SRC TR-10-070

HPV1 TEST RULE STUDY – DATA ADEQUACY ASSESSMENT

2,4-Hexadienoic acid, CAS No. 110-44-1

Acute Toxicity to Daphnia

Prepared by:

Ashlee Aldridge, Justine Von Runnen, Teresa Manyin 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, 2010HPV1 Test Rule – Data Adequacy Assessment

2,4-Hexadienoic acid, CAS No. 110-44-1

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 22202Draft HPV1 Test Rule – Data Adequacy – Acut e Toxicity to Daphnia CAS No. 110-44-1

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 – Acut e Toxicity to Daphnia CAS No. 110-44-1

ACUTE TOXICITY TO DAPHNIA

Title: Daphnia magna 48-h Acute Survival Test

This study is: Adequate Not Adequate

The study is considered to be inadequate becaus e the test report lacked study details including test methods/procedures, test substance purity, GLP co mpliance, weight/length and acclimatization of test organisms, loading rate and whether or not analytical monitoring was

conducted.

TEST SUBSTANCE

Identity: 2,4-Hexadienoic acid, CAS No. 110-44-1

Remarks: Sorbic Acid, Lot # 200710051; Water Solubility: slightly soluble (1000 mg/L at 25 oC); Log Kow: 0.62; Melting Point: 133.9 oC; Boiling Point: 228 oC

METHOD

Method/guideline followed: The study was performed according to ASTM E 729-96 (2002) and EPA-821-R-02-012 (2002).

Test type: Acute Invertebrate Toxicity- Static Renewal

GLP compliant?: Not Specified

Year study was performed: 2008

Test species: Daphnia magna (< 24 hrs old at study initiation)

Analytical monitoring: Not Specified

Exposure duration: 48 hours

Concentrations tested: 0 (dilution water), 25, 50, 100, 175 and 250 mg/L

Test method/conditions remarks:

Statistical methods used: The LC 50 value was determined using the Trimmed Spearman-Karber

method. Dilution water: Lab water; moderately hard reconstituted water Acclimation: Cultured in moderately hard reconstituted water Feeding: None Exposure Chamber: 30 mL disposable plastic cup Volume of exposure chamber: 25 mL Number of animals exposed/chamber: 5

2Draft HPV1 Test Rule – Data Adequacy – Acut e Toxicity to Daphnia CAS No. 110-44-1

Number of replicates/treatment: 4

Test Conditions:

Dissolved O 2 : 7.0 – 7.1 mg/L pH: 4.84 – 8.11 Conductivity (μmho/cm): 338 – 359 Test Temperature: 19.5 – 20.5 oC

Water chemistry (analysis of control and 250 mg/L samples): Analysis Temperature: 25 oC (reconstitute d water); 20 oC (250 mg/L sorbic acid) Total Hardness: 93 – 102 mg CaCO 3 /L pH: 4.80 – 8.08 Alkalinity: 1.8 – 63 mg CaCO 3 /L Conductivity: 314 – 325 μmho/cm Total Dissolved Solids: 154 mg /L (250 mg/L sample only) Dissolved Oxygen: 7.0 – 7.5 mg O 2 /L Ammonia: 0.03 mg NH 3 /L (250 mg/L sample only)

Un-ionized Ammonia: < 0.1 (250 mg/L sample only) Total Residual Chlorine: 0.03 mg/L (250 mg/L sample only)

Control performance criteria : 90% survival required

Observation time: 24 and 48 hours Endpoints investigated: Mortality

RESULTS

Endpoint value(s): The 48-hour LC 50 value for daphnids was 128.2 mg/L (95% C.I. 120.6 – 136.3 mg/L).

Results remarks: Percent survival at concentrations of 0, 25, 50, 100, 175 and 250 mg/L was 100%, 100%, 100%, 95%, 0% and 0%, respectively. Control performance criteria were met.

CONCLUSIONS

48-hour LC 50 = 128.2 mg/L

STUDY RELIABILITY

[3] Not reliable

− The test report lacked study deta ils including test methods/procedures, test substance purity, GLP compliance, weight/length and acclimatization of test organisms, loading rate and whether or not analyt ical monitoring was conducted.

3Draft HPV1 Test Rule – Data Adequacy – Acut e Toxicity to Daphnia CAS No. 110-44-1

REFERENCE

EPA-HQ-OPPT-2005-0033-0313. Wego Chemi cal & Mineral Corp (WEGO). Daphnia magna

48-h Acute Survival Test. Pe rforming Laboratory: GEI Consu ltants Inc./Chadwick Ecological Division. Report Compiled by Garden Stat e Analytix. Test No. 09-0619-0082 (2008).

4Review of TSCA Section 4 Final Study Report

CITB Project Manager: Catherine Roman

Chemical Name: 2,4-Hexadienoic acid

CAS Registry Number: 110-44-1

Report Title: Daphnia magna 48-h Acute Survival Test

Test Sponsor: Wego Chemical & Mineral Corp (WEGO)

Testing Lab: GEI Consultants Inc./Chadwick Ecological Division

Test Standard: ASTM E 729

Date Study Received by EPA: August 13, 2008 Date Study Due to EPA: May 17, 2007

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.

[C] 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.

[A] 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.

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

[C] 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.

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

[A] 10. The final study report inadequately describes the test system.

[A] 11. The final study report inadequately describes the dosage information.[A] 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. [ ] 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 [X] reasons in notes below).

Data Reliability 4. The study results are precise and accurate. [ ] 5. Appropriate methods were used to determine [X] statistical significance. 6. If repeated, the test results would be expected to be the [ ]

same.

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

Reviewer Comments (continue on separate page, if needed): Part I, Question 2: In this study, the loading rate wa s 200 daphnids/L. Although ASTM E 729 does not specify a maximum loading rate based on number of daphnids/L, OPPTS 850.1010 recommends that the loading rate should not exceed 40 daphnids/L. Analytical monitoring of the test substance concentration is not described and measured concentrations are not provided; theref ore, it cannot be determined whether or not the requirements for analytical monitoring were violated. Part I, Question 5: The Statement of Quality Assurance is not signed. Part I, Question 6: GLP compliance is not stated Part I, Question 7: Purity of the test substance is not provided. Part I, Questions 9 and 10: The study report does not adequately describe the study method or test system, as

described in other comments. Part I, Question 11: Analytical measurements of the test substance are not provided. Part I, Question 12: Water quality data are not provided for each replicate. Part II, Study Conduct: The test report does not prov ide sufficient information, as described above. Part II, Data Reliability: Given that the total dissolved solids concentration is 154 mg/L at the treatment concentration of 250 mg sorbic acid/L, it appears that the test substance did not dissolve completely in solution. Additionally, because insufficient test details have been provided, it is unclear if the test results would be the same if the test was repeated. Part II, Data Utility: Because the abse nce of analytical chemistry data prev ents the determination of an accurate

LC 50 , the data are not useful.

Reviewer Name/Title/Bran ch/Division/Office: Justine Von Runnen, SRC, Inc.

Reviewer Signature: David Brooks/HPVCB/RAD/OPPTSPart 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).