Alcohols, C14-18 and C16-18-unsatd.

Endpoint summary

Description of key information

EpiSkin™ human epidermis skin constructs test was used to assess the skin irritation potential, in vitro

The test substance, elicited a mean tissue viability of 120.6 ± 11.6% and was predicted as non-irritant to the skin.

The Bovine Corneal Opacity and Permeability Assay (BCOP)

The test substance, elicited an In Vitro Irritancy Score of -1.0 ± 0.6 and was predicted to have a classification of No Category according to the UN GHS.

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

Study period:

19 Nov 2014 - 20 Nov 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to OECD and EC guidelines and in compliance with GLPStudy conducted to OECD and EC guidelines and in compliance with GLP

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)

Species:

other: not applicable, in vitro test, human epidermis model

Strain:

other: Not applicable.

Details on test animals or test system and environmental conditions:

-Not applicable, as an human epideris model was used.
On receipt, the kit contents were checked and the inserts with tissues on agarose were stored at room temperature until use. The kit was used within the expiry date indicated by the supplier (expiry date: 22 December 2014). The maintenance medium was pre-warmed to 37ºC. The tissues were removed from the agar and placed into wells of 12 well plates containing 2 mL pre-warmed maintenance medium per well. The tissues were incubated for a minimum of 24 hours at 37 ± 2 ºC in a humidified atmosphere of 5% CO2 in air.

Type of coverage:

other: Not applicable for In-Vitro tests

Preparation of test site:

other: Not applicable for In-Vitro tests

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

10 μL neat test substance

Duration of treatment / exposure:

24 hr incubation in maintenance medium, doses for 15 min with test substance, negative/positive controls at RT, then rinsing, followed by 42 h of post-incubation period at 37 °C, transfered to wells containing 2mL 0.3 mg/mL MTT, incubated for 3 hours

Observation period:

cell viability measurement: 3 h

Number of animals:

N/A - In-Vitro assay

Details on study design:

The test substance was applied to EpiSkin™ human epidermis skin constructs. The constructs consisted of normal, human-derived epidermal keratinocytes, which had been cultured to form a multilayered, highly differentiated model of the human epidermis with a functional multilayered stratum corneum. The constructs were treated with the neat test substance for 15 minutes. After rinsing of the test substance the constructs were incubated for 42 hours. The cell viability was determined by mitochondrial dehydrogenase activity, assessed by the reduction of MTT (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) to a soluble, coloured, formazan product. The prediction model uses the percentage viability values (compared tonegative control viability) to identify irritant and non-irritant substances.

This assay was valid with negative and positive controls showing results within the acceptable
range.

Controls
The negative control was sterile Dulbecco’s Phosphate Buffered Saline (DPBS) with magnesium and calcium.
The positive control was 5% Sodium Dodecyl Sulphate (SDS) in purified water.
Preparation/application of samples

The test substance, Unjecol 85AN, positive and negative controls were in liquid form and were applied by dispensing a volume of 10 µL over each tissue using a positive displacement pipette.

Irritation / corrosion parameter:

% tissue viability

Value:

132.5

Vehicle controls validity:

not applicable

Negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation / corrosion parameter:

% tissue viability

Value:

120.2

Vehicle controls validity:

not applicable

Negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation / corrosion parameter:

% tissue viability

Value:

109.2

Vehicle controls validity:

not applicable

Negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The test substance elicited a mean tissue viability of 120.6 ± 11.6% and was predicted as non-irritant to the skin.

Results from assay

Sample	Tissue viability as percentage of mean OD negative control				Prediction MTT endpoint
	Replicate Tissues			Mean ± SD
	a	b	c
Negative Control	88.6	95.4	116.1	100.0 ± 14.3	Not applicable
Positive Control	16.0	14.0	14.3	14.7 ± 1.1	Irritant
Unjecol 85AN	132.5	120.2	109.2	120.6 ± 11.6	Non-Irritant

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

It was concluded that the test substance, Unjecol 85AN, with a mean tissue viability of 120.6 ± 11.6%, was predicted as non-irritant to the skin.

