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

Endpoint summary

Description of key information

Rosin acid derivative has been tested in in vitro eye and in vitro skin irritation assays. The results of the studies are:

Skin: Not irritating when tested according to OECD 439.

Eye: Not irritating when tested according to OECD 437.

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:

2020

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)

Version / remarks:

2019

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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: no data
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: not applicable
- 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: not applicable
- 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): unchanged
- 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): unchanged

FORM AS APPLIED IN THE TEST (if different from that of starting material) : unchanged

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Justification for test system used:

The test system, when used according to the OECD 439 protocol, may be used to determine change in tissue vaibility following exposure to a test article which allows for an estimation of skin irritation potential.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: MatTek EpiDerm Reconstructed Human Epidermic (RhE)
- Tissue batch number(s): Lot 33722
- Production date: - Shipping date: no data
- Delivery date: no data
- Date of initiation of testing: 2020-09-16

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37C
- Temperature of post-treatment incubation (if applicable): 37C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: tissues were washed with a stream of DPBS and blotted with a sterile cotton swab and paper towel
- Observable damage in the tissue due to washing: no data
- Modifications to validated SOP: no data

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1mg/mL in DMEM
- Incubation time: 3 hours
- Spectrophotometer: no data
- Wavelength: 570nm
- Filter: no data
- Filter bandwidth: no data
- Linear OD range of spectrophotometer: no data


NUMBER OF REPLICATE TISSUES: three


NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: one

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be irritating to skin if the mean tissue viability is less than or equal to 50% following a 1 hour exposure.
- The test substance is considered to be non-irritating to skin if the mean tissue viability is greater than 50% following a 1 hour exposure.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25mg
- Concentration (if solution): applied unchanged

VEHICLE : test article applied unchanged

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30uL
- Concentration (if solution): applied unchanged

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30uL
- Concentration (if solution): applied unchanged

Duration of treatment / exposure:

1 hour

Duration of post-treatment incubation (if applicable):

3 hours

Number of replicates:

three

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Lot K02007

Value:

96

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Lot W90402

Value:

101

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Interpretation of results:

GHS criteria not met

Conclusions:

Rosin acid derivative was found to be non-irritating in a GLP-compliant, OECD Guideline 439 study.

Executive summary:

The skin irritation potential of rosin acid derivative was assessed in a GLP-compliant, OECD 439. The test substance was applied to EpiDerm Reconstructed Human Epidermic (RhE) tissue unchanged for a 1 hour exposure. DPBS was used as a negative control and 5% SDS was used as positive control. The tissues were washed with DPBS and analyzed with an MTT assay. The percent tissue viability of the two lots were 96% and 101%. Based on the OECD Guideline 439 prediction model, rosin acid derivative is not irritating 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

Study period:

2020

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)

Version / remarks:

2019

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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

Species:

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

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

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

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

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Justification for classification or non-classification

Based on the results of the studies, rosin acid derivative is not classified for skin or eye irritation/corrosion.