SRC TR-10-061

HPV1 TEST RULE STUDY – DATA ADEQUACY ASSESSMENT

Methyl Heptanone, CAS No. 409-02-9

Acute Fish Toxicity

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 Wo rk Assignment Manager

March 5, 2010Draft HPV1 Test Rule – Data Adequacy – Acute Toxicity to Fish CAS No. 409-02-9

HPV1 Test Rule – Data Adequacy Assessment

Methyl Heptanone, CAS No. 409-02-9

Acute Toxicity to Fish

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 22202Draft HPV1 Test Rule – Data Adequacy – Acute Toxicity to Fish CAS No. 409-02-9

PREPARATION OF THE DAT A ADEQUACY ASSESSMENT

This document contains the review of a toxi city 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 guide s, the study was reviewed for adequacy and evaluated for compliance with the test guideline. (T he applicable test guid eline 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 assi gned a Klimisch reliability code. If the study was determined to be inadequate, a basis for th at conclusion is supplie d. For studies already assigned Klimisch codes, an explanation is prov ided if the reviewer’s assessment of data adequacy differed from the pre-assigned Klimisch code.

A detailed evaluation of the test report a nd study conduct is provided at the end of the summary document in a section entitled “Revie w 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 Procedur es, Good Laboratory Prac tice Standards, and Quality Assurance procedures, as well as the use of appropriate controls and test substances. Any deficiencies in the descri ptions of the study method, test system, animals studied, physical conditions, dosage information, indi vidual 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.

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

ACUTE TOXICITY TO FISH

Title: Fish Acute Toxicity Test

This study is: Adequate Not Adequate

TEST SUBSTANCE

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

Remarks: The test substance, Baton Rouge MEK H eavy Ketone Heart Cut C8 Ketone Fraction, was characterized as a complex mixture.

METHOD

Method/guideline followed: OECD TG 203 (1992)

Test type: Static Renewal (24-hr renewal period)

GLP compliant?: Yes

Year study was performed: 2007

Test species: Oncorhynchus mykiss . Rainbow trout used in this study were obtained from the Thomas Fish Company (Anderson, CA Lot 512, Recei pt date: 31-Oct-07). A test initiation fish were approximately 56 days post hatch, fish mean weight was 0.749 g, and mean total length was 4.5 cm. Total loading was 0.666 g of fish/L test solution.

Analytical monitoring: Yes

Exposure duration: 96-hr

Concentrations tested: 0 (control), 4.2, 8.5, 20, 45, and 101 mg /L test substance water accomadated fractions (WAF), which corresponde d to mean measured concentrations of 0 (control), 3.2, 5.4, 13, 36, and 103 mg/L.

Test method/conditions remarks: Individual Water Accommoda ted Fractions (WAF's) were prepared for each treatment. The test substance wa s added to 12 L of reconstituted water in glass aspirator bottles (capacity 13.5 L). The solutions were mixe d for 23 to 24 hours using a 5% vortex (of the static liquid depth). The test solution was removed th rough the outlet at the bottom of each mixing vessel (following a one hour settling period) into two replicates of approximately 4.5 L in 4.5 L size aspirator bottles ( no headspace). Four fish were added to each replicate and the replicates were closed with foil covered stoppers. Daily renewals were performed by removing approximately 90% of th e test solution through th e outlet at the bottom of the aspirator bottle an d refilling with fresh solution. The fi sh were not fed during the study. Mean test temperature ranged from 13.8 - 14.9°C. Diurnal light was approximately 16 hours

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

light and 8 hours dark with 433 to 438 Lux dur ing full daylight periods. Dissolved oxygen ranged from 4.5 to 9.3 mg/L and pH ranged from 6.3 to 7.9 during the study. Water hardness was 92 mg/L as CaCO3.

Due to the 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; 2) It was deemed more appropriate to prepare individual treatment solutions by adding the test substance to dilution water and removing the WAF of each mixture for testing than to prepare dilu tions of a stock solution; 3) The fish used in the study were held/acclimated for 10 days instead of at least 12 da ys as stated in the protocol; and 4) There were no culls or mortality during acclimation and the fish appeared healthy and

normal. This is not believed to have adversely impacted the study.

RESULTS

Endpoint value(s): The 96-hour LC 50 value for rainbow trout was 68 mg/L based on WAF loading rates and 61 mg/L based on mean meas ured concentrations. The maximum measured concentration causing no mort ality after 96-hours was 36 mg/L, which corresponded to a maximum WAF loading rate of 45 mg/L. The mi nimum measured concentration causing 100% mortality after 96-hours was 103 mg/L, which corr esponded to a minimum WAF loading rate of 101 mg/L.

Results remarks: Sub-lethal effects (e.g., loss of equili brium and dark pigmentation) were observed in all fish exposed to 36 mg/L te st substance (mean measured) by the 96-hour

observation. No sub-lethal eff ects were observed in lower trea tment concentrations, and 100% mortality was observed in fish exposed to 103 mg/L test substance (mean measured).

CONCLUSIONS

After Oncorhynchus mykiss were exposed to WA Fs prepared from Baton Rouge MEK Heavy Ketone Heart Cut - C8 Ketone Fraction for 96-hours, the LL 50 based on WAF loading rates was 68 mg/L and the LC 50 based on mean measured concentrations was 61 mg/L.

STUDY RELIABILITY

[1] Reliable without restrictions.

REFERENCE

ExxonMobil Biomedical Sciences, Inc. 2008. FI SH, ACUTE TOXICITY TEST on Baton Rouge MEK Heavy Ketone Heart Cut - C8 Ketone Fraction. Study # 0700858.

3Review of TSCA Section 4 Final Study Report

CITB Project Manager: Catherine Roman

Chemical Name: Methyl Heptanone

CAS Registry Number: 409-02-9

Report Title: Fish Acute Toxicity Test

Test Sponsor: ExxonMobil Chemical Company

Testing Lab: ExxonMobil Biomedical Sciences Inc.

Test Standard: ASTM E 729

Date Study Received by EPA: 06/23/09

Date Study Due to EPA: 05/17/07

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) inspect ions, their dates, and findings were missing or not signed. There is evidence that QA pr ocedures 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 di fferent 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 summar y 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/Bran ch/Division/Office: Kendra Moran/SRC, Inc.

Reviewer Signature: David Brooks/HPVCB/RAD/OPPTS

Part B. To be filled out by CITB PM

Tiered Testing Decision 1. The tiered testing trigger was me t, 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).