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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 not to be in compliance with 

the test guidelines specified by the test rule (71 FR 13807, March 16, 2006: 

-~-, 

Chronic Toxicity to 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 below is 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 ASTM E 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 ASTM E1193 : 

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 

C.'3 

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 7 review of chronic daphnia study_6-1-08 . doc 

David Brooks 

Risk Assessment Division (7403M) 

High Production Volume Chemical Branch 

EPA East Room 6308N 

(202) 564-8603 
HPV 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 study was 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 that the test was considered valid until day 10 . 

Conclusions : Primarily, the test is inadequate because it lasted only 10 days, 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 order that 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 that the 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 of the saturated solution. The test was useful in providing a high concentration from which to 

start, and in showing that the test substance is rather stable after ten 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 generated by a generator column system (semi-static). NOTOX B.V., NOTOX Project 486848. `s 

Hertogenbosch, The Netherlands.