[No public or meaningful name is available]

Genetic toxicity: in vitro

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant study conducted to recognised international test guidelines

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

Diundecyl phthalate

EC Number:

222-884-9

EC Name:

Diundecyl phthalate

Cas Number:

3648-20-2

IUPAC Name:

diundecyl phthalate

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

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

Controls

Controls 1

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

Controls 2

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 homogenate 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.