DODECANEDIOIC ACID, 1,12-BIS(2-OCTYLDODECYL) ESTER

Endpoint summary

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Sept - Oct 2020

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Principles of method if other than guideline:

- Principle of test:
The acute toxicity potential of Dioctyldodecyl Dodecanedioate was assessed in vitro using XCellR8's Internally Validated Human Cell-Based Screen (Non-Regulatory Method).

- Short description of test conditions:
The test uses cultured human dermal fibroplasts in animal product-free culture, Neutral Red Uptake (NRU) endpoint measurement and a prediction model, based on the GHS classification system for acute toxicity.
After a 24 hour (+/- 1 hour) exposure of 8 concentrations of the test substance in cell culture medium containing 1% DMSO to Human Dermal Fibroblasts (HDFn-XF), cytotoxicity was evaluated.

- Parameters analysed / observed:
IC50 values were >20 ug/ml (max dose tested) in each of the replicates (reps 3,4 and 5) for the test item as the cell viability did not fall below 50%.

GLP compliance:

yes

Test type:

other: In vitro

Limit test:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source : Croda Europe. Supplier batch: P-4697
- Purity: 100%


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room Temperature
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: Test item was stable, test item formulations prepared using solvent DMSO and used on the day of dosing

Species:

other: Human Dermal Fibroblasts in Xeno-Free Culture (HDFn-XF)

Route of administration:

other: The cells were incubated with the test item (concentration 20ug/ml) for 24 +/- 1h in a humidified incubator set at 37C, 5% CO2

Vehicle:

other: Cell culture medium with 1% DMSO

Doses:

0.02 mg/mL, 0.01 mg/mL, 0.005 mg/mL, 0.0025 mg/mL, 0.00125 mg/mL, 0.00063 mg/mL, 0.00031 mg/mL, 0.00016 mg/mL

No. of animals per sex per dose:

Five main experiments were carried out allowing the determination of the IC50.

Control animals:

other: A single application of culture medium was applied as the negative control. A single application of culture medium containing 1% DMSO was applied as the solvent control

Details on study design:

Preliminary testing: Determination of the top concentration by solubility testing
Range Finding Experiment: To assess test item cytotoxicity and set a more precise dosing range for the main experiment
Main Experiment: Final classification of the test item. 5 repetitions were performed.

Sex:

not specified

Dose descriptor:

other: IC50

Effect level:

> 20 other: ug/ml

Based on:

test mat.

Five main experiments were performed with a top test concentration of 20 ug/ml and a dilution factor of 2. Data from Reps 1 and 2 was not used to obtain the result due to not meeting the required acceptance criteria. All acceptance criteria was met for Reps 3, 4 and 5. 

A positive control plate with a top concentration of 100ug/ml and a dilution factor of 1.2 was run in parallel with each test item replicate to validate the assay. 

The test item did not cross the 50% viability threshold at any  of the concentrations tested and as a result the IC50 was not able to be calculated and therefore is >20ug/ml (0.02 mg/ml). 

Due to this, and the solubility limitiations not enabling the concentration of the test item to be increased in the Main Experiment, the maximum dose used in the assay was 20 ug/ml.

 

Interpretation of results:

Category 4 based on GHS criteria

Remarks:

A prediction model was used to convert the IC50 value to a corresponding GHS classification for acute oral toxicity

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

discriminating dose

Value:

> 20