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David Brooks/DC/USEPA/US 07/15/2008 03:01 PM

Hi Mike/Catherine: To Mike Mattheisen/DC/USEPA/US@EPA, Catherine Roman/DC/USEPA/US@EPA cc Mark Townsend/DC/USEPA/US@EPA, opreskodm@oml .gov bcc Subject Chronic Toxicity to Daphnia test requirment for Solvent Black

7 We have reviewed our contractor's report and are in agreement with their evaluation of information submitted for Solvent Black 7. This report covers chronic Daphnia toxicity endpoint. 

### The following chronic aquatic information was reviewed and found notto be in compliancewith the test guidelines specified bythetest rule (71 FR 13807, March 16, 2006 : -~-, ChronicToxicityto Daphnia In your April 29, 2008 email you indicated the following: "At issue in this request is whether a study conducted in response to TSCA section 4 test rule, but which primarily followed procedures in test guidelines not specified by the test rule, still meets the data objectives of the test rule as defined by the test guideline specified-- in the rule. Attached to the message belowis a study that SOCMA had conducted with the intention of satisfying a TSCA section 4 test rule requirement of chronic toxicity to Daphnia for the HPV-1 test rule chemical, Solvent Black 7. The test rule requires that this test be performed according to ASTME 1193 (Standard Guide for Conducting Daphnia Magna Life-Cycle Toxicity Tests .) The title of the submitted study report is "Daphnia Magna, Reproduction test with a saturated solution of C.I . Solvent Black 7 generated by a generator column system (semi-static) ." The report, on page 6, states that the study procedures were based on the following three guidelines, and that the procedures generally comply with ASTME1193: 1) OECD guideline No . 211 ("Daphnia Magna, Reproduction Test", adopted 9/21/1998) 2) OECD 23, Guidance document on aquatic toxicity testing of difficult substances and mixtures (12/14/2000) . 3) ISO 10706,"Determination of long term toxicity of substances to Daphnia magna Straus (Cladocera, Crustacea) ." (3/30/2000) Please review the submitted study and determine if it satisfies the requirments of ASTM E 1193, or in your professional opinion is an adequate study that satisfies the data objectives of a chronic toxicity to Daphnia study ." --:~ E

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Again, the submitted study does not satify the requirements of ASTM E 1193 and in our professional opinion is not an adequate study that satisfies the data objectives of a chronic Daphnia toxicity study as indicated in the attached report .Solvent Black 7reviewof chronic daphnia study_6-1-08.doc



### David Brooks Risk Assessment Division (7403M) High Production Volume Chemical Branch EPAEast Room

6308N (202) 564-8603HPV Data Summary Review Sheet CAS No.: 8005-02-5 Chemical : Solvent Black 7 Endpoint: ChronicToxicitv to Daphnia Discipline coordinator Other instructions: Reviewer's Overall Judgment: Data appear -Adequate X Inadequate Reviewer's Comments Study description: Three test solutions were used. These included a blank control, a treatment control, and a saturated solution of C .I. Solvent Black 7. The saturated solution was prepared by using a column generator system

. The treatment control was prepared the same way, but without the test substance. The daphnids were fed once a day, and the test solutions were renewed three times a week. The daphnids were checked daily for immobility and number of young. The studywas interrupted after 10 days due to a nearly 50%death rate in the C .I. Solvent Black test chambers. No live young were released by the Daphnia magna that were in the Solvent Black test chambers. No significant mortality was observed in the blank and treatment controls. Reproduction was also normal in the controls, however the results are not given. The study authors state thatthe test was considered valid until day 10. Conclusions: Primarily, the test is inadequate because it lasted only 10days, when it should have lasted 21 days, and furthermore, the concentration of the solvent black 7 solution was lethal . For a valid chronic test there needs to be concentrations that are not lethal in orderthat the sublethal endpoint (in this case reproduction) can be observed. In this study only one test concentration was used, which is reported in the summary as having an initial concentration of 8.7 mg/L. The raw data indicates that the initial saturated solution had a concentration of 9.73 mg/L, and thatthe concentration after ten days was 8.70 mg/L. In order to determine adequate chronic mortality and reproductive effects, the test needs to last 21 days (which will allow

the control Daphnia magna to have three broods), and there needs to be at least four dilutions ofthe saturated solution. The testwas useful in providing a high concentration fromwhich to start, and in showing that the test substance is rather stable afterten days. However, because the test was halted prematurely, the test is not valid.

References: Migchielsen, I .M .H.J 2008. Daphnia magna, Reproduction test with a saturated solution of C .I. Solvent Black 7 generatedby a generator column system(semi-static) . NOTOXB .V., NOTOXProject 486848. `s Hertogenbosch, The Netherlands.