SRC TR - 12 - 043

HPV3 REPRODUCTI ON/DEVELOPMENTAL TOXICITY STUDY PLAN REVIEW

( FINAL

– DOES NOT CONTAIN CBI) Benzenesulfonic acid, 3 - nitro - , sodium salt (1:1), CASRN 127 - 68 - 4 Prepared b y: Teresa Man yi n, William L. Richards, and Laura Morlacci Ch emical, Biological and Environmental Center

SRC, Inc. 7502 Round Pond Road North Sy racuse, NY 13212 Contract No. EP - W - 09 - 027 Task FG007.2.009.006 Submitted to: OCSPP/OPPT/RAD (7403M) U.S. Environmental Protection Agency 1200 Pennsylvania Avenue Washin gton, D.C. 20460 Debra Milligan, Project Officer Meena Sonawane, Work Assignment Manager David Brooks, Alternate Work Assignment Manager

June 5 , 2012HPV3 Reproduction/Developmental Toxicity Study Plan Review ( Final ) CASRN 127- 68- 4

1 **HPV3 Reproduction/Developmental Toxicity Study Plan**  **Review: Benzenesulfonic acid, 3**  **- nitro**  **- , sodium sal**  **t (1:1), CASRN**  **127 -**  **68 -**  **4** The test rule, “Testing of Certain High Production Volume Chemicals; Third Group of Chemicals” (76 FR 65385, October 21, 2011), requires F2 toxicity testing for benzenesulfonic acid, 3 - nitro - , sodium salt (1:1) (CASRN 127 - 68 - 4) . T he F2 requirement consists of a Reproduction/Developmental Toxicity Screening Test following 40 CFR 799.9355. A study plan for this test, to be conducted b y J ai Research Foundation, was submitted to EPA by Tiarco Chemical on February

24, 2012. Given that the study plan stated that the purity of the test substance would be 96.9%, which does not meet the test rule requirement for a purity of at least 99%, EPA requested that Tiarco Chemical submit a detailed description of the test substance’s composition to determine if the proposed test substance would yi eld toxicity test data that were representative of the test rule chemical. The composition of the proposed test substance was submitted to EPA by

Tiarco Chemical in a CBI submission (DCN 40120000059) recei ved b y EPA on April 11, 2012. The study plan is reviewed herein for compliance with the test rule guideline, 40 CFR 799.9355. The composition of the proposed test substance is also evaluated. **Review of Study Plan :**  The submitted study

plan , “Reproduction/Developmental Toxicity Screening Test of Sodium 3 - Nitrobenzene Sulfonate (CAS # 127 - 68 - 4; US EPA HPV3) in Wistar Rats” is considered adequate to satisfy

the mammalian reproduction/developmental toxicity testing requirement with the following stipulations and modifications : 1. In section 5.11: Mating Procedure (p. 10), the study plan states: “ If mating has not occurred during two weeks mating period, re - mating of females with proven males of the same group may

be performed. ” This appears to indicate that the mating period could be longer than two weeks, whereas the 40 CFR 799.9355 indicates that the mating period should be no longer than two weeks. This statement should be revised to more clearl y explain the mating procedure, while adhering to the test rule guideline. 2. The study

plan does not indicate when unmated females will be sacrificed. According to 40 CFR 799.9355 , females showing no evidence of copulation are sacrificed 24 - 26 day s after the last day

of the mating period. These details should be added to the study plan. 3. The test guideline cited in section 2.2: Regulatory Guidelines (p. 5 of the study plan) is OPPTS 870.3550. This section should also cite the test rule guideline, 40 CFR 799.9355 (“TSCA Reproduction/Developmental Toxicity Screening Test” , December 15, 2000) ; the laboratory

conducting this test should ensure compliance with 40 CFR 799.9355 . 4. In section 5.7: Housing (p. 9), the study plan states: “ Throughout experiment period, the rats will be housed individually . Except during the mating p eriod, the rats will be housed in groups of 2 rats/cage (one male plus one female). ” This is unclear. This section should be revised to state: “Throughout the experiment period, the rats will be housed individually , with the exception that during the mating period, the rats will be housed in groups of 2 rats/cage (one male plus one female).”HPV3 Reproduction/Developmental Toxicity Study Plan Review ( Final ) CASRN 127- 68- 4

2 5. In section 7.2: Pups (p. 12), the study plan should be modified to include the identification of runts (pups that are significantl y smaller than corresponding control pups) during the examination of pups at birth. 6. In section 7.2.1: Pup Body Weight (p. 12), the study plan states that individual pup body weights will be recorded. Additionally , the total weight of pups in each litter should be calculated (b y summing th e individual pup body weights for each litter); litter weights should be statistically

analyzed to determine treatment effects. 7. In section 7.3: Necrops y (p. 13), the word “scarified” should be replaced with “sacrificed”. 8. In section 8.2: Data Evaluation (p. 14), a statement should be added to indicate that statistical analy ses on pup data (e.g., number of pups born, pup body weights, pup survival) will be conducted on a per - litter or per - dam basis. **Review of Test Substance Composition**  **:** The composition of th e proposed test substance , which was submitted to EPA by Tiarco Chemical in a CBI submission (DCN 40120000059), is considered to be acceptable, with the stipulation that the dose levels must be expressed as the concentration of active ingredient (i.e., CAS RN 127 - 68 - 4). A more detailed review of the test substance composition, which includes CBI, is contained in DCN 40120000064 . **Conclusion:** The submitted study

plan and proposed test substance are acceptable with the stipulations described above.