Registration Dossier

Administrative data

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was rated a "1" because it was conducted in accordance with international guidelines, used appropriate testing procedures, and applied GLP.

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
Di(isononyl) Phthalate (DINP) and Di(isodecyl) Phathalate (DIDP) Are Not Mutagenic
Author:
McKee RH, Przygoda RT, Chirdon MA, Engelhardt G and Stanley M
Year:
2000
Bibliographic source:
Journal of Applied Toxicology 20:491-497
Reference Type:
study report
Title:
Unnamed
Year:
1994
Report Date:
1994

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Deviations:
yes
Remarks:
1000 polychromatic erythrocytes were scored per animal.
Principles of method if other than guideline:
Method: other: as described by Schmid (1975). Mutat. Res. 31:9-15 and modified from Heddle et al. (1983) Mutat Res. 123:61-118.
GLP compliance:
yes
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
IUCLID4 Test substance: other TS

TS-Freetext:
CAS #68515-49-1; 1,2-benzenedicarboxylic acid, di-C9,C10 and C11 branched alkyl ester, C10 Rich

Test animals

Species:
mouse
Strain:
CD-1
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Portage, MI
- Age at study initiation: 6-9 weeks of age
- Weight at study initiation: 29.9-37.6g males and 21.1-27.9 g females
- Assigned to test groups randomly: [no/yes, under following basis: ] yes
- Fasting period before study:
- Housing: group housed by sex up to 7/cage
- Diet (e.g. ad libitum): ad libitum for the duration of the study
- Water (e.g. ad libitum): ad libitum for the duration of the study
- Acclimation period: 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 72+/- 6 deg F
- Humidity (%): 55 +/- 15%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12 hour light/12 hour dark cycle


IN-LIFE DATES: From: 1994-3-1 To: 1994-3-31

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
- Vehicle(s)/solvent(s) used: Duke's Corn Oil
- Justification for choice of solvent/vehicle: In the solubility test 1.0072 g of the test substance was added to 2.0 ml of corn oil. This resulted in a clear, light yellow solution upon mixing, at a final concentration of 503.6 mg/ml. This solution passed readily through a 20 G needle.
- Concentration of test material in vehicle: Stock solution prepared by adding 19ml corn oil to 11.000 g of test substance with a final concentration of 500 mg/kg. Dilutions of this stock were prepared for the 2500 and 1250 mg/kg dose levels.
- Lot/batch no. (if required): Lot #2B
Details on exposure:
The vehicle control consisted of corn oil and was administered concurrently with the test substance at a volume of 10 ml/kg. The postive control, cyclophosphamide, was solubilized in sterile deionized water and was administerd by oral gavage at 80 mg/kg.
Duration of treatment / exposure:
Mice were sacrificed 24, 48, and 72 hours following dosing.
Mice administered the positive control were sacrificed 24 hours after administration.
Frequency of treatment:
Single administration of test substance.
Doses / concentrations
Remarks:
Doses / Concentrations:
1250, 2500, and 5000 mg/kg
Basis:
actual ingested
No. of animals per sex per dose:
5 animals per sex per dose
Positive control(s):
cyclophosphamide
- Route of administration: oral gavage
- Doses / concentrations: 80 mg/kg

Examinations

Tissues and cell types examined:
Polychromatic and normochromatic erythrocytes from mouse bone marrow were counted.
Details of tissue and slide preparation:
At 24, 48, and 72 hours after dosing, animals from each dose group were sacrificed by CO2 asphyxiation. Bone marrow from each femur was aspirated with fetal bovine serum and and suspended. Cells were placed on slides, air dried, fixed in methanol, and stained with May-Grunwald solution then Giemsa. Slides were air dried and coverslipped, then coded prior to scoring.
Evaluation criteria:
The criteria for a positive response were a statistically significant dose-related increase in micronucleated PCEs or the detection of a reproducible and statistically significant increase at at least one dose level.
Statistics:
Analysis of variance was performed on the proportion of cells with micronuclei per animal (square root arscine proportion). Tukey's Studentized range test, with adjustment for multiple comparisons, was used at each harvest time to determine which dose groups were significantly different (p<0.05) from vehicle controls.

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
no effects
Remarks:
. A range finding study was used to select doses for each of the definitive micronucleus tests, but because there was no evidence of toxicity, the highest doses utilized were based on limit tests. The study followed EPA Toxic Substances Control Act guide
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
All animals were observed immediately after dosing and periodically throughout the duration of the assay for signs of toxicity and mortalities. All animals in the vehicle and positive control groups appeared normal after dosing and remained healthy until sacrifice. All experimentally dosed groups appeared normal immediately after dosing and remained healthy until sacrifice.

The test substance induced no significant increases in micronucleated polychromatic erythrocytes over the levels observed in the vehicle controls in either sex or at any time point. The positive control induced significant increases in micronucleated polychromatic erythrocytes in both sexes with means and standard errors of 2.04% +/- 0.34% and 1.76% +/- 0.28% for the males and females, respectively.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
The test substance did not induce a significant increase in micronuclei in bone marrow polychromatic erythrocytes under the conditions of this assay and is considered negative in the mouse bone marrow micronucleus test.
Executive summary:

In an in vivo mouse bone marrow micronucleus assay, groups of 15 male and 15 female CD-1 mice received a single oral gavage dose of 1,250, 2,500, or 5,000 mg/kg test substance. Bone marrow cells were harvested at 24, 48, or 72 hours post-treatment. The test material was administered in corn oil.

There were no clinical signs of toxicity during the study. The test substance was not cytotoxic to the target cell. The positive control induced significant increases in micronucleated polychromatic erythrocytes in both sexes. The test substance did not produce a significant increase in the frequency of micronucleated polychromatic erythrocytes in bone marrow after any treatment time.