1,3-bis(2,4,4-trimethylpentan-2-yl)trisulfane

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Study period:

01 Feb - 03 May 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

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:

bis(2,4,4-trimethylpentan-2-yl)trisulfane

EC Number:

941-787-9

Cas Number:

98222-50-5

Molecular formula:

C16 H34 S3

IUPAC Name:

bis(2,4,4-trimethylpentan-2-yl)trisulfane

Test material form:

liquid

Details on test material:

Name: DAILUBE IS-30
Chemical Name: 1, 3-bis (2, 4, 4-trimethylpentan-2yl) trisulfane
Use: Lubricant Additive
Molecular Formula: CH3-C(CH3)2-CH2-C(CH3)2-SSS-C(CH3)2-CH2-C(CH3)2-CH3
Molecular Weight: 322 g/mol
Appearance Pale yellow
Physical State: Liquid
Batch Number: L002
Purity: 95.4% by weight
Storage: Room temperature, in the dark
Expiry Date: 2016/8/15

Method

Species / strain

Species / strain / cell type:

lymphocytes:

Remarks:

human

Details on mammalian cell type (if applicable):

- Sex, age and number of blood donors if applicable:
Preliminary Toxicity Test: male, aged 25 years
Main Experiment: male, aged 26 years

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Test concentrations with justification for top dose:

Preliminary Toxicity Test
Concentratin (μg/mL): 0, 7.81, 15.63, 31.25, 62.5, 125, 250, 500, 1000 and 2000 μg/mL.
The maximum dose was the maximum recommended dose level.

Main Study
4(20)-hour without S9 (μg/mL): 7.81, 15.63, 31.25, 46.87, 62.5, 125
4(20)-hour with S9 (2%) (μg/mL): 7.81, 15.63, 31.25, 46.87, 62.5, 125
24-hour without S9 (μg/mL): 7.81, 15.63, 31.25, 46.87, 62.5, 125
The dose levels used in the Main Experiment were selected using data from the preliminary toxicity test where the results indicated that the maximum concentration should be limited by precipitate, and toxicity also being considered in the 24-hour exposure group.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: acetone;
- Justification for choice of solvent/vehicle: solubility

Controls

Negative solvent / vehicle controls:

yes

Remarks:

acetone

Positive controls:

yes

Positive control substance:

cyclophosphamide

mitomycin C

Details on test system and experimental conditions:

DURATION
- Exposure duration:
1. 4 hours exposure in the presence of an induced rat liver homogenate metabolizing system (S9), at a 2% final concentration with cell harvest after a 20-hour expression period,
2. 4 hours exposure in the absence of metabolic activation (S9) with a 20-hour expression period and
3. a 24-hour exposure in the absence of metabolic activation.

NUMBER OF REPLICATIONS: Duplicate cultures of human lymphocytes

METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED:
The lymphocytes were re-suspended in several mL of fresh fixative before centrifugation and re-suspension in a small amount of fixative. Several drops of this suspension were dropped onto clean, wet microscope slides and left to air dry. When the slides were dry they were stained in 5% Giemsa for 5 minutes, rinsed, dried and a cover slip applied using mounting medium.

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index;

Statistics:

Fisher's Exact test.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

lymphocytes: human

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

lymphocytes: human

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

The test item was considered to be non-clastogenic to human lymphocytes in vitro.