Resin acids and Rosin acids, esters with glycerol and diethylene glycol

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

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)

Version / remarks:

2019

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Resin acids and Rosin acids, esters with glycerol and diethylene glycol

EC Number:

288-327-7

EC Name:

Resin acids and Rosin acids, esters with glycerol and diethylene glycol

Cas Number:

85711-66-6

Molecular formula:

UVCB substance

IUPAC Name:

esterification product of (1R,4aR,4bR,10aR)-1,4a-dimethyl-7-(propan-2-yl)-1,2,3,4,4a,4b,5,6,10,10a-decahydrophenanthrene-1-carboxylic acid; (1R,4aR,4bS,10aR)-1,4a-dimethyl-7-(propan-2-ylidene)-1,2,3,4,4a,4b,5,6,7,9,10,10a-dodecahydrophenanthrene-1-carboxylic acid; (1R,4aS,10aR)-1,4a-dimethyl-7-(propan-2-yl)-1,2,3,4,4a,5,6,9,10,10a-decahydrophenanthrene-1-carboxylic acid; (1R,4aS,10aR)-1,4a-dimethyl-7-(propan-2-yl)-1,2,3,4,4a,9,10,10a-octahydrophenanthrene-1-carboxylic acid and 2,2'-oxydiethanol and propane-1,2,3-triol

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: Arakawa Chemical Lot K02007 and Lot W90402
- Purity, including information on contaminants, isomers, etc.: no data

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: no data
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis: no data
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: no data
- Reactivity of the test material with the incubation material used (e.g. plastic ware): no data

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): material was powdered using a mortar and pestle
- Preliminary purification step (if any): no data
- Final concentration of a dissolved solid, stock liquid or gel: not applicable
- Final preparation of a solid (e.g. stock crystals ground to fine powder using a mortar and pestle): material was powdered using a mortar and pestle

FORM AS APPLIED IN THE TEST (if different from that of starting material) : material was powdered using a mortar and pestle

Test animals / tissue source

Species:

other: EpiOcular Reconstituted Human Cornea-like Epithelium (RhCE)

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50mg
- Concentration (if solution): not applicable

VEHICLE
- Amount(s) applied (volume or weight with unit): 50mL
- Concentration (if solution): not applicable
- Lot/batch no. (if required): sterile water lot no. 2121448; methyl acetate lot no. 090120ALA
- Purity: ≥ 99.9%

Duration of treatment / exposure:

6 hours

Duration of post- treatment incubation (in vitro):

post-exposure incubatation of 18 hours

Number of animals or in vitro replicates:

three

Details on study design:

- Details of the test procedure used : per guideline OECD 492
- Doses of test chemical and control substances used : 50 mg of test article and 50mL of control substances
- Duration and temperature of exposure, post-exposure immersion and post-exposure incubation periods (where applicable) : 6 hour incubation exposure
- Justification for the use of a different negative control than ultrapure H2O (if applicable) : not applicable
- Justification for the use of a different positive control than neat methyl acetate (if applicable): not applicable
- Description of any modifications to the test procedure : not applicable
- Indication of controls used for direct MTT-reducers and/or colouring test chemicals (if applicable) : no data
- Number of tissue replicates used per test chemical and controls (positive control, negative control, NSMTT, NSCliving and NSCkilled, if applicable) : three
- For hCE cells: number of runs and of hCE models used within each run : not applicable
- Wavelength and band pass (if applicable) used for quantifying MTT formazan, and linearity range of measuring device (e.g. spectrophotometer) : 570nm
- Description of the method used to quantify MTT formazan, if applicable : no data
- For hCE cells: data of QC check for TEER measuring system : not applicable
- Description of the qualification of the HPLC/UPLC-spectrophotometry system (if applicable) : no data
- Description of evaluation criteria used including the justification for the selection of the cut-off point for the prediction model : no data
- Reference to historical positive and negative control results demonstrating suitable run acceptance criteria : see study report
- Complete supporting information for the specific RhCE tissue construct or hCE cells used : see study report
- Reference to historical data of the RhCE tissue construct / hCE cells : see study report
- Demonstration of proficiency in performing the test method before routine use by testing of the proficiency chemicals : no data
- Positive and negative control means and acceptance ranges based on historical data : see study report
- Acceptable variability between tissue replicates for positive and negative controls : no data
- Acceptable variability between tissue replicates for the test chemical: no data

REMOVAL OF TEST SUBSTANCE

- Washing (if done): three washes following 6 hour exposure

SCORING SYSTEM: Prediction model per OECD 492

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

mean percent tissue viability 

Run / experiment:

lot K02007

Value:

89

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

mean percent tissue viability 

Run / experiment:

lot W90402

Value:

103

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the OECD Guideline 492 prediction model, rosin acid derivative is not classified for eye irritation.

Executive summary:

A GLP-compliant, OECD Guideline 492 in vitro eye irritation test was performed with rosin acid derivative. 50mg of the test article was applied to EpiOcular Reconstituted Human Cornea-like Epithelium (RhCE) and the treated tissue was incubated for 6-hours. Three replicates were performed. 50mL of sterile was was used as a negative control and 50mL of methyl acetate was used as a positive control. Following treatment, an MTT assay was performed to assess tissue viability as percent of the negative control. The data for the lots was 89% and 103% tissue viability. A mean tissue viability percentage of greater than 60% indicates that no classification is required. Based on the OECD Guideline 492 prediction model, rosin acid derivative is not classified for eye irritation.