Bis[2-(diethylamino)ethyl] adipate

Endpoint summary

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: 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

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

other: epidermal keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

The test system is a commercially available EpiDermTM-Kit, procured by MatTek. The EpiDermTM tissue consists of human-derived epidermal keratinocytes which have been cultured to form a multi-layered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDermTM tissues are cultured on specially prepared cell cultures inserts.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

50 µL

Duration of treatment / exposure:

3 minutes and 1 hour

Duration of post-treatment incubation (if applicable):

3 hours

Number of replicates:

2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3min

Value:

ca. 79.9

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1 hour

Value:

ca. 8.7

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

Negative Control		Test Item		Positive Control		Incubation
Tissue 1	Tissue 2	Tissue 1	Tissue 2	Tissue 1	Tissue 2
1.906	1.857	1.479	1.509	0.394	0.392	3 min
1.875	1.815	1.465	1.509	0.386	0.389
1.869	1.807	1.464	1.467	0.389	0.386
1.987	1.956	0.806	0.806	0.176	0.165	1 h
1.975	1.929	0.793	0.797	0.178	0.163
1.984	1.928	0.800	0.788	0.176	0.162
Mean		Mean		Mean
1.855		1.482		0.389		3 min
2.2%		1.2%		0.1%		SD
1.960		0.798		0.170		1 h
1.6%		0.2%		5.5%		SD

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

The test item is considered non-corrosive to skin. After 3 minutes treatment, the relative absorbance values were decreased to 79.9%. This value is well above the threshold for corrosivity (50%). After 1 hour treatment relative ab-sorbance values were reduced to 40.7%. This value is well above the threshold for corrosivity (15%). In the guideline values greater or equal to the threshold are considered as “non-corrosive to skin”. The values of the negative control met the required acceptability criterion of mean OD ≥ 0.8 and ≤ 2.8 for both treatment intervals, thus showing the quality of the tissues. The positive control has met the validity criterion too, thus ensuring the validity of the test system. For these reasons, the result of the test is considered valid.

Executive summary:

One valid experiment was performed. Two tissues of the human skin model EpiDerm^TMwere treated with Bis[2-(diethylamino)ethyl]adipate for 3 minutes and 1 hour, respectively. The test item was applied to each tissue and spread to match the tissue size. Demineralised water was used as negative control. 8 M KOH was used as positive control. After treatment, the respective substance was rinsed from the tissues. Then, cell viability of the tissues was evaluated by addition of MTT which can be reduced to a blue formazan. Formazan production was evaluated by measuring the optical density (OD) of the resulting solution.

After treatment with the negative control, the absorbance values were within the required acceptability criterion of mean OD ≥ 0.8 and ≤ 2.8 for both treatment intervals thus showing the quality of the tissues. The OD was 1.9 (3 minutes experiment) and 2.0 (1 hour experiment). The positive control showed clear corrosive effects for both treatment intervals. The relative absorbance value was reduced to 8.7 % for the 1 hour treatment. After 3 minutes treatment with the test item, the relative absorbance value was reduced to 79.9 %. This value is above the threshold for corrosion potential (50%). After 1 hour treatment, relative absorbance value was reduced to 40.7 %. This value, , is also above the threshold for corrosion potential (15%). In the guideline, values greater or equal to the threshold are considered as “non-corrosive to skin”. Therefore, Bis[2-(diethylamino)ethyl]adipate is considered as non-corrosive to skin in the Reconstructed human Epidermis (RhE) Test Method.

Reference 2

Endpoint:

skin 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

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

other: epidermal keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

The test system is a commercially available EpiDermTM-Kit, procured by MatTek. The EpiDermTM tissue consists of human-derived epidermal keratinocytes which have been cultured to form a multi-layered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers representing main lipid classes analogous to those found in vivo. The EpiDermTM tissues are cultured on specially prepared cell culture inserts.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

30 µL

Duration of treatment / exposure:

Tissues were dosed in 1-minute-intervals. After dosing the last tissue, all plates were transferred into the incubator for 35 minutes at 37 ± 1°C and 5.0 ± 0.5% CO2. 1 hour after the first application, the inserts were removed from the plates using sterile forceps and rinsed immediately in 1-minute-intervals. After rinsing, each tissue was blotted with sterile cellulose tissue and then transferred into a new 6-well-plate with fresh assay medium (0.9 mL).

Duration of post-treatment incubation (if applicable):

Then, the tissues were set in the incubator for 23 hours and 27 minutes at 37 ± 1°C and 5.0 ± 0.5% CO2.

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1

Value:

ca. 2.6

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

2

Value:

ca. 2.8

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3

Value:

ca. 2.6

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

All validity criteria were met. Values for negative control and for positive control were within the range of historical data of the test facility. Therefore, the experiment is considered valid.

Designation	Bis[2-(diethylamino)ethyl] adipate	Positive Control
% tissue viability (tissue 1)	2.6%	3.1%
% tissue viability (tissue 2)	2.8%	3.4%
% tissue viability (tissue 3)	2.6%	2.6%
% tissue viability (mean)	2.6%	3.0%
± SD of mean tissue viability (%)	0.1%	0.5%

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

The test item is considered as irritant to skin. After the treatment, the relative absorbance values were reduced to 2.6%. This value is below the threshold for skin irritation (50%). The optical density of the negative control was well within the required acceptability criterion of 0.8 ≤ mean OD ≤ 2.8. The positive control has met the acceptance criterion too, for thus ensuring the validity of the test system. Variation within replicates was within the accepted range for negative control, positive control and test item (required: ≤ 18%). For these reasons, the result of the test is considered valid.

Executive summary:

Three tissues of the human skin model EpiDerm^TMwere treated withBis[2-(diethylamino)ethyl]adipatefor 60 minutes.

The test item was applied directly to each tissue and spread to match the tissue size (0.63 cm²; as indicated by the supplier).

DPBS-buffer was used as negative control, 5% SDS solution was used as positive control.

After treatment with the negative control, the absorbance values were within the required acceptability criterion of 0.8 ≤ mean OD ≤ 2.8, OD was 1.7. The positive control showed clear irritating effects. Relative absorbance was reduced to 3.0% (required: ≤ 20%).

Variation within the tissue replicates was acceptable (required: ≤ 18%).

 

After the treatment with the test item, the relative absorbance value was reduced to 2.6 %. This value is below the threshold for skin irritation potential (50%).

 

 

Therefore,Bis[2-(diethylamino)ethyl]adipateis considered as

irritant to skin in the Reconstructed Human Epidermis (RhE) Test Method.

 

The OECD guideline 439 addresses the human health endpoint skin irritation. Therefore further testing on skin corrosion potential with another suitablein vitrostudy (e.g. OECD 431) may be required.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Two in vitro skin irritation/corrosion studies were performed. The skin irritation study was positive, the skin corrosion study negative. It is therefore classified accoring to ECHA Guidance on Information Requirements and Chemical Safety Assessment Chapter R.7a: Endpoint specific guidance Version 6.0 July2017 the substance is classified as "skin irritant".

Two in vitro eye irritation/corrosion studies were performed. Both studies showed positive results. It is therefore classified accoring to ECHAGuidance on Information Requirements and Chemical Safety Assessment Chapter R.7a: Endpoint specific guidance Version 6.0 July2017 the substance is classified as "eye damage".