[No public or meaningful name is available]

Endpoint summary

Description of key information

The substance was not irritant to the skin ad eye in vitro.

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:

From February 2022 to March 2022

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)

Specific details on test material used for the study:

Batch No.: OP.1501019

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

foreskin from multiple donors

Justification for test system used:

It was decided to use this model as it is cited among the suitable kits both by the OECD 439: 2021 and by Reg. CE 440/2008 Annex Part B, B .46. Furthermore, the test proposed by MatTek was validated in 2007 in an international validation study involving BASF SE (Ludwigshafen, Germany), IIVS Inc. (Gaithersburg, USA) ZEBET at BfR (Berlin, Germany) and Zet-LSL (Linz , Austria)

Details on test system:

The EpiDermTM model was used which involves corneal epithelium tissue.
The kit used for the test belongs to lot #36122, derived from the keratinocyte strain 00267. In the certificate sent by the supplier, it is stated that no HIV-1, hepatitis B and hepatitis C viruses were found in the batch. Furthermore, no bacteria, yeasts or other fungi were detected.
In the lot quality certificate, the supplier reported that the functionality of the cellular barrier of the lot has passed all the required checks.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

Amount of sample tested in three replicates: Rep1 = 25.7 mg, Rep 2 = 24.8 mg, Rep 3 = 25.1 mg.

Duration of treatment / exposure:

Tissues were topically exposed to the sample for 1 hour.

Duration of post-treatment incubation (if applicable):

After exposure fresh medium was added and tissues were incubated at 37 °C and 5% CO2 for 24h. The medium was replaced with fresh one for 18h incubation. Then, tissues were transferred to MTT medium and incubated at the same conditions. After 3 hours, tissues were washed with DPBS and blotted.

Number of replicates:

3 replicates were tested

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

The result is an average of three measurements derived by three replicates.

Value:

ca. 52.5

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Interpretation of results:

GHS criteria not met

Conclusions:

Under test conditions, the test item was not irritant to 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

Study period:

From February 2022 to March 2022

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Batch No.: OP.1501019

Species:

human

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

Amount of sample tested in two replicates: Rep 1 = 50.3 mg, Rep.2 = 50.9 mg

Duration of treatment / exposure:

Tissues were exposed for 6 hours to the raw substance by direct contact.

Duration of post- treatment incubation (in vitro):

The exposure is followed by washing with DPBS and an 18-hour post-incubation. and an incubation with MTT medium for 3 hours.

Number of animals or in vitro replicates:

Two replicates were tested

Details on study design:

In vitro "EpiOcular Eye Irritation Test" was used:
the tissues are conditioned by pre-incubation (1 hour or overnight) to release compounds related to transport stress and debris. Then they undergo a further pre-incubation phase for 18 hours. After the pre-incubation phase, the tissues are transferred to a fresh culture medium and exposed for 6 hours to the raw substance by direct contact. The exposure is followed by washing with DPBS and an 18-hour post-incubation. Two tissues are used respectively for the sample, for the negative control (C-) and for the positive control (C+). After exposure, the tissues are rinsed and dried, the test medium is replaced with the MTT medium. After 3 hours of incubation, the tissues are washed with DPBS and dried, then the blue formazan salts are extracted with isopropanol. The optical density (OD) of the formazan extract is determined by a spectrophotometer set at λ = 570 nm and the cell viability is calculated for each tissue as the percentage of the mean of the negative control tissues. The potential eye irritation caused by the test substance is determined according to the residual cell viability obtained after exposure.

Irritation parameter:

percent tissue viability 

Run / experiment:

The result is an average of two measurements derived by two replicates.

Value:

ca. 94.9

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Interpretation of results:

GHS criteria not met

Conclusions:

Under test conditions, the test item was not irritant to eyes.

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

Based on the study available, the substance has not to be classified under CLP regulation.