Fatty acids, C16-18 and C18-unsatd., reaction products with glycidyl neodecanoate and 7-oxabicyclo[4.1.0]hept-3-ylmethyl 7-oxabicyclo[4.1.0]heptane-3-carboxylate

Endpoint summary

Description of key information

Non-irritating in an in vitro EpiDerm(R) OECD 439 study, and non-irritating in an ex vivo BCOP OECD 437 study.

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

Justification for type of information:

Guideline study under GLP

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

other: Human derived transformed keratinocytes.

Cell source:

other: EpiDerm(R)

Details on animal used as source of test system:

Origin:
EpiDerm(TM) tissues were procured from MatTek In Vitro Life Science Laboratories, Bratislava.
Designation of the kit: EPI-200-SIT
Day of delivery: 08. Mar. 2016
Batch no.: 23320

Justification for test system used:

validated test system

Vehicle:

unchanged (no vehicle)

Details on test system:

Specification:
Commercially available EpiDerm (TM)-Kit.
The EpiDerm(TM) 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 EpiDerm(TM) tissues are cultured on specially prepared cell culture inserts.

Control samples:

yes, concurrent positive control

other: Dulbecco PBS

Amount/concentration applied:

The test item was applied directly to each tissue and spread to match the tissue size (0.63 cm2)

Duration of treatment / exposure:

60 minutes

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

main

Value:

>= 105.3 - <= 106.2

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Remarks:

Results given as % of negative control

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: no
- Direct-MTT reduction: tested and found to be inactive
- Colour interference with MTT: tested and found to be inactive

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes
- Range of historical values if different from the ones specified in the test guideline: within range

Interpretation of results:

GHS criteria not met

Conclusions:

Skin irritation of the test substance was tested in vitro using the EpiDerm (R) system in OECD 439 guideline under GLP. No decrease in cell viability was observed, as measured by MTT reduction to formazan. The substance is not classified as a skin irritant.

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

Justification for type of information:

Guideline study under GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

GLP compliance:

yes (incl. QA statement)

Species:

other: cattle

Strain:

other: Bos primigenius Taurus

Details on test animals or tissues and environmental conditions:

Bovine corneas were obtained from slaughtered cattle which were bettween 12 and 60 months old.

Vehicle:

physiological saline

Controls:

yes, concurrent vehicle

yes, concurrent positive control

Amount / concentration applied:

750 ml applied directly on the entire surface of the cornea

Duration of treatment / exposure:

10 minutes

Duration of post- treatment incubation (in vitro):

2 hour incubation, then washed, and incubated for an additional 90 min.

Details on study design:

SELECTION AND PREPARATION OF CORNEAS: examined and only corneas which were free from damages were used. The corneas were excised with a scalpel and cut from the globe with a 2-3 mm ring of sclera around the outside. Each cornea was transferred to a cornea holder in which pre-warmed cMEM (32 ± 1 °C) without phenol red was filled. The holders were then incubated for 1 h in the incubation chamber at 32 ± 1 °C.

QUALITY CHECK OF THE ISOLATED CORNEAS: performed and checked for tissue damage.

NUMBER OF REPLICATES: 3

NEGATIVE CONTROL USED: 0.9% sodium chloride

SOLVENT CONTROL USED (if applicable)

POSITIVE CONTROL USED: DMF

APPLICATION DOSE AND EXPOSURE TIME: see above

TREATMENT METHOD: closed chamber

POST-INCUBATION PERIOD: yes. If YES please specify duration: see above

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: 1 set of multiple rinses, using cMEM without phenol red
- POST-EXPOSURE INCUBATION: yes, 90 min

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: measured by placing the cornea in a spectral photometer and recording the absorbance at 570 nm.
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of [UV/VIS spectrophotometry / microtiter plate reader] (OD490): yes
-

SCORING SYSTEM: In Vitro Irritancy Score (IVIS), see above

DECISION CRITERIA: please specify if the decision criteria as indicated in the TG was used. yes

Irritation parameter:

in vitro irritation score

Run / experiment:

mean

Value:

0.41

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

The test substance caused no effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) is 0.41. All controls were valid and within historical control ranges.

Interpretation of results:

GHS criteria not met

Conclusions:

The test item showed no effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) is 0.41. As the value is below 3, the substance is not an eye irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The test item showed no effects on viability of human epidermal cells (EpiDerm(R)) detected through the MTT assay, with viability at 105% of negative control values.

The test item showed no effects on the cornea of the bovine eye, with a calculated IVIS (in vitro irritancy score) is 0.41. As the value is below 3, the substance is not an eye irritant.

Justification for classification or non-classification

According to the OECD 439 Guideline, substances which result in a decrease in skin cell viability of 50% are considered skin irritants. The viability of test substance was not decreased, but was slightly higher than negative control values. According to the OECD 437 Guideline, an in vitro irritancy score (IVIS) of less than or equal to 3 is evidence that the test substance is not an eye irritant. As the IVIS for the substance is 0.41, the criterion for classification is not met. The substance is not classified for skin or eye irritation.