cultured in-house. Age was <24 

hours old from 16-day old parents. 

 

Analytical monitoring: Yes 

Exposure duration: 48 hrs

Concentrations tested: 0 (control), 0.75, 2.0, 3.9, 9.1, and 22 mg/L test substance water 

accomadated fraction (WAF), which corresponded to mean measured concentrations of 0 

(control), 0.755, 1.78, 3.34, 7.99, and 16.6 mg/L.

Test method/conditions remarks: Individual Water Accommodated Fractions (WAFs) were 

prepared. The test substance was added to 4 L of reconstituted water in glass aspirator bottles 

(capacity 4.5 L). The test solutions and a control were mixed for 23.5 hours using a 4% vortex 

(of the static liquid depth). The mixtures were removed through an outlet at the bottom of each 

mixing vessel into four replicates of approximately 140 mL in 125 mL Erlenmeyer flasks (no 

headspace). Five daphnids were added to each replicate and the replicates were closed. The test 

was performed under static conditions with no aeration. 

Dissolved oxygen ranged from 8.2 to 8.4 mg/L and pH ranged from 7.6 to 7.8 during the study. 

Water hardness was 142 mg/L as CaCO3. The TOC of the dilution water was 0.54 ppm. The test 

water temperatures ranged from 20.7°C to 20.9°C. The environmental condition daily average 

temperatures ranged from 19.6°C to 20.6°C. Diurnal light was 16 hours light and 8 hours dark 

2 
Draft HPV1 Test Rule – Data Adequacy – Acute Toxicity to Daphnia CAS No. 409-02-9

with light intensity ranging from 444 to 456 Lux during full light hours. 

Due to the relatively complex nature and limited water solubility of the test substance, the 

following exceptions to the guideline apply for this study: 1) The concentration of the test 

substance in solution was not determined prior to use.; and 2) It was deemed more appropriate to 

prepare an individual treatment solution by adding the test substance to dilution water and 

removing the WAF for testing, rather than preparing a dilution of the stock solution as outlined 

in the guideline. 

RESULTS

Endpoint value(s): The 48-hour EC50 value for daphnids was 5.2 mg/L based on mean 

measured concentrations and the EL50 was 6.0 mg/L based on WAF loading rates. 

Results remarks: 

The maximum actual loading rate causing no immobilization was 3.9 mg/L. The maximum 

measured concentration causing no immobilization was 3.3 mg/L. The minimum actual loading 

rate causing 100% immobilization was 9.1 mg/L. The minimum measured concentration causing 

100% immobilization was 8.0 mg/L. Control survival was 100%. 

CONCLUSIONS

After Daphnia magna were exposed to a water accommodated fraction (WAF) prepared from 

Baton Rouge MEK Heavy Ketone Heart Cut - C8 Ketone Fraction for 48 hours, the EC50 was 5.2 

mg/L based on mean measured concentrations. The EL50 was 6.0 mg/L based on WAF loading 

rates. 

STUDY RELIABILITY

[1] Reliable without restrictions. 

REFERENCE

ExxonMobil Biomedical Sciences, Inc. (2007). Daphnia sp., Acute Immobilization 

Test, Study # 0700842 

3 
Review of TSCA Section 4 Final Study Report 

CITB Project Manager: Catherine Roman 

Chemical Name: Methyl Heptanone 

CAS Registry Number: 409-02-9 

Report Title: Daphnid Acute Immobilization Test 

Test Sponsor: Exxon Mobil Chemical Company 

Testing Lab: Exxon Mobil Biomedical Sciences, Inc. 

Test Standard: ASTM E 729 

Date Study Received by EPA: 6/23/09 (date posted to http://www.regulations.gov) 

Date Study Due to EPA: 5/17/07 

Special Instructions (e.g., regarding approved protocol modifications, etc.): 

 

Part I. Test Report and Study Conduct

Enter one of the following codes into each of the checkboxes 1 through 13: Agree [A], Disagree [D], Not 

Applicable [NA], or Cannot Determine [C], and add comment, below, as needed

 [D] 1. The final study report indicates evidence of gross, unauthorized or undocumented deviation from 

 the EPA test standard requirements, including “shall,” “must,” or “will” violations. List 

 significant test standard deviations and “shall,” “must,” or “will” violations in comments below. 

 [D] 2. The final study report indicates evidence of deviations from the “should” statements in the study protocol 

that may have a bearing on the study conclusions. List these in comments below. 

 [D] 3. Approved modifications of test standard or testing protocol (CITB PM describe or attach) were not 

followed or are inadequately documented. 

 [D] 4. There is evidence that laboratory Standard Operating Procedures (SOPs) were inadequately followed. 

 [D] 5. The Statement of Quality Assurance (QA) inspections, their dates, and findings were missing or not signed. 

There is evidence that QA procedures may be inadequate. 

[D] 6. There is evidence that Good Laboratory Practice Standards (GLPS) were inadequately followed. 

[D] 7. The test, control, or reference substances are different from that specified in the section 4 test rule or 

Enforceable Consent Agreement (ECA). 

[D] 8. Animal identification is inadequate or unclear; use of unhealthy animals or other factors (e.g., deviations 

from temperature, lighting, dosing) are apparent. 

[D] 9. The final study report inadequately describes the study method. 

[D] 10. The final study report inadequately describes the test system. 
[D] 11. The final study report inadequately describes the dosage information. 

[D] 12. The final study report does not accurately reflect the individual animal data (or that of other discrete 

observations pertinent to the type of study conducted), or individual data were not provided. 

[D] 13. The final study report inadequately describes the calculations, interpolations, data transformations, 

statistical analyses, or modeling parameters used. 

Part II. Conclusions

Place a check mark beside the summary statement(s) that apply to this review and add comments below as 

appropriate

Part A. To be filled out by Reviewer

 

Study Conduct 1. The study was well-conducted. [x] 

 2. The study was generally well-conducted, with some minor [ ] 

 deficiencies (list in notes below). 

 3. The study conduct was significantly deficient or questionable, (give [ ] 

 reasons in notes below). 

Data Reliability 4. The study results are precise and accurate. [x] 

 5. Appropriate methods were used to determine [x] 

 statistical significance. 

 6. If repeated, the test results would be expected to be the [x] 

 same. 

Data Utility 7. The data are useful for risk assessment for the endpoint tested. [x] 

 8. The data are useful for some other purpose (e.g., dose-setting). [ ] 

 9. The data are not useful for any purpose (describe in notes below). [ ] 

Reviewer Comments (continue on separate page, if needed): 

Reviewer Name/Title/Branch/Division/Office: Kendra Moran, SRC 

Reviewer Signature: David Brooks/HPVB/RAD/OPPTS 

Part B. To be filled out by CITB PM

Tiered Testing Decision 1. The tiered testing trigger was met, additional testing is indicated. [ ] 

(when applicable) 2. The trigger for additional (tiered) testing was not met. [ ] 

 

Study Acceptability 3. The study is acceptable. [ ] 

 4. The study may not be acceptable [ ] 

 (provide rationale in memo to CITB chief).