Diisodecyl adipate

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Well documented study report, comparable to guideline study with acceptable restrictions (no data on analytical purity, non-GLP, no positive control)

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Reference 2

Reference Type:

secondary source

Title:

Unnamed

Year:

2010

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

no positive control

Principles of method if other than guideline:

The present study (Study ID 40553) was part of a multiple endpoint study (Study ID 40552). Details on methods for dosing, animal care etc. were derived from the 13-weeks-repeated dose dermal study.

GLP compliance:

no

Remarks:

The study was conducted according to GLP regulations, but the report was not reviewed by the quality assurance unit.

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

Bis(tridecyl) adipate

EC Number:

241-029-0

EC Name:

Bis(tridecyl) adipate

Cas Number:

16958-92-2

IUPAC Name:

ditridecyl adipate

Details on test material:

- Name of test material (as cited in study report): [Trade name]
- Chemical name: Di-tridecyl adipate
- Physical state: liquid
- Composition of test material, percentage of components: no data
- CRU No.: #84083
- Lot/batch No.: 830 5530

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Details on the strain: Rat/crl COBS CD[SD] BR/Charles River, Lakeview, New Jersey
- Source: Charles River, Lakeview, New Jersey
- Age at study initiation: 49 days
- Weight at study initiation: males: 161.9 - 166.8 g; females: 149.2 - 150.5 g
- Housing: individually, in suspended, stainless steel cages, with wire mesh bottoms and fronts
- Diet: Purina Certified Lab Chow #5002 in pellet form; ad libitum
- Water: tap water, delivered by an automatic watering system; ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 50
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Route of administration:

dermal

Vehicle:

- Vehicle(s)/solvent(s) used: none

Details on exposure:

TEST SITE
- Area of exposure: on the backs of the animals, beginning at the scapula and continuing laterally and posteriorly
- Type of wrap if used: no wrap. Animals were fitted with cardboard "Elisabethan" collars to minimize ingestion of test material.
- Time intervals for shavings or clipplings: approx. 24 h before the start of dosing, hair was then reclipped as necessary, but at least once per week

REMOVAL OF TEST SUBSTANCE
- Washing: approx. 24 h after the last dose each week, as much residual test substance as practical was wiped off with gauze pads.

TEST MATERIAL
- Constant volume or concentration used: yes

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

24 h, 5 days/week

Post exposure period:

none

Doses / concentrations

Remarks:

Doses / Concentrations:
800 and 2000 mg/kg bw/day
Basis:
nominal conc.

No. of animals per sex per dose:

10

Control animals:

yes, concurrent no treatment

Positive control(s):

None.
Since the study was part of a 13-weeks repeated dose dermal study, no positive control was run in parallel.

Examinations

Tissues and cell types examined:

Peripheral blood smears and bone marrow were taken from the femurs of 5 animals/sex/dose.

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION:
The doses were the same as chosen for the 13-weeks repeated dermal dose study.

TREATMENT AND SAMPLING TIMES:
To ascertain if the bone marrow and peripheral blood were equally sensitive for micronuclei detection, each target tissue was scored as an independent phase of the study and the results were compared. Three peripheral blood slides and four bone marrow slides were made for each animal.

DETAILS OF SLIDE PREPARATION:
Slides were air dried and fixed in absolute methanol for 15 min. After the slides had dried, they were placed in an acridine orange (0.125 mg/mL in Giordano's buffer, pH 5.4-6.5) solution for 7 to 10 min, rinsed in Giordano's buffer and washed in fresh Giordano's buffer for an additional 5-10 min.

METHOD OF ANALYSIS:
1000 polychromatic erythrocytes and 1000 normochromatic erythrocytes were scored in each target tissue to determine the incidence of micronuclei.

Statistics:

Several statistical methods, including ANOVA (analysis of variance) and SAS GLM (general linear model) models, were applied to the data. The ANOVA F test was used to determine significant differences between mean values. The Tukey's Studentized Range Test and the Scheffe's Test were performed to determine which means were different. SAS GLM is a studentized linear regression analysis that can be used to determine dose responsiveness.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

not applicable

Any other information on results incl. tables

Table 1: Micronucleus assay of bone marrow from rats treated dermally with the substance for 13 weeks (averaged data)

 

Dose [mg/kg bw/day]	Sex	No. of animals	PCE/NCE	% PCE with MN	% NCE with MN
0	F	5	0.38 (0.19)	0.12 (0.11)	0.04 (0.05)
0	M	5	0.44 (0.17)	0.04 (0.05)	0.02 (0.04)
0	M+F	10	0.41 (0.17)	0.08 (0.09)	0.03 (0.05)
800	F	5	0.40 (0.12)	0.14 (0.11)	0.04 (0.05)
800	M	5	0.40 (0.08)	0.08 (0.13)	0.00 (0.00)
800	M+F	10	0.40 (0.10)	0.11 (0.12)	0.02 (0.04)
2000	F	5	0.46 (0.16)	0.06 (0.05)	0.02 (0.04)
2000	M	5	0.41 (0.09)	0.02 (0.04)	0.08 (0.08)
2000	M+F	10	0.44 (0.12)	0.04 (0.05)	0.05 (0.07)

Averaged data: means (standard deviation)

PCE: polychromatic erythrocytes

NCE: normochromatic erythrocytes

MN: micronuclei

 

 

Table 2: Micronucleus assay of peripheral red blood cells from rats treated dermally with the substance for 13 weeks (averaged data)

 

Dose [mg/kg bw/day]	Sex	No. of animals	PCE/NCE	% PCE with MN	% NCE with MN
0	F	5	0.02 (0.00)	0.02 (0.04)	0.02 (0.04)
0	M	5	0.02 (0.00)	0.14 (0.11)	0.02 (0.04)
0	M+F	10	0.02 (0.00)	0.08 (0.10)	0.02 (0.04)
800	F	5	0.02 (0.00)	0.10 (0.12)	0.04 (0.05)
800	M	5	0.03 (0.01)	0.08 (0.18)	0.00 (0.00)
800	M+F	10	0.02 (0.01)	0.09 (0.14)	0.02 (0.04)
2000	F	5	0.02 (0.01)	0.08 (0.08)	0.02 (0.04)
2000	M	5	0.03 (0.01)	0.06 (0.09)	0.00 (0.00)
2000	M+F	10	0.02 (0.00)	0.07 (0.08)	0.01 (0.03)

Averaged data: means (standard deviation)

PCE: polychromatic erythrocytes

NCE: normochromatic erythrocytes

MN: micronuclei

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative