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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
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
Study period:
From October 14,2009 to 24 January,2010
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP compliant study conducted to recognised international test guidelines. For read-across justification see Section 13.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
Deviations:
yes
Qualifier:
according to
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Deviations:
yes
GLP compliance:
yes (incl. certificate)
Type of assay:
in vitro mammalian chromosome aberration test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: Diplast L11
- Physical state: Light yellowish clear undefined liquid
- Analytical purity: 99.97%
- Lot/batch No.:Partita n°5-6 foglio di marcia n°2 del 24/05/09
- Expiration date of the lot/batch:08/07/2010
- Amount received:3000 g
- Container: Colourless liquid
- Storage: room temperature

- Molecular formula (if other than submission substance): C30 H50 O4
- Molecular weight (if other than submission substance): 474.73
- Smiles notation (if other than submission substance): O=C(OCCCCCCCCCCC)c(c(ccc1)C(=O)OCCCCCCCCCCC)c1

Method

Species / strain
Species / strain / cell type:
lymphocytes: human
Details on mammalian cell type (if applicable):
Whole blood was collected from two healthy male volunteer donors. The volunteers are non smokers and were receiving any medication prior to sampling
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
4740, 2370, 1190, 593, 296, 148 , 74.1 and 37.0 µg/ml

Vehicle / solvent:
acetone
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
mitomycin C
Remarks:
Migrated to IUCLID6: without S9
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
cyclophosphamide
Remarks:
Migrated to IUCLID6: with S9
Details on test system and experimental conditions:
S9 tissue omogenate has the following characteristics:

Inducing Agents: Phenobarbital – 5,6-Benzoflavone
Strain : Sprague Dawley
Sex of donors : Male
Protein content: 34.0 mg/ml

In the first experiment, the cells were treated for 3 hours in the presence and absence of S9 metabolism, respectively. The harvest time of 24 hours corresponding to approximately 1.5 cell cycle was used

Appropriate negative and positive control cultures were included in each experiment. Positive control treated cultures received Mitomycin-C in the absence of S9 metabolism at dose levels of 0.75 and 0.50 µg/ml in the first main experiment. For the second experiment cultures received Mitomycin-C at dose levels of 0.45 and 0.30 µg/ml.
In the presence of S9 metabolism cultures received Cyclophosphamide at dose levels of 18.0 and 23.0 µg/ml.

Two cultures were prepared at each test point. Air-dried slides were prepared from each culture and stained 3% Giemsa.

Following treatment, for both experiments the pH and osmolality of the treatment media at the higher dose levels were determined.
No remarkable variation of pH was observed in the absence or presence of S9 metabolism



Evaluation criteria:
Criterion for outcome
(i) Statistically significant increases in the incidence of cells bearing aberrations are observed at any dose-level over the concurrent control.
(ii) The increases are reproduced in both replicate cultures and must be observed in both experiments.
(iii) The increases must exceed historical controls. Any significant increase over the concurrent negative controls is therefore compared with
historical control.
Statistics:
Fisher's Exact Test used to compare the number of cells bearing aberrations (assumed to be Poisson distributed) in control and treated cultures. Analysis performed using sets of data either including or excluding gaps.

Results and discussion

Test results
Species / strain:
lymphocytes: human
Metabolic activation:
with and without
Genotoxicity:
negative
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: no relevant change
- Effects of osmolality:For the first experiment, slight dose related reduction of osmolality was observed at the two higher dose levels selected for treatment in the absence and presence of S9 metabolism (4740 and 2370 µg/ml). In the absence of S9 slight reductions were also observed at the next two lower concentrations (1190 and 593 µg/ml). For the second experiment slight reductions were observed at the two higher dose levels.


Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Statistically significant increases in aberrant cells compared with the relevant control values were seen in cultures treated with the positive controls Mitomycin-C and Cyclophosphamide, indicating the correct functioning of the assay system.

Following treatment with the test item, no statistically significant increase in the incidence of cells bearing aberrations including or excluding gaps over the control values, was observed in any experiment both in the absence or presence of S9 metabolism.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

On the basis of these results it is concluded that 1,2-BENZENEDICARBOXYLIC ACID, DIUNDECYL ALKYL ESTER does not induce chromosomal aberrations in human lymphocytes after in vitro treatment, under the reported experimental conditions.
Executive summary:

The ability to cause chromosomal damage in cultured human lymphocytes, following in vitro treatment in the absence and presence of S9 metabolic activation has been investigated according to OECD/EU test methods.

No statistically significant increase in the incidence of cells bearing aberrations, including or excluding gaps, was observed at any dose level or any sampling time.