Bis[2-(diethylamino)ethyl] adipate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Deviations:

yes

Remarks:

The MTT concentrate was diluted with MTT solvent directly before use and not stored in the fridge until usage. This deviation can be seen as uncritical, because the MTT solution was freshly prepared and no storage was necessary.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Bis[2-(diethylamino)ethyl] adipate

EC Number:

240-610-6

EC Name:

Bis[2-(diethylamino)ethyl] adipate

Cas Number:

16545-00-9

Molecular formula:

C18H36N2O4

IUPAC Name:

1,6-bis[2-(diethylamino)ethyl] hexanedioate

impurity 1

Reference substance name:

2-diethylaminoethanol

EC Number:

202-845-2

EC Name:

2-diethylaminoethanol

Cas Number:

100-37-8

Molecular formula:

C6H15NO

IUPAC Name:

2-(diethylamino)ethanol

Test material form:

liquid

Test animals / tissue source

Species:

human

Details on test animals or tissues and environmental conditions:

Commercially available EpiOcularTM kit. The EpiOcularTM tissue consists of normal, human-derived keratinocytes which have been cultured to form a stratified squamous epithelium similar to that found in the human cornea. It consists of highly organized basal cells. These cells are not transformed or transfected with genes to induce an extended life span. The EpiOcularTM tissues are cul-tured in specially prepared cell culture inserts with a porous membrane through which nutrients can pass to the cells. The tissue surface is 0.6 cm2.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

50 µL

Duration of treatment / exposure:

28 minutes

Duration of post- treatment incubation (in vitro):

120 minutes

Number of animals or in vitro replicates:

2

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

%Viability Mean

Value:

ca. 13.3

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Designation	Positive Control	Bis[2-(diethylamino)ethyl]adipate
% Viability (Tissue 1)	32.2%	12.2%
% Viability (Tissue 2)	31.3%	14.5%
% Viability Mean	31.8%	13.3%

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

Under the conditions of the test system, Bis[2-(diethylamino)ethyl]adipate is considered as eye irritant in the EpiOcularTM Eye Irritation Test. After treatment with the test item, the relative absorbance values were reduced to 13.3%.
This value is below the threshold for eye irritation potential (≤ 60%). All validity criteria were met. The criterion for optical density of the negative control was fulfilled: The OD value was 1.6 (> 0.8 and < 2.5). The positive control induced a decrease in the relative absorbance as compared to the negative control to 31.8%. Variation within the replicates was acceptable (≤20 %). For these reasons, the result of the test is considered as valid.

Executive summary:

One valid experiment was performed. The test item Bis[2-(diethylamino)ethyl]adipate was applied to a three-dimensional human cornea tissue model in duplicate for an exposure time of 28 minutes. 50 µL of the liquid test item was applied to each tissue. After treatment, the respective substance was rinsed from the tissue; then, cell viability of the tissues was evaluated by addition of MTT, which can be reduced to formazan. The formazan production was evaluated by measuring the optical density (OD) of the resulting solution. Demineralised water was used as negative control, Methyl acetate was used as positive control.

The controls showed the following results: After treatment with the negative control, the absorbance values were within the required acceptability criterion of 0.8 ≤ mean OD ≤ 2.5, OD was 1.6. The positive control showed clear eye irritating effects, the relative absorbance value was reduced to 31.8 % (< 50%). Variation within tissue replicates was acceptable (≤ 20%). After treatment with the test item, the relative absorbance values were reduced to 13.3 %. This value is below the threshold for eye irritation potential (≤ 60%). According to the OECD Guideline 492, the EpiOcular^TMEye Irritation Test does not allow discrimination between eye irritation/reversible effects on the eye (Category 2) and serious eye damage/irreversible effects on the eye (Category 1). For these purposes, further testing with other suitable test methods is required. Under the conditions of the test system, Bis[2-(diethylamino)ethyl]adipate is considered as eye irritant / inducing serious eye damage in the EpiOcular^TMEye Irritation Test.