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:

other: Study conducted to OECD and EC guidelines and in compliance with GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: Bovine eyes

Strain:

not specified

Details on test animals or tissues and environmental conditions:

- No animals required for In-Vitro test
-Bovine eyes supplied and excised by abottoir, collected soon after slaughter.
-Eyes obtained from cattle approximately less than 30 months.
_Excised eyes maintained and transported to laboratory, at ambient temperarture, sufficient HBSS containing 1% (v/v) penicillin/streptomycin solution to cover eyes.
-Used witnin 4 hours of slaughter.

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

-Undiluted substance used.
-750 µl applied

Duration of treatment / exposure:

10 minutes ± 30 seconds at 32 ºC ± 1ºC , post incubation 2 hrs with cMEM at 32ºC ± 1ºC

Observation period (in vivo):

- 1 mL sodium fluorescein solution added and incubated for 90 ± 5 minutes in a waterbath at 32ºC ± 1ºC.
- Aliquote of medium removed and transfered to cuvette for spectrophotometric read at 490 nm.

Number of animals or in vitro replicates:

Corneas were treated in triplicate with either the test substance, positive control (ethanol) or negative control (0.9% saline).

Details on study design:

- No animals required for In-Vitro test
The test substance and the positive control were tested undiluted.
-All eyes examined macroscopically, for defects.
-Surrounding tissue around the eyeball pulled away from the cornea, leaving 2-3mm of sclera present around the cornea.
-Stored in HBSS plus 1% penicillin/streptomycin solution until all the corneas had been dissected. Once all the corneas had been dissected, they were rinsed in fresh HBSS plus 1% penicillin/streptomycin solution prior to mounting.
-Mounted in the cornea holders with the endothelial side against the O-ring of the posterior half of the holder, cornea flattened, holders screwed into position.
-Compartments filled with HBSS plus 1% penicillin/streptomycin, incubated overnight, in upright position at room temperature.
- HBSS plus 1% penicillin/streptomycin removed and replaced with cMEM, incubated for 60mins in upright position at 32 ºC ± 1°C in waterbath, medium removed, compartments refilled with cMEM.
-Posterior compartment plugged and basal opacity measurements performed.
-Opacitometer used to measure light transmission through centre of each mounted cornea.
-pH of test substance diluents recorded.
- Corneas treated in triplicates with either the test substance, positive control (ethanol) or negative control (0.9% saline).

Irritation parameter:

cornea opacity score

Value:

-1

Vehicle controls validity:

not applicable

Negative controls validity:

not applicable

Positive controls validity:

other: No prediction can be made

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Throughout the assay the corneas were examined for opaque spots or other irregularities. Following treatment with test substance, Unjecol 85AN, the corneas were noted as clear. The corneas treated with the positive control, ethanol, were opaque and the corneas treated with the negative control, 0.9% saline, were clear.

Sample	Opacity ± SD	Permeability ± SD	In vitro irritancy Score ± SD	In vitro classification
Unjecol 85AN	-1.000 ± 0.577	-0.003 ±0.004	-1.0 ±0.6	No category
Ethanol	19.667 ±1.732	2.225 ±2.438	53.0 ±35.7	No prediction can be made
0.9% Saline	2.333 ±1.155	0.025 ± 0.018	Not applicable	Not applicable

Interpretation of results:

study cannot be used for classification

Remarks:

Migrated information

Conclusions:

The test substance, Unjecol 85AN, elicited an In Vitro Irritancy Score of -1.0 ± 0.6 and was predicted to have a classification of No Category according to the UN GHS.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

The test substance had a mean tissue viability of 120.6 ± 11.6%, and was predicted as non-irritant to the skin, therefore it is classified as No Category for skin irritation.

The test substance elicited an In Vitro Irritancy Score of -1.0 ± 0.6 for eye irritation, therefore it is classified as No Category for eye irritation.