Reaction mass of bis(2-ethylhexyl) adipate and cyclohexyl 2-ethylhexyl adipate and dicyclohexyl adipate

Endpoint summary

Description of key information

Eye irritation in vitro: not irritant

Skin irritation in vitro: not irritant

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

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)

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 30869
- Expiration date of the lot/batch: 2019-01-31

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

Skin model: API-200
Supplier: MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia

Control samples:

yes, concurrent negative control

yes, concurrent positive control

other: MTT reduction control

Amount/concentration applied:

30µl

Duration of treatment / exposure:

1h total (25min at room temperature and 35 minutes in the incubator)

Duration of post-treatment incubation (if applicable):

The tissues were incubated in the incubator at 37°C for 24 ± 2 hours. After 24 ± 2 hours, the tissues were transferred into new 6-well plates pre-filled with 0.9 mL fresh medium and placed into the incubator for an additional 18 ± 2-hour post-incubation period.

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Value:

107

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

The substance is considered not to require classification as a skin irritant, if the mean tissue viability exceeds 50%.

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results observed and by applying the evaluation criteria, it was concluded that Reaction mass of Hexanedioic acid, 1-cyclohexyl 6-(2-ethylhexyl) ester, Hexanedioic acid, 1,6-dicyclohexyl ester and Hexanedioic acid, 1,6-bis(2-ethylhexyl) ester does not show a skin irritation potential in the EpiDermTM in vitro skin irritation test under the test conditions chosen.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 30869
- Expiration date of the lot/batch: 2019-01-31

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature

Species:

human

Details on test animals or tissues and environmental conditions:

EpiOcular OCL-200 kit from MatTek In Vitro Life Sciences, Bratislave, Slovakia
Reconstructed cornea: three-dimensional non-keratinized tissue construct composed of normal human-derived epidermal keratinocytes

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

other: MTT reduction control

Amount / concentration applied:

50µl

Duration of treatment / exposure:

30min

Duration of post- treatment incubation (in vitro):

2h

Number of animals or in vitro replicates:

2

Irritation parameter:

other: tissue viability

Value:

103

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Test substance		tissue 1	tissue 2	mean	inter-tissue variability [%]
NC	mean OD570	2.205	2.233	2.219
	viability [% of NC]	99.4	100.6	100.0	1.2
18/0104-1	mean OD570	2.308	2.242	2.275
	viability [% of NC]	104.0	101.0	102.5	3.0
PC	mean OD570	0.810	0.671	0.741
	viability [% of NC]	36.5	30.2	33.4	6.3

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results observed and by applying the evaluation criteria, it was concluded that Reaction mass of Hexanedioic acid, 1-cyclohexyl 6-(2-ethylhexyl) ester, Hexanedioic acid, 1,6-dicyclohexyl ester and Hexanedioic acid, 1,6-bis(2-ethylhexyl) ester does not show an eye irritation potential in the EpiOcular™ in vitro eye irritation test under the test conditions chosen.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The potential to cause dermal irritation was assessed by a single topical application of 30 μL of the undiluted test substance to a reconstructed three-dimensional human epidermis model (EpiDerm™). The irritation test was performed with three EpiDerm™ tissues, which were incubated with the test substance for 1 hour followed by a 24 -hours post-incubation period. Tissue destruction was determined by measuring the metabolic activity of the tissue after exposure/post-incubation using a colorimetric test (MTT) to measure relative viability. The mean viability of the test-substance treated tissues was 107%. Based on the observed results, it was concluded, that the test substance does not show a skin irritation potential in the EpiDerm™ in vitro skin irritation and corrosion test strategy under the test conditions chosen. No further test for skin corrosion is required.

The potential to cause ocular irritation was assessed by a single topical application of 50 µL of the undiluted test substance to a reconstructed three-dimensional human cornea model (EpiOcular™). Two EpiOcular™ tissues were incubated with the test substance for 30 minutes followed by a 2-hours post-incubation period. Tissue destruction was determined by measuring the metabolic activity of the tissue after exposure/post-incubation using a colorimetric test (MTT). The mean viability of the test-substance treated tissues was 103%. Consequently, the test substance does not show an eye irritation potential in the in vitro eye irritation test strategy under the test conditions chosen. No further in vitro assays (BCOP) were required, since for non-irritant substances the differentiation between irritant and corrosive properties is unnecessary.

Justification for classification or non-classification

Since no indication of skin or eye irriation was detected in the corresponding in vitro assays, no classification according to Regulation (EC) No 1272/2008 (CLP) is required